Supplement I to JP XV by dfgh4bnmu

VIEWS: 175 PAGES: 254

									  The Ministry of Health, Labour and
 Welfare Ministerial Notification No. 316

   Pursuant to Paragraph 1, Article 41 of the Pharmaceutical AŠairs Law (Law No.
145, 1960), we hereby revise a part of the Japanese Pharmacopoeia (Ministerial
Notiˆcation No. 285, 2006) as follows*, and the revised Japanese Pharmacopoeia
shall come into eŠect on October 1, 2007. However, in the case of drugs which are
listed in the Japanese Pharmacopoeia (hereinafter referred to as ``previous Phar-
macopoeia'') [limited to those listed in the Japanese Pharmacopoeia whose standards
are changed in accordance with this notiˆcation (hereinafter referred to as ``new
Pharmacopoeia'')] and drugs which have been approved as of October 1, 2007 as
prescribed under Paragraph 1, Article 14 of the same law [including drugs the
Minister of Health, Labour and Welfare speciˆes (the Ministry of Health and Welfare
Ministerial Notiˆcation No. 104, 1994) as those exempted from marketing approval
pursuant to Paragraph 1, Article 14 of the Pharmaceutical AŠairs Law (hereinafter
referred to as ``drugs exempted from approval'')], the Name and Standards estab-
lished in the previous Pharmacopoeia (limited to part of the Name and Standards for
the drugs concerned) may be accepted to conform to the Name and Standards estab-
lished in the new Pharmacopoeia before and on March 31, 2009. In the case of drugs
which are listed in the new Pharmacopoeia (excluding those listed in the previous
Pharmacopoeia) and drugs which have been approved as of October 1, 2007 as
prescribed under Paragraph 1, Article 14 of the same law (including those exempted
from approval), they may be accepted as those being not listed in the new Phar-
macopoeia before and on March 31, 2009. Further, Standards listed in the section
9.01 Reference Standards of the General Tests, Processes and Apparatus of the previ-
ous Pharmacopoeia at the end of application of this notiˆcation may be treated under
the previous regulation irrespective of the prescription of the section 9.01(1) Refer-
ence Standards of the General Tests, Processes and Apparatus of the new Phar-
macopoeia.

                                                                        Yoichi Masuzoe
                                             The Minister of Health, Labour and Welfare

  September 28, 2007

(The text referred to by the term ``as follows'' are omitted here. All of them are made
available for public exhibition at the Evaluation and Licensing Division, Pharmaceu-
tical and Food Safety Bureau, Ministry of Health, Labour and Welfare, at each
Regional Bureau of Health and Welfare, and at each Prefectural Office in Japan).

*The term ``as follows'' here indicates the contents of Supplement I to the Japanese Pharmacopoeia
Fifteenth Edition from General Notice to Ultraviolet-visible Reference Spectra (pp. 1789 – 1997).
                                             CONTENTS
Preface ...................................................... i     9.22 Standard Solutions ........................ 1817
Supplement I to The Japanese Pharmacopoeia,                          9.41 Reagents, Test Solutions................. 1817
Fifteenth Edition ............................. 1789–1997            9.42 Solid Supports/Column Packings for
  General Notices ................................... 1789                Chromatography........................... 1824
  General Rules for Crude Drugs ............... 1791
                                                                   O‹cial Monographs ................................ 1825
  General Rules for Preparations ............... 1793
                                                                    Crude Drugs ....................................... 1937
  General Tests, Processes and Apparatus ... 1795
  1.09 Qualitative Tests ........................... 1795
                                                                   Infrared Reference Spectra ................ 1971–1986
  2.01 Liquid Chromatography ................. 1795
  2.02 Gas Chromatography..................... 1799
                                                                   Ultraviolet-visible Reference Spectra .... 1987–1997
  2.48 Water Determination (Karl Fischer
        Method) ...................................... 1801
                                                                   General Information
  2.49 Optical Rotation Determination ....... 1801
                                                                      8. International Harmonization Implemented
  4.01 Bacterial Endotoxins Test ............... 1802
                                                                         in the Japanese Pharmacopoeia Fifteenth
  4.05 Microbiological Examination of Non-
                                                                         Edition ......................................... 1999
        sterile Products............................. 1802
                                                                     12. Microbial Attributes of Non-sterile
  6.01 Test for Metal Particles in Opthalmic
                                                                         Pharmaceutical Products.................. 2004
        Ointments.................................... 1813
                                                                     21. Quality Control of Water for
  6.08 Insoluble Particulate Matter Test for
                                                                         Pharmaceutical Use......................... 2006
        Ophthalmic Solutions..................... 1813
                                                                     31. Purity Tests on Crude Drugs Using
  6.10 Dissolution Test............................ 1813
                                                                         Genetic Information........................ 2007
  6.11 Foreign Insoluble Matter Test for
        Ophthalmic Solutions..................... 1814
                                                                   Index.................................................... 2011
  9.01 Reference Standards ...................... 1814
                                                                   Index in Latin Name................................ 2027
  9.21 Standard Solutions for Volumetric
                                                                   Index in Japanese.................................... 2029
        Analysis ...................................... 1817
                                           PREFACE
   The 15th Edition of the Japanese Pharmacopoeia            drugs, which are important from the viewpoint of
(JP) was promulgated by Ministerial Notification No.         health care and medical treatment, clinical results and
285 of the Ministry of Health, Labour and Welfare            frequency of use, as soon as possible after they reach
(MHLW) on March 31, 2006.                                    the market.
   In July 2006, the Committee on JP established the            The target date for the publication of JP 16th Edi-
basic principles for the preparation of the JP 16th Edi-     tion (the Japanese edition) was set as April 2011.
tion, setting out the roles and characteristics of the JP,      JP Expert Committees are organized with the fol-
the definite measures for the revision, and the date of      lowing panels: Panel on the Principles of Revisions;
the revision.                                                Sub-committee on the Principles of Revisions; Panel
   At the above Committee, the five basic principles of      on Medicinal Chemicals; Panel on Antibiotics; Panel
JP, which we refer to as the ``five pillars'' were estab-    on Biologicals; Panel on Crude Drugs; Panel on Phar-
lished as follows: 1) Including all drugs which are im-      maceutical Excipients; Panel on Physico-Chemical
portant from the viewpoint of health care and medical        Methods; Panel on Preparations; Panel on Physical
treatment; 2) Making qualitative improvement by in-          Methods; Panel on Biological Tests; Panel on Nomen-
troducing the latest science and technology; 3)              clature; Panel on International Harmonization; Panel
Promoting internationalization; 4) Making prompt             on Pharmaceutical Water; and Panel on Reference
partial revision as necessary and facilitating smooth        Standards. Furthermore, three working groups under
administrative operation; and 5) Ensuring transparen-        the Panel on Medicinal Chemicals are established to
cy regarding the revision, and disseminating the JP to       expedite discussion of revision drafts of Monographs.
the public. It was agreed that the Committee on JP              In the Committee on JP, Takao Hayakawa took the
should make efforts, on the basis of these principles,       role of chairman from March 2006 to September 2007.
to ensure that the JP is used more effectively in the           In addition to the regular revision every five years in
fields of health care and medical treatment by taking        line with the basic principles for the preparation of the
appropriate measurements, including getting the un-          JP it was agreed that partial revision should be done as
derstanding and cooperation of other parties con-            necessary to take account of recent progress of science
cerned.                                                      and in the interests of international harmonization.
   It was agreed that the JP should provide an official         In accordance with the above principles, the panels
standard, being required to assure the quality of medi-      initiated deliberations on selection of articles, and on
cines in Japan in response to the progress of science        revisions for General Notices, General Rules for
and technology and medical demands at the time. It           Crude Drugs, General Rules for Preparations, Gener-
should define the standards for specifications, as well      al Tests, Monographs and so on.
as the methods of testing to assure overall quality of          Draft revisions covering subjects in General No-
all drugs in principle, and it should have a role in         tices, General Rules for Crude Drugs, General Rules
clarifying the criteria for quality assurance of drugs       for Preparations, General Tests and Monographs, for
that are recognized to be essential for public health        which discussions were finished between September
and medical treatment.                                       2005 and March 2007, were prepared for a supplement
   The JP has been prepared with the aid of the              to the JP 15. They were examined by the Committee
knowledge and experience of many professionals in            on JP in April 2007, followed by the Pharmaceutical
the pharmaceutical field. Therefore, the JP should           Affairs and Food Sanitation Council (PAFSC) in June
have the characteristics of an official standard, which      2007, and then submitted to the Minister of MHLW.
might be widely used by all parties concerned. It               Numbers of discussions in the panels to prepare the
should provide information and understanding about           supplement drafts were as follows: Panel on Principles
the quality of drugs to the public, and it should be         of Revisions (7); Sub-committee on the Principles of
conducive to smooth and effective regulatory control         Revisions (6); Panel on Medicinal Chemicals (33, in-
of the quality of drugs, as well as promoting and            cluding the working groups); Panel on Antibiotics (9);
maintaining international consistency and harmoniza-         Panel on Biologicals (8); Panel on Crude Drugs (17);
tion of technical requirements.                              Panel on Pharmaceutical Excipients (7); Panel on
   It was also agreed that JP articles should cover          Physico-Chemical Methods (12); Panel on Prepara-
                                                                                                                      i
ii        Preface                                                                          Supplement I, JP XV

tions (10); Panel on Physical Methods (8); Panel on        (15) Purity tests
Biological Tests (7); Panel on Nomenclature (9); Panel     (16) Loss on drying or ignition, or water
on International Harmonization (2); and Panel on           (17) Residue on ignition, total ash or acid-insoluble
Pharmaceutical Water (7).                                       ash
   It should be noted that in the preparation of the       (18) Tests being required for pharmaceutical prepa-
drafts for the supplement, generous cooperation was             rations and other special tests
given by the Technical Committee of the Pharmaceuti-       (19) Isomer ratio
cal Manufacturer's Association of Osaka and of             (20) Assay or the content of the ingredient(s)
Tokyo, the Tokyo Crude Drugs Association, the              (21) Containers and storage
Japan Pharmaceutical Excipients Council, the Japan         (22) Expiration date
Kampo Medicine Manufacturers' Association, the             (23) Others
Japan Flavor and Fragrance Materials Association,
                                                              4. In each monograph, the following physical and
the Japan Medical Plants Federation, the Japan Phar-
                                                           chemical values representing the properties and quali-
maceutical Manufacturers Association, and the Japan
                                                           ty of the drug are given in the order indicated below,
Oilseeds Processors Association.
                                                           except that unnecessary items are omitted depending
   In consequence of this revision, the JP 15th Edition
                                                           on the nature of the drug:
carries 1567 articles, owing to the addition of 90 arti-
                                                            (1) Alcohol number
cles and the deletion of 6 articles.
                                                            (2) Absorbance
   The principles of description and the salient points     (3) Congealing point
of the revision in this Supplement are as follows:          (4) Refractive index
   1. The Supplement I to JP 15th Edition comprises         (5) Osmolarity
the following items, in order: Notification of MHLW;        (6) Optical rotation
Contents; Preface; General Notices; General Rules for       (7) Viscosity
Crude Drugs; General Rules for Preparations; Gener-         (8) pH
al Tests, Processes and Apparatus; Official Mono-           (9) Specific gravity
graphs; Infrared Reference Spectra; and Ultraviolet-       (10) Boiling point
visible Reference Spectra; then followed by General        (11) Melting point
Information; and as an appendix a Cumulative Index         (12) Acid value
containing references to the main volume and the Sup-      (13) Saponification value
plement I.                                                 (14) Ester value
                                                           (15) Hydroxyl value
  2. The articles in General Rules for Preparations,
                                                           (16) Iodine value
Official Monographs, Infrared Reference Spectra and
Ultraviolet-visible Reference Spectra are respectively        5. Identification tests comprise the following
placed in alphabetical order.                              items, which are generally put in the order given below:
                                                            (1) Coloration reactions
  3. The following items in each monograph are put
                                                            (2) Precipitation reactions
in the order shown below, except that unnecessary i-
                                                            (3) Decomposition reactions
tems are omitted depending on the nature of the drug:
                                                            (4) Derivatives
 (1) English title
                                                            (5) Infrared and/or ultraviolet-visible absorption
 (2) Commonly used name(s)
                                                                 spectrometry
 (3) Latin title (only for crude drugs)
                                                            (6) Special reactions
 (4) Title in Japanese
                                                            (7) Cations
 (5) Structural formula or empirical formula
                                                            (8) Anions
 (6) Molecular formula and molecular mass
 (7) Chemical name                                           6. Purity tests comprise the following items, which
 (8) Origin                                                are generally put in the order given below, except that
 (9) Limits of the content of the ingredient(s) and/or     unnecessary items are omitted depending on the na-
      the unit of potency                                  ture of the drug:
(10) Labeling requirements                                  (1) Color
(11) Method of preparation                                  (2) Odor
(12) Description/Description of crude drugs                 (3) Clarity and/or color of solution
(13) Identification tests                                   (4) Acidity or alkalinity
(14) Specific physical and/or chemical values               (5) Acidity
Supplement I, JP XV                                                                         Preface        iii

 (6)   Alkalinity                                        (1) 1.09 Qualitative Tests
 (7)   Chloride                                          (2) 2.01 Liquid Chromatography
 (8)   Sulfate                                           (3) 2.02 Gas Chromatography
 (9)   Sulfite                                           (4) 2.48 Water Determination (Karl Fisher Method)
(10)   Nitrate                                           (5) 2.49 Optical Rotation Determination
(11)   Nitrite                                           (6) 4.01 Bacterial Endotoxins Test
(12)   Carbonate                                         (7) 4.05 Microbial Limit Test
(13)   Bromide                                           (8) 6.01 Test for Metal Particles in Ophthalmic
(14)   Iodide                                                Ointments
(15)   Soluble halide                                    (9) 6.08 Insoluble Particulate Matter Test for
(16)   Thiocyanide                                           Ophthalmic Solutions
(17)   Selenium                                         (10) 6.10 Dissolution Test
(18)   Cationic salts
                                                           11. The following Reference Standards were new-
(19)   Ammonium
                                                        ly added:
(20)   Heavy metals
                                                           Amlexanox
(21)   Iron
                                                           Amlodipine Besilate
(22)   Manganese
                                                           Clobetasol Propionate
(23)   Chromium
                                                           Enalapril Maleate
(24)   Bismuth
                                                           Manidipine Hydrochloride
(25)   Tin
                                                           Mizoribine
(26)   Aluminum
                                                           Nabumetone
(27)   Zinc
                                                           Nizatidine
(28)   Cadmium
                                                           Ozagrel Sodium
(29)   Mercury
                                                           Vincristine Sulfate
(30)   Copper
                                                           Zidovudine
(31)   Lead
(32)   Silver                                             12. The following Reference Standards were delet-
(33)   Alkaline earth metals                            ed.
(34)   Arsenic                                            Fosfestrol
(35)   Foreign matter                                     Hypromellose Phthalate
(36)   Related substances                                 Sulfinpyrazone
(37)   Residual solvent                                   Tubocurarine Chloride Hydrochloride
(38)   Other impurities
                                                          13. English and Latin titles of drugs were based, in
(39)   Readily carbonizable substances
                                                        principle, on the International Nonproprietary Names
  7. The abbreviations used for the principal units,    for Pharmaceutical Substances, and the chemical
``mol'', ``mmol'', ``mmol/L'' and ``Pa・s'' were ad-     names were based on the Rules of the International
ded to and ``pH'' was deleted from the paragraph 9 of   Union of Pure and Applied Chemistry (IUPAC).
the General Notices.
                                                           14. Molecular formulas of organic compounds be-
  8. The following items of the General Rules for       gin with C and then H, followed by other involved ele-
Preparations were revised:                              ments in the alphabetical order of the symbols of the
 (1) Extracts                                           elements.
 (2) Ophthalmic Ointments
                                                          15. Structural formula of drug represents, as far
 (3) Tinctures
                                                        as possible, the steric configuration.
 (4) Ophthalmic Solutions
 (5) Fluidextracts                                         16. The test procedures in monographs were writ-
                                                        ten in full, except within the same monograph and in
  9. The following item was added to the General
                                                        the monographs for preparations having a cor-
Tests, Processes and Apparatus:
                                                        responding monograph of their principal material sub-
  6.11 Foreign Insoluble Matter Test for Ophthalmic
                                                        stances.
Solutions
                                                          17. The following monographs were added:
  10. The following items of the General Tests,
                                                          Acemetacin
Processes and Apparatus were revised:
                                                          Alminoprofen
iv         Preface                                                             Supplement I, JP XV

     Alminoprofen Tablets                            Josamycin Tablets
     Alprostadil Injection                           Labetalol Hydrochloride
     Amikacin Sulfate Injection                      Labetalol Hydrochloride Tablets
     Amlexanox                                       Manidipine Hydrochloride
     Amlexanox Tablets                               Manidipine Hydrochloride Tablets
     Amlodipine Besilate                             Minocycline Hydrochloride for Injection
     Amosulalol Hydrochloride                        Mitomycin C for Injection
     Amosulalol Hydrochloride Tablets                Mizoribine
     Ampicillin Sodium for Injection                 Mizoribine Tablets
     Azelastine Hydrochloride                        Nabumetone
     Aztreonam for Injection                         Nabumetone Tablets
     Benzylpenicillin Potassium for Injection        Nafamostat Mesilate
     Biotin                                          Nizatidine
     Bisoprolol Fumarate                             Nizatidine Capsules
     Bisoprolol Fumarate Tablets                     Omeprazole
     Bucillamine Tablets                             Ozagrel Sodium
     Buformin Hydrochloride                          Ozagrel Sodium for Injection
     Buformin Hydrochloride Enteric-coated Tablets   Peplomycin Sulfate for Injection
     Buformin Hydrochloride Tablets                  Piperacillin Hydrate
     Buprenorphine Hydrochloride                     Rokitamycin Tablets
     Cefadroxil Capsules                             L-Serine
     Cefadroxil for Syrup                            Sodium Starch Glycolate
     Cefazolin Sodium for Injection                  Tobramycin Injection
     Cefmetazole Sodium for Injection                L-Tyrosine
     Ceftazidime for Injection                       Ubenimex
     Cetirizine Hydrochloride                        Zidovudine
     Cetirizine Hydrochloride Tablets                Aralia Rhizome
     Chlorphenesin Carbamate Tablets                 Powdered Corydalis Tuber
     Cibenzoline Succinate                           Crataegus Fruit
     Cibenzoline Succinate Tablets                   Hangekobokuto Extract
     Cilazapril Hydrate                              Keishibukuryogan Extract
     Cilazapril Tablets                              Leonurus Herb
     Clindamycin Phosphate Injection                 Lilium Bulb
     Clobetasol Propionate                           Peucedanum Root
     Clorazepate Dipotassium                         Powdered Turmeric
     Clorazepate Dipotassium Capsules
                                                     18. The following monographs were revised:
     L-Cysteine
                                                     Acetylcholine Chloride for Injection
     L-Cysteine Hydrochloride Hydrate
                                                     Ajimaline Tablets
     Domperidone
                                                     Aminophylline Injection
     Doxorubicin Hydrochloride for Injection
                                                     Amitriptyline Hydrochloride Tablets
     Emorfazone
                                                     L-Arginine Hydrochloride Injection
     Enalapril Maleate
                                                     Ascorbic Acid Injection
     Enalapril Maleate Tablets
                                                     Baclofen Tablets
     Erythromycin Enteric-Coated Tablets
                                                     Betahistine Mesilate
     Etizolam Fine Granules
                                                     Bisacodyl Suppositories
     Etizolam Tablets
                                                     Calcium Chloride Injection
     Felbinac
                                                     Calcium Folinate
     L-Glutamine
                                                     Camostat Mesilate
     Griseofulvin Tablets
                                                     Cefalotin Sodium
     Ibudilast
                                                     Cefatrizine Propylene Glycolate
     Isoxsuprine Hydrochloride
                                                     Chlordiazepoxide Tablets
     Isoxsuprine Hydrochloride Tablets
                                                     Chlorphenesin Carbamate
     Itraconazole
                                                     Chlorpromazine Hydrochloride Injection
Supplement I, JP XV                                                            Preface         v

 Chlorpromazine Hydrochloride Tablets          Roxithromycin
 Chlorpropamide Tablets                        Salicylic Acid
 Cilostazol Tablets                            Sodium Bicarbonate Injection
 Creosote                                      10z Sodium Chloride Injection
 Cyanocobalamin                                Sodium Citrate Injection for Transfusion
 Cyanocobalamin Injection                      Sodium Thiosulfate Injection
 Deferoxamine Mesilate                         Sulbactam Sodium
 Dehydrocholic Acid Injection                  Sulpyrine Injection
 Deslanoside Injection                         Sultamicillin Tosilate Hydrate
 Dextran 40                                    Suxamethonium Chloride for Injection
 Anhydrous Dibasic Calcium Phosphate           Suxamethonium Chloride Injection
 Dibasic Calcium Phosphate Hydrate             Talc
 Dopamine Hydrochloride Injection              Teceleukin for Injection (Genetical Recombination)
 Edrophonium Chloride Injection                Thiamine Chloride Hydrochloride Injection
 Ephedrine Hydrochloride Injection             Thiopental Sodium for Injection
 Ephedrine Hydrochloride Tablets               Tipepidine Hibenzate Tablets
 Famotidine for Injection                      Trimetazidine Hydrochloride
 Faropenem Sodium Hydrate                      Vincristine Sulfate
 Faropenem Sodium Tablets                      Water for Injection
 Faropenem Sodium for Syrup                    Xylitol Injection
 Folic Acid Injection                          Alpinia Offcinarum Rhizome
 Folic Acid Tablets                            Anemarrhena Rhizome
 Fructose Injection                            Angelica Dahurica Root
 Gabexate Mesilate                             Apricot Kernel
 Glucose Injection                             Asiasarum Root
 Hydralazine Hydrochloride Tablets             Asparagus Tuber
 Hypromellose Phthalate                        Atractylodes Rhizome
 Idoxuridine Ophthalmic Solution               Powdered Atractylodes Rhizome
 Imipramine Hydrochloride Tablets              Belladonna Extract
 Indometacin Capsules                          Calumba
 Isotonic Sodium Chloride Solution             Powdered Calumba
 Anhydrous Lactose                             Cimicifuga Rhizome
 Levallorphan Tartrate Injection               Clematis Root
 Magnesium Sulfate Injection                   Cnidium Rhizome
 D-Mannitol Injection                          Powdered Cnidium Rhizome
 Medazepam                                     Condurango Fluidextract
 Mefruside Tablets                             Coptis Rhizome
 Methyldopa Tablets                            Powdered Coptis Rhizome
 Morphine Hydrochloride Tablets                Corydalis Tuber
 Neostigmine Methylsulfate Injection           Cyperus Rhizome
 Nicardipine Hydrochloride Injection           Powdered Cyperus Rhizome
 Nicorandil                                    Dioscorea Rhizome
 Nicotinic Acid Injection                      Powdered Dioscorea Rhizome
 Noradrenaline Injection                       Fritillaria Bulb
 Papaverine Hydrochloride Injection            Gastrodia Tuber
 Pethidine Hydrochloride Injection             Gentian
 Prednisolone Sodium Succinate for Injection   Powdered Gentian
 Protamine Sulfate                             Glehnia Root
 Protamine Sulfate Injection                   Glycyrrhiza Extract
 Pyridoxine Hydrochloride Injection            Crude Glycyrrhiza Extract
 Reserpine Injection                           Hochuekkito Extract
 Riboflavin Sodium Phosphate Injection         Imperata Rhizome
 Ringer's Solution                             Ipecac
vi         Preface                                                              Supplement I, JP XV

     Powdered Ipecac                                 Tubocurarine Chloride Hydrochloride Injection
     Japanese Gentian
                                                     Those who were engaged in the preparation of Sup-
     Powdered Japanese Gentian
                                                   plement I to JP 15 are as follows:
     Japanese Valerian
     Powdered Japanese Valerian
                                                   Norio Aimi                 Yuuichi Kikuchi
     Kakkonto Extract
                                                   Fumiaki Akahori            Mitsukazu Kitada
     Kamishoyosan Extract
                                                   Mitsuo Aoki                Fumiyuki Kiuchi
     Lindera Root
                                                   Kiichi Aonuki              Junko Kizu
     Lithospermum Root
                                                   Nobuo Aoyagi**             Takashi Kobayashi
     Lycium Bark
                                                   Keiko Arimoto              Masayoshi Kohase
     Magnolia Bark
                                                   Hiroshi Asama              Shigeo Kojima
     Powdered Magnolia Bark
                                                   Kazuhide Ashizawa          Hiroyasu Kokubo
     Mulberry Bark
                                                   Shinichiro Aso             Katsuko Komatsu
     Notopterygium Rhizome
                                                   Yukio Aso                  Akio Komura
     Nuphar Rhizome
                                                   Takashi Bamba              Tetsuya Konda
     Nux Vomica Extract
                                                   Makoto Emura               Toshifumi Konda
     Panax Japonicus Rhizome
                                                   Hiroyuki Fuchino           Kenji Kondo
     Powdered Panax Japonicus Rhizome
                                                   Shigeyuki Fujikura         Seizo Kondo
     Peach Kernel
                                                   Akihiko Fujise             Takao Kunisada
     Powdered Peach Kernel
                                                   Goro Funamoto              Masaaki Kurihara
     Perilla Herb
                                                   Yukihiro Goda              Fumiyo Kusu
     Platycodon Fluidextract
                                                   Ruri Hanajiri              Kumiko Kusuyama
     Polygala Root
                                                   Kouji Hasegawa             Masako Maeda
     Powdered Polygala Root
                                                   Mitsuru Hashida            Midori Makita
     Polygonum Root
                                                   Rika Hatano                Yoshihisa Matsuda
     Polyporus Sclerotium
                                                   Takao Hayakawa*            Norio Matsuki
     Powdered Polyporus Sclerotium
                                                   Masahiro Hayashi           Eiichi Mikami
     Processed Aconite Root
                                                   Yoshinori Hayashi          Tsuyoshi Miyagawa
     Powdered Processed Aconite Root
                                                   Kenji Higuchi              Satoshi Minobe
     Processed Ginger
                                                   Fusayoshi Hirayama         Tsuyoshi Miura
     Rehmannia Root
                                                   Yukio Hiyama               Hiroto Miyamoto
     Ryokeijutsukanto Extract
                                                   Kunimoto Hotta             Naoki Miyata
     Saposhnikovia Root
                                                   Masashi Hyuga              Michinao Mizugaki
     Saussurea Root
                                                   Nobukazu Igoshi            Taiichi Mizuta
     Scopolia Extract
                                                   Kenichi Inui               Kaoru Morikawa
     Scopolia Rhizome
                                                   Akiko Ishii                Osamu Morita
     Scutellaria Root
                                                   Yuji Ito                   Takashi Morita
     Powdered Scutellaria Root
                                                   Takashi Itoh               Toshimi Murai
     Senega
                                                   Masao Izaki                Masashi Muroi
     Powdered Senega
                                                   Kenichi Izutsu             Hiroaki Nagashige
     Smilax Rhizome
                                                   Akemi Kai                  Shinsaku Nakagawa
     Powdered Smilax Rhizome
                                                   Kazuaki Kakehi             Emi Nakajima
     Sophora Root
                                                   Takemine Kanai             Hiroshi Nakamura
     Powdered Sophora Root
                                                   Motoko Kanke               Tatsuya Nakano
     Turmeric
                                                   Hirohito Katayama          Tatsumi Nakashima
     Uva Ursi Fluidextract
                                                   Noriko Katori              Mitsuo Nanaura
     Zedoary
                                                   Nobuo Kawahara             Masaaki Naotsuka
     19. The following monographs were deleted:    Toru Kawanishi             Masao Nasu
     Fosfestrol                                    Yoshihiko Kawarasaki       Shingo Niimi
     Fosfestrol Tablets                            Nana Kawasaki              Hiroshi Nishimura
     Sulfinpyrazone                                Toshisuke Kawasaki         Shukichi Ochiai
     Sulfinpyrazone Tablets                        Yoshiaki Kawashima         Atsuyuki Ohtsuki
     Tubocurarine Chloride Hydrochloride Hydrate   Keiji Kijima               Ryozo Oishi
Supplement I, JP XV                                                         Preface       vii

Minoru Okada          Michiko Sekiguchi     Haruo Tanaka             Yoshimasa Uehara
Satoshi Okada         Setsuko Sekita        Toshihiro Tanaka         Kazuichi Umemoto
Kimiya Okazaki        Yasuo Shimada         Kenichi Tanamoto**       Haruo Watanabe
Tsuneo Okubo          Kesamitsu Shimizu     Tsuyoshi Tanimoto        Takehiko Yajima
Haruhiro Okuda        Kyoko Shimura         Susumu Terabayashi       Teruhide Yamaguchi
Masami Otsuka         Osamu Shirota         Reiko Teraoka            Keiichi Yamamoto
Tadashi Ouchi         Hisashi Sonobe        Keijiro Terashita        Keiji Yamamoto
Kazuhiro Owada        Shoko Sueyoshi        Masafumi Teshigawara     Tosuke Yamamoto
Kenji Saiki           Shinji Sugaya         Hiroshi Tokunaga         Takeshi Yamazaki
Yoshikazu Sakagami    Hisakazu Sunada       Kiyoshi Tomioka          Masato Yasuhara
Eiji Sakai            Hideyo Suzuki         Motowo Tomita            Hikaru Yoden
Tomoaki Sakamoto      Yoshikazu Takahashi   Hideya Tsuge             Chikako Yomota
Hideki Sasaki         Toshio Takachi        Yosuke Tsuji             Hitoo Yoshida
Hiroshi Sasaki        Kikuo Takatera        Eriko Uchida             Sumie Yoshioka
Tsuguo Sasaki         Tadahiro Takeda
                                            *: Chairman, the Committee on JP
Motoyoshi Satake      Yasushi Takeda
                                            **: Acting Chairman, the Committee on JP
Kyoko Sato            Toyoshige Tanabe
                     GENERAL NOTICES
Change the paragraph 9 to read:                           per centimeter                    cm-1
                                                          newton                            N
  9. The following abbreviations are used for the
                                                          kilopascal                        kPa
principal units.
                                                          pascal                            Pa
       meter                          m
                                                          mole per liter                    mol/L
       centimeter                     cm
                                                          millimole per liter               mmol/L
       millimeter                     mm
                                                          pascal second                     Pa・s
       micrometer                     mm
                                                          millipascal second                mPa・s
       nanometer                      nm
                                                          square millimeter per second      mm2/s
       kilogram                       kg
                                                          lux                               lx
       gram                           g
                                                          mass per cent                     z
       milligram                      mg
                                                          mass parts per million            ppm
       microgram                      mg
                                                          mass parts per billion            ppb
       nanogram                       ng
                                                          volume per cent                   volz
       picogram                       pg
                                                          volume parts per million          vol ppm
       mole                           mol
                                                          mass per volume per cent          w/vz
       millimole                      mmol
                                                          endotoxin unit                    EU
       Celsius degree                 9 C
       square centimeter              cm2
                                                    Note: ``ppm'' used in the Nuclear Magnetic
       liter                          L
                                                    Resonance Spectroscopy indicates the chemical shift,
       milliliter                     mL
                                                    and ``w/vz'' is used in the formula or composition of
       microliter                     mL
                                                    preparations.
       megahertz                      MHz




                                                                                                    1789
                     GENERAL RULES FOR
                       CRUDE DRUGS
Change the paragraph 1 to read:                              bitis Seed, Phellodendron Bark, Picrasma Wood, Pinellia
                                                             Tuber, Plantago Herb, Plantago Seed, Platycodon Root,
   1. Crude drugs in the monographs include medicinal
                                                             Polygala Root, Polygonatum Rhizome, Polygonum Root,
parts obtained from plants or animals, cell inclusions and
                                                             Polyporus Sclerotium, Poria Sclerotium, Powdered Acacia,
secretes separated from the origins, their extracts, and
                                                             Powdered Agar, Powdered Alisma Rhizome, Powdered
minerals. General Rules for Crude Drugs and Crude Drugs
                                                             Aloe, Powdered Amomum Seed, Powdered Atractylodes
Test are applicable to the following:
                                                             Lancea Rhizome, Powdered Atractylodes Rhizome, Pow-
   Acacia, Achyranthes Root, Agar, Akebia Stem, Alisma
                                                             dered Calumba, Powdered Capsicum, Powdered Cinnamon
Rhizome, Aloe, Alpinia Officinarum Rhizome, Amomum
                                                             Bark, Powdered Clove, Powdered Cnidium Rhizome,
Seed, Anemarrhena Rhizome, Angelica Dahurica Root,
                                                             Powdered Coix Seed, Powdered Coptis Rhizome, Powdered
Apricot Kernel, Aralia Rhizome, Areca, Artemisia Capil-
                                                             Corydalis Tuber, Powdered Cyperus Rhizome, Powdered
laris Flower, Asiasarum Root, Asparagus Tuber, Astragalus
                                                             Dioscorea Rhizome, Powdered Fennel, Powdered Gambir,
Root, Atractylodes Lancea Rhizome, Atractylodes Rhi-
                                                             Powdered Gardenia Fruit, Powdered Gentian, Powdered
zome, Bear Bile, Bearberry Leaf, Belladonna Root, Benin-
                                                             Geranium Herb, Powdered Ginger, Powdered Ginseng,
casa Seed, Benzoin, Bitter Cardamon, Bitter Orange Peel,
                                                             Powdered Glycyrrhiza, Powdered Ipecac, Powdered
Bupleurum Root, Burdock Fruit, Calumba, Capsicum,
                                                             Japanese Angelica Root, Powdered Japanese Gentian,
Cardamon, Cassia Seed, Catalpa Fruit, Chrysanthemum
                                                             Powdered Japanese Valerian, Powdered Magnolia Bark,
Flower, Cimicifuga Rhizome, Cinnamon Bark, Citrus
                                                             Powdered Moutan Bark, Powdered Oyster Shell, Powdered
Unshiu Peel, Clematis Root, Clove, Cnidium Monnieri
                                                             Panax Japonicus Rhizome, Powdered Peach Kernel,
Fruit, Cnidium Rhizome, Coix Seed, Condurango, Coptis
                                                             Powdered Peony Root, Powdered Phellodendron Bark,
Rhizome, Cornus Fruit, Corydalis Tuber, Crataegus Fruit,
                                                             Powdered Picrasma Wood, Powdered Platycodon Root,
Cyperus Rhizome, Digenea, Dioscorea Rhizome, Dolichos
                                                             Powdered Polygala Root, Powdered Polypourus Scleroti-
Seed, Eleutherococcus Senticosus Rhizome, Ephedra Herb,
                                                             um, Powderd Poria Sclerotium, Powdered Processed
Epimedium Herb, Eucommia Bark, Evodia Fruit, Fennel,
                                                             Aconite Root, Powdered Rhubarb, Powdered Rose Fruit,
Forsythia Fruit, Fritillaria Bulb, Gambir, Gardenia Fruit,
                                                             Powdered Scutellaria Root, Powdered Senega, Powdered
Gastrodia Tuber, Gentian, Geranium Herb, Ginger,
                                                             Senna Leaf, Powdered Smilax Rhizome, Powdered Sophora
Ginseng, Glehnia Root, Glycyrrhiza, Gypsum, Hemp Fruit,
                                                             Root, Powdered Sweet Hydrangea Leaf, Powdered Swertia
Honey, Houttuynia Herb, Immature Orange, Imperata
                                                             Herb, Powdered Tragacanth, Powdered Turmeric, Pow-
Rhizome, Ipecac, Japanese Angelica Root, Japanese Genti-
                                                             dered Zanthoxylum Fruit, Processed Aconite Root,
an, Japanese Valerian, Jujube, Jujube Seed, Leonurus
                                                             Processed Ginger, Prunella Spike, Pueraria Root, Red Gin-
Herb, Lilium Bulb, Lindera Root, Lithospermum Root,
                                                             seng, Rehmannia Root, Rhubarb, Rice Starch, Rose Fruit,
Longgu, Lonicera Leaf and Stem, Loquat Leaf, Lycium
                                                             Rosin, Safflower, Saffron, Saposhnikovia Root, Sappan
Bark, Lycium Fruit, Magnolia Bark, Magnolia Flower,
                                                             Wood, Saussurea Root, Schisandra Fruit, Schizonepeta
Mallotus Bark, Mentha Herb, Moutan Bark, Mulberry
                                                             Spike, Scopolia Rhizome, Scutellaria Root, Senega, Senna
Bark, Nelumbo Seed, Notopterygium Rhizome, Nuphar
                                                             Leaf, Sinomenium Stem, Smilax Rhizome, Sophora Root,
Rhizome, Nux Vomica, Ophiopogon Tuber, Oriental
                                                             Sweet Hydrangea Leaf, Swertia Herb, Turmeric, Toad
Bezoar, Oyster Shell, Panax Japonicus Rhizome, Peach
                                                             Venom, Tragacanth, Tribulus Fruit, Trichosanthes Root,
Kernel, Peony Root, Perilla Herb, Peucedanum Root, Phar-
                                                             Uncaria Hook, Zanthoxylum Fruit, Zedoary.




                                                                                                                1791
                           GENERAL RULES
                          FOR PREPARATIONS
                                                                 a transparency which does not interfere with the test for
                     7.    Extracts                              foreign matter.
                                                                   (9) Unless otherwise specified, Ophthalmic Solutions
                                                                 meet the Insoluble Particulate Matter Test for Ophthalmic
Change (1) to read:
                                                                 Solutions <6.08>.
  (1) Extracts are prepared by evaporating the extractives
of crude drugs. Generally, there are two kinds of Extracts
which are:                                                                         27.      Tinctures
  (i) viscous extracts (ii) dry extracts

                                                                 Change (2) to read:
                8.     Fluidextracts                                (2) Unless otherwise specified, Tinctures are usually pre-
                                                                 pared from coarse powder or fine cuttings of crude drug
                                                                 substance(s) either by maceration or by percolation as
Change (4) to read:
                                                                 described below.
   (4) Unless otherwise specified, Fluidextracts meet the           Maceration: Place crude drugs in a suitable container, and
requirements of the Heavy Metals Limit Test <1.07> when          add the total volume or about three-fourths of the total
the test solution and the control solution are prepared as       volume of a solvent to be used. Stopper, and allow the
follows.                                                         container to stand at ordinary temperature with occasional
   Test solution: Ignite 1.0 g of Fluidextracts to ash, warm     stirring for about 5 days or until the soluble constituents
with 3 mL of dilute hydrochloric acid, filter, and wash the      have satisfactorily dissolved. Filter the liquid through cloth.
residue with two 5 mL portions of water. Add 1 drop of           In the case where about three-fourths of the total volume of
phenolphthalein TS to the combined filtrate and washings,        the solvent is added, wash the residue with a suitable quanti-
add ammonia TS dropwise until the solution becomes a pale        ty of the solvent, press, and combine the filtrate and wash-
red, filter, if necessary, and add 2 mL of dilute acetic acid    ings to make up the volume. In the case where the total
and water to make 50 mL.                                         volume of the solvent is added, sufficient solvent may be ad-
   Control solution: Proceed with 3 mL of dilute                 ded, if necessary, to make up for the decreasing amount. Al-
hydrochloric acid as directed in the preparation of test solu-   low the mixture to stand for about 2 days, and obtain a clear
tion, and add 3.0 mL of Standard Lead Solution and water         liquid by decantation or filtration.
to make 50 mL.                                                      Percolation: Pour the solvent in small portions on crude
                                                                 drugs placed in a container, and mix well to moisten the
                                                                 crude drugs. Stopper the container, and allow it to stand for
      17.      Ophthalmic Ointments                              about 2 hours at room temperature. Pack the contents as
                                                                 tightly as possible in a suitable percolator, open the lower
Change (5) to read:                                              opening, and slowly pour sufficient solvent to cover the
                                                                 crude drugs. When the percolate begins to drip, close the
 (5) Unless otherwise specified, Ophthalmic Ointments            opening, and allow the mixture to stand for 2 to 3 days at
meet the requirements of the Test for Metal Particles in         room temperature. Open the opening, and allow the perco-
Ophthalmic Ointments <6.01>.                                     late to drip at a rate of 1 to 3 mL per minute. Add an
                                                                 appropriate quantity of the solvent, and continue to perco-
                                                                 late until the desired volume has passed. Mix thoroughly,
       18.      Ophthalmic Solutions                             allow standing for 2 days, and obtain a clear liquid by
                                                                 decantation or filtration. The time of standing and the flow
Change (8), (9) to read:                                         rate may be varied depending on the kind and amount of
                                                                 crude drugs to be percolated.
  (8) Ophthalmic Solutions prepared as aqueous solution             Tinctures prepared by either of the above methods for
and aqueous vehicles attached to Ophthalmic Solutions,           which the content of the drug substance is specified are
unless otherwise specified, meet the requirement of the          prepared by assaying the drug substance using a portion of
Foreign Insoluble Matter Test for Ophthalmic Solutions           the sample and adjusting, if necessary, with the percolate or
<6.11>. The containers of Ophthalmic Solutions should have       with the solvent to the specified content.


                                                                                                                          1793
        GENERAL TESTS, PROCESSES
            AND APPARATUS
Change the introduction to read:                                   examination, purity test, loss on drying, total ash, acid-
                                                                   insoluble ash, extract content, and essential oil content of
   General Tests, Processes and Apparatus includes common
                                                                   crude drugs are performed as directed in the corresponding
methods for tests, useful test methods for quality recogni-
                                                                   items under Crude Drugs Test.
tion of drugs and other articles related to them. Unless
                                                                      The number of each test method is a category number
otherwise specified, the procedures for acid-neutralizing
                                                                   given individually. The number in blackets (< >) appeared
capacity determination of gastrointestinal medicines,
                                                                   in monograph indicates the number corresponding to the
alcohol number determination, ammonium determination,
                                                                   general test method.
arsenic determination, atomic absorption spectrophotomet-
ry, test for bacterial endotoxins, boiling point determina-
tion, distilling range determination, chloride determination,
conductivity measurement, congealing point determination,                    1.09        Qualitative Tests
determination of bulk and tapped densities, digestion test,
                                                                   Add the following next to Mercurous salt:
disintegration test, dissolution test, endpoint detection in
titrimetry, test of extractable volume for injection, flame        Mesilate
coloration, fluorometry, foreign insoluble matter test for           (1) To mesilates add twice its mass of sodium hydroxide,
injections, foreign insoluble matter test for ophthalmic           heat gently to melt, and continue heating for 20 to 30
solutions, gas chromatography, heavy metals determination,         seconds. After cooling, add a little amount of water, then
test for glass containers for injections, infrared spec-           add dilute hydrochloric acid, and warm: the gas evolved
trophotometry, insoluble particulate matter test for injec-        changes moistened potassium iodate-starch paper to blue.
tions, insoluble particulate matter test for ophthalmic              (2) To mesilates add threefold its mass of sodium nitrate
solutions, iron determination, liquid chromatography, loss         and anhydrous sodium carbonate, mix, and heat gradually.
on drying determination, loss on ignition determination,           After cooling, dissolve the residue in diluted hydrochloric
melting point determination, test for metal particles in           acid (1 in 5), and filter if necessary. The filtrate yields a
ophthalmic ointments, methanol determination, microbial            white precipitate upon addition of barium chloride TS.
assay for antibiotics, test for microbial limit, test for
microbial limit for crude drugs, mineral oil determination,
nitrogen determination, nuclear magnetic resonance spec-               2.01       Liquid Chromatography
troscopy, optical rotation determination, osmolarity deter-
mination, oxygen flask combustion method, particle size
                                                                   Change to read:
distribution test for preparations, pH determination, test for
plastic containers, powder particle density determination,            Liquid Chromatography is a method to develop a mixture
powder particle size determination, test for pyrogen,              injected into a column prepared with a suitable stationary
qualitative test, test for readily carbonizable substances,        phase by passing a liquid as a mobile phase through the
refractive index determination, residual solvents test, residue    column, in order to separate the mixture into its components
on ignition determination, test for rubber closure for aque-       by making use of the difference of retention capacity against
ous infusions, specific gravity and density determination,         the stationary phase, and to determine the components. This
specific surface area determination, test for sterility, sulfate   method can be applied to a liquid or soluble sample, and is
determination, thermal analysis, thin-layer chromatogra-           used for identification, purity test, and quantitative determi-
phy, test for total organic carbon, ultravioletvisible spec-       nation.
trophotometry, uniformity of dosage units (test for content           A mixture injected into the column is distributed between
uniformity, mass variation test), viscosity determination,         the mobile phase and the stationary phase with a characteris-
vitamin A assay, water determination, and X-ray powder             tic ratio (k) for each component.
diffraction are performed as directed in the corresponding
                                                                            amount of compound in the stationary phase
articles under the General Tests, Processes and Apparatus.             k=
                                                                             amount of compound in the mobile phase
The tests for melting point of fats, congealing point of fatty
acids, specific gravity, acid value, saponification value, ester     The ratio k represents the mass distribution ratio k? in liq-
value, hydroxyl value, unsaponifiable matter and iodine            uid chromatography.
value of fats and fatty oils are performed as directed in the        Since the relation given below exists among the ratio (k),
corresponding items under Fats and Fatty Oils Test, and            the time for which the mobile phase is passed through the
sampling, preparation of sample for analysis, microscopic          column (t0: time measured from the time of injection of a

                                                                                                                            1795
1796       General Tests, Processes and Apparatus                                                  Supplement I, JP XV

compound with k=0 to the time of elution at the peak maxi-      the peak shape of the sample is unchanged after mixing the
mum), and the retention time (tR: time measured from the        sample with an authentic specimen.
time of injection of a compound to be determined to the            In general, the purity of the sample is determined by com-
time of elution at the peak maximum), the retention time for    paring the sample solution with a standard solution which is
a compound on a column has a characteristic value under         prepared by diluting the sample solution to a concentration
fixed chromatographic conditions.                               corresponding to the specified limit amount of the impurity,
                                                                or by the peak area percentage method. Unless otherwise
                        tR=(1+k) t0
                                                                specified, if a sample is separated into isomers in the chro-
Apparatus                                                       matogram, the isomer ratio is calculated by using the peak
   Basically, the apparatus required for the liquid chromato-   area percentage method.
graphic procedure consists of a pumping system for the             The peak area percentage method is a method to calculate
mobile phase, a sample injection port, a column, a detector     the proportion of the components from the ratio of the peak
and a recorder. A mobile phase component regulator, a ther-     area of each component to the sum of the peak areas of
mostat for the column, a pumping system for reaction            every peak recorded in the chromatogram. In order to
reagents and a chemical reaction chamber are also used, if      obtain accurate results in evaluating the proportion of the
necessary. The pumping system serves to deliver the mobile      components, it is necessary to correct the area of each
phase and the reagents into the column and connecting tube      component based on the relative response factor to the prin-
at a constant flow rate. The sample injection port is used to   cipal component.
deliver a quantity of the sample to the apparatus with high
                                                                Assay
reproducibility. The column is a tube with a smooth interior,
                                                                   (1) Internal standard method—In the internal standard
made of inert metal, etc., in which a packing material for
                                                                method, choose a stable compound as an internal standard
liquid chromatography is uniformly packed. A column with
                                                                which shows a retention time close to that of the compound
a stationary phase chemically bound on the inside wall
                                                                to be assayed, and whose peak is well separated from all
instead of the column packed with the packing material may
                                                                other peaks in the chromatogram. Prepare several kinds of
be used. The detector is used to detect a property of the
                                                                standard solutions containing a fixed amount of the internal
samples which is different from that of the mobile phase,
                                                                standard and several graded amounts of the authentic speci-
and may be an ultraviolet or visible spectrophotometer,
                                                                men specified in the individual monograph. Based on the
fluorometric detector, differential refractometer, elec-
                                                                chromatogram obtained by injection of a fixed volume of
trochemical detector, chemiluminescence detector, electric
                                                                individual standard solutions, calculate the ratio of peak
conductivity detector, mass spectrophotometer, etc. The
                                                                area or peak height of the authentic specimen to that of the
output signal is usually proportional to the concentration of
                                                                internal standard, and prepare a calibration curve by plot-
samples at amounts of less than a few mg. The recorder is
                                                                ting these ratios on the ordinate against the amount of the
used to record the output signals of the detector. As
                                                                authentic specimen or the ratio of the amount of the authen-
required, a data processor may be used as the recorder to
                                                                tic specimen to that of the internal standard on the abscissa.
record or output the chromatogram, retention times or
                                                                The calibration curve is usually obtained as a straight line
amounts of the components. The mobile phase component
                                                                passing through the origin. Then, prepare a sample solution
regulator is used to vary the ratio of the mobile phase
                                                                containing the internal standard in the same amount as in
components in a stepwise or gradient fashion.
                                                                the standard solutions used for the preparation of the
Procedure                                                       calibration curve according to the method specified in the
  Fix the detector, column and mobile phase to the appara-      individual monograph, perform the liquid chromatography
tus, and adjust the flow rate and the column temperature to     under the same operating conditions as for the preparation
the values described in the operating conditions specified in   of the calibration curve, calculate the ratio of the peak area
the individual monograph. Inject a volume of the sample so-     or peak height of the objective compound to that of the in-
lution or the standard solution specified in the individual     ternal standard, and read the amount of the compound from
monograph with the sample injector into the column              the calibration curve.
through the sample injection port. The separated compo-            In an individual monograph, generally one of the stan-
nents are detected by the detector, and recorded by the         dard solutions with a concentration within the linear range
recorder as a chromatogram. If the components to be             of the calibration curve and a sample solution with a concen-
analyzed have no readily detectable physical properties such    tration close to that of the standard solution are prepared,
as absorbance or fluorescence, the detection is achieved by     and the chromatography is performed with these solutions
changing the components to suitable derivatives. Usually,       under fixed conditions to determine the amount of the
the derivatization is performed as a pre- or post-column        objective compound.
labeling.                                                          (2) Absolute calibration curve method—Prepare stan-
                                                                dard solutions with several graded amounts of the authentic
Identification and purity test
                                                                specimen, and inject accurately a fixed volume of these stan-
  Identification of a component of a sample is performed by
                                                                dard solutions. With the chromatogram obtained, prepare a
confirming agreement of the retention time of the sample
                                                                calibration curve by plotting the peak areas or peak heights
with that of an authentic specimen, or by confirming that
Supplement I, JP XV                                                  General Tests, Processes and Apparatus                    1797

on the ordinate against the amount of the authentic speci-          tability'' is usually required, and in order to confirm, in
men on the abscissa. The calibration curve is generally             some degree, the linearity of response near its specification
obtained as a straight line passing through the origin. Then,       limit, the range of expected response to the injection of a
prepare a sample solution according to the method specified         certain volume of target impurity solution at the concentra-
in the individual monograph, perform the liquid chro-               tion of its specification limit should be prescribed. For limit
matography under the same conditions as for the prepara-            test, ``Test for required detectability'' is not required, if the
tion of the calibration curve, measure the peak area or peak        test is performed by comparing the response from sample so-
height of the objective compound, and read the amount of            lution with that from standard solution at the concentration
the compound from the calibration curve.                            of its specification limit. ``Test for required detectability'' is
   In an individual monograph, generally one of the stan-           also not required, if it is confirmed that the impurity can be
dard solutions with a concentration within the linear range         detected at its specification limit by the evaluation of ``Sys-
of the calibration curve and a sample solution with a concen-       tem repeatability'' or some other procedure.
tration close to that of the standard solution are prepared,           (2) System performance
and the chromatography is performed with these solutions               When it is confirmed that the specificity for determining
under a fixed condition to obtain the amount of the compo-          the test ingredient is ensured, it is considered verified that
nent. In this method, all procedures, such as the injection         the system used has adequate performance to achieve its in-
procedure, must be carried out under a strictly constant            tended use.
condition.                                                             In assay, ``System performance'' should be defined by the
                                                                    resolution between the test ingredient and a target substance
Method for peak measuring
                                                                    to be separated (a closely eluting compound is preferable),
  Generally, the following methods are used.
                                                                    and when appropriate, by their order of elution. In purity
  (1) Peak height measuring method
                                                                    tests, both the resolution and the order of elution between
  (i) Peak height method: Measure the distance between
                                                                    the test ingredient and a target substance to be separated (a
the maximum of the peak and the intersecting point of a
                                                                    closely eluting compound is preferable) should be
perpendicular line from the maximum of the peak to the
                                                                    prescribed. In addition, if necessary, the symmetry factor of
horizontal axis of recording paper with a tangent linking the
                                                                    the test ingredient should be prescribed together with them.
baselines on both sides of the peak.
                                                                    However, if there is no suitable target substance to be sepa-
  (ii) Automatic peak height method: Measure the signals
                                                                    rated, it is acceptable to define ``System performance'' using
from the detector as the peak height using a data processing
                                                                    the number of theoretical plates and the symmetry factor of
system.
                                                                    the test ingredient.
  (2) Peak area measuring method
                                                                       (3) System repeatability
  (i) Width at half-height method: Multiply the peak
                                                                       When it is confirmed that the degree of variation (preci-
width at the half-height by the peak height.
                                                                    sion) of the response of the test ingredient is at a level that
  (ii) Automatic integration method: Measure the signals
                                                                    meets the requirement of ``System repeatability'', it is consi-
from the detector as the peak area using a data processing
                                                                    dered verified that the system used has adequate perfor-
system.
                                                                    mance to achieve its intended use.
System suitability                                                     The allowable limit of ``System repeatability'' is normally
   System suitability is an integral part of analytical methods     defined as the relative standard deviation (RSD) of the
using chromatography, and is used to ensure that the perfor-        response of the test ingredient in replicate injections of stan-
mance of the chromatographic systems used is adequate for           dard solution. It is acceptable to confirm the repeatability of
the analysis of the drug to be tested, as the suitability of the    the system not only by replicate injections of standard solu-
method for the evaluation of the quality of the drug was            tion before sample injections, but also by divided injections
verified. System suitability test should be carried out at every    of standard solution before and after sample injections, or
series of drug analysis. The test procedures and acceptance         by interspersed injections of standard solution among sam-
criteria of system suitability must be prescribed in the test       ple injections.
method of the drug. Sample analysis is not acceptable unless           In principle, total number of replicate injections should be
the requirements of system suitability have been met.               6. However, in the case that a long time is necessary for one
   In system suitability test of the chromatographic systems,       analysis, such as the analysis using the gradient method, or
the evaluation of ``System performance'' and ``System               the analysis of samples containing late eluting components,
repeatability'' is usually required. For purity tests, the evalu-   it may be acceptable to decrease the number of replicate in-
ation of ``Test for required detectability'' may also be re-        jections by adopting new allowable limit of ``System repeat-
quired.                                                             ability'' which can guarantee a level of ``System repeatabili-
   (1) Test for required detectability                              ty'' equivalent to that at 6 replicate injections.
   For purity tests, when it is confirmed that the target im-          The allowable limit of ``System repeatability'' should be
purity is distinctly detected at the concentration of its           set at an appropriate level based on the validation data when
specification limit, it is considered verified that the system      the suitability of the method for the evaluation of the quality
used has adequate performance to achieve its intended use.          of the drug was verified.
   For quantitative purity tests, ``Test for required detec-
1798       General Tests, Processes and Apparatus                                                   Supplement I, JP XV

Point to consider on changing the operating conditions            Relative standard deviation: Generally, it is defined as
   Among the operating conditions specified in the individ-      RSD (z) in the following equation.
ual monograph, inside diameter and length of the column,                                                 n
particle size of the packing material, column temperature,
                                                                                     100
                                                                                                        S(x -X)
                                                                                                        i= 1
                                                                                                             ˜  i
                                                                                                                     2

composition ratio of the mobile phase, composition of                       RSD (z)=     ×
                                                                                      X
                                                                                      ˜                        n-1
buffer solutions in the mobile phase, pH of the mobile
phase, concentration of ion-pair forming agents in the mo-         xi: Observed value,
bile phase, ionic strength of the mobile phase, flow rate of       X: Mean of observed values,
                                                                    ˜
the mobile phase, number and timing of mobile phase com-           n: Number of replicate measurements.
position changes in gradient program, flow rate of mobile
                                                                    Complete separation of peak: It means that the resolution
phase in gradient program, composition and flow rate of
                                                                 between two peaks is not less than 1.5. It is also called as
derivatizing reagents, and reaction time and chamber tem-
                                                                 ``baseline separation''.
perature in chemical reaction may be modified within the
ranges in which the liquid chromatographic system used con-        Peak-valley ratio: It indicates the degree of separation be-
forms to the requirements of system suitability.                 tween 2 peaks on a chromatogram when baseline separation
                                                                 cannot be attained, and is defined as p/v by the following
Terminology
                                                                 formula.
  S/N ratio: It is defined by the following formula.
                                                                                                   Hp
                                 2H                                                       p/ v =
                         S/N=                                                                      Hv
                                  h
                                                                   Hp: peak height from the baseline of the minor peak,
  H: Peak height of the target ingredient peak from the
                                                                   Hv: height from the baseline of the lowest point (peak
      baseline (the median value of background noise),
                                                                       valley) of the curve between major and minor peaks.
  h: Width of background noise of the chromatogram of
     sample solution or solvent blank around the peak of
     the target ingredient.

  The baseline and background noise are measured over a
range 20 times of peak width at the center point of peak
height of the target ingredient. When a solvent blank is used,
measure over almost the same range as mentioned above
around the point where the target ingredient elutes.




                                                                    Separation factor: It shows the relation between the reten-
                                                                 tion times of peaks in the chromatogram, and is defined as a
                                                                 in the following equation.

                                                                                              tR2-t0
                                                                                         a=
                                                                                              tR1-t0

                                                                   tR1, tR2: Retention times of two compounds used for the
                                                                             resolution measurement (tR1ºtR2),
                                                                   t0: Time of passage of the mobile phase through the
  Symmetry factor: It shows the degree of symmetry of a                column (time measured from the time of injection of a
peak in the chromatogram, and is defined as S in the follow-           compound with k=0 to the time of elution at the peak
ing equation.                                                          maximum).
                             W0.05 h                               The separation factor (a) indicates thermodynamic differ-
                        S=
                              2f                                 ence in partition of two compounds. It is basically the ratio
  W0.05 h: Width of the peak at one-twentieth of the peak        of their partition equilibrium coefficients or of their mass-
           height,                                               distribution ratios, and is obtained from the chromatogram
  f: Distance between the perpendicular from the peak max-       as the ratio of the retention times of the two compounds.
     imum and the leading edge of the peak at one-twentieth         Resolution: It shows the relation between the retention
     of the peak height,                                         time and the peak width of peaks in the chromatogram, and
where W0.05 h and f have the same unit.                          is defined as RS in the following equation.
Supplement I, JP XV                                              General Tests, Processes and Apparatus                  1799

                                                                port and flow regulator for a combustion gas, a burning
                                tR2-tR1
                RS=1.18×                                        supporting gas and an accessory gas and sample injection
                             W0.5 h1+W0.5 h2
                                                                port for headspace are also used, if necessary. The carrier
  tR1, tR2: Retention times of two compounds used for the       gas-introducing port and flow regulator serves to deliver the
            measurement of resolution (tR1ºtR2),                carrier gas into the column at a constant flow rate, and
  W0.5 h1, W0.5 h2: Peak widths at half peak height,            usually consist of a pressure regulation valve, a flow rate
                                                                regulation valve and a pressure gauge. The sample injection
where tR1, tR2, W0.5 h1 and W0.5 h2 have the same unit.
                                                                port is used to deliver a quantity of the sample to the flow
  Number of theoretical plates: It indicates the extent of      line of carrier gas with high reproducibility. There are
band broadening of a compound in the column, and is             sample injection ports for packed column and for capillary
generally defined as N in the following equation.               column. There are both divided injection mode and non-
                                                                divided injection mode to sample injection port for capillary
                                  tR2
                     N=5.54×                                    column. The columns are usually classified as packed
                                 W0.5 h2
                                                                column or capillary column. The packed column is a tube
  tR: Retention time of compound,                               made of inert metal, glass or synthetic resin, in which a
  W0.5 h: Width of the peak at half peak height,                packing material for gas chromatography is uniformly pack-
                                                                ed. The packed column with not more than 1 mm in inside
where tR and W0.5 h have the same unit.
                                                                diameter is also called a packed capillary column (micro
Note Avoid the use of authentic specimens, internal stan-       packed column). A capillary column is a tube made of inert
dards, reagents or solvents containing substances that may      metal, glass, quartz or synthetic resin, whose inside wall is
interfere with the determination.                               bound chemically with stationary phase for gas chro-
                                                                matography. The column oven has the setting capacity for a
                                                                column with required length and the temperature regulation
      2.02       Gas Chromatography                             system for keeping the constant column temperature. The
                                                                detector is used to detect a component separated on the
                                                                column, and may be an alkaline thermal ionization detector,
Change to read:
                                                                a flame photometry detector, mass spectrophotometer,
   Gas Chromatography is a method to develop a mixture          hydrogen flame-ionization detector, an electron capture
injected into a column prepared with a suitable stationary      detector, a thermal conductivity detector, etc. The recorder
phase by passing a gas (carrier gas) as a mobile phase          is used to record the output signals of the detector.
through the column, in order to separate the mixture into its
                                                                Procedure
components by making use of the difference of retention
                                                                   Unless otherwise specified, proceed by the following
capacity against the stationary phase, and to determine the
                                                                method. Fix the detector, column and carrier gas to the
components. This method can be applied to a gaseous or
                                                                apparatus, and adjust the flow rate and the column tempera-
vaporizable sample, and is used for identification, purity
                                                                ture to the values described in the operating conditions speci-
test, and quantitative determination.
                                                                fied in the individual monograph. Inject a volume of the
   A mixture injected into the column is distributed between
                                                                sample solution or the standard solution specified in the
the mobile phase and the stationary phase with a characteris-
                                                                individual monograph with the sample injector into the
tic ratio (k) for each component.
                                                                column system through the sample injection port. The sepa-
         amount of compound in the stationary phase             rated components are detected by the detector, and recorded
    k=
          amount of compound in the mobile phase                by the recorder as a chromatogram.
   Since the relation given below exists among the ratio (k),   Identification and purity test
the time for which the mobile phase is passed through the         Identification of a component of a sample is performed by
column (t0: time measured from the time of injection of a       confirming agreement of the retention time of the sample
compound with k=0 to the time of elution at the peak maxi-      with that of an authentic specimen, or by confirming that
mum), and the retention time (tR: time measured from the        the peak shape of the sample is unchanged after mixing the
time of injection of a compound to be determined to the         sample with an authentic specimen.
time of elution at the peak maximum), the retention time for      In general, the purity of the sample is determined by com-
a compound on a column has a characteristic value under         paring the sample solution with a standard solution which is
fixed chromatographic conditions.                               prepared by diluting the sample solution to a concentration
                                                                corresponding to the specified limit amount of the impurity,
                        tR=(1+k)t0
                                                                or by the peak area percentage method. Unless otherwise
Apparatus                                                       specified, if a sample is separated into isomers in the chro-
  Basically, the apparatus required for the gas chromato-       matogram, the isomer ratio is calculated by using the peak
graphic procedure consists of a carrier gas-introducing port    area percentage method.
and flow regulator, a sample injection port, a column, a          The peak area percentage method is a method to calculate
column oven, a detector and a recorder. Gas introducing         the proportion of the components from the ratio of the peak
1800       General Tests, Processes and Apparatus                                                     Supplement I, JP XV

area of each component to the sum of the peak areas of           graphy under the same conditions as for the preparation of
every peak recorded in the chromatogram. In order to             the calibration curve, measure the peak area or peak height
obtain accurate results in evaluating the proportion of the      of the objective compound, and read the amount of the
components, it is necessary to correct the area of each          compound from the calibration curve.
component based on its relative response factor to the prin-        In an individual monograph, generally one of the stan-
cipal component.                                                 dard solutions with a concentration within the linear range
                                                                 of the calibration curve and a sample solution with a concen-
Assay
                                                                 tration close to that of the standard solution are prepared,
   In general, perform the assay by using the internal stan-
                                                                 and the chromatography is performed with these solutions
dard method. The absolute calibration curve method is used
                                                                 under a fixed condition to obtain the amount of the compo-
when a suitable internal standard is not available. Perform
                                                                 nent. In this method, all procedures, such as the injection
the assay by using the standard addition method when the
                                                                 procedure, must be carried out under a strictly constant
effect of the component other than the compound to be
                                                                 condition.
assayed on the quantitative determination is not negligible
                                                                    (3) Standard addition method—Pipet a fixed volume of
against a result of the determination.
                                                                 more than 4 sample solutions, add exactly the standard
   (1) Internal standard method—In the internal standard
                                                                 solution so that stepwise increasing amounts of the object
method, choose a stable compound as an internal standard
                                                                 compound are contained in the solutions except 1 sample
which shows a retention time close to that of the compound
                                                                 solution, diluted exactly each solution with and without
to be assayed, and whose peak is well separated from all
                                                                 standard solution to a definite volume, and use each solution
other peaks in the chromatogram. Prepare several kinds of
                                                                 as the sample solution. Based on the chromatogram ob-
standard solutions containing a fixed amount of the internal
                                                                 tained by exact injection of a fixed volume of individual
standard and several graded amounts of the authentic speci-
                                                                 sample solutions, measure the peak area or peak height of
men specified in the individual monograph. Based on the
                                                                 individual sample solutions. Calculate the concentration of
chromatogram obtained by injection of a fixed volume of
                                                                 standard objective compound added into each sample solu-
individual standard solutions, calculate the ratio of peak
                                                                 tion, plot the amounts (concentration) of added standard
area or peak height of the authentic specimen to that of the
                                                                 object compound on the abscissa and the peak area or peak
internal standard, and prepare a calibration curve by plot-
                                                                 height on the ordinate on the graph, extend the calibration
ting these ratios on the ordinate against the amount of the
                                                                 curve obtained by linking the plots, and determine the
authentic specimen or the ratio of the amount of the authen-
                                                                 amount of object compound to be assayed from the distance
tic specimen to that of the internal standard on the abscissa.
                                                                 between the origin and the intersecting point of the calibra-
The calibration curve is usually obtained as a straight line
                                                                 tion curve with the abscissa. This method is available only in
passing through the origin. Then, prepare a sample solution
                                                                 the case that the calibration curve is a straight line, and pass-
containing the internal standard in the same amount as in
                                                                 es through the origin when the absolute calibration curve
the standard solutions used for the preparation of the
                                                                 method is employed. In this method, all procedures must be
calibration curve according to the method specified in the
                                                                 carried out under a strictly constant condition.
individual monograph, perform the gas chromatography
under the same operating conditions as for the preparation       Method for peak measuring
of the calibration curve, calculate the ratio of the peak area     Generally, the following methods are used.
or peak height of the objective compound to that of the            (1) Peak height measuring method
internal standard, and read the amount of the compound             (i) Peak height method: Measure the distance between
from the calibration curve.                                      the maximum of the peak and the intersecting point of a per-
   In an individual monograph, generally one of the stan-        pendicular line from the maximum of the peak to the
dard solutions with a concentration within the linear range      horizontal axis of recording paper with a tangent linking the
of the calibration curve and a sample solution with a concen-    baselines on either side of the peak.
tration close to that of the standard solution are prepared,       (ii) Automatic peak height method: Measure the signals
and the chromatography is performed with these solutions         from the detector as the peak height using a data processing
under fixed conditions to determine the amount of the            system.
objective compound.                                                (2) Peak area measuring method
   (2) Absolute calibration curve method—Prepare stan-             (i) Width at half-height method: Multiply the peak
dard solutions with several graded amounts of the authentic      width at the half-height by the peak height.
specimen, and inject accurately a fixed volume of these            (ii) Automatic integration method: Measure the signals
standard solutions. With the chromatogram obtained, pre-         from the detector as the peak area using a data processing
pare a calibration curve by plotting the peak areas or peak      system.
heights on the ordinate against the amount of the authentic
                                                                 System suitability
specimen on the abscissa. The calibration curve is generally
                                                                   Refer to ``System suitability'' described under 2.01 Liquid
obtained as a straight line passing through the origin. Then,
                                                                 Chromatography.
prepare a sample solution according to the method specified
in the individual monograph, perform the gas chromato-
Supplement I, JP XV                                                 General Tests, Processes and Apparatus                   1801

Point to consider in changing the operating conditions             tion, and allow to stand for more than 24 hours before use.
   Among the operating conditions specified in the individ-           These Karl Fischer TSs, prepared by any one of the above
ual monograph, inside diameter and length of column, parti-        methods, must be standardized before every use, because of
cle size of packing material, concentration or thickness of        its activity change with the lapse of time. Further preserve
stationary phase, column temperature, temperature rising-          the TS in a cold place, protecting it from light and moisture.
rate, kind and flow rate of carrier gas, and split ratio may be       Standardization—According to the procedure described
modified within the ranges in which the gas chromatograph-         below, take a suitable quantity of methanol for Karl Fischer
ic system used conforms to the requirements of system              method in a dried titration flask, and titrate the solvent with
suitability. Headspace sample injection device and its oper-       a Karl Fischer TS to make the inside of the flask anhydrous.
ating conditions may be also modified, provided that they          Then, weigh about 30 mg of water accurately and put it in
give equivalent or more accuracy and precision.                    the titration flask quickly, and titrate the water dissolved in
                                                                   the solvent with a Karl Fischer TS to the end point, under
Terminology
                                                                   vigorous stirring. Calculate the water equivalence factor,
  The definition of terms described under 2.01 Liquid Chro-
                                                                   f (mg/mL), corresponding to the amount of water (H2O) in
matography shall apply in 2.02 Gas Chromatography.
                                                                   mg per 1 mL of the Karl Fischer TS by using the following
Note Avoid the use of authentic specimens, internal stan-          equation:
dards, reagents or solvents containing substances that may
                                                                                      Amount of water taken (H2O) (mg)
interfere with the determination.                                     f (mg/mL)=
                                                                                     Volume of Karl Fischer TS consumed
                                                                                     for titration of water (H2O) (mL)

       2.48 Water Determination
          (Karl Fischer Method)                                    Change to read:

Change to read the following part under 1.                                    2.49     Optical Rotation
Volumetric titration:
                                                                                      Determination
Preparation of test solutions and standard solutions
   (1) Karl Fischer TS for water determination                        Optical Rotation Determination is a method for the
   Prepare according to the following method (i), (ii) or (iii).   measurement of the angular rotation of the sample using a
Additives may be added for the purpose of improving the            polarimeter.
stability or other performances if it is confirmed that they          Generally, the vibrations of light take place on planes
give almost the same results as those obtained from the spe-       perpendicular to the direction of the beam. In case of the or-
cified method.                                                     dinary light, the directions of the planes are unrestricted,
   (i) Preparation 1                                               while in case of the plane polarized light, commonly called
   Dissolve 63 g of iodine in 100 mL of pyridine for Karl          as polarized light, the vibrations take place on only one
Fischer method, cool the solution in ice bath, and pass dried      plane that includes the advancing direction of the beam.
sulfur dioxide gas through this solution until the mass in-        And it is called that these beams have plane of polarization.
crease of the solution reaches 32 g. Then make up to 500 mL        Some drugs in the liquid state or in solution have a property
by adding chloroform for Karl Fischer method or methanol           of rotating the plane of the polarized light either to the right
for Karl Fischer method, and allow to stand for more than          or to the left. This property is referred to as optical activity
24 hours before use.                                               or optical rotation, and is inherently related to the chemical
   (ii) Preparation 2                                              constitution of the substance.
   Dissolve 102 g of imidazole for Karl Fischer method in 350         The optical rotation is a degree of rotation of polarized
mL of diethylene glycol monoethyl ether for water determi-         plane, caused by the optically active substance or its solu-
nation, cool the solution in ice bath, and pass dried sulfur di-   tion, and it is measured by the polarimeter. This value is
oxide gas through this solution until the mass increase of the     proportional to the length of the polarimeter tube, and is
solution reaches 64 g, keeping the temperature between 259    C    also related to the solution concentration, the temperature
and 309 Then dissolve 50 g of iodine in this solution, and
         C.                                                        and the measurement wavelength. The character of the rota-
allow to stand for more than 24 hours before use.                  tion is indicated by the direction of the rotation, when facing
   (iii) Preparation 3                                             to the advancing direction of the polarized light. Thus in
   Pass dried sulfur dioxide gas through 220 mL of propy-          case of rotation to the right, it is called dextrorotatory and
lene carbonate for water determination until the mass in-          expressed by placing plus sign (+), while in case of rotation
crease of the solvent reaches 32 g. To this solution, add 180      to the left, it is called levorotatory and expressed by placing
mL of propylene carbonate, or diethylene glycol monoethyl          minus sign (-) before the figure of the angular rotation. For
ether for water determination, in which 81 g of 2-methyl-          example, +209means 209of rotation to the right, while
aminopyridine for Karl Fischer method is dissolved and             -209means 209of rotation to the left.
cooled in ice bath. Then dissolve 36 g of iodine in this solu-        The angular rotation at means degree of rotation, when it
                                                                                                x
1802       General Tests, Processes and Apparatus                                                    Supplement I, JP XV

is measured at t9 by using specific monochromatic light x
                 C                                               from the contents of a sufficient number of containers. If
(expressed by wavelength of light source or the specific beam    test specimens are diluted with fluid medium, the test should
name). Usually, the measurement is performed at 209 or    C      be performed quickly. In performing the test, precautions
259 with a polarimeter tube of 100 mm in length, and with
    C,                                                           must be taken to prevent biohazard.
the D line of sodium lamp.
                                                                 I. Microbiological Examination of Non-sterile Products:
   The specific rotation is expressed by the following
                                                                 Microbial Enumeration Tests
equation:
                                                                   These tests are harmonized with the European Phar-
                                  100 a                          macopoeia and the U.S. Pharmacopeia.
                        [a ]t =
                            x
                                    lc
                                                                 1 Introduction
  t: The temperature of measurement.                               The tests described hereafter will allow quantitative
  x: The wavelength or the name of the specific monochro-        enumeration of mesophilic bacteria and fungi which may
matic light (in the case of the Sodium D line, it is described   grow under aerobic conditions.
as D).                                                             The tests are designed primarily to determine whether a
  a: The angle, in degrees, of rotation of the plane of the      substance or preparation complies with an established
polarized light.                                                 specification for microbiological quality. When used for
  l: The thickness of the layer of sample solution, i.e., the    such purposes follow the instructions given below, including
length of the polarimeter tube (mm).                             the number of samples to be taken and interpret the results
  c: Drug concentration in g/mL. When an intact liquid           as stated below.
drug is used for the direct measurement without dilution by        The methods are not applicable to products containing
an appropriate solvent, c equals to its density (g/mL).          viable micro-organisms as active ingredients.
However, unless otherwise specified, the specific gravity is       Alternative microbiological procedures, including auto-
conventionally used in stead of the density.                     mated methods, may be used, provided that their equiva-
                                                                 lence to the Pharmacopoeial method has been demonstrat-
   The description in the monograph, for example, ``[a]20: D
                                                                 ed.
-33.0 – -36.09 (after drying, 1 g, water, 20 mL, 100
mm),'' means the measured specific rotation [a]20 should be
                                                 D               2 General Procedures
in the range of -33.09and -36.09 when 1 g of accurately
                                     ,                              Carry out the determination under conditions designed to
weighed sample dried under the conditions, specified in the      avoid extrinsic microbial contamination of the product to be
test item of Loss on drying, is taken, and dissolved in water    examined. The precautions taken to avoid contamination
to make exactly 20 mL, then put in the polarimeter tube of       must be such that they do not affect any micro-organisms
100 mm length, of which temperature is kept at 209    C.         which are to be revealed in the test.
                                                                    If the product to be examined has antimicrobial activity,
                                                                 this is insofar as possible removed or neutralized. If inactiva-
  4.01        Bacterial Endotoxins Test                          tors are used for this purpose their efficacy and their absence
                                                                 of toxicity for micro-organisms must be demonstrated.
                                                                    If surface-active substances are used for sample prepara-
Change to read the Preparation of Standard
                                                                 tion, their absence of toxicity for micro-organisms and their
Endotoxin Stock Solution as follows:
                                                                 compatibility with inactivators used must be demonstrated.
Preparation of Standard Endotoxin Stock Solution
                                                                 3 Enumeration Methods
  Prepare Standard Endotoxin Stock Solution by dissolving
                                                                   Use the membrane filtration method, or the plate-count
Endotoxin Reference Standard in water for bacterial
                                                                 methods, as prescribed. The most probable number (MPN)
endotoxins test (BET). Endotoxin is expressed in Endotoxin
                                                                 method is generally the least accurate method for microbial
Units (EU). One EU is equal to one International Unit (IU)
                                                                 counts, however, for certain product groups with very low
of endotoxin.
                                                                 bioburden, it may be the most appropriate method.
                                                                   The choice of a method is based on factors such as the
                                                                 nature of the product and the required limit of micro-organ-
       4.05       Microbial Limit Test                           isms. The method chosen must allow testing of a sufficient
                                                                 sample size to judge compliance with the specification. The
Change to read as follows:                                       suitability of the chosen method must be established.

                                                                 4 Growth Promotion Test and Suitability of the Counting
4.05      Microbiological Examination                            Method
          of Non-sterile Products                                4-1 General considerations
                                                                   The ability of the test to detect micro-organisms in the
   This chapter includes microbial enumeration tests and         presence of product to be tested must be established.
tests for specified micro-organisms. For the test, use a mix-      Suitability must be confirmed if a change in testing perfor-
ture of several portions selected at random from the bulk or     mance, or the product, which may affect the outcome of the
Supplement I, JP XV                                               General Tests, Processes and Apparatus                 1803

                                Table 4.05-I-1   Preparation and use of test micro-organisms

                                                                                       Suitability of counting method in
                                                      Growth promotion
                                                                                          the presence of the product
                          Preparation of
  Micro-organism
                            test strain
                                                Total aerobic    Total yeasts and       Total aerobic       Total yeasts and
                                               microbial count    moulds count         microbial count       moulds count

 Staphylococcus        Casein soya bean       Casein soya bean                      Casein soya bean
 aureus                digest agar or         digest agar and                       digest agar/
                       casein soya bean       casein soya bean                      MPN casein soya
 such as ATCC          digest broth           digest broth                          bean digest broth
 6538, NCIMB           30 – 359C              ≦100 CFU                              ≦100 CFU
 9518, CIP 4.83 or     18 – 24 h              30 – 359C                             30 – 359C
 NBRC 13276                                   ≦3 days                               ≦3 days

 Pseudomonas           Casein soya bean       Casein soya bean                      Casein soya bean
 aeruginosa            digest agar or         digest agar and                       digest agar/MPN
                       casein soya bean       casein soya bean                      casein soya bean
 such as ATCC          digest broth           digest broth                          digest broth
 9027, NCIMB           30 – 359C              ≦100 CFU                              ≦100 CFU
 8626, CIP 82.118      18 – 24 h              30 – 359C                             30 – 359C
 or NBRC 13275                                ≦3 days                               ≦3 days

 Bacillus subtilis  Casein soya bean          Casein soya bean                      Casein soya bean
                    digest agar or            digest agar and                       digest agar/MPN
 such as ATCC       casein soya bean          casein soya bean                      casein soya bean
 6633, NCIMB        digest broth              digest broth                          digest broth
 8054, CIP 52.62 or 30 – 359C                 ≦100 CFU                              ≦100 CFU
 NBRC 3134          18 – 24 h                 30 – 359C                             30 – 359C
                                              ≦3 days                               ≦3 days

 Candida albicans      Sabouraud-dextrose     Casein soya bean   Sabouraud-dex-     Casein soya bean        Sabouraud-
                       agar or Sabouraud-     digest agar        trose agar         digest agar             dextrose agar
 such as ATCC          dextrose broth         ≦100 CFU           ≦100 CFU           ≦100 CFU                ≦100 CFU
 10231, NCPF           20 – 259C              30 – 359C          20 – 259C          30 – 359C               20 – 259C
 3179, IP 48.72 or     2 – 3 days             ≦5 days            ≦5 days            ≦5 days                 ≦5 days
 NBRC 1594                                                                          MPN: not applicable

 Aspergillus niger     Sabouraud-dextrose     Casein soya bean   Sabouraud-dex-     Casein soya bean        Sabouraud-
                       agar or potato-dex-    digest agar        trose agar         digest agar             dextrose agar
 such as ATCC          trose agar             ≦100 CFU           ≦100 CFU           ≦100 CFU                ≦100 CFU
 16404, IMI            20 – 259 C             30 – 359C          20 – 259C          30 – 359C               20 – 259C
 149007, IP            5 – 7 days, or         ≦5 days            ≦5 days            ≦5 days                 ≦5 days
 1431.83 or NBRC       until good sporula-                                          MPN: not applicable
 9455                  tion is achieved



test is introduced.                                              prepared and then an appropriate volume of the spore
4-2 Preparation of test strains                                  suspension is used for test inoculation. The stable spore sus-
   Use standardised stable suspensions of test strains or        pension may be maintained at 2 – 89 for a validated period
                                                                                                       C
prepare as stated below.                                         of time.
   Seed lot culture maintenance techniques (seed-lot systems)    4-3 Negative control
are used so that the viable micro-organisms used for inocula-       To verify testing conditions a negative control is per-
tion are not more than 5 passages removed from the original      formed using the chosen diluent in place of the test prepara-
master seed-lot. Grow each of the bacterial and fungal test      tion. There must be no growth of micro-organisms.
strains separately as described in Table 4.05-I-1.               4-4 Growth promotion of the media
   Use buffered sodium chloride-peptone solution pH 7.0 or          Test each batch of ready-prepared medium and each batch
phosphate buffer solution pH 7.2 to make test suspensions;       of medium, prepared either from dehydrated medium or
to suspend A. niger spores, 0.05 per cent of polysorbate 80      from the ingredients described.
may be added to the buffer. Use the suspensions within 2 h          Inoculate portions/plates of casein soya bean digest broth
or within 24 h if stored at 2 – 89 As an alternative to
                                     C.                          and casein soya bean digest agar with a small number (not
preparing and then diluting a fresh suspension of vegetative     more than 100 CFU) of the micro-organisms indicated in
cells of A. niger or B. subtilis, a stable spore suspension is   Table 4.05-I-1, using a separate portion/plate of medium for
1804       General Tests, Processes and Apparatus                                                     Supplement I, JP XV

each. Inoculate plates of Sabouraud-dextrose agar with a          the adhesive surface with sterile porous material, for exam-
small number (not more than 100 CFU) of the micro-organ-          ple sterile gauze, to prevent the patches from sticking
isms indicated in Table 4.05-I-1, using a separate plate of       together, and transfer the patches to a suitable volume of the
medium for each. Incubate in the conditions described in          chosen diluent containing inactivators such as polysorbate
Table 4.05-I-1.                                                   80 and/or lecithin. Shake the preparation vigorously for at
   For solid media, growth obtained must not differ by a          least 30 min.
factor greater than 2 from the calculated value for a stan-       4-5-2 Inoculation and dilution
dardized inoculum. For a freshly prepared inoculum,                  Add to the sample prepared as described above (4-5-1) and
growth of the micro-organisms comparable to that previous-        to a control (with no test material included) a sufficient
ly obtained with a previously tested and approved batch of        volume of the microbial suspension to obtain an inoculum
medium occurs.                                                    of not more than 100 CFU. The volume of the suspension of
   Liquid media are suitable if clearly visible growth of the     the inoculum should not exceed 1 per cent of the volume of
micro-organisms comparable to that previously obtained            diluted product.
with a previously tested and approved batch of medium oc-            To demonstrate acceptable microbial recovery from the
curs.                                                             product, the lowest possible dilution factor of the prepared
4-5 Suitability of the counting method in the presence of         sample must be used for the test. Where this is not possible
product                                                           due to antimicrobial activity or poor solubility, further
4-5-1 Preparation of the sample                                   appropriate protocols must be developed.
   The method for sample preparation depends on the physi-           If inhibition of growth by the sample cannot otherwise be
cal characteristics of the product to be tested. If none of the   avoided, the aliquot of the microbial suspension may be ad-
procedures described below can be demonstrated to be satis-       ded after neutralization, dilution or filtration.
factory, an alternative procedure must be developed.              4-5-3 Neutralization/removal of antimicrobial activity
Water-soluble products—Dissolve or dilute (usually a 1 in 10         The number of micro-organisms recovered from the
dilution is prepared) the product to be examined in buffered      prepared sample diluted as described in 4-5-2 and incubated
sodium chloride-peptone solution pH 7.0, phosphate buffer         following the procedure described in 4-5-4, is compared to
solution pH 7.2 or casein soya bean digest broth. If necessa-     the number of micro-organisms recovered from the control
ry adjust to pH 6 – 8. Further dilutions, where necessary, are    preparation.
prepared with the same diluent.                                      If growth is inhibited (reduction by a factor greater than
Non-fatty products insoluble in water—Suspend the product         2), then modify the procedure for the particular enumera-
to be examined (usually a 1 in 10 dilution is prepared) in        tion test to ensure the validity of the results. Modification of
buffered sodium chloride-peptone solution pH 7.0, phos-           the procedure may include, for example, (1) an increase in
phate buffer solution pH 7.2 or casein soya bean digest           the volume of the diluent or culture medium, (2) incorpora-
broth. A surface-active agent such as 1 g/L of polysorbate        tion of a specific or general neutralizing agents into the dil-
80 may be added to assist the suspension of poorly wettable       uent, (3) membrane filtration or (4) a combination of the
substances. If necessary adjust to pH 6 – 8. Further dilu-        above measures.
tions, where necessary, are prepared with the same diluent.       Neutralizing agents—Neutralizing agents may be used to
Fatty products—Dissolve in isopropyl myristate, sterilised        neutralize the activity of antimicrobial agents (Table
by filtration or mix the product to be examined with the          4.05-I-2). They may be added to the chosen diluent or the
minimum necessary quantity of sterile polysorbate 80 or           medium preferably before sterilization. If used, their effica-
another non-inhibitory sterile surface-active reagent, heated     cy and their absence of toxicity for micro-organisms must be
if necessary to not more than 409 or in exceptional cases
                                    C,                            demonstrated by carrying out a blank with neutralizer and
to not more than 459 Mix carefully and if necessary main-
                        C.                                        without product.
tain the temperature in a water-bath. Add sufficient of the          If no suitable neutralizing method can be found, it can be
pre-warmed chosen diluent to make a 1 in 10 dilution of the       assumed that the failure to isolate the inoculated organism is
original product. Mix carefully whilst maintaining the            attributable to the microbicidal activity of the product. This
temperature for the shortest time necessary for the forma-        information serves to indicate that the article is not likely to
tion of an emulsion. Further serial tenfold dilutions may be      be contaminated with the given species of the micro-organ-
prepared using the chosen diluent containing a suitable con-      ism. However, it is possible that the product only inhibits
centration of sterile polysorbate 80 or another non-inhibito-     some of the micro-organisms specified herein, but does not
ry sterile surface-active reagent.                                inhibit others not included amongst the test strains or for
Fluids or solids in aerosol form—Aseptically transfer the         which the latter are not representative. Then, perform the
product into a membrane filter apparatus or a sterile con-        test with the highest dilution factor compatible with microbi-
tainer for further sampling. Use either the total contents or a   al growth and the specific acceptance criterion.
defined number of metered doses from each of the contain-         4-5-4 Recovery of micro-organism in the presence of
ers tested.                                                       product
Transdermal patches—Remove the protective cover sheets               For each of the micro-organisms listed in Table 4.05-I-1,
(``release liner'') of the transdermal patches and place them,    separate tests are performed. Only micro-organisms of the
adhesive side upwards, on sterile glass or plastic trays. Cover   added test strain are counted.
Supplement I, JP XV                                               General Tests, Processes and Apparatus                    1805

                      Table 4.05-I-2    Common neutralizing agents/method for interfering substances

                      Interfering substance                                 Potential neutralizing agents/method

        Glutaraldehyde, Mercurials                                      Sodium hydrogen sulfite (Sodium bisulfite)

        Phenolics, Alcohol, Aldehydes, Sorbate                          Dilution

        Aldehydes                                                       Glycine

        Quaternary Ammonium Compounds (QACs),                           Lecithin
        Parahydroxybenzoates (Parabens),
        Bis-biguanides

        QAC, Parabens, Iodine                                           Polysorbate

        Mercurials                                                      Thioglycollate

        Mercurials, Halogens, Aldehydes                                 Thiosulfate

        EDTA (edetate)                                                  Mg or Ca ions



4-5-4-1 Membrane filtration                                      Petri dishes are used, the volume of the agar is increased
   Use membrane filters having a nominal pore size not           accordingly. Dry the plates, for example in a laminar-air-
greater than 0.45 mm. The type of filter material is chosen in   flow cabinet or in an incubator. For each of the micro-or-
such a way that the bacteria-retaining efficiency is not af-     ganisms listed in Table 4.05-I-1, at least 2 Petri dishes are
fected by the components of the sample to be investigated.       used. Spread a measured volume of not less than 0.1 mL of
For each of the micro-organisms listed in Table 4.05-I-1, one    the sample prepared as described under 4-5-1 to 4-5-3 over
membrane filter is used.                                         the surface of the medium. Incubate and count as prescribed
   Transfer a suitable amount of the sample prepared as          under 4-5-4-2-1.
described under 4-5-1 to 4-5-3 (preferably representing 1 g of   4-5-4-3 Most-probable-number (MPN) method
the product, or less if large numbers of CFU are expected) to       The precision and accuracy of the MPN method is less
the membrane filter, filter immediately and rinse the mem-       than that of the membrane filtration method or the plate-
brane filter with an appropriate volume of diluent.              count method. Unreliable results are obtained particularly
   For the determination of total aerobic microbial count        for the enumeration of moulds. For these reasons the MPN
(TAMC), transfer the membrane filter to the surface of           method is reserved for the enumeration of TAMC in situa-
casein soya bean digest agar. For the determination of total     tions where no other method is available. If the use of the
combined yeasts/moulds count (TYMC) transfer the mem-            method is justified, proceed as follows.
brane to the surface of Sabouraud-dextrose agar. Incubate           Prepare a series of at least 3 serial tenfold dilutions of the
the plates as indicated in Table 4.05-I-1. Perform the count-    product as described under 4-5-1 to 4-5-3. From each level of
ing.                                                             dilution, 3 aliquots of 1 g or 1 mL are used to inoculate 3
4-5-4-2 Plate-count methods                                      tubes with 9 – 10 mL of casein soya bean digest broth. If
   Perform plate-count methods at least in duplicate for each    necessary a surface-active agent such as polysorbate 80, or
medium and use the mean count of the result.                     an inactivator of antimicrobial agents may be added to the
4-5-4-2-1 Pour-plate method                                      medium. Thus, if 3 levels of dilution are prepared 9 tubes are
   For Petri dishes 9 cm in diameter, add to the dish 1 mL of    inoculated.
the sample prepared as described under 4-5-1 to 4-5-3 and 15        Incubate all tubes at 30 – 359 for not more than 3 days.
                                                                                                    C
– 20 mL of casein soya bean digest agar or Sabouraud-dex-        If reading of the results is difficult or uncertain owing to the
trose agar, both media being at not more than 459 If larg-
                                                    C.           nature of the product to be examined, subculture in the same
er Petri dishes are used, the amount of agar medium is in-       broth, or casein soya bean digest agar, for 1 – 2 days at the
creased accordingly. For each of the micro-organisms listed      same temperature and use these results. Determine the most
in Table 4.05-I-1, at least 2 Petri dishes are used.             probable number of micro-organisms per gram or millilitre
   Incubate the plates as indicated in Table 4.05-I-1. Take      of the product to be examined from Table 4.05-I-3.
the arithmetic mean of the counts per medium and calculate       4-6 Results and interpretation
the number of CFU in the original inoculum.                         When verifying the suitability of the membrane filtration
4-5-4-2-2 Surface-spread method                                  method or the plate-count method, a mean count of any of
   For Petri dishes 9 cm in diameter, add 15 – 20 mL of          the test organisms not differing by a factor greater than 2
casein soya bean digest agar or Sabouraud-dextrose agar at       from the value of the control defined in 4-5-2 in the absence
about 459 to each Petri dish and allow to solidify. If larger
           C                                                     of the product must be obtained. When verifying the
1806       General Tests, Processes and Apparatus                                                      Supplement I, JP XV

suitability of the MPN method the calculated value from the         5-2-2 Plate-count methods
inoculum must be within 95 per cent confidence limits of the        5-2-2-1 Pour-plate method
results obtained with the control.                                     Prepare the sample using a method that has been shown to
  If the above criteria cannot be met for one or more of the        be suitable as described in section 4. Prepare for each medi-
organisms tested with any of the described methods, the             um at least 2 Petri dishes for each level of dilution. Incubate
method and test conditions that come closest to the criteria        the plates of casein soya bean digest agar at 30 – 359 forC
are used to test the product.                                       3 – 5 days and the plates of Sabouraud-dextrose agar at 20 –
                                                                    259 for 5 – 7 days. Select the plates corresponding to a
                                                                         C
5 Testing of Products
                                                                    given dilution and showing the highest number of colonies
5-1 Amount used for the test
                                                                    less than 250 for TAMC and 50 for TYMC. Take the
    Unless otherwise prescribed, use 10 g or 10 mL of the
                                                                    arithmetic mean per culture medium of the counts and calcu-
product to be examined taken with the precautions referred
                                                                    late the number of CFU per gram or per millilitre of
to above. For fluids or solids in aerosol form, sample 10
                                                                    product.
containers. For transdermal patches, sample 10 patches.
                                                                    5-2-2-2 Surface-spread method
    The amount to be tested may be reduced for active sub-
                                                                       Prepare the sample using a method that has been shown to
stances that will be formulated in the following conditions:
                                                                    be suitable as described in section 4. Prepare at least 2 Petri
the amount per dosage unit (e.g. tablet, capsule, injection) is
                                                                    dishes for each medium and each level of dilution. For incu-
less than or equal to 1 mg or the amount per gram or mil-
                                                                    bation and calculation of the number of CFU proceed as
lilitre (for preparations not presented in dose units) is less
                                                                    described for the pour-plate method.
than 1 mg. In these cases, the amount of sample to be tested
                                                                    5-2-2-3 Most-probable-number method
is not less than the amount present in 10 dosage units or 10 g
                                                                       Prepare and dilute the sample using a method that has
or 10 mL of the product.
                                                                    been shown to be suitable as described in section 4. Incubate
    For materials used as active substances where sample
                                                                    all tubes for 3 – 5 days at 30 – 359 Subculture if necessary,
                                                                                                        C.
quantity is limited or batch size is extremely small (i.e. less
                                                                    using the procedure shown to be suitable. Record for each
than 1000 mL or 1000 g), the amount tested shall be 1 per
                                                                    level of dilution the number of tubes showing microbial
cent of the batch unless a lesser amount is prescribed or
                                                                    growth. Determine the most probable number of micro-or-
justified and authorised.
                                                                    ganisms per gram or millilitre of the product to be examined
    For products where the total number of entities in a batch
                                                                    from Table 4.05-I-3.
is less than 200 (e.g. samples used in clinical trials), the sam-
                                                                    5-3 Interpretation of the results
ple size may be reduced to 2 units, or 1 unit if the size is less
                                                                       The total aerobic microbial count (TAMC) is considered
than 100.
                                                                    to be equal to the number of CFU found using casein soya
    Select the sample(s) at random from the bulk material or
                                                                    bean digest agar; if colonies of fungi are detected on this
from the available containers of the preparation. To obtain
                                                                    medium, they are counted as part of TAMC. The total com-
the required quantity, mix the contents of a sufficient
                                                                    bined yeasts/mould count (TYMC) is considered to be equal
number of containers to provide the sample.
                                                                    to the number of CFU found using Sabouraud-dextrose
5-2 Examination of the product
                                                                    agar; if colonies of bacteria are detected on this medium,
5-2-1 Membrane filtration
                                                                    they are counted as part of TYMC. When the TYMC is
    Use a filtration apparatus designed to allow the transfer of
                                                                    expected to exceed the acceptance criterion due to the bac-
the filter to the medium. Prepare the sample using a method
                                                                    terial growth, Sabouraud-dextrose agar containing antibiot-
that has been shown suitable as described in section 4 and
                                                                    ics may be used. If the count is carried out by the MPN
transfer the appropriate amount to each of 2 membrane
                                                                    method the calculated value is the TAMC.
filters and filter immediately. Wash each filter following the
                                                                       When an acceptance criterion for microbiological quality
procedure shown to be suitable.
                                                                    is prescribed it is interpreted as follows:
    For the determination of TAMC, transfer one of the
                                                                       -101 CFU: maximum acceptable count=20,
membrane filters to the surface of casein soya bean digest
                                                                       -102 CFU: maximum acceptable count=200,
agar. For the determination of TYMC, transfer the other
                                                                       -103 CFU: maximum acceptable count=2000, and so
membrane to the surface of Sabouraud-dextrose agar. Incu-
                                                                    forth.
bate the plate of casein soya bean digest agar at 30 – 359      C
                                                                       The recommended solutions and media are described in
for 3 – 5 days and the plate of Sabouraud-dextrose agar at
                                                                    Tests for specified micro-organisms.
20 – 259 for 5 – 7 days. Calculate the number of CFU per
          C
gram or per millilitre of product.
    When examining transdermal patches, filter 10 per cent of       II. Microbiological Examination of Non-sterile Products:
the volume of the preparation described under 4-5-1                 Tests for Specified Micro-organisms
separately through each of 2 sterile filter membranes. Trans-
                                                                     These tests are harmonized with the European Phar-
fer one membrane to casein soya bean digest agar for TAMC
                                                                    macopoeia and the U.S. Pharmacopeia.
and the other membrane to Sabouraud-dextrose agar for
TYMC.                                                               1 Introduction
                                                                      The tests described hereafter will allow determination of
Supplement I, JP XV                                              General Tests, Processes and Apparatus              1807

                             Table 4.05-I-3   Most-probable-number values of micro-organisms

              Observed combinations of numbers of tubes
                      showing growth in each set
                                                                          MPN per g or                  95 per cent
               Number of g or mL of product per tube                    per mL of product            confidence limits
         0.1                    0.01                  0.001
          0                       0                       0                Less than 3                    0 – 9.4
          0                       0                       1                      3                       0.1 – 9.5
          0                       1                       0                      3                       0.1 – 10
          0                       1                       1                     6.1                      1.2 – 17
          0                       2                       0                     6.2                      1.2 – 17
          0                       3                       0                     9.4                      3.5 – 35
          1                       0                       0                     3.6                      0.2 – 17
          1                       0                       1                     7.2                      1.2 – 17
          1                       0                       2                     11                        4 – 35
          1                       1                       0                     7.4                      1.3 – 20
          1                       1                       1                     11                        4 – 35
          1                       2                       0                     11                        4 – 35
          1                       2                       1                     15                        5 – 38
          1                       3                       0                     16                        5 – 38
          2                       0                       0                     9.2                      1.5 – 35
          2                       0                       1                     14                        4 – 35
          2                       0                       2                     20                        5 – 38
          2                       1                       0                     15                        4 – 38
          2                       1                       1                     20                        5 – 38
          2                       1                       2                     27                        9 – 94
          2                       2                       0                     21                        5 – 40
          2                       2                       1                     28                        9 – 94
          2                       2                       2                     35                        9 – 94
          2                       3                       0                     29                        9 – 94
          2                       3                       1                     36                        9 – 94
          3                       0                       0                     23                        5 – 94
          3                       0                       1                     38                        9 – 104
          3                       0                       2                     64                       16 – 181
          3                       1                       0                     43                        9 – 181
          3                       1                       1                     75                       17 – 199
          3                       1                       2                     120                      30 – 360
          3                       1                       3                     160                      30 – 380
          3                       2                       0                     93                       18 – 360
          3                       2                       1                     150                      30 – 380
          3                       2                       2                     210                      30 – 400
          3                       2                       3                     290                      90 – 990
          3                       3                       0                     240                      40 – 990
          3                       3                       1                     460                     90 – 1980
          3                       3                       2                    1100                     200 – 4000
          3                       3                       3              More than 1100

the absence of, or limited occurrence of specified micro-      substance or preparation complies with an established
organisms which may be detected under the conditions           specification for microbiological quality. When used for
described.                                                     such purposes follow the instructions given below, including
  The tests are designed primarily to determine whether a      the number of samples to be taken and interpret the results
1808       General Tests, Processes and Apparatus                                                   Supplement I, JP XV

as stated below.                                                 and then diluting down a fresh suspension of vegetative cells
  Alternative microbiological procedures, including auto-        of Cl. sporogenes, a stable spore suspension is used for test
mated methods may be used, provided that their equivalence       inoculation. The stable spore suspension may be maintained
to the Pharmacopoeial method has been demonstrated.              at 2 – 89 for a validated period.
                                                                           C
                                                                 3-2 Negative control
2 General Procedures
                                                                    To verify testing conditions a negative control is per-
   The preparation of samples is carried out as described in
                                                                 formed using the chosen diluent in place of the test prepara-
Microbial enumeration tests.
                                                                 tion. There must be no growth of micro-organisms.
   If the product to be examined has antimicrobial activity,
                                                                 3-3 Growth promotion and inhibitory properties of the
this is insofar as possible removed or neutralized as
                                                                 media
described in Microbial enumeration tests.
                                                                    Test each batch of ready-prepared medium and each batch
   If surface-active substances are used for sample prepara-
                                                                 of medium prepared either from dehydrated medium or
tion, their absence of toxicity for micro-organisms and their
                                                                 from ingredients.
compatibility with inactivators used must be demonstrated
                                                                    Verify suitable properties of relevant media as described
as described in Microbial enumeration tests.
                                                                 in Table 4.05-II-1.
3 Growth Promoting and Inhibitory Properties of the                 Test for growth promoting properties, liquid media: in-
Media and Suitability of the Test                                oculate a portion of the appropriate medium with a small
   The ability of the test to detect micro-organisms in the      number (not more than 100 CFU) of the appropriate micro-
presence of the product to be tested must be established.        organism. Incubate at the specified temperature for not
Suitability must be confirmed if a change in testing perfor-     more than the shortest period of time specified in the test.
mance, or the product, which may affect the outcome of the       Clearly visible growth of the micro-organism comparable to
test is introduced.                                              that previously obtained with a previously tested and ap-
3-1 Preparation of test strains                                  proved batch of medium occurs.
   Use standardised stable suspensions of test strains or pre-      Test for growth promoting properties, solid media: per-
pare as stated below. Seed lot culture maintenance tech-         form surface-spread method, inoculating each plate with a
niques (seed-lot systems) are used so that the viable micro-     small number (not more than 100 CFU) of the appropriate
organisms used for inoculation are not more than 5 passages      micro-organism. Incubate at the specified temperature for
removed from the original master seed-lot.                       not more than the shortest period of time specified in the
3-1-1 Aerobic micro-organisms                                    test. Growth of the micro-organism comparable to that
   Grow each of the bacterial test strains separately in con-    previously obtained with a previously tested and approved
tainers containing casein soya bean digest broth or on casein    batch of medium occurs.
soya bean digest agar at 30 – 359 for 18 – 24 hours. Grow
                                  C                                 Test for inhibitory properties, liquid or solid media: in-
the test strain for Candida albicans separately on               oculate the appropriate medium with at least 100 CFU of the
Sabouraud-dextrose agar or in Sabouraud-dextrose broth at        appropriate micro-organism. Incubate at the specified tem-
20 – 259 for 2–3 days.
          C                                                      perature for not less than the longest period of time specified
   Staphylococcus aureus such as ATCC 6538, NCIMB                in the test. No growth of the test micro-organism occurs.
9518, CIP 4.83 or NBRC 13276,                                       Test for indicative properties: perform surface-spread
   Pseudomonas aeruginosa such as ATCC 9027, NCIMB               method, inoculating each plate with a small number (not
8626, CIP 82.118 or NBRC 13275,                                  more than 100 CFU) of the appropriate micro-organism.
   Escherichia coli such as ATCC 8739, NCIMB 8545, CIP           Incubate at the specified temperature for a period of time
53.126 or NBRC 3972,                                             within the range specified in the test. Colonies are compara-
   Salmonella enterica subsp. enterica serovar Typhimurium       ble in appearance and indication reactions to those previous-
such as ATCC 14028                                               ly obtained with a previously tested and approved batch of
or, as an alternative,                                           medium.
   Salmonella enterica subsp. enterica serovar Abony such as     3-4 Suitability of the test method
NBRC 100797, NCTC 6017 or CIP 80.39,                                For each product to be tested perform sample preparation
   Candida albicans such as ATCC 10231, NCPF 3179, IP            as described in the relevant paragraph in section 4. Add each
48.72 or NBRC 1594.                                              test strain at the time of mixing, in the prescribed growth
   Use buffered sodium chloride-peptone solution pH 7.0 or       medium. Inoculate the test strains individually. Use a num-
phosphate buffer solution pH 7.2 to make test suspensions.       ber of micro-organisms equivalent to not more than 100
Use the suspensions within 2 hours or within 24 hours if         CFU in the inoculated test preparation.
stored at 2 – 89C.                                                  Perform the test as described in the relevant paragraph in
3-1-2 Clostridia                                                 section 4 using the shortest incubation period prescribed.
   Use Clostridium sporogenes such as ATCC 11437 (NBRC              The specified micro-organisms must be detected with the
14293, NCIMB 12343, CIP 100651) or ATCC 19404 (NCTC              indication reactions as described in section 4.
532 or CIP 79.3). Grow the clostridial test strain under            Any antimicrobial activity of the product necessitates a
anaerobic conditions in reinforced medium for Clostridia at      modification of the test procedure (see 4-5-3 of Microbial
30 – 359 for 24 – 48 hours. As an alternative to preparing
          C                                                      Enumeration Tests).
Supplement I, JP XV                                                General Tests, Processes and Apparatus                1809

                     Table 4.05-II-1    Growth promoting, inhibitory and indicative properties of media

    Medium                                      Property                    Test strains

    Test for bile-tolerant gram-negative bacteria

    Enterobacteria enrichment                   Growth promoting            E. coli
    broth-Mossel                                                            P. aeruginosa
                                                Inhibitory                  S. aureus

    Violet red bile glucose agar                Growth promoting+           E. coli
                                                Indicative                  P. aeruginosa

    Test for Escherichia coli

    MacConkey broth                             Growth promoting            E. coli
                                                Inhibitory                  S. aureus

    MacConkey agar                              Growth promoting+           E. coli
                                                Indicative

    Test for Salmonella

    Rappaport Vassiliadis Salmonella            Growth promoting            Salmonella enterica subsp. enterica serovar
    enrichment broth                                                        Typhimurium or
                                                                            Salmonella enterica subsp. enterica serovar
                                                                            Abony

                                                Inhibitory                  S. aureus

    Xylose, lysine, deoxycholate agar           Growth promoting+           Salmonella enterica subsp. enterica serovar
                                                Indicative                  Typhimurium or
                                                                            Salmonella enterica subsp. enterica serovar
                                                                            Abony

                                                Indicative                  E. coli
    Test for Pseudomonas aeruginosa

    Cetrimide agar                              Growth promoting            P. aeruginosa
                                                Inhibitory                  E. coli

    Test for Staphylococcus aureus

    Mannitol salt agar                          Growth promoting+           S. aureus
                                                Indicative

                                                Inhibitory                  E. coli
    Test for Clostridia

    Reinforced medium for Clostridia            Growth promoting            Cl. sporogenes

    Columbia agar                               Growth promoting            Cl. sporogenes
    Test for Candida albicans

    Sabouraud dextrose broth                    Growth promoting            C. albicans
    Sabouraud dextrose agar                     Growth promoting+           C. albicans
                                                Indicative




  If for a given product the antimicrobial activity with         4 Testing of Products
respect to a micro-organism for which testing is prescribed      4-1 Bile-tolerant gram-negative bacteria
cannot be neutralised, then it is to be assumed that the in-     4-1-1 Sample preparation and pre-incubation
hibited micro-organism will not be present in the product.         Prepare a sample using a 1 in 10 dilution of not less than 1
1810       General Tests, Processes and Apparatus                                                       Supplement I, JP XV

                                             Table 4.05-II-2   Interpretation of results

                         Results for each quantity of product
                                                                                              Probable number of bacteria
                                                                                              per gram or mL of product
     0.1 g or 0.1 mL            0.01 g or 0.01 mL              0.001 g or 0.001 mL

            +                            +                             +                     more than 103

            +                            +                             -                     less than 103 and more than 102

            +                            -                             -                     less than 102 and more than 10

            -                            -                             -                     less than 10




g of the product to be examined as described in Microbial            4-2-3 Interpretation
enumeration tests, but using casein soya bean digest broth as           Growth of colonies indicates the possible presence of
the chosen diluent, mix and incubate at 20 – 259 for a time
                                                   C                 E. coli. This is confirmed by identification tests.
sufficient to resuscitate the bacteria but not sufficient to en-        The product complies with the test if no colonies are
courage multiplication of the organisms (usually 2 hours but         present or if the identification tests are negative.
not more than 5 hours).                                              4-3 Salmonella
4-1-2 Test for absence                                               4-3-1 Sample preparation and pre-incubation
   Unless otherwise prescribed use the volume corresponding             Prepare the product to be examined as described in
to 1 g of the product, as prepared in 4-1-1 to inoculate             Microbial enumeration tests and use the quantity cor-
enterobacteria enrichment broth-Mossel. Incubate at 30 –             responding to not less than 10 g or 10 mL to inoculate a suit-
359 for 24 – 48 hours. Subculture on plates of violet red
    C                                                                able amount (determined as described under 3-4) of casein
bile glucose agar. Incubate at 30 – 359 for 18 – 24 hours.
                                         C                           soya bean digest broth, mix and incubate at 30 – 359 forC
   The product complies with the test if there is no growth of       18 – 24 hours.
colonies.                                                            4-3-2 Selection and subculture
4-1-3 Quantitative test                                                 Transfer 0.1 mL of casein soya bean digest broth to 10 mL
4-1-3-1 Selection and subculture                                     of Rappaport Vassiliadis Salmonella enrichment broth and
   Inoculate suitable quantities of enterobacteria enrichment        incubate at 30 – 359 for 18 – 24 hours. Subculture on plates
                                                                                          C
broth-Mossel with the preparation as described under 4-1-1           of xylose, lysine, deoxycholate agar. Incubate at 30 – 359  C
and/or dilutions of it containing respectively 0.1 g, 0.01 g         for 18 – 48 hours.
and 0.001 g (or 0.1 mL, 0.01 mL and 0.001 mL) of the                 4-3-3 Interpretation
product to be examined. Incubate at 30 – 359 for 24 – 48
                                                 C                      The possible presence of Salmonella is indicated by the
hours. Subculture each of the cultures on a plate of violet          growth of well-developed, red colonies, with or without
red bile glucose agar. Incubate at 30 – 359 for 18 – 24
                                                 C                   black centres. This is confirmed by identification tests.
hours.                                                                  The product complies with the test if colonies of the types
4-1-3-2 Interpretation                                               described are not present or if the confirmatory identifica-
   Growth of colonies constitutes a positive result. Note the        tion tests are negative.
smallest quantity of the product that gives a positive result        4-4 Pseudomonas aeruginosa
and the largest quantity that gives a negative result. Deter-        4-4-1 Sample preparation and pre-incubation
mine from Table 4.05-II-2 the probable number of bacteria.              Prepare a sample using a 1 in 10 dilution of not less than
4-2 Escherichia coli                                                 1 g of the product to be examined as described in Microbial
4-2-1 Sample preparation and pre-incubation                          enumeration tests and use 10 mL or the quantity correspond-
   Prepare a sample using a 1 in 10 dilution of not less than        ing to 1 g or 1 mL to inoculate a suitable amount (deter-
1 g of the product to be examined as described in Microbial          mined as described under 3-4) of casein soya bean digest
enumeration tests and use 10 mL or the quantity correspond-          broth and mix. When testing transdermal patches, filter the
ing to 1 g or 1 mL to inoculate a suitable amount (deter-            volume of sample corresponding to 1 patch of the prepara-
mined as described under 3-4) of casein soya bean digest             tion described in Microbial enumeration tests (4-5-1)
broth, mix and incubate at 30 – 359 for 18 – 24 hours.
                                       C                             through a sterile filter membrane and place in 100 mL of
4-2-2 Selection and subculture                                       casein soya bean digest broth. Incubate at 30 – 359 for 18 –
                                                                                                                          C
   Shake the container, transfer 1 mL of casein soya bean            24 hours.
digest broth to 100 mL of MacConkey broth and incubate at            4-4-2 Selection and subculture
42 – 449 for 24 – 48 hours. Subculture on a plate of Mac-
         C                                                              Subculture on a plate of cetrimide agar and incubate at
Conkey agar at 30 – 359 for 18 – 72 hours.
                           C                                         30 – 359 for 18 – 72 hours.
                                                                              C
Supplement I, JP XV                                                General Tests, Processes and Apparatus                 1811

4-4-3 Interpretation                                              mL of Sabouraud-dextrose broth and mix. Incubate at 30 –
   Growth of colonies indicates the possible presence of          359 for 3-5 days.
                                                                      C
P. aeruginosa. This is confirmed by identification tests.         4-7-2 Selection and subculture
   The product complies with the test if colonies are not            Subculture on a plate of Sabouraud-dextrose agar and
present or if the confirmatory identification tests are nega-     incubate at 30 – 359 for 24 – 48 hours.
                                                                                       C
tive.                                                             4-7-3 Interpretation
4-5 Staphylococcus aureus                                            Growth of white colonies may indicate the presence of C.
4-5-1 Sample preparation and pre-incubation                       albicans. This is confirmed by identification tests.
   Prepare a sample using a 1 in 10 dilution of not less than 1      The product complies with the test if such colonies are not
g of the product to be examined as described in Microbial         present or if the confirmatory identification tests are nega-
enumeration tests and use 10 mL or the quantity correspond-       tive.
ing to 1 g or 1 mL to inoculate a suitable amount (deter-            The following section is given for information.
mined as described under 3-4) of casein soya bean digest
                                                                  5 Recommended Solutions and Culture Media
broth and homogenise. When testing transdermal patches,
                                                                     The following solutions and culture media have been
filter the volume of sample corresponding to 1 patch of the
                                                                  found satisfactory for the purposes for which they are
preparation described in Microbial enumeration tests (4-5-1)
                                                                  prescribed in the test for microbial contamination in the
through a sterile filter membrane and place in 100 mL of
                                                                  Pharmacopoeia. Other media may be used if they have simi-
casein soya bean digest broth. Incubate at 30 – 359 for 18 –
                                                     C
                                                                  lar growth promoting and inhibitory properties.
24 hours.
4-5-2 Selection and subculture                                    Stock buffer solution. Transfer 34 g of potassium dihydro-
   Subculture on a plate of mannitol salt agar and incubate       gen phosphate to a 1000 mL volumetric flask, dissolve in 500
at 30 – 359 for 18 – 72 hours.
            C                                                     mL of purified water, adjust to pH 7.2±0.2 with sodium
4-5-3 Interpretation                                              hydroxide, add purified water to volume and mix. Dispense
   The possible presence of S. aureus is indicated by the         in containers and sterilize. Store at a temperature of 2 – 89
                                                                                                                              C.
growth of yellow/white colonies surrounded by a yellow
                                                                  Phosphate buffer solution pH 7.2
zone. This is confirmed by identification tests.
                                                                  Prepare a mixture of purified water and stock buffer solu-
   The product complies with the test if colonies of the types
                                                                  tion (800:1 V/V) and sterilize.
described are not present or if the confirmatory identifica-
tion tests are negative.                                          Buffered sodium chloride-peptone solution pH 7.0
4-6 Clostridia                                                    Potassium dihydrogen phosphate                          3.6 g
4-6-1 Sample preparation and heat treatment                       Disodium hydrogen phosphate dihydrate                   7.2 g
   Prepare the product to be examined as described in                                                             equivalent to
Microbial enumeration tests.                                                                              0.067 mol phosphate
   Take 2 equal portions corresponding to not less than 1 g       Sodium chloride                                         4.3 g
or 1 mL of the product to be examined. Heat 1 portion at          Peptone (meat or casein)                                1.0 g
809 for 10 min and cool rapidly. Do not heat the other por-
     C                                                            Purified water                                       1000 mL
tion.                                                               Sterilize in an autoclave using a validated cycle.
4-6-2 Selection and subculture
   Transfer the quantity corresponding to 1 g or 1 mL of the      Casein soya bean digest broth
product to be examined from each of the mixed portions to 2       Pancreatic digest of casein                            17.0 g
containers (38 mm×200 mm) or other containers containing          Papaic digest of soya bean                              3.0 g
100 mL of reinforced medium for Clostridia. Incubate              Sodium chloride                                         5.0 g
under anaerobic conditions at 30 – 359 for 48 hours. After
                                        C                         Dipotassium hydrogen phosphate                          2.5 g
incubation, make subcultures from each tube on Columbia           Glucose monohydrate                                     2.5 g
agar and incubate under anaerobic conditions at 30 – 359     C    Purified water                                       1000 mL
for 48 hours.                                                       Adjust the pH so that after sterilization it is 7.3±0.2 at
4-6-3 Interpretation                                              259 Sterilize in an autoclave using a validated cycle.
                                                                     C.
   The occurrence of anaerobic growth of rods (with or
without endospores) giving a negative catalase reaction indi-     Casein soya bean digest agar
cates the presence of Clostridia.                                 Pancreatic digest of casein                            15.0 g
   If no anaerobic growth of micro-organisms is detected on       Papaic digest of soya bean                              5.0 g
Columbia agar or the catalase test is positive, the product       Sodium chloride                                         5.0 g
complies with the test.                                           Agar                                                   15.0 g
4-7 Candida albicans                                              Purified water                                       1000 mL
4-7-1 Sample preparation and pre-incubation                         Adjust the pH so that after sterilization it is 7.3±0.2 at
   Prepare the product to be examined as described in             259 Sterilize in an autoclave using a validated cycle.
                                                                     C.
Microbial enumeration tests and use 10 mL or the quantity
corresponding to not less than 1 g or 1 mL to inoculate 100
1812       General Tests, Processes and Apparatus                                                   Supplement I, JP XV

Sabouraud-dextrose agar                                         MacConkey agar
Glucose                                                40.0 g   Pancreatic digest of gelatin                           17.0 g
Mixture of peptic digest of animal tissue and pan-              Peptones (meat and casein)                              3.0 g
creatic digest of casein (1:1)                         10.0 g   Lactose monohydrate                                    10.0 g
Agar                                                   15.0 g   Sodium chloride                                         5.0 g
Purified water                                       1000 mL    Bile salts                                              1.5 g
  Adjust the pH so that after sterilization it is 5.6±0.2 at    Agar                                                   13.5 g
259 Sterilize in an autoclave using a validated cycle.
   C.                                                           Neutral red                                            30 mg
                                                                Crystal violet                                          1 mg
Potato dextrose agar                                            Purified water                                       1000 mL
Infusion from potatoes                                  200 g     Adjust the pH so that after sterilization it is 7.1±0.2 at
Glucose                                                20.0 g   259 Boil for 1 min with constant shaking then sterilize in
                                                                    C.
Agar                                                   15.0 g   an autoclave using a validated cycle.
Purified water                                       1000 mL
  Adjust the pH so that after sterilization it is 5.6±0.2 at    Rappaport Vassiliadis Salmonella enrichment broth
259 Sterilize in an autoclave using a validated cycle.
   C.                                                           Soya peptone                                              4.5 g
                                                                Magnesium chloride hexahydrate                           29.0 g
Sabouraud-dextrose broth                                        Sodium chloride                                           8.0 g
Glucose                                                20.0 g   Dipotassium hydrogen phosphate                            0.4 g
Mixture of peptic digest of animal tissue and pan-              Potassium dihydrogen phosphate                            0.6 g
creatic digest of casein (1:1)                         10.0 g   Malachite green                                          36 mg
Purified water                                       1000 mL    Purified water                                        1000 mL
  Adjust the pH so that after sterilization it is 5.6±0.2 at       Dissolve, warming slightly. Sterilize in an autoclave using
259 Sterilize in an autoclave using a validated cycle.
   C.                                                           a validated cycle, at a temperature not exceeding 1159 The
                                                                                                                        C.
                                                                pH is to be 5.2±0.2 at 259 after heating and autoclaving.
                                                                                            C
Enterobacteria enrichment broth-Mossel
Pancreatic digest of gelatin                           10.0 g   Xylose, lysine, deoxycholate agar
Glucose monohydrate                                     5.0 g   Xylose                                                  3.5 g
Dehydrated ox bile                                     20.0 g   L-Lysine                                                5.0 g
Potassium dihydrogen phosphate                          2.0 g   Lactose monohydrate                                     7.5 g
Disodium hydrogen phosphate dihydrate                   8.0 g   Sucrose                                                 7.5 g
Brilliant green                                        15 mg    Sodium chloride                                         5.0 g
Purified water                                      1000 mL     Yeast extract                                           3.0 g
  Adjust the pH so that after heating it is 7.2±0.2 at 259C.    Phenol red                                             80 mg
Heat at 1009 for 30 min and cool immediately.
              C                                                 Agar                                                   13.5 g
                                                                Sodium deoxycholate                                     2.5 g
Violet red bile glucose agar                                    Sodium thiosulfate                                      6.8 g
Yeast extract                                           3.0 g   Ammonium iron (III) citrate                             0.8 g
Pancreatic digest of gelatin                            7.0 g   Purified water                                      1000 mL
Bile salts                                              1.5 g     Adjust the pH so that after heating it is 7.4±0.2 at 259C.
Sodium chloride                                         5.0 g   Heat to boiling, cool to 509 and pour into Petri dishes. Do
                                                                                           C
Glucose monohydrate                                    10.0 g   not heat in an autoclave.
Agar                                                   15.0 g
Neutral red                                            30 mg    Cetrimide agar
Crystal violet                                          2 mg    Pancreatic digest of gelatin                           20.0 g
Purified water                                      1000 mL     Magnesium chloride                                       1.4 g
  Adjust the pH so that after heating it is 7.4±0.2 at 259C.    Dipotassium sulfate                                    10.0 g
Heat to boiling; do not heat in an autoclave.                   Cetrimide                                                0.3 g
                                                                Agar                                                   13.6 g
MacConkey broth                                                 Purified water                                      1000 mL
Pancreatic digest of gelatin                           20.0 g   Glycerol                                             10.0 mL
Lactose monohydrate                                    10.0 g     Heat to boiling for 1 min with shaking. Adjust the pH so
Dehydrated ox bile                                      5.0 g   that after sterilization it is 7.2±0.2 at 259 Sterilize in an
                                                                                                             C.
Bromocresol purple                                     10 mg    autoclave using a validated cycle.
Purified water                                       1000 mL
  Adjust the pH so that after sterilization it is 7.3±0.2 at    Mannitol salt agar
259 Sterilize in an autoclave using a validated cycle.
   C.                                                           Pancreatic digest of casein                              5.0 g
                                                                Peptic digest of animal tissue                           5.0 g
Supplement I, JP XV                                               General Tests, Processes and Apparatus                  1813

Beef extract                                             1.0 g
D-Mannitol                                             10.0 g    Add the following next to Procedure:
Sodium chloride                                        75.0 g
                                                                 Evaluation
Agar                                                   15.0 g
                                                                   The preparation complies with the test if the total number
Phenol red                                             25 mg
                                                                 of metal particles of a size equal to or greater than 50 mm
Purified water                                      1000 mL
                                                                 found in 10 units tested, is not more than 50, and also the
  Heat to boiling for 1 min with shaking. Adjust the pH so
                                                                 number of dishes containing more than 8 particles is not
that after sterilization it is 7.4±0.2 at 259 Sterilize in an
                                             C.
                                                                 more than 1. If this requirement is snot met, repeat the test
autoclave using a validated cycle.
                                                                 with a further 20 units in the same manner, and if the total
                                                                 number of the particles found in the total of 30 units is not
Reinforced medium for Clostridia
                                                                 more than 150, and also the number of dishes containing
Beef extract                                           10.0 g
                                                                 more than 8 particles is not more than 3, the preparation
Peptone                                                10.0 g
                                                                 complies with the test.
Yeast extract                                           3.0 g
Soluble starch                                          1.0 g
Glucose monohydrate                                     5.0 g
Cysteine hydrochloride                                  0.5 g    6.08 Insoluble Particulate Matter
Sodium chloride                                         5.0 g      Test for Ophthalmic Solutions
Sodium acetate                                          3.0 g
Agar                                                    0.5 g    Add the following next to Procedure:
Purified water                                      1000 mL
                                                                 Evaluation
   Hydrate the agar, dissolve by heating to boiling with con-
                                                                   The preparation complies with the test if the calculated
tinuous stirring. If necessary, adjust the pH so that after
                                                                 number per mL of insoluble particles of a size equal to or
sterilization it is about 6.8±0.2 at 259 Sterilize in an
                                          C.
                                                                 greater than 300 mm is not more than 1.
autoclave using a validated cycle.

Columbia agar
Pancreatic digest of casein                            10.0 g
                                                                            6.10       Dissolution Test
Meat peptic digest                                      5.0 g
                                                                 Change the Apparatus for Paddle Method
Heart pancreatic digest                                 3.0 g
                                                                 (Apparatus 2) to read:
Yeast extract                                           5.0 g
Corn starch                                             1.0 g       Apparatus for Paddle Method (Apparatus 2)—Use the
Sodium chloride                                         5.0 g    assembly from Apparatus 1, except that a paddle formed
Agar, according to gelling power             10.0 g to 15.0 g    from a blade and a shaft is used as the stirring element. The
Purified water                                      1000 mL      shaft is positioned so that its axis is not more than 2 mm
   Hydrate the agar, dissolve by heating to boiling with con-    from the vertical axis of the vessel, at any point, and rotates
tinuous stirring. If necessary, adjust the pH so that after      smoothly without significant wobble that could affect the
sterilization it is 7.3±0.2 at 259 Sterilize in an autoclave
                                  C.                             results. The vertical center line of the blade passes through
using a validated cycle. Allow to cool to 45 – 509 add,
                                                     C;          the axis of the shaft so that the bottom of the blade is flush
where necessary, gentamicin sulfate corresponding to 20 mg       with the bottom of the shaft. The paddle conforms to the
of gentamicin base and pour into Petri dishes.                   specifications shown in Fig. 6.10-2. The distance of 25±2
                                                                 mm between the bottom of the blade and the inside bottom
                                                                 of the vessel is maintained during the test. The metallic or
  6.01      Test for Metal Particles in                          suitably inert, rigid blade and shaft comprise a single entity.
                                                                 A suitable two-part detachable design may be used provided
           Ophthalmic Ointments                                  the assembly remains firmly engaged during the test. The
                                                                 paddle blade and shaft may be coated with a suitable coating
Change the Preparation of test sample to read:
                                                                 so as to make them inert. The dosage unit is allowed to sink
Preparation of test sample                                       to the bottom of the vessel before rotation of the blade is
  The test should be carried out in a clean place. Take 10       started. A small, loose piece of nonreactive material, such as
ophthalmic ointments to be tested, and extrude 5 g each of       not more than a few turns of wire helix or such one shown in
their contents into separate flat-bottomed petri dishes 60       Fig. 6.10-2a, may be attached to the dosage unit that would
mm in diameter. Cover the dishes, and heat between 859     C     otherwise float. Other validated sinker devices may also be
and 1109 for 2 hours to dissolve bases completely. Allow
          C                                                      used. ◆If the use of sinker is specified, unless otherwise
the samples to cool to room temperature without agitation        specified, use the sinker device shown in Fig. 6.10-2a.◆
to solidify the contents. When the amount of the content is 5
g or less, extrude the contents as completely as practicable,
and proceed in the same manner as described above.
1814       General Tests, Processes and Apparatus                                                  Supplement I, JP XV


                                                                       9.01        Reference Standards
                                                                Change to read:
                                                                  Reference Standards are the reference substances pre-
                                                                pared to a specified quality necessary with regard to their in-
                                                                tended use as prescribed in monographs of the Phar-
                                                                macopoeia.
                                                                  The Japanese Pharmacopoeia Reference Standards are as
                                                                follows:

                                                                * A: Assay
                                                                  AF: Anti-factor IIa activity
                                                                  B: Bacterial Endotoxins Test <4.01>
                                                                  C: Content ratio of active principle
                                                                  D: Dissolution
                                                                  DG: Digestion Test <4.03>
                                                                  HB: Heparin-binding capacity
                                                                  I: Identification
                                                                  IS: Isomer ratio
                                                                  M: Melting Point Determination <2.60>
                                                                  P: Purity
       Fig. 6. 10–2   Apparatus 2, Paddle stirring element        T: Thermal Analysis <2.52>
                                                                  U: Uniformity of dosage units
                                                                  V: Vitamin A Assay <2.55>

                                                                (1) The reference standards which are prepared by those who
                                                                have been registered to prepare them by the Minister of
                                                                Health, Labour and Welfare, according to the Ministerial
                                                                ordinance established by the Minister separately.

                                                                Reference Standard                             Intended Use*

                                                                Aceglutamide                                   I, P, A
                                                                Acetaminophen                                  I, A
                                                                Adrenaline Bitartrate                          P
                                                                Alprostadil                                    I, P, A
                Fig. 6. 10–2a   Alternative sinker
                                                                p-Aminobenzoyl Glutamic Acid                   P
                                                                Amitriptyline Hydrochloride                    I, U, D, A
                                                                Amlexanox                                      I, U, D, A
                                                                Amlodipine Besilate                            I, A
Add the following:                                              Anhydrous Lactose                              I
                                                                Ascorbic Acid                                  A
   6.11 Foreign Insoluble Matter                                Aspirin                                        A
                                                                Atropine Sulfate                               I, A
   Test for Ophthalmic Solutions                                Azathioprine                                   I, A
                                                                Baclofen                                       I, U, D, A
   Foreign Insoluble Matter Test for Ophthalmic Solutions is    Baicalin                                       I, A
a test method to examine foreign insoluble matters in           Beclometasone Dipropionate                     I, A
ophthalmic solutions.                                           Berberine Chloride                             I, A
   When inspect with the unaided eyes at a position of          Betamethasone                                  I, P, U, D, A
luminous intensity of 3000 – 5000 lx under an incandescent      Betamethasone Sodium Phosphate                 I, A
lamp after cleaning the exterior of containers, Ophthalmic      Betamethasone Valerate                         I, A
Solutions must be clear and free from readily detectable for-   Bisacodyl                                      I, U, A
eign insoluble matters.                                         Caffeine                                       A
                                                                Calcium Folinate                               I, A
                                                                Calcium Oxalate Monohydrate                    T
                                                                Camostat Mesilate                              I, A
Supplement I, JP XV                            General Tests, Processes and Apparatus              1815

d-Camphor                        A            Hydrochlorothiazide                       I, A
dl-Camphor                       A            Hydrocortisone                            I, P, A
Carbidopa                        I, P, A      Hydrocortisone Acetate                    I, A
Cellacefate                      I            Hydrocortisone Sodium Phosphate           I, A
Chlordiazepoxide                 I, P, U, A   Hydrocortisone Succinate                  I, A
Chlormadinone Acetate            I, A         Idoxuridine                               I, A
Chlorpheniramine Maleate         I, U, A      Imipramine Hydrochloride                  I, U, D, A
Cholecalciferol                  I, A         Indomethacin                              I, P, U, D, A
Ciclosporin                      I, P, A      Insulin                                   P, A
Cilostazol                       I, U, D, A   Interleukin-2                             A
Cisplatin                        I, A         Isoflurane                                I, P, A
Clobetasol Propionate            I, A         Kallidinogenase                           A
Clofibrate                       I, A         Lactose                                   I
Clomifene Citrate                I, A         Lactulose                                 P, A
Cortisone Acetate                I, A         Lanatoside C                              I, P, U, D, A
Cyanocobalamin                   I, P, A      Limaprost                                 P, A
Deferoxamine Mesilate            I, A         Low-molecular Mass Heparin                AF, A
Deslanoside                      I, P, A      Loxoprofen                                A
Dexamethasone                    I, A         Lysozyme                                  A
Diclofenamide                    I, P, D, A   Maltose                                   A
Diethylcarbamazine Citrate       A            Manidipine Hydrochloride                  I, U, D, A
Digitoxin                        I, U, D, A   Mecobalamin                               I, A
Digoxin                          I, U, D, A   Melting Point Standard-Acetanilide        M
Dihydroergotoxine Mesilate       A            Melting Point Standard-Acetopheneti-
Dobutamine Hydrochloride         I, A            dine                                   M
Edrophonium Chloride             I, A         Melting Point Standard-Caffeine           M
Elcatonin                        A            Melting Point Standard-Sulfanilamide      M
Enalapril Maleate                I, U, D, A   Melting Point Standard-Sulfapyridine      M
Endotoxin                        B            Melting Point Standard-Vanillin           M
Epitiostanol                     P, A         Menatetrenone                             I, P, A
Ergocalciferol                   I, A         Mestranol                                 I, A
Ergometrine Maleate              P, U, A      Methotrexate                              I, A
Estradiol Benzoate               I, P, A      Methoxsalen                               I, A
Estriol                          I, U, D, A   Methyldopa                                I, U, A
Ethenzamide                      A            Methylergometrine Maleate                 I, U, D,   A
Ethinylestradiol                 I, U, D, A   Methylprednisolone Succinate              I, P, A
Ethyl Aminobenzoate              A            Methyltestosterone                        I, U, A
Ethyl Icosapentate               I, P, A      Metildigoxin                              I, A
Etoposide                        I, A         Mexiletine Hydrochloride                  I, P, A
Fluocinolone Acetonide           I, A         Mizoribine                                I, U, D,   A
Fluocinonide                     I, A         Nabumetone                                I, D, A
Fluorometholone                  I, A         Neostigmine Methylsulfate                 I, A
Fluoxymesterone                  I, A         Nicotinamide                              I, A
Folic Acid                       I, U, A      Nicotinic Acid                            I, A
Furosemide                       I, U, D, A   Nilvadipine                               I, U, D,   A
Fursultiamine Hydrochloride      I, A         Nizatidine                                I, U, D,   A
Gabexate Mesilate                I, P, A      Noradrenaline Bitartrate                  P, A
Ginsenoside Rb1                  I, A         Norgestrel                                I, U, D,   A
Ginsenoside Rg1                  I, A         Oxytocin                                  P, A
Gitoxin                          P            Ozagrel Sodium                            I, A
Glycyrrhizinic Acid              I, A         Paeoniflorin                              I, A
Gonadorelin Acetate              I, A         Pentobarbital                             P, A
Guaifenesin                      I, A         Perphenazine                              I, U, D,   A
Heparin Sodium                   HB, A        Phytonadione                              A
High-molecular Mass Urokinase    A            Potassium Sucrose Octasulfate             P, A
Human Chorionic Gonadotrophin    A            Povidone                                  I
Human Insulin                    I, A         Pravastatin 1,1,3,3-tetramethylbutylam-
Human Menopausal Gonadotrophin   P, A           monium                                  I, A
1816      General Tests, Processes and Apparatus                                             Supplement I, JP XV

Prednisolone                                I, U, D, A       Amoxicillin                             I, A
Prednisolone Acetate                        I, A             Amphotericin B                          I, P, A
Prednisolone Succinate                      I, A             Ampicillin                              I, P, A
Primidone                                   A                Arbekacin Sulfate                       I, A
Probenecid                                  I, D, A          Aspoxicillin                            I, P, A
Prochlorperazine Maleate                    I, A             Astromicin Sulfate                      I, A
Progesterone                                I, A             Azithromycin                            I, A
Protamine Sulfate                           P                Aztreonam                               I, P, A
Puerarin                                    I, A             Bacampicillin Hydrochloride             I, A
Pyridoxine Hydrochloride                    I, A             Bacitracin                              I, A
Ranitidine Hydrochloride                    I, A             Bekanamycin Sulfate                     I, A
Reserpine                                   I, U, D, A       Benzylpenicillin Potassium              I, A
Retinol Acetate                             I, V             Bleomycin A2 Hydrochloride              A
Retinol Palmitate                           I, V             Carumonam Sodium                        I, P, A
Riboflavin                                  I, A             Cefaclor                                I, P, U, D, A
Ritodrine Hydrochloride                     I, U, D, A       Cefadroxil                              I, U, D, A
Roxatidine Acetate Hydrochloride            I, U, D, A       Cefalexin                               A
Saccharated Pepsin                          A                Cefalotin Sodium                        I, P, A
Scopolamine Hydrobromide                    I, A             Cefapirin Sodium                        I, A
Sennoside A                                 I, A             Cefatrizine Propylene Glycolate         I, A
Sennoside B                                 A                Cefazolin                               P, A
Serum Gonadotrophin                         A                Cefbuperazone                           A
Spironolactone                              I, A             Cefcapene Pivoxil Hydrochloride         I, U, A
Sulfadiazine Silver                         I, A             Cefdinir                                I, P, D, A
Swertiamarin                                I, A             Cefditoren Pivoxil                      I, U, D, A
Testosterone Propionate                     I, A             Cefepime Dihydrochloride                I, A
Thiamine Chloride Hydrochloride             I, P, A          Cefixime                                I, P, A
Thiamylal                                   A                Cefmenoxime Hydrochloride               I, P, A
Thrombin                                    A                Cefmetazole                             A
Tocopherol                                  I, P, A          Cefminox Sodium                         I, A
Tocopherol Acetate                          I, A             Cefodizime Sodium                       I, P, A
Tocopherol Nicotinate                       I, A             Cefoperazone                            A
Tocopherol Succinate                        A                Cefotaxime                              P, A
Tolazamide                                  I, A             Cefotetan                               I, P, A
Tolbutamide                                 D                Cefotiam Hexetil Hydrochloride          I, P, IS, A
Tolnaftate                                  I, A             Cefotiam Hydrochloride                  I, P, A
Tranexamic Acid                             I, P, D, A       Cefozopran Hydrochloride                I, A
Triamcinolone                               I, A             Cefpiramide                             P, A
Triamcinolone Acetonide                     I, A             Cefpirome Sulfate                       I, A
Trichlormethiazide                          I, U, D, A       Cefpodoxime Proxetil                    I, IS, A
Trihexyphenidyl Hydrochloride               I, U, D, A       Cefroxadine                             I, A
Tyrosine                                    A, DG            Cefsulodin Sodium                       I, P, A
Ubidecarenone                               I, A             Ceftazidime                             I, A
Ulinastatin                                 A                Cefteram Pivoxil Mesitylene Sulfonate   A
Vasopressin                                 A                Ceftibuten Hydrochloride                A
Vinblastine Sulfate                         I, U, A          Ceftizoxime                             P, A
Vincristine Sulfate                         I, A             Ceftriaxone Sodium                      I, A
Warfarin Potassium                          I, U, A          Cefuroxime Axetil                       I, P, IS, A
Zidovudine                                  I, A             Cefuroxime Sodium                       I, A
                                                             Chloramphenicol                         I, A
(2) The reference standards which are prepared by National   Chloramphenicol Palmitate               I, A
Institute of Infectious Diseases.                            Chloramphenicol Succinate               A
                                                             Ciclacillin                             I, A
Reference Standard                          Intended Use*    Clarithromycin                          I, P, U, D, A
                                                             Clindamycin Hydrochloride               I, U, D, A
Aclarubicin                                 A                Clindamycin Phosphate                   I, P, A
Actinomycin D                               I, A             Cloxacillin Sodium                      I, P, A
Amikacin Sulfate                            I, A             Colistin Sodium Methanesulfonate        I, A
Supplement I, JP XV                                  General Tests, Processes and Apparatus                 1817

Colistin Sulfate                       A            Streptomycin Sulfate                          I, A
Cycloserine                            I, A         Sulbactam                                     P, A
Daunorubicin Hydrochloride             I, A         Sulbenicillin Sodium                          I, A
Demethylchlortetracycline Hydrochlo-                Sultamicillin Tosilate                        I, A
   ride                                I, P, A      Talampicillin Hydrochloride                   I, A
Dibekacin Sulfate                      I, P, A      Teicoplanin                                   I, A
Dicloxacillin Sodium                   I, A         Tetracycline Hydrochloride                    I, P, A
Diethanolammonium Fusidate             A            Tobramycin                                    I, A
Doxorubicin Hydrochloride              I, A         Trichomycin                                   A
Doxycycline Hydrochloride              I, A         Vancomycin Hydrochloride                      I, A
Enviomycin Sulfate                     A            Zinostatin Stimalamer                         I, A
Epirubicin Hydrochloride               I, P, A
Erythromycin                           I, P, A
Faropenem Sodium                       I, P, U, A        9.21     Standard Solutions for
Flomoxef Triethylammonium              P, A
Fosfomycin Phenethylammonium           A
                                                                 Volumetric Analysis
Fradiomycin Sulfate                    I, A
                                                    Add the following:
Gentamicin Sulfate                     I, A
Gramicidin                             I, A                         Zinc sulfate, 0.02 mol/L
Griseofulvin                           I, P, U, A     1000 mL of this solution contains 5.7516 g of zinc sulfate
Idarubicin Hydrochloride               I, U, A      heptahydrate (ZnSO4.7H2O: 287.58).
Imipenem                               I, U, A        Preparation—Before use, dilute 0.1 mol/L zinc sulfate VS
Isepamicin Sulfate                     I, P, A      with water to make exactly 5 times the initial volume.
Josamycin                              I, C, U, A
Josamycin Propionate                   I, A
Kanamycin Monosulfate                  I, P, A              9.22       Standard Solutions
Latamoxef Ammonium                     P, A
Lenampicillin Hydrochloride            I, A         Add the following:
Leucomycin A5                          C, A
                                                      Standard Aluminum Solution for Atomic Absorption
Lincomycin Hydrochloride               I, P, A
                                                    Spectrophotometry To exactly 10 mL of Standard Alumi-
Lithium Clavulanate                    A
                                                    num Stock Solution add water to make exactly 100 mL. Pre-
Meropenem                              I, A
                                                    pare before use. Each mL of this solution contains 0.100 mg
Micronomicin Sulfate                   I, A
                                                    of aluminum (Al).
Midecamycin                            I, A
Midecamycin Acetate                    I, A           Standard Iron Stock Solution Dissolve exactly 4.840 g
Minocycline Hydrochloride              I, P, A      of iron (III) chloride hexahydrate in diluted hydrochloric
Mitomycin C                            I, U, A      acid (9 in 25) to make exactly 100 mL.
Mupirocin Lithium                      P, A
                                                      Standard Iron Solution for Atomic Absorption Spec-
Netilmicin Sulfate                     I, A
                                                    trophotometry To exactly 5 mL of Standard Iron Stock
Nystatin                               I, P, A
                                                    Solution add water to make exactly 200 mL. Prepare before
Oxytetracycline Hydrochloride          I, A
                                                    use. Each mL of this solution contains 0.250 mg of iron (Fe).
Panipenem                              A
Peplomycin Sulfate                     I, A           Standard Magnesium Stock Solution Dissolve exactly
Phenethicillin Potassium               A            8.365 g of magnesium chloride hexahydrate in 2 mol/L
Pimaricin                              I, A         hydrochloric acid TS to make exactly 1000 mL.
Piperacillin                           I, A
                                                      Standard Magnesium Solution for Atomic Absorption
Pirarubicin                            I, P, A
                                                    Spectrophotometry To exactly 1 mL of Standard Magnesi-
Pivmecillinam Hydrochloride            I, P, A
                                                    um Stock Solution add water to make exactly 100 mL. Pre-
Polymixin B Sulfate                    I, A
                                                    pare before use. Each mL of this solution contains 0.0100
Pyrrolnitrin                           I, A
                                                    mg of magnesium (Mg).
Ribostamycin Sulfate                   I, A
Rifampicin                             I, P, A
Rokitamycin                            I, U, D, A
Roxithromycin                          I, P, A
                                                       9.41       Reagents, Test Solutions
Siccanin                               I, A
                                                    Change the following to read:
Sisomicin Sulfate                      I, A
Spectinomycin Hydrochloride            I, A           Albiflorin C23H28O11.xH2O White powder having no
Spiramycin Acetate II                  C, A         odor. Freely soluble in water, in methanol and in ethanol
1818       General Tests, Processes and Apparatus                                                   Supplement I, JP XV

(99.5).                                                           dehydrocorydaline nitrate for component determination in 1
  Identification Determine the absorption spectrum of a           mL of a mixture of water and methanol (1:1), and use this
solution of albiflorin in diluted methanol (1 in 2) (1 in         solution as the sample solution. Pipet 0.5 mL of the sample
100,000) as directed under Ultraviolet-visible Spectrophoto-      solution, add a mixture of water and methanol (1:1) to make
metry <2.24>: it exhibits a maximum between 230 nm and            exactly 50 mL, and use this solution as the standard solu-
234 nm.                                                           tion. Perform the test with these solutions as directed under
  Purity (1) Related substances 1—Dissolve 1 mg of al-            Thin-layer Chromatography <2.03>. Spot 5 mL of the sample
biflorin in 1 mL of mehthanol, and perform the test with 10       solution and standard solution on a plate of silica gel for
mL of this solution as directed in the Identification (2) under   thin-layer chromatography. Develop immediately with a
Peony Root: any spot other than the principal spot which          mixture of methanol, a solution of ammonium acetate (3 in
appears at around Rf 0.2 does not appear.                         10) and acetic acid (100) (20:1:1) to a distance of about 10
  (2) Related substances 2—Dissolve 1 mg of albiflorin in         cm, and air-dry the plate. Spray Dragendorff's TS on the
10 mL of diluted methanol (1 in 2), and use this solution as      plate, air-dry, and spray sodium nitrite TS: the spots other
the sample solution. Perform the test with 10 mL of the sam-      than the principal spot from the sample solution are not
ple solution as directed in the Assay under Peony Root, and       more intense than the spot from the standard solution.
measure the peak areas about 2 times as long as the retention        (2) Related substances 2—Dissolve 5.0 mg of de-
time of peoniflorin: the total area of the peaks other than al-   hydrocorydaline nitrate for component determination in 10
biflorin from the sample solution is not larger than 1/10         mL of the mobile phase, and use this solution as the sample
times the total area of the peaks other than the solvent peak.    solution. Pipet 1 mL of the sample solution, add the mobile
                                                                  phase to make exactly 100 mL, and use this solution as the
   Amygdalin for thin-layer chromatography C20H27NO11
                                                                  standard solution. Perform the test with exactly 5 mL each of
A white, odorless powder. Soluble in water, sparingly solu-
                                                                  the sample solution and standard solution as directed under
ble in methanol, and practically insoluble in ethanol (99.5).
                                                                  Liquid Chromatography <2.01> according to the following
   Identification Determine the absorption spectrum of a
                                                                  conditions, and measure each peak area from these solutions
solution of amygdalin for thin-layer chromatography in
                                                                  by the automatic integration method: the total area of peaks
methanol (1 in 1000) as directed under Ultraviolet-visible
                                                                  other than dehydrocorydaline from the sample solution is
Spectrophotometry <2.24>: it exhibits maxima between 250
                                                                  not larger than the peak area of dehydrocorydaline from the
nm and 254 nm, between 255 nm and 259 nm, between 261
                                                                  standard solution.
nm and 265 nm, and between 267 nm and 271 nm.
                                                                  Operating conditions
   Purity Related substances—Dissolve 5 mg of amygdalin
                                                                     Column, column temperature, mobile phase, and flow
for thin-layer chromatography in 2 mL of methanol, and use
                                                                  rate: Proceed as directed in the operating conditions in the
this solution as the sample solution. Pipet 1 mL of the sam-
                                                                  Component determination under Corydalis Tuber.
ple solution, add methanol to make exactly 100 mL, and use
                                                                     Detector: Ultraviolet absorption photometer (wavelength:
this solution as the standard solution. Perform the test with
                                                                  230 nm)
10 mL each of the sample solution and standard solution as
                                                                     Time span of measurement: About 3 times as long as the
directed in the Identification under Peach Kernel: any spot
                                                                  retention time of dehydrocorydaline, beginning after the
other than the principal spot at the Rf value of about 0.3 ob-
                                                                  peak of nitric acid.
tained from the sample solution is not more intense than the
                                                                  System suitability
spot from the standard solution.
                                                                     System performance and system repeatability: Proceed as
  Capsaicin for component determination        See (E)-capsai-    directed in the system suitability in the Component determi-
cin for component determination.                                  nation under Corydalis Tuber.
                                                                     Test for required detectability: To exactly 1 mL of the
  Capsaicin for thin-layer chromatography      See (E)-capsai-
                                                                  standard solution add the mobile phase to make exactly 20
cin for thin-layer chromatography.
                                                                  mL. Confirm that the peak area of dehydrocorydaline
  Chlorogenic acid for thin-layer chromatography           See    obtained from 5 mL of this solution is equivalent to 3.5 to
(E)-chlorogenic acid for thin-layer chromatography.               6.5z of that from 5 mL of the standard solution.

  Cinnamaldehyde for thin-layer chromatography             See       [6]-Gingerol for thin-layer chromatography C17H26O4
(E)-cinnamaldehyde for thin-layer chromatography.                 A yellow-white to yellow, liquid or solid. Freely soluble in
                                                                  methanol, in ethanol (99.5) and in diethyl ether, and practi-
  Dehydrocorydaline nitrate for component determination
                                                                  cally insoluble in water.
C22H24N2O7 Yellow, crystals or crystalline powder. It is
                                                                     Identification Determine the absorption spectrum of a
sparingly soluble in methanol, and slightly soluble in water
                                                                  solution of [6]-gingerol for thin-layer chromatography in
and in ethanol (99.5). Melting point: about 2409 (with
                                                    C
                                                                  ethanol (99.5) (7 in 200,000) as directed under Ultraviolet-
decomposition).
                                                                  visible Spectrophotometry <2.24>: it exhibits a maximum be-
  Absorbance <2.24> E1z (333 nm): 577 – 642 (3 mg,
                           1 cm
                                                                  tween 279 nm and 283 nm.
water, 500 mL). Use the sample dried in a desiccator (silica
                                                                     Purity Related substances—Dissolve 1.0 mg of [6]-gin-
gel) for not less than 1 hour for the test.
                                                                  gerol for thin-layer chromatography in exactly 2 mL of
  Purity (1) Related substances 1—Dissolve 5.0 mg of
Supplement I, JP XV                                              General Tests, Processes and Apparatus                  1819

methanol. Perform the test with 10 mL of this solution as       (C18H24N2O5S.HCl), calculated on the anhydrous basis.]
directed in the Identification under Ginger: any spot other
                                                                   Amygdalin for component determination Amygdalin
than the principal spot at the Rf value of about 0.3 does not
                                                                for thin-layer chromatography. However, it meets the fol-
appear.
                                                                lowing requirements:
   Magnolol for component determination Use magnolol               Absorbance <2.24> E1z (263 nm): 55 – 58 [20 mg,
                                                                                            1 cm
for thin-layer chromatography meeting the following addi-       methanol, 20 mL; separately determine the water <2.48> (5
tional specifications.                                          mg, coulometric titration) and calculate on the anhydrous
   Absorbance <2.24> E1z (290 nm): 270 – 293 (10 mg,
                           1 cm                                 basis].
methanol, 500 mL). Use the sample dried in a desiccator (sil-      Purity Related substances—Dissolve 5 mg of amygdalin
ica gel) for not less than 1 hour for the sample.               for component determination in 10 mL of the mobile phase,
   Purity Related substances—Dissolve 5.0 mg of mag-            and use this as the sample solution. Pipet 1 mL of the sample
nolol for component determination in 10 mL of the mobile        solution, add the mobile phase to make exactly 100 mL, and
phase, and use this solution as the sample solution. Pipet 1    use this as the standard solution. Perform the test with ex-
mL of the sample solution, add the mobile phase to make         actly 10 mL each of the sample solution and standard solu-
exactly 100 mL, and use this solution as the standard solu-     tion as directed under Liquid Chromatography <2.01> ac-
tion. Perform the test with exactly 10 mL each of the sample    cording to the following conditions, and determine each
solution and standard solution as directed under Liquid         peak area by the automatic integration method: the total
Chromatography <2.01> according to the following condi-         area of the peaks other than amygdalin from the sample so-
tions, and determine the area of each peak from these solu-     lution is not larger than the peak area of amygdalin from the
tions by the automatic integration method: the total area of    standard solution.
peaks other than the peak of magnolol from the sample solu-     Operating conditions
tion is not larger than the peak area of magnolol from the         Detector, column, column temperature, mobile phase,
standard solution.                                              and flow rate: Proceed as directed in the operating condi-
Operating conditions                                            tions in the Assay (3) under Keishibukuryogan Extract.
   Detector, column, column temperature, mobile phase,             Time span of measurement: About 3 times as long as the
and flow rate: Proceed as directed in the operating condi-      retention time of amygdalin.
tions in the component determination under Magnolia Bark.       System suitability
   Time span of measurement: About 3 times as long as the          Test for required detectability: Pipet 1 mL of the standard
retention time of magnolol.                                     solution, and add the mobile phase to make exactly 20 mL.
System suitability                                              Confirm that the peak area of amygdalin obtained with 10
   System performance, and system repeatability: Proceed as     mL of this solution is equivalent to 3.5 to 6.5z of that with
directed in the system suitability in the component determi-    10 mL of the standard solution.
nation under Magnolia Bark.                                        System performance and system repeatability: Proceed as
   Test for required detectability: To exactly 1 mL of the      directed in the system suitability in the Assay (3) under
standard solution add the mobile phase to make exactly 20       Keishibukuryogan Extract.
mL. Confirm that the peak area of magnolol obtained with
                                                                  Bisoprolol fumarate for assay (C18H31NO4)2.C4H4O4
10 mL of this solution is equivalent to 3.5 to 6.5z of that
                                                                [Same as the monograph Bisoprolol Fumarate. However,
with 10 mL of the standard solution.
                                                                when dried, it contains not less than 99.0z of bisoprolol
                                                                fumarate [(C18H31NO4)2.C4H4O4]. Also, when performing
Add the following:
                                                                the Purity (2) under Bisoprolol Fumarate, the total area of
  Alminoprofen for assay C13H17NO2 [Same as the                 the peaks other than bisoprolol is not greater than 1/5 times
monograph Alminoprofen. When dried, it contains not less        the peak area of bisoprolol from the standard solution].
than 99.5z of alminoprofen (C13H17NO2).]                          Purify as follows if needed.
                                                                  Purification method—Dissolve, with heating, 2 g of Bi-
  6-Amidino-2-naphthol methanesulfonate
                                                                soprolol Fumarate in 200 mL of ethyl acetate, add 0.5 g of
C11H10N2O.CH4O3S A white to pale yellow crystalline
                                                                activated carbon, shake well, and filter using a glass filter
powder. Melting point: about 2339 (with decomposition).
                                C
                                                                (G4). Place the filtrate in ice water for 2 hours while oc-
  Purity A solution obtained by dissolving 0.5 g of 6-
                                                                casional shaking. Collect the crystals that precipitate out us-
amidino-2-naphthol methanesulfonate in 10 mL of
                                                                ing a glass filter (G3). Dry the crystals obtained in vacuum at
methanol is clear.
                                                                809 for 5 hours using phosphorus (V) oxide as a dessicant.
                                                                    C
  Aminopyrine C13H17N3O White to pale yellow crystals
                                                                  Bromothymol blue-sodium hydroxide-ethanol TS Dis-
or crystalline powder.
                                                                solve 50 mg of bromothymol blue in 4 mL of diluted 0.2
  Melting point <2.60>: 107 – 1099C
                                                                mol/L sodium hydroxide TS (1 in 10) and 20 mL of ethanol
   Amosulalol hydrochloride for assay C18H24N2O5S.HCl           (99.5), and add water to make 100 mL.
[Same as the monograph Amosulalol Hydrochloride. It con-
                                                                  Bucillamine for assay C7H13NO3S2 [Same as the mono-
tains not less than 99.0z of amosulalol hydrochloride
                                                                graph Bucillamine. However, when dried, it contains not
1820       General Tests, Processes and Apparatus                                                    Supplement I, JP XV

less than 99.0z of bucillamine (C7H13NO3S2). Furthermore,           (E)-Capsaicin for thin-layer chromatography
it conforms to the following test.]                              C18H27NO3 White crystals, having a strong irritative odor.
   Purity Related substances—Dissolve 60 mg of bucilla-          Very soluble in methanol, freely soluble in ethanol (95) and
mine for assay in 20 mL of a mixture of water and methanol       in diethyl ether, and practically insoluble in water.
(1:1) and use this solution as the sample solution. Pipet 1 mL      Melting point <2.60>: 64.5 – 66.59 C
of this solution, add the mixture of water and methanol (1:1)       Purity Related substances—Dissolve 20 mg of capsaicin
to make exactly 100 mL, and use this solution as the stan-       for thin-layer chromatography in 2 mL of methanol, and use
dard solution. When the test is performed according to the       this solution as the sample solution. Pipet 1 mL of the sam-
Purity (3) under Bucillamine, the total area of the peaks        ple solution, add methanol to make exactly 100 mL, and use
other than the bucillamine peak from the sample solution is      this solution as the standard solution. Perform the test with
not larger than the peak area of bucillamine from the stan-      10 mL each of the sample solution and standard solution as
dard solution.                                                   directed in the Identification under Capsicum: any spot
                                                                 other than the principal spot at the Rf value of about 0.5
  Buformin hydrochloride for assay C6H15N5.HCl
                                                                 from the sample solution is not more intense than the spot
[Same as the monograph Buformin Hydrochloride. When
                                                                 from the standard solution.
dried, it contains not less than 99.5z of buformin hydro-
chloride (C6H15N5.HCl).]                                           Cesium chloride CsCl White crystals or crystalline
                                                                 powder. Very soluble in water, and freely soluble in ethanol
  Butyl benzoate C6H5COOCH2CH2CH2CH3                  A clear
                                                                 (99.5).
and colorless liquid.
                                                                   Loss on drying <2.41>: Not more than 1.0z (1 g, 1109 2  C,
  Refractive index <2.45> n20: 1.495 – 1.500
                             D
                                                                 hours).
  Specific gravity <2.56> d 20: 1.006 – 1.013
                            20
                                                                   Content: not less than 99.0z. Assay—Weigh accurately
  n-Butylboronic acid C4H11BO2        White flakes.              about 0.5 g, previously dried, and dissolve in water to make
  Melting point <2.60>: 90 – 929
                               C                                 exactly 200 mL. Pipet 20 mL of this solution, add 30 mL of
                                                                 water, and titrate <2.50> with 0.1 mol/L silver nitrate VS (in-
   (E)-Capsaicin for component determination Use (E)-
                                                                 dicator: fluorescein sodium TS).
capsaicin for thin-layer chromatography meeting the follow-
ing additional specifications.                                             Each mL of 0.1 mol/L silver nitrate VS
   Absorbance <2.24> E1z (281 nm): 97 – 105 (10 mg,
                            1 cm                                             =16.84 mg of CsCl
methanol, 200 mL). Use the sample dried in a desiccator (in
                                                                   Cesium chloride TS To 25.34 g of cesium chloride add
vacuum, phosphorus (V) oxide, 409 for 5 hours for the
                                       C)
                                                                 water to make 1000 mL.
test.
   Purity Related substances—Dissolve 10 mg of capsaicin           Cetirizine hydrochloride for assay C21H25ClN2O3.2HCl
for component determination in 50 mL of methanol, and            [Same as the monograph Cetirizine Hydrochloride. When
use this solution as the sample solution. Pipet 1 mL of the      dried, it contains not less than 99.5z of cetirizine
sample solution, add methanol to make exactly 100 mL, and        hydrochloride (C21H25ClN2O3.2HCl).]
use this solution as the standard solution. Perform the test
                                                                    3?-Chloro-3?-deoxythymidine for liquid chromatography
with exactly 20 mL each of the sample solution and standard
                                                                 C10H13N2O4Cl Occurs as a white powder.
solution as directed under Liquid Chromatography <2.01>
                                                                    Purity—Dissolve 10 mg of 3?-chloro-3?-deoxythymidine
according to the following conditions, and measure each
                                                                 for liquid chromatography in the mobile phase to make 100
peak area from these solutions by the automatic integration
                                                                 mL. Perform the test with 10 mL of this solution as directed
method: the total area of the peaks other than capsaicin
                                                                 in the Purity (3) under Zidovudine: a peak is not observed at
from the sample solution is not larger than the peak area of
                                                                 the retention time for zidovudine.
capsaicin from the standard solution.
Operating conditions                                               (E)-Chlorogenic acid for thin-layer chromatography
   Detector, column, column temperature, mobile phase,           C16H18O9.xH2O A white powder. Freely soluble in
and flow rate: Proceed the operating conditions in the Com-      methanol and in ethanol (99.5), and sparingly soluble in
ponent determination under Capsicum.                             water. Melting point: about 2059 (with decomposition).
                                                                                                    C
   Time span of measurement: About 3 times as long as the          Purity Related substances—Dissolve 1.0 mg of chloro-
retention time of capsaicin beginning after the solvent peak.    genic acid for thin-layer chromatography in 2 mL of
System suitability                                               methanol, and use this solution as the sample solution. Per-
   System performance, and system repeatability: Proceed         form the test with this solution as directed under Thin-layer
the system suitability in the Component determination under      Chromatography <2.03>. Spot 10 mL of the sample solution
Capsicum.                                                        on a plate of silica gel for thin-layer chromatography, de-
   Test for required detectability: Pipet 1 mL of the standard   velop the plate with a mixture of ethyl acetate, water and
solution, and add methanol to make exactly 20 mL. Con-           formic acid (6:1:1) to a distance of about 10 cm, and air-dry
firm that the peak area of capsaicin from 20 mL of this solu-    the plate. Examine under ultraviolet light (main wavelength:
tion is equivalent to 3.5 to 6.5z of that of capsaicin from 20   365 nm): no spot other than the principal spot at around Rf
mL of the standard solution.                                     0.5 appears.
Supplement I, JP XV                                               General Tests, Processes and Apparatus                 1821

  Chlorphenesin carbamate for assay C10H12ClNO4                  matography in 100 mL of methanol and perform the test as
[Same as the monograph Chlorphenesin Carbamate. When             directed in the Purity (2) under Zidovudine: spots other than
dried, it contains not less than 99.0z of chlorphenesin car-     the principal spot with an Rf value of about 0.23 are not
bamate (C10H12ClNO4).]                                           observed.

   Cibenzoline succinate for assay C18H18N2.C4H6O4                 Dilute sodium pentacyanonitrosylferrate (III)-potassium
[Same as the monograph Cibenzoline Succinate. When               hexacyanoferrate (III) TS To 5 mL of a solution of sodium
dried, it contains not less than 99.0z of cibenzoline suc-       pentacyanonitrosylferrate (III) dihydrate (3 in 50), 5 mL of a
cinate (C18H18N2.C4H6O4) and meets the following require-        solution of potassium hexacyanoferrate (III) (13 in 200) and
ment.]                                                           2.5 mL of a solution of sodium hydroxide (1 in 10) add water
   Purity: Related substances—Dissolve 0.10 g in 2 mL of         to make 25 mL, and mix. Use after the color of the solution
methanol, and use this solution as the sample solution. Pipet    changes from dark red to light yellow. Prepare at the time of
1 mL of the sample solution, and add methanol to make ex-        use.
actly 100 mL. To exactly 1 mL of this solution add methanol
                                                                   1,2-Dinitrobenzene C6H4(NO2)2 Occurs as yellowish
to make exactly 10 mL, and use this solution as the standard
                                                                 white to brownish yellow crystals or a crystalline powder.
solution. Perform the test with these solutions as directed
                                                                   Identification: Determine the infrared absorption spec-
under Thin-layer Chromatography <2.03>. Spot 10 mL each
                                                                 trum of 1,2-dinitrobenzene as directed in the paste method
of the sample solution and standard solution on a plate of
                                                                 under Infrared Spectrophotometry <2.25>: it exhibits ab-
silica gel with fluorescent indicator for thin-layer chro-
                                                                 sorption at the wave numbers of about 3100 cm-1, 1585
matography. Develop the plate with a mixture of ethyl
                                                                 cm-1, 1526 cm-1, 1352 cm-1, and 793 cm-1.
acetate, methanol and ammonia solution (28) (20:3:2) to a
                                                                   Melting point <2.60>: 116 – 1199C
distance of about 10 cm, air-dry the plate, and dry at 809   C
for 30 minutes. After cooling, examine under ultraviolet           Enalapril maleate C20H28N2O5.C4H4O4          [Same as the
light (main wavelength: 254 nm): the spot other than the         namesake monograph]
principal spot obtained with the sample solution is not more
                                                                   2-Ethylhexyl parahydroxybenzoate C15H22O3 Pale yel-
intense than the spot with the standard solution. On stand-
                                                                 low, clear viscous liquid. Miscible with methanol (99.5).
ing the plate for 30 minutes in the tank saturated with iodine
                                                                 Practically insoluble in water.
vapor, the spot other than the principal spot obtained with
                                                                   Content: not less than 98.0z. Assay—Proceed as
the sample solution is not more intense than the spot with
                                                                 directed in the Assay under Ethyl Parahydroxybenzoate.
the standard solution.
                                                                         Each mL of 1 mol/L sodium hydroxide VS
  Cilazapril C22H31N3O5.H2O       [Same as the monograph
                                                                           =250.3 mg of C15H22O3
Cilazapril Hydrate]
                                                                   Etizolam for assay C17H15ClN4S [Same as the mono-
  Cilazapril for assay C22H31N3O5.H2O [Same as the
                                                                 graph Etizolam. When dried, it contains not less than 99.0z
monograph Cilazapril Hydrate. It contains not less than
                                                                 of etizolam (C17H15ClN4S).]
99.0z of cilazapril (C22H31N3O5), calculated on the anhy-
drous basis.]                                                       [6]-Gingerol for component determination [6]-Gingerol
                                                                 for thin-layer chromatography. However, it meets the fol-
   (E)-Cinnamaldehyde for thin-layer chromatography
                                                                 lowing requirements:
C9H8O A colorless or light yellow liquid, having a charac-
                                                                    Absorbance <2.24> E1z (281 nm): 101 – 112 [7 mg,
                                                                                            1 cm
teristic aromatic odor. Very soluble in methanol and in
                                                                 ethanol (99.5), 200 mL].
ethanol (99.5), and practically insoluble in water.
                                                                    Purity Related substances—Dissolve 5 mg of [6]-gin-
   Absorbance <2.24> E1z (285 nm): 1679 – 1943 (5 mg,
                            1 cm
                                                                 gerol for component determination in 5 mL of methanol,
methanol, 2000 mL)
                                                                 and use this as the sample solution. Pipet 1 mL of the sample
   Purity Related substances—Dissolve 10 mg in 2 mL of
                                                                 solution, add methanol to make exactly 50 mL, and use this
methanol. Perform the test with 1 mL of this solution as
                                                                 as the standard solution. Perform the test with exactly 10 mL
directed in the Identification (3) under Kakkonto Extract: no
                                                                 each of the sample solution and standard solution as direct-
spot other than the principal spot (Rf value is about 0.4)
                                                                 ed under Liquid Chromatography <2.01> according to the
appears.
                                                                 following conditions, and determine each peak area by the
  Clorazepate dipotassium for assay                              automatic integration method: the total area of the peaks
C16H10ClKN2O3.KOH [Same as the monograph Cloraze-                other than [6]-gingerol from the sample solution is not larger
pate Dipotassium. When dried it contains not less than           than the peak area of [6]-gingerol from the standard solu-
99.0z of clorazepate dipotassium (C16H10ClKN2O3.KOH).]           tion.
                                                                 Operating conditions
  1-[(2R,5S)-2,5-Dihydro-5-(hydroxymethyl)-2-furyl] thy-
                                                                    Detector, column, column temperature, mobile phase,
mine for thin-layer chromatography C10H12N2O4 Occurs
                                                                 and flow rate: Proceed as directed in the operating condi-
as a white powder.
                                                                 tions in the Assay (3) under Hangekobokuto Extract.
  Purity—Dissolve 0.1 g of 1-[(2R,5S)-2,5-dihydro-5-
                                                                    Time span of measurement: About 6 times as long as the
(hydroxymethyl)-2-furyl]thymine for thin-layer chro-
1822       General Tests, Processes and Apparatus                                                   Supplement I, JP XV

retention time of [6]-gingerol.                                     Purity Related substances—Dissolve 1.0 mg of mag-
System suitability                                               nolol for thin-layer chromatography in 1 mL of methanol,
   Test for required detectability: Pipet 1 mL of the standard   and use this solution as the sample solution. Perform the test
solution, and add methanol to make exactly 20 mL. Con-           with the sample solution as directed under Thin-layer Chro-
firm that the peak area of [6]-gingerol obtained with 10 mL      matography <2.03>. Spot 10 mL of the sample solution on a
of this solution is equivalent to 3.5 to 6.5z of that with 10    plate of silica gel with fluorescent indicator for thin-layer
mL of the standard solution.                                     chromatography, develop the plate with a mixture of
   System performance and system repeatability: Proceed as       hexane, acetone and acetic acid (100) (20:15:1) to a distance
directed in the system suitability in the Assay (3) under Han-   of about 10 cm, and air-dry the plate. Examine under ultrav-
gekobokuto Extract.                                              iolet light (main wavelength: 254 nm): any spot other than
                                                                 the principal spot of around Rf 0.5 does not appear.
   4-Hydroxyisophthalic acid HOC6H3(COOH)2 White
crystals or powder.                                                Miconazole nitrate C18H14Cl4N2O.HNO3          [Same as the
   Content: not less than 98.0z. Assay—Weigh accurately          namesake monograph]
about 0.14 g, dissolve in 50 mL of ethanol (95), and titrate
                                                                   Minocycline hydrochloride     C23H27N3O7.HCl      [Same as
<2.50> with 0.1 mol/L sodium hydroxide VS (potentiometric
                                                                 the namesake monograph]
titration). Perform a blank determination in the same man-
ner, and make any necessary correction.                            Nile blue   C20H20ClN3O     Blue-green powder.

       Each mL of 0.1 mol/L sodium hydroxide VS                    3-Nitroaniline C6H6N2O2 Yellow crystals or crystalline
         =9.106 mg of C8H6O5                                     powder.
                                                                   Melting point <2.60>: 112 – 1169C
  Hyperoside for thin-layer chromatography C21H20O12
Yellow crystals or crystalline powder. Slightly soluble in         Nodakenin for thin-layer chromatography C20H24O9
methanol, very slightly soluble in ethanol (99.5), and practi-   White powder. Slightly soluble in water and in methanol,
cally insoluble in water. Melting point: about 2209 (with
                                                      C          and very slightly soluble in ethanol (99.5). Melting point:
decomposition).                                                  about 2209 (with decomposition).
                                                                             C
  Identification: Determine the absorption spectrum of a           Identification: Determine the absorption spectrum of a
solution of hyperoside for thin-layer chromatography in          solution of nodakenin for thin-layer chromatography in
methanol (1 in 100,000) as directed under Ultraviolet-visible    methanol (1 in 100,000) as directed under Ultraviolet-visible
Spectrophotometry <2.24>: it exhibits a maximum between          Spectrophotometry <2.24>: it exhibits a maximum between
255 nm and 259 nm.                                               333 nm and 337 nm.
  Purity Related substances—Dissolve 1 mg of hyperoside            Optical rotation <2.49>: [a]20: +50 – +689 (5 mg,
                                                                                                    D
for thin-layer chromatography in 20 mL of methanol. Per-         methanol, 10 mL, 100 mm).
form the test with 10 mL of this solution as directed in the       Purity Related substances—Dissolve 1 mg of nodakenin
Identification under Crataegus Fruit: any spot other than        for thin-layer chromatography in 3 mL of methanol, and use
the principal spot of around Rf 0.5 does not appear.             this solution as the sample solution. Pipet 1 mL of the sam-
                                                                 ple solution, add methanol to make exactly 100 mL, and use
  Isoxsuprine hydrochloride for assay C18H23NO3.HCl
                                                                 this solution as the standard solution. Proceed with 5 mL
[Same as the monograph Isoxsuprine Hydrochloride]
                                                                 each of these solutions as directed in the Identification (2)
  Labetalol hydrochloride      C19H24N2O3.HCl      [Same as      under Peucedanum Root: the spot other than the principal
the namesake monograph]                                          spot of around Rf 0.3 from the sample solution is not more
                                                                 intense than the spot from the standard solution.
  Labetalol hydrochloride for assay C19H24N2O3.HCl
[Same as the monograph Labetalol Hydrochloride.                     Oleic acid C18H34O2 Occurs as a colorless or pale yel-
However, when dried, it contains not less than 99.0z of          low transparent liquid and has a slightly distinct odor. It is
labetalol hydrochloride (C19H24N2O3.HCl).]                       miscible with ethanol (95) and with diethyl ether, and practi-
                                                                 cally insoluble in water.
  Lanthanum chloride TS To 58.65 g of lanthanum (III)
                                                                    Specific gravity <2.56> d 20: about 0.9
                                                                                              20
oxide add 100 mL of hydrochloric acid, and boil. After cool-
                                                                    Content: not less than 99.0z. Assay—To 40 mL of oleic
ing, add water to make 1000 mL.
                                                                 acid to be examined add 1 mL of a solution of boron trifluo-
   Magnolol for thin-layer chromatography C18H18O2               ride in methanol (3 in 20), mix, and heat on a water bath for
Odorless, white crystals or crystalline powder. Freely soluble   3 minutes. After cooling, add 10 mL of petroleum ether and
in methanol and in ethanol (99.5), and practically insoluble     10 mL of water, shake, collect the ether layer after allowing
in water. Melting point: about 1029   C.                         to stand, and use as the sample solution. Perform the test
   Identification: Determine the absorption spectrum of a        with 0.2 mL of the sample solution as directed under Gas
solution of magnolol for thin-layer chromatography in            Chromatography <2.02> according to the following condi-
methanol (1 in 50,000) as directed under Ultraviolet-visible     tions, determine each peak area by the automatic integration
Spectrophotometry <2.24>: it exhibits a maximum between          method, and calculate the amount of methyl oleate by the
287 nm and 291 nm.                                               area percentage method.
Supplement I, JP XV                                               General Tests, Processes and Apparatus                 1823

Operating conditions                                             ameter).
   Detector: A hydrogen flame-ionization detector                   Column temperature: A constant temperature of about
   Column: A glass column 3 mm in inside diameter and 2 m        409 C.
in length, packed with siliceous earth for gas chro-                Mobile phase: A mixture of diluted phosphoric acid (1 in
matography (149 – 177 mm) coated with methyl polyacrylate        1000) and acetonitrile (4:1).
in a rate of 5 – 10z.                                               Flow rate: Adjust the flow rate so that the retention time
   Column temperature: A constant temperature of about           of rosmarinic acid is about 14 minutes.
2209 C.                                                             Time span of measurement: About 4 times as long as the
   Carrier gas: Helium                                           retention time of rosmarinic acid.
   Flow rate: Adjust the flow rate so that the retention time    System suitability
of methyl oleate is about 10 minutes.                               Test for required detectability: Pipet 1 mL of the standard
Time span of measurement: About 2 times as long as the           solution, and add methanol to make exactly 20 mL. Con-
retention time of methyl oleate, beginning after the solvent     firm that the peak area of rosmarinic acid obtained with 10
peak.                                                            mL of this solution is equivalent to 3.5 to 6.5z of that with
                                                                 10 mL of the standard solution.
   (±)-Praeruptorin A for thin-layer chromatography
                                                                    System performance: When the procedure is run with 10
C21H22O7 White crystals or crystalline powder. Soluble in
                                                                 mL of the standard solution under the above operating con-
methanol, sparingly slightly soluble in ethanol (99.5), and
                                                                 ditions, the number of theoretical plates and the symmetry
practically insoluble in water.
                                                                 factor of the peak of rosmarinic acid are not less than 5000
   Identification: Determine the absorption spectrum of a
                                                                 and not more than 1.5, respectively.
solution of (±)-praeruptorin A for thin-layer chro-
                                                                    System repeatability: When the test is repeated 6 times
matography in methanol (1 in 100,000) as directed under
                                                                 with 10 mL of the standard solution under the above operat-
Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a
                                                                 ing conditions, the relative standard deviation of the peak
maximum between 320 nm and 324 nm.
                                                                 area of rosmarinic acid is not more than 1.5z.
   Melting point <2.60>: 152 – 1569 C
   Purity Related       substances—Dissolve       2 mg      of      Rosmarinic acid for thin-layer chromatography
(±)-praeruptorin A for thin-layer chromatography in 2 mL         C18H16O8 White to pale yellow crystals or crystalline pow-
of methanol, and use this solution as the sample solution.       der. Freely soluble in ethanol (99.5), and slightly soluble in
Pipet 1 mL of the sample solution, add methanol to make          water. Melting point: about 2059 (with decomposition).
                                                                                                    C
exactly 100 mL, and use this solution as the standard solu-         Identification: Determine the absorption spectrum of a
tion. Proceed with 5 mL each of these solutions as directed in   solution of rosmarinic acid for thin-layer chromatography in
the Identification (1) under Peucedanum Root: the spot           ethanol (99.5) (1 in 100,000) as directed under Ultraviolet-
other than the principal spot of around Rf 0.3 from the sam-     visible Spectrophotometry <2.24>: it exhibits maxima be-
ple solution is not more intense than the spot from the stan-    tween 217 nm and 221 nm, between 290 nm and 294 nm, and
dard solution.                                                   between 330 nm and 334 nm.
                                                                    Purity Related substances—Dissolve 10 mg of rosmarin-
   Rosmarinic acid for component determination Rosmar-
                                                                 ic acid for thin-layer chromatography in 2 mL of ethanol
inic acid for thin-layer chromatography. However, it meets
                                                                 (99.5), and use this solution as the sample solution. Pipet 1
the following requirements:
                                                                 mL of the sample solution, add ethanol (99.5) to make
   Absorbance <2.24> E1z (332 nm): 526 – 559 [5 mg,
                            1 cm
                                                                 exactly 50 mL, and use this solution as the standard solu-
ethanol (99.5), 500 mL].
                                                                 tion. Proceed with 10 mL each of the sample solution and
   Purity Related substances—Dissolve 5 mg of rosmarinic
                                                                 standard solution as directed in the Identification (2) under
acid for component determination in 20 mL of the mobile
                                                                 Hangekobokuto Extract: the spot other than the principal
phase, and use this as the sample solution. Pipet 1 mL of the
                                                                 spot of around Rf 0.5 from the sample solution is not more
sample solution, add methanol to make exactly 50 mL, and
                                                                 intense than the spot from the standard solution.
use this as the standard solution. Perform the test with ex-
actly 10 mL each of the sample solution and standard solu-         Sodium di-2-ethylhexyl sulfosuccinate
tion as directed under Liquid Chromatography <2.01> ac-          C8H17COOCH2(C8H17COO)CHSO3Na White or translu-
cording to the following conditions, and determine each          cent white mucilaginous soft masses. Sparingly soluble in
peak area by the automatic integration method: the total         water.
area of the peaks other than rosmarinic acid from the sample       Purity Clarity and color of solution: A solution pre-
solution is not larger than the peak area of rosmarinic acid     pared by dissolving 1.0 g in 100 mL of water is clear and
from the standard solution.                                      colorless.
Operating conditions                                               Loss on drying <2.41>: not more than 5.0z (1 g, 1059 2
                                                                                                                       C,
   Detector: An ultraviolet absorption photometer (wave-         hours).
length: 240 nm).
                                                                   Sodium dihydrogen phosphate TS, pH 2.2 Dissolve 1.56
   Column: A stainless steel column 4.6 mm in inside di-
                                                                 g of sodium dihydrogen phosphate dihydrate in 800 mL of
ameter and 15 cm in length, packed with octadecylsilanized
                                                                 water, adjust the pH to 2.2 with phosphoric acid, and add
silica gel for liquid chromatography (5 mm in particle di-
1824       General Tests, Processes and Apparatus                                                    Supplement I, JP XV

water to make 1000 mL.                                              Melting point <2.60>: 49 – 529  C
                                                                    Purity: Other phenols—Shake vigorously 1.0 g of the sub-
  1 mol/L Sulfuric acid TS Add 60 mL of sulfuric acid in
                                                                 stance to be examined with 20 mL of warm water for 1
1000 mL of water slowly with stirring, then allow to cool.
                                                                 minute, and filter. To 5 mL of the filtrate add 1 drop of a so-
  5 mol/L Sulfuric acid TS Add 300 mL of sulfuric acid in        lution of iron (III) chloride hexahydrate (27 in 100): the solu-
1000 mL of water slowly with stirring, then allow to cool.       tion reveals a green but not a blue to purple color.

   Thymine for liquid chromatography C5H6N2O2 Oc-                  Thymol-sulfuric acid-methanol TS for spraying Dissolve
curs as a white powder.                                          1.5 g of thymol for spraying test solution in 100 mL of
   Purity—Dissolve 10 mg of the substance to be examined         methanol, and add 5.7 mL of sulfuric acid.
in 100 mL of methanol, add the mobile phase to make exact-
                                                                    Triphenylmethanol for thin-layer chromatography
ly 250 mL, and use this solution as the sample solution.
                                                                 C19H15OH Occurs as a white powder.
Pipet 5 mL of this solution, add the mobile phase to make
                                                                    Purity—Dissolve 0.1 g of triphenylmethanol for thin-lay-
exactly 100 mL, and use this solution as the standard solu-
                                                                 er chromatography in 100 mL of methanol and perform the
tion. Pipet 10 mL each of these solutions and perform the
                                                                 test as directed in the Purity (2) under Zidovudine: spots
test as directed in the Purity (3) under Zidovudine. Deter-
                                                                 other than the principal spot with an Rf value of about 0.73
mine the area of each peak in the sample and standard solu-
                                                                 are not observed.
tions by the automatic integration method: the total area of
peaks other than thymine from the sample solution is not
larger than that from the standard solution. However, the
time span of measurement is about 10 times the retention             9.42 Solid Supports/Column
time of thymine, beginning after the solvent peak.                   Packings for Chromatography
   Thymol for spraying test solution C10H14O White crys-
tals or crystalline powder, having an aromatic odor. Very        Add the following:
soluble in methanol and in ethanol (99.5), and practically in-     Porous styrene-divinylbenzene copolymer for liquid chro-
soluble in water.                                                matography A porous styrene-divinylbenzen copolymer
   Identification: Determine the infrared absorption spec-       prepared for liquid chromatography.
trum as directed in the potassium bromide disk method un-
der Infrared Spectrophotometry <2.25>: it exhibits absorp-        Strongly basic ion exchange resin for column chro-
tion at the wave numbers of about 2960 cm-1, 1420 cm-1,          matography Prepared for column chromatography.
1290 cm-1, 1090 cm-1 and 810 cm-1.
                               Official Monographs
Add the following:                                                 the standard solution. Perform the test with these solutions
                                                                   as directed under Thin Layer Chromatography <2.03>. Spot
Acemetacin                                                         5 mL each of the sample solution and standard solution on a
                                                                   plate of silica gel with fluorescent indicator for thin layer
アセメタシン                                                             chromatography. Develop the plate with a mixture of
                                                                   hexane, 4-methyl-2-pentanone and acetic acid (100) (3:2:1)
                                                                   to a distance of about 10 cm, and air-dry the plate. Examine
                                                                   under ultraviolet light (main wavelength: 254 nm): not more
                                                                   than 2 spots other than the principal spot appear from the
                                                                   sample solution, and these spots are not more intense than
                                                                   the spot obtained from the standard solution.

                                                                   Loss on drying <2.41>                                C,
                                                                                            Not more than 0.5z (1 g, 1059 2
C21H18ClNO6: 415.82
                                                                   hours).
2-{2-[1-(4-Chlorobenzoyl)-5-methoxy-2-methyl-1H-
indol-3-yl]acetyloxy}acetic acid [53164-05-9]                      Residue on ignition <2.44>   Not more than 0.1z (1 g).

                                                                   Assay Weigh accurately about 0.35 g of Acemetacin,
  Acemetacin, when dried, contains not less than
                                                                   previously dried, dissolve in 20 mL of acetone, add 10 mL of
99.0z and not more than 101.0z of C21H18ClNO6.
                                                                   water, and then titrate <2.50> with 0.1 mol/L sodium
Description Acemetacin occurs as a light yellow crystalline        hydroxide VS (potentiometric titration). Perform a blank
powder.                                                            determination in the same method, and make any necessary
   It is soluble in acetone, sparingly soluble in methanol,        correction.
slightly soluble in ethanol (99.5), and practically insoluble in
                                                                          Each mL of 0.1 mol/L sodium hydroxide VS
water.
                                                                            =41.58 mg of C21H18ClNO6
Identification (1) To 1 mg of Acemetacin add 1 mL of
                                                                   Containers and storage    Containers—Tight containers.
concentrated chromotropic acid TS, and heat in a water bath
for 5 minutes: a red-purple color develops.
  (2) Determine the absorption spectrum of a solution of
Acemetacin in methanol (1 in 50,000) as directed under             Acetylcholine Chloride for
Ultraviolet-visible Spectrophotometry <2.24>, and compare          Injection
the spectrum with the Reference Spectrum: both spectra
exhibit similar intensities of absorption at the same              注射用アセチルコリン塩化物
wavelengths.
  (3) Determine the infrared absorption spectrum of                Add the following next to Residue on ignition:
Acemetacin as directed in the potassium bromide disk
                                                                   Uniformity of dosage units <6.02>   It meets the requirement
method under Infrared Spectrometry <2.25>, and compare
                                                                   of the Mass variation test.
the spectrum with the Reference Spectrum: both spectra
exhibit similar intensities of absorption at the same wave         Foreign insoluble matter <6.06> Perform the test according
numbers.                                                           to Method 2: it meets the requirement.
  (4) Perform the test with Acemetacin as directed under
                                                                   Insoluble particulate matter <6.07>   It meets the require-
Flame Coloration Test <1.04> (2): a green color appears.
                                                                   ment.
Melting point <2.60>             C
                        151 – 1549
                                                                   Sterility <4.06> Perform the test according to the Mem-
Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of               brane filtration method: it meets the requirement.
Acemetacin according to Method 4, and perform the test.
Prepare the control solution with 2.0 mL of Standard Lead
Solution (not more than 20 ppm).
  (2) Related substances—Dissolve 0.40 g of Acemetacin
in 10 mL of acetone, and use this solution as the sample
solution. Pipet 1 mL of the sample solution, and add
acetone to make exactly 50 mL. Pipet 1 mL of this solution,
add acetone to make exactly 10 mL, and use this solution as

                                                                                                                          1825
1826       Official Monographs                                                                        Supplement I, JP XV

                                                                   <2.24>, and compare the spectrum with the Reference Spec-
Ajimaline Tablets                                                  trum: both spectra exhibit similar intensities of absorption at
                                                                   the same wavelengths.
アジマリン錠                                                                (2) Determine the infrared absorption spectrum of
                                                                   Alminoprofen as directed in the potassium bromide disk
Add the following next to Identification:                          method under Infrared Spectrophotometry <2.25>, and com-
                                                                   pare the spectrum with the Reference Spectrum: both spec-
Uniformity of dosage units <6.02> Perform the test accord-
                                                                   tra exhibit similar intensities of absorption at the same wave
ing to the following method: it meets the requirement of the
                                                                   numbers.
Content uniformity test.
   To 1 tablet of Ajimaline Tablets add 150 mL of 2nd fluid        Melting point <2.60>             C
                                                                                           106 – 1089
for dissolution test, shake to disintegrate the tablet, then add
                                                                   Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of
2nd fluid for dissolution test to make exactly 200 mL, and
                                                                   Alminoprofen according to Method 2, and perform the test.
filter this solution through a membrane filter with a pore size
                                                                   Prepare the control solution with 2.0 mL of Standard Lead
not exceeding 0.8 mm. Discard the first 10 mL of the filtrate,
                                                                   Solution (not more than 10 ppm).
pipet V mL of the subsequent filtrate equivalent to about 0.5
                                                                      (2) Arsenic <1.11>—Prepare the test solution with 1.0 g
mg of ajimaline (C20H26N2O2), add 2nd fluid for dissolution
                                                                   of Alminoprofen according to Method 3, and perform the
test to make exactly 10 mL, and use this solution as the sam-
                                                                   test (not more than 2 ppm).
ple solution. Separately, weigh accurately about 25 mg of
                                                                      (3) Related substances—Conduct this procedure using
ajimaline for assay, previously dried in vacuum at 809 for C
                                                                   light-resistant vessels. Dissolve 50 mg of Alminoprofen in
3 hours, dissolve in 2nd fluid for dissolution test to make ex-
                                                                   100 mL of the mobile phase, and use this solution as the
actly 500 mL, and use this solution as the standard solution.
                                                                   sample solution. Pipet 2 mL of the sample solution, add the
Determine the absorbances at 288 nm, AT and AS, of the
                                                                   mobile phase to make exactly 200 mL, and use this solution
sample solution and standard solution as directed under
                                                                   as the standard solution. Perform the test with exactly 5 mL
Ultraviolet-visible Spectrophotometry <2.24>.
                                                                   each of the sample solution and standard solution as direct-
          Amount (mg) of ajimaline (C20H26N2O2)                    ed under Liquid Chromatography <2.01> according to the
           =WS×(AT/AS)×(1/V)×4                                     following conditions. Determine each peak area of these so-
                                                                   lutions by the automatic integration method: the area of the
  WS: Amount (mg) of ajimaline for assay
                                                                   peak other than alminoprofen obtained from the sample so-
                                                                   lution is not larger than 1/5 times the peak area of
                                                                   alminoprofen from the standard solution. Furthermore, the
Add the following:
                                                                   total area of the peaks other than alminoprofen from the
                                                                   sample solution is not larger than the peak area of
Alminoprofen                                                       alminoprofen from the standard solution.
アルミノプロフェン                                                          Operating conditions—
                                                                      Detector: An ultraviolet absorption photometer (wave-
                                                                   length: 254 nm).
                                                                      Column: A stainless steel column 6.0 mm in inside di-
                                                                   ameter and 15 cm in length, packed with octadecylsilanized
                                                                   silica gel for liquid chromatography (5 mm in particle di-
                                                                   ameter).
C13H17NO2: 219.28                                                     Column temperature: A constant temperature of about
(2RS)-2-{[4-(2-Methylprop-2-en-1-                                  259  C.
yl)amino]phenyl}propanoic acid [39718-89-3]                           Mobile phase: A mixture of methanol and diluted acetic
                                                                   acid (100) (1 in 1000) (4:1).
  Alminoprofen, when dried, contains not less than                    Flow rate: Adjust the flow rate so that the retention time
99.0z and not more than 101.0z of C13H17NO2.                       of alminoprofen is about 5 minutes.
Description Alminoprofen occurs as white to pale yellow               Time span of measurement: About 5 times as long as the
crystals or crystalline powder.                                    retention time of alminoprofen, beginning after the solvent
  It is freely soluble in ethanol (99.5) and in acetic acid        peak.
(100), and very slightly soluble in water.                         System suitability—
  It gradually turns brown on exposure to light.                      Test for required detectability: Pipet 1 mL of the standard
  A solution of Alminoprofen in ethanol (99.5) (1 in 10)           solution, and add the mobile phase to make exactly 10 mL.
shows no optical rotation.                                         Confirm that the peak area of alminoprofen obtained from
                                                                   5 mL of this solution is equivalent to 7 to 13z of that from 5
Identification (1) Determine the absorption spectrum of            mL of the standard solution.
a solution of Alminoprofen in ethanol (99.5) (3 in 500,000)           System performance: Dissolve 10 mg each of
as directed under Ultraviolet-visible Spectrophotometry            Alminoprofen and butyl parahydroxybenzoate in 100 mL of
Supplement I, JP XV                                                                      Official Monographs            1827

methanol. Pipet 10 mL of this solution, and add methanol        the test with exactly 5 mL each of the sample solution and
to make exactly 50 mL. When the procedure is run with 5 mL      standard solution as directed under Liquid Chromatography
of this solution under the above operating conditions,          <2.01> according to the conditions described in the Purity (3)
alminoprofen and butyl parahydroxybenzoate are eluted in        under Alminoprofen. Determine each peak area of each so-
this order with the resolution between these peaks being not    lution by the automatic integration method: the area of the
less than 2.0.                                                  peak other than alminoprofen obtained from the sample so-
   System repeatability: When the test is repeated 6 times      lution is not larger than 1/2 times the peak area of
with 5 mL of the standard solution under the above operat-      alminoprofen from the standard solution. Furthermore, the
ing conditions, the relative standard deviation of the peak     total area of the peaks other than alminoprofen from the
area of alminoprofen is not more than 2.0z.                     sample solution is not larger than 2 times the peak area of
                                                                alminoprofen from the standard solution.
Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-
um, phosphorus (V) oxide, 1 hour).                              Uniformity of dosage units <6.02> Perform the test accord-
                                                                ing to the following method: it meets the requirement of the
Residue on ignition <2.44>   Not more than 0.1z (1 g).
                                                                Content uniformity test.
Assay Weigh accurately about 0.3 g of Alminoprofen,               To 1 tablet of Alminoprofen Tablets add 5 mL of water,
previously dried, dissolve in 50 mL of acetic acid (100), and   shake until the tablet is disintegrated, add 50 mL of ethanol
titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio-     (99.5), shake for 20 minutes, then add ethanol (99.5) to
metric titration). Perform a blank determination in the same    make exactly 100 mL, and centrifuge. Pipet 3 mL of the
manner, and make any necessary correction.                      supernatant liquid, add ethanol (99.5) to make exactly 50
                                                                mL. Pipet V mL of this solution, add ethanol (99.5) to make
        Each mL of 0.1 mol/L perchloric acid VS
                                                                exactly V? mL so that each mL contains about 6 mg of
          =21.93 mg of C13H17NO2
                                                                alminoprofen (C13H17NO2), and use this solution as the sam-
Containers and storage Containers—Well-closed contain-          ple solution. Then, proceed as directed in the Assay.
ers.
                                                                        Amount (mg) of alminoprofen (C13H17NO2)
  Storage—Light-resistant.
                                                                         =WS×(AT/AS)×(V?/V)×(1/3)

                                                                  WS: Amount (mg) of alminoprofen for assay
Add the following:
                                                                Dissolution <6.10> When the test is performed at 50 revolu-
                                                                tions per minute according to the Paddle method, using 900
Alminoprofen Tablets                                            mL of 2nd fluid for dissolution test as the dissolution medi-
                                                                um, the dissolution rate in 45 minutes of Alminoprofen
アルミノプロフェン錠
                                                                Tablets is not less than 80z.
                                                                   Start the test with 1 tablet of Alminoprofen Tablets,
  Alminoprofen Tablets contain not less than 93.0z
                                                                withdraw not less than 20 mL of the medium at specified
and not more than 107.0z of the labeled amount of
                                                                minute after starting the test, and filter through a membrane
alminoprofen (C13H17NO2: 219.28).
                                                                filter with a pore size not exceeding 0.45 mm. Discard the
Method of preparation    Prepare as directed under Tablets,     first 10 mL of the filtrate, pipet V mL of the subsequent
with Alminoprofen.                                              filtrate, add 0.05 mol/L sodium hydroxide TS to make
                                                                exactly V? mL so that each mL contains about 8.9 mg of
Identification Take an amount of powdered Alminoprofen
                                                                alminoprofen (C13H17NO2) according to the labeled amount,
Tablets, equivalent to 30 mg of Alminoprofen according to
                                                                and use this solution as the sample solution. Separately,
the labeled amount, add ethanol (99.5) to make 100 mL,
                                                                weigh accurately about 30 mg of alminoprofen for assay,
shake thoroughly, and centrifuge. To 2 mL of the super-
                                                                previously dried in vacuum for 1 hour using phosphorus (V)
natant liquid add ethanol (99.5) to make 100 mL, and deter-
                                                                oxide as the dessicant, and dissolve in 0.05 mol/L sodium
mine the absorption spectrum of this solution as directed un-
                                                                hydroxide TS to make exactly 100 mL. Pipet 3 mL of this
der Ultraviolet-visible Spectrophotometry <2.24>: it exhibits
                                                                solution, add 0.05 mol/L sodium hydroxide TS to make
maxima between 253 nm and 257 nm, and between 298 nm
                                                                exactly 100 mL, and use this solution as the standard solu-
and 302 nm.
                                                                tion. Determine the absorbances, AT and AS, at 245 nm of
Purity Related substances—Conduct this procedure using          the sample solution and standard solution as directed under
light-resistant vessels. Powder 10 tablets of Alminoprofen      Ultraviolet-visible Spectrophotometry <2.24>.
Tablets, weigh a portion of the powder equivalent to 50 mg
                                                                Dissolution rate (z) with respect to the labeled amount of
of Alminoprofen according to the labeled amount, add 50
                                                                alminoprofen (C13H17NO2)
mL of the mobile phase, shake for 15 minutes, add the mo-
                                                                  =WS×(AT/AS)×(V?/V)×(1/C)×27
bile phase to make exactly 100 mL, centrifuge, and use the
supernatant liquid as the sample solution. Pipet 2 mL of the      WS: Amount (mg) of alminoprofen for assay
sample solution, add the mobile phase to make exactly 200         C: Labeled amount (mg) of alminoprofen (C13H17NO2) in
mL, and use this solution as the standard solution. Perform          1 tablet
1828       Official Monographs                                                                       Supplement I, JP XV

Assay Weigh accurately the mass of not less than 20 tablets       entire volume of the sample solution and 100 mL of the
of Alminoprofen Tablets, and powder. Weigh accurately an          standard solution on a plate of silica gel for thin-layer chro-
amount equivalent to about 60 mg of alminoprofen                  matography. Then, develop the plate with a mixture of ethyl
(C13H17NO2), add ethanol (99.5) and shake well, add ethanol       acetate, ethanol (99.5) and acetic acid (100) (100:5:1) to a
(99.5) to make exactly 200 mL, and centrifuge. Pipet 2 mL         distance of about 10 cm, and air-dry the plate. Spray evenly
of the supernatant liquid, add ethanol (99.5) to make exactly     a solution of phosphomolybdic acid n-hydrate in ethanol
100 mL, and use this solution as the sample solution.             (99.5) (1 in 10) on the plate, and heat at 1009 for 5 minutes:
                                                                                                                 C
Separately, weigh accurately about 30 mg of alminoprofen          the color of the spot obtained from the standard solution
for assay, previously dried in vacuum for 1 hour using phos-      and the spot corresponding to that location obtained from
phorus (V) oxide as the dessicant, dissolve in ethanol (99.5)     the sample solution is dark blue.
to make exactly 100 mL. Pipet 2 mL of this solution, add
                                                                  pH   Being specified separately.
ethanol (99.5) to make exactly 100 mL, and use this solution
as the standard solution. Determine the absorbances, AT and       Purity (1) Heavy metals <1.07>—Proceed with 4.0 mL of
AS, at the wavelength of maximum absorption at about 255          Alprostadil Injection according to Method 2, and perform
nm of the sample solution and standard solution as directed       the test. Prepare the control solution with 2.0 mL of Stan-
under Ultraviolet-visible Spectrophotometry <2.24>.               dard Lead Solution (not more than 5 ppm).
                                                                    (2) Prostaglandin A1—Use the sample solution obtained
        Amount (mg) of alminoprofen (C13H17NO2)
                                                                  in the Assay as the sample solution. Separately, weigh ac-
         =WS×(AT/AS)×2
                                                                  curately about 10 mg of prostaglandin A1, previously dried
  WS: Amount (mg) of alminoprofen for assay                       for 4 hours in a desiccator (in vacuum, phosphorus (V)
                                                                  oxide), and dissolve in ethanol (99.5) to make exactly 100
Containers and storage     Containers—Well-closed contain-
                                                                  mL. Pipet 2.5 mL of this solution, and add the mobile phase
ers.
                                                                  to make exactly 50 mL. Pipet 1 mL of this solution, add ex-
                                                                  actly 1 mL of the internal standard solution, and use this so-
                                                                  lution as the standard solution. Perform the test with 40 mL
Add the following:
                                                                  each of the sample solution and standard solution as direct-
                                                                  ed under Liquid Chromatography <2.01> according to the
Alprostadil Injection                                             following conditions, determine the ratios, QT and QS, of the
                                                                  peak area of prostaglandin A1 to that of the internal stan-
アルプロスタジル注射液
                                                                  dard, and calculate the amount of prostaglandin A1 convert-
                                                                  ed to alprostadil using the following equation: not more
  Alprostadil Injection is an emulsion-type injection.
                                                                  than 3.0 mg per a volume, equivalent to 5 mg of alprostadil
  It contains not less than 80.0z and not more than
                                                                  (C20H34O5).
125.0z of the labeled amount of alprostadil
(C20H34O5: 354.48).                                               Amount (mg) of prostaglandin A1 (C20H32O4), converted to
                                                                  alprostadil
Method of preparation      Prepare as directed under Injec-
                                                                    =WS×(QT/QS)×(1/2)×1.054
tions, with Alprostadil.
                                                                    WS: Amount (mg) of prostaglandin A1
Description Alprostadil Injection occurs as a white emul-
sion and is slightly viscous. It has a distinctive odor.          Internal standard solution—Dissolve 50 mg of 1-naphthol in
                                                                  20 mL of ethanol (99.5). To 3 mL of this solution add the
Identification To a quantity of Alprostadil Injection, cor-
                                                                  mobile phase to make 100 mL.
responding to 10 mg of Alprostadil according to the labeled
                                                                  Operating conditions—
amount, add 2 mL of acetonitrile, shake well, and cen-
                                                                     Proceed as directed in the operating conditions in the
trifuge. To 3.5 mL of the supernatant liquid add 7 mL of
                                                                  Assay.
diluted phosphoric acid (1 in 1000), and then run this solu-
                                                                  System suitability—
tion on a column (prepared by filling a 10 mm inside di-
                                                                     Test for required detectability: To exactly 1 mL of the
ameter, 9 mm long chromatography tube with 0.4 g of 70
                                                                  standard solution add the mobile phase to make exactly 5
mm octadecylsilanized silica gel for pretreatment) prewashed
                                                                  mL. Confirm that the peak area of prostaglandin A1 ob-
with 10 mL of methanol and then 10 mL of water. Wash the
                                                                  tained with 40 mL of this solution is equivalent to 14 to 26z
column with 10 mL of water and then 20 mL of petroleum
                                                                  of that with 40 mL of the standard solution.
ether, followed by elution with 2.5 mL of a mixture of
                                                                     System performance, and system repeatability: Proceed as
methanol and water (4:1). Remove the solvent from the ef-
                                                                  directed in the system suitability in the Assay.
fluent under reduced pressure, dissolve the residue in 100 mL
                                                                     (3) Peroxide—Pipet 4 mL of Alprostadil Injection,
of ethyl acetate, and use this solution as the sample solution.
                                                                  place in a glass-stoppered flask, add 15 mL of a mixture of
Separately, dissolve 1 mg of Alprostadil Reference Standard
                                                                  acetic acid (100) and isooctane (3:2), previously having
in 10 mL of ethyl acetate, and use this solution as the
                                                                  undergone a 30 minute nitrogen substitution, and dissolve
standard solution. Perform the test with these solutions as
                                                                  with gentle shaking. To this solution add 0.5 mL of saturat-
directed under Thin-layer Chromatography <2.03>. Spot the
Supplement I, JP XV                                                                      Official Monographs            1829

ed potassium iodide TS, replace the inside of the vessel with   1 mL of the internal standard solution, shake, and use this
nitrogen, and shake for exactly 5 minutes. Then, add 0.5 mL     solution as the sample solution. Separately, weigh accurately
of starch TS, shake vigorously, add 15 mL of water, and         about 5 mg of Alprostadil Reference Standard, previously
shake vigorously. Under a stream of nitrogen, titrate <2.50>    dried in a desiccator (in vacuum, phosphorus (V) oxide) for
with 0.01 mol/L sodium thiosulfate VS until the color of the    4 hours, dissolve in ethanol (99.5) to make exactly 50 mL,
solution disappears. Separately, perform a blank determina-     and use this solution as standard stock solution. Pipet 2.5
tion using 4 mL of water, and make any necessary correc-        mL of the standard stock solution, add the mobile phase to
tion. Calculate the amount of peroxides using the following     make exactly 50 mL, pipet 1 mL, add exactly 1 mL of the
equation: not more than 0.5 meq/L.                              internal standard solution, and use this solution as the stan-
                                                                dard solution. Perform the test with 40 mL each of the sam-
          Amount (meq/L) of peroxides=V×2.5
                                                                ple solution and standard solution as directed under Liquid
  V: Amount (mL) of 0.01 mol/L sodium thiosulfate VS            Chromatography <2.01> according to the following condi-
     consumed                                                   tions using an apparatus equipped with an automatic
                                                                pretreatment device (using a postcolumn reaction), and cal-
   (4) Free fatty acids—Pipet 3 mL of Alprostadil Injec-
                                                                culate the ratios, QT and QS, of the peak area of alprostadil
tion, add exactly 15 mL of a mixture of 2-propanol, heptane
                                                                to that of the internal standard.
and 0.5 mol/L sulfuric acid TS (40:10:1), and shake for 1
minute. After leaving for 10 minutes, add exactly 9 mL of                 Amount (mg) of alprostadil (C20H34O5)
heptane and exactly 9 mL of water, shake the test tube by in-              =WS×(QT/QS)×(1/2)
verting 10 times, leave for 15 minutes, and pipet 9 mL of the
                                                                  WS: Amount (mg) of Alprostadil Reference Standard
supernatant liquid. To this solution, add 3 mL of a solution
prepared by combining 1 volume of Nile blue solution (1 in      Internal standard solution—Dissolve 50 mg of 1-naphthol in
5000) washed 5 times with heptane and 9 volumes of ethanol      20 mL of ethanol (99.5). To 3 mL of this solution add the
(99.5), and use this solution as the sample solution. Titrate   mobile phase to make 100 mL.
<2.50> this solution with 0.02 mol/L sodium hydroxide VS        Operating conditions—
under a stream of nitrogen. Separately, dissolve 5.65 g of         Equipment: Liquid chromatograph consisting of 2 pumps
oleic acid in heptane to make exactly 200 mL, and use this      for pumping the mobile phase and the reaction reagent, an
solution as the standard solution. Pipet 25 mL of the stan-     automatic pretreatment device, column, reaction coil, detec-
dard solution, add 2 drops of phenolphthalein TS, titrate       tor, and recording apparatus. Use a reaction coil that is
<2.50> with 0.1 mol/L potassium hydroxide-ethanol VS until      maintained at a constant temperature.
a light red color develops, and determine the correction fac-      Detector: An ultraviolet absorption photometer (wave-
tor f. Pipet 30 mL of the standard solution and add heptane     length: 278 nm).
to make exactly 200 mL. Pipet 3 mL of this solution, add ex-       Column: A stainless steel column 4.6 mm in inside di-
actly 15 mL of a mixture of 2-propanol, heptane and 0.5         ameter and 15 cm in length, packed with octadecylsilanized
mol/L sulfuric acid TS (40:10:1), and shake for 1 minute.       silica gel for liquid chromatography (5 mm in particle di-
After leaving for 10 minutes, add exactly 6 mL of heptane       ameter).
and exactly 12 mL of water, shake the test tube by inverting       Column temperature: A constant temperature of about
10 times, and then titrate <2.50> in the same manner as for     609  C.
the sample solution. Determine the volume (mL), VT and VS,         Reaction coil: Polytetrafluoroethylene tube 0.5 mm in
of 0.02 mol/L sodium hydroxide VS consumed by the sam-          inside diameter and 10 m in length.
ple and standard solutions: the amount of free fatty acid is       Mobile phase: Dissolve 9.07 g of potassium dihydrogen
not more than 12.0 meq/L.                                       phosphate in water to make 1000 mL and adjust the pH to
                                                                6.3 by adding a solution prepared by dissolving 9.46 g of dis-
  Amount (meq/L) of free fatty acids=(VT/VS)×f×15
                                                                odium hydrogen phosphate in water to make 1000 mL. To 1
Bacterial endotoxins <4.01>    Less than 10 EU/mL.              volume of this solution add 9 volumes of water. To 3
                                                                volumes of this solution add 1 volume of acetonitrile for liq-
Extractable volume <6.05>     It meets the requirement.
                                                                uid chromatography.
Foreign insoluble matter <6.06> Perform the test according         Reaction reagent: Potassium hydroxide TS.
to Method 1: no easily detectable foreign matter is observed.      Reaction temperature: A constant temperature of about
                                                                609  C.
Sterility <4.06> Perform the test according to the Mem-
                                                                   Mobile phase flow rate: Adjust the flow rate so that the
brane filter method: it meets the requirement. However, use
                                                                retention time of alprostadil is about 7 minutes.
the sample solution consisting of equal volume of Al-
                                                                   Reaction reagent flow rate: 0.5 mL per minute.
prostadil Injection and a solution prepared by adding water
                                                                   Automatic pretreatment device: Composed of a pretreat-
to 0.1 g of polysorbate 80 to make 100 mL.
                                                                ment column, pump for pumping pretreatment column wash
Particle diameter   Being specified separately.                 solution, and routing valve for 2 high pressure flow paths.
                                                                   Pretreatment column: A stainless steel column 4 mm in in-
Assay Measure exactly a volume of Alprostadil Injection
                                                                side diameter and 2.5 cm in length, packed with octadecyl-
corresponding to 5 mg of alprostadil (C20H34O5), add exactly
1830       Official Monographs                                                                       Supplement I, JP XV

silanized silica gel for liquid chromatography (5 mm in parti-    area of alprostadil to that of the internal standard is not
cle diameter).                                                    more than 2.0z.
   Pretreatment column wash solution: Ethanol (99.5).
                                                                  Containers and storage Containers—Hermetic containers.
   Flow rate of wash solution: A constant flow rate of about
                                                                    Storage—Light-resistant, not exceeding 59 avoiding
                                                                                                             C,
2.0 mL per minute.
                                                                  freezing.
   Flow path operating conditions: Change the flow path
operating conditions at the times shown in the table below
using the valves shown in the figure.
                                                                  Add the following:

                      Time of switchover (minutes)                Amikacin Sulfate Injection
   Valve       0        9.0        9.1       *1)        *2)       アミカシン硫酸塩注射液
   RVA         0         0          1         0          0
                                                                    Amikacin Sulfate Injection is an aqueous injection.
   RVB         0         1          1         1          0          It contains not less than 90.0z and not more than
*1) After the internal standard has completely eluted             115.0 z of the labeled amount of amikacin
*2) 0.1 minutes after *1)                                         (C22H43N5O13: 585.60).
                                                                  Method of preparation Prepare as directed under Injec-
                                                                  tions, with Amikacin Sulfate.

                                                                  Description Amikacin Sulfate Injection occurs as a color-
                                                                  less or pale yellow clear liquid.

                                                                  Identification To a volume of Amikacin Sulfate Injection,
                                                                  equivalent to 0.1 g (potency) of Amikacin Sulfate according
                                                                  to the labeled amount, add water to make 4 mL, and use this
                                                                  solution as the sample solution. Separately, dissolve 25 mg
                                                                  (potency) of Amikacin Sulfate Reference Standard in 1 mL
  A: RVA valve                                                    of water, and use this solution as the standard solution.
  B: RVB valve                                                    Then, proceed as directed in the Identifiction (2) under
  C: Sample injector                                              Amikacin Sulfate.
  D: Mobile phase                                                 Osmotic pressure ratio   Being specified separately.
  E: Column for pressure correction
  F: Column                                                       pH <2.54>   6.0 – 7.5
  G: Pretreatment column                                          Bacterial endotoxins <4.01>   Less than 0.50 EU/mg (poten-
  H: Wash solution                                                cy).
  I: Drain
  J: Pump                                                         Extractable volume <6.05>     It meets the requirement.

                                                                  Foreign insoluble matter <6.06> Perform the test according
 Figure    Components of automatic pretreatment system            to Method 1: it meets the requirement.

System suitability—                                               Insoluble particulate matter <6.07>    It meets the require-
   System performance: Dissolve 10 mg of prostaglandin A1,        ment.
previously dried in a desiccator (in vacuum, phosphorus (V)       Sterility <4.06> Perform the test according to the Mem-
oxide) for 4 hours, in ethanol (99.5) to make 100 mL. To 2.5      brane filtration method: it meets the requirement.
mL of this solution add 2.5 mL of the standard stock solu-
tion, and add the mobile phase to make 50 mL. To 1 mL of          Assay Take exactly a volume of Amikacin Sulfate Injec-
this solution add 1 mL of the internal standard solution,         tion, equivalent to about 0.1 g (potency) of Amikacin Sul-
shake, and perform the test under the above conditions with       fate, and add water to make exactly 100 mL. Separately,
40 mL of the solution. Alprostadil, prostaglandin A1 and the      weigh accurately an amount of Amikacin Sulfate Reference
internal standard are eluted in this order, and the resolution    Standard, equivalent to about 50 mg (potency), and add
between the peaks of alprostadil and prostaglandin A1 is not      water to make exactly 50 mL. Take exactly 200 mL each of
less than 10, and that between prostaglandin A1 and the           these solutions into stoppered test tubes, then proceed as
internal standard is not less than 2.0.                           directed in the Assay under Amikacin Sulfate.
   System repeatability: When the test is repeated 6 times            Amount [mg (potency)] of amikacin (C22H43N5O13)
with 40 mL of the standard solution under the above condi-             =WS×(HT/HS)×2
tions, the relative standard deviation of the ratio of the peak
Supplement I, JP XV                                                                     Official Monographs              1831

  WS: Amount [mg (potency)] of Amikacin Sulfate Refer-         Add the following:
      ence Standard

Container and storage   Containers—Hermetic containers.        Amlexanox
                                                               アンレキサノクス

Aminophylline Injection
アミノフィリン注射液

Add the following next to Identification:                      C16H14N2O4: 298.29
                                                               2-Amino-7-(1-methylethyl)-5-oxo-
Bacterial endotoxins <4.01>   Less than 0.6 EU/mg.
                                                               5H-[1]benzopyrano[2,3-b]pyridine-3-carboxylic acid
                                                               [68302-57-8]
Add the following next to Extractable volume:
Foreign insoluble matter <6.06> Perform the test according      Amlexanox, when dried, contains not less than 98.0
to Method 1: it meets the requirement.                         z and not more than 102.0z of C16H14N2O4.
Insoluble particulate matter <6.07>   It meets the require-    Description Amlexanox occurs as white to yellowish white
ment.                                                          crystals or crystalline powder.
                                                                 It is very slightly soluble in ethanol (99.5), and practically
Sterility <4.06> Perform the test according to the Mem-
                                                               insoluble in water.
brane filtration method: it meets the requirement.
                                                                 It dissolves in diluted sodium hydroxide TS (1 in 3).

                                                               Identification (1) Determine the absorption spectrum of
Amitriptyline Hydrochloride                                    a solution of Amlexanox in ethanol (99.5) (1 in 250,000) as
                                                               directed under Ultraviolet-visible Spectrophotometry <2.24>,
Tablets                                                        and compare the spectrum with the Reference Spectrum or
アミトリプチリン塩酸塩錠                                                   the spectrum of a solution of Amlexanox Reference Stan-
                                                               dard prepared in the same manner as the sample solution:
Add the following next to Identification:                      both spectra exhibit similar intensities of absorption at the
                                                               same wavelengths.
Uniformity of dosage units <6.02> Perform the test accord-        (2) Determine the infrared absorption spectrum of Am-
ing to the following method: it meets the requirement of the   lexanox as directed in the potassium bromide disk method
Content uniformity test.                                       under Infrared Spectrophotometry <2.25>, and compare the
  To 1 tablet of Amitriptyline Hydrochloride Tablets add 50    spectrum with the Reference Spectrum or the spectrum of
mL of diluted methanol (1 in 2), shake to disintegrate the     Amlexanox Reference Standard: both spectra exhibit similar
tablet, then add diluted methanol (1 in 2) to make exactly     intensities of absorption at the same wave numbers.
100 mL, and filter. Discard the first 20 mL of the filtrate,
pipet V mL of the subsequent filtrate, add methanol to make    Purity (1) Chloride <1.03>—Dissolve 1.0 g of Amlexanox
exactly V? mL so that each mL contains about 10 mg of          in 20 mL of water and 10 mL of sodium hydroxide TS, add
amitriptyline hydrochloride (C20H23N.HCl), and use this so-    15 mL of dilute nitric acid and water to make 50 mL, cen-
lution as the sample solution. Then, proceed as directed in    trifuge, and then filter the supernatant liquid. To 25 mL of
the Assay.                                                     this filtrate add water to make 50 mL. Perform the test using
                                                               this solution as the test solution. The control solution con-
Amount (mg) of amitriptyline hydrochloride (C20H23N.HCl)       sists of 5 mL of sodium hydroxide TS, 7.5 mL of dilute
 =WS×(AT/AS)×(V?/V)×(1/20)                                     nitric acid, 0.30 mL of 0.01 mol/L hydrochloric acid VS,
  WS: Amount (mg) of          Amitriptyline   Hydrochloride    and water added to make 50 mL (not more than 0.021z).
      Reference Standard                                          (2) Heavy metals <1.07>—Proceed with 1.0 g of Amlexa-
                                                               nox according to Method 2, and perform the test. Prepare
                                                               the control solution with 2.0 mL of Standard Lead Solution
                                                               (not more than 20 ppm).
                                                                  (3) Related substances—(i) Dissolve 30 mg of Amlexa-
                                                               nox in 50 mL of the mobile phase, and use this solution as
                                                               the sample solution. Pipet 1 mL of the sample solution, and
                                                               add the mobile phase to make exactly 50 mL. Pipet 1 mL of
                                                               this solution, add the mobile phase to make exactly 20 mL,
                                                               and use this solution as the standard solution. Perform the
                                                               test with exactly 10 mL each of the sample solution and stan-
                                                               dard solution as directed under Liquid Chromatography
1832       Official Monographs                                                                     Supplement I, JP XV

<2.01> according to the following conditions. Determine          mL of this solution is equivalent to 7 to 13z of that from 10
each peak area of these solutions by the automatic integra-      mL of the standard solution.
tion method: the area of the peak other than amlexanox ob-          System performance: Pipet 1 mL of the sample solution,
tained from the sample solution is not larger than 2 times the   and add the mobile phase to make 100 mL. To 5 mL of this
peak area of amlexanox from the standard solution.               solution add 15 mL of the solution of benzophenone in the
Operating conditions—                                            mobile phase (3 in 1,000,000). When perform the test with
   The detector, column, column temperature, mobile phase,       10 mL of this solution according to the above conditions,
and flow rate: Proceed as directed in the operating              amlexanox and benzophenone are eluted in this order with
conditions in the Assay.                                         the resolution between these peaks being not less than 10.
   Time span of measurement: Until completion of the                System repeatability: When the test is repeated 6 times
elution of amlexanox beginning after the solvent peak.           with 10 mL of the standard solution under the above operat-
System suitability—                                              ing conditions, the relative standard deviation of the peak
   Test for required detectability: Pipet 10 mL of the stan-     area of amlexanox is not more than 2.0z.
dard solution, and add the mobile phase to make exactly 100         (iii) The total amount of related substances, when calcu-
mL. Confirm that the peak area of amlexanox obtained             lated according to the following formula, is not more than
from 10 mL of this solution is equivalent to 7 to 13z of that    0.5z.
from 10 mL of the standard solution.
                                                                          Total amount (z) of related substances
   System performance: Proceed as directed in the system
                                                                            ={(AT1/AS1)+(AT2/AS2)}×(1/10)
suitability in the Assay.
   System repeatability: When the test is repeated 6 times       AT1: Total area of the peaks other than amlexanox from the
with 10 mL of the standard solution under the above operat-           sample solution obtained in (i)
ing conditions, the relative standard deviation of the peak      AT2: Total area of the peaks other than amlexanox from the
area of amlexanox is not more than 2.0z.                              sample solution obtained in (ii)
   (ii) Dissolve 30 mg of Amlexanox in 50 mL of the mobile       AS1: Peak area of amlexanox from the standard solution
phase, and use this solution as the sample solution. Pipet 1          obtained in (i)
mL of the sample solution, and add the mobile phase to           AS2: Peak area of amlexanox from the standard solution
make exactly 50 mL. Pipet 1 mL of this solution, add the              obtained in (ii)
mobile phase to make exactly 20 mL, and use this solution as
                                                                 Loss on drying <2.41>                               C,
                                                                                         Not more than 0.3z (1 g, 1059 2
the standard solution. Perform the test with exactly 10 mL
                                                                 hours).
each of the sample solution and standard solution as direct-
ed under Liquid Chromatography <2.01> according to the           Residue on ignition <2.44>   Not more than 0.1z (1 g).
following conditions, and determine each peak area of these
                                                                 Assay Weigh accurately about 30 mg each of Amlexanox
solutions by the automatic integration method: the area of
                                                                 and Amlexanox Reference Standard, both dried, and dis-
the peak other than amlexanox obtained from the sample
                                                                 solve them separately in the mobile phase to make exactly 50
solution is not larger than 2 times the peak area of amlexa-
                                                                 mL. Pipet 5 mL each of these solutions, and add exactly 15
nox from the standard solution.
                                                                 mL of the internal standard solution, and use these solutions
Operating conditions—
                                                                 as the sample solution and the standard solution, respective-
   Detector, column, and column temperature: Proceed as
                                                                 ly. Perform the test with 10 mL each of the sample solution
directed in the operating conditions in the Assay.
                                                                 and standard solution as directed under Liquid Chro-
   Mobile phase: Dissolve 7.2 g of disodium hydrogen phos-
                                                                 matography <2.01> according to the following conditions,
phate dodecahydrate in water to make 1000 mL. Adjust the
                                                                 and calculate the ratios, QT and QS, of the peak area of am-
pH of this solution to 8.0 by adding a solution prepared by
                                                                 lexanox to that of the internal standard, respectively.
dissolving 3.1 g of sodium dihydrogen phosphate dihydrate
in 1000 mL of water. To 400 mL of this solution add 600 mL              Amount (mg) of C16H14N2O4=WS×(QT/QS)
of acetonitrile.
                                                                   WS: Amount (mg) of Amlexanox Reference Standard
   Flow rate: To 15 mL of a solution of benzophenone in the
mobile phase (3 in 1,000,000) add the mobile phase to make       Internal standard solution—A solution of 3-nitroaniline in
20 mL. Adjust the flow rate so that the retention time of        the mobile phase (1 in 4000).
benzophenone is about 6.5 minutes when perform the test          Operating conditions—
with 10 mL of this solution under the conditions described          Detector: An ultraviolet absorption photometer (wave-
above.                                                           length: 254 nm).
   Time span of measurement: About 3 times as long as the           Column: A stainless steel column 4.0 mm in inside di-
retention time of benzophenone, beginning after the peak of      ameter and 15 cm in length, packed with octadecylsilanized
amlexanox.                                                       silica gel for liquid chromatography (5 mm in particle di-
System suitability—                                              ameter).
   Test for required detectability: Pipet 5 mL of the standard      Column temperature: A constant temperature of about
solution, and add the mobile phase to make exactly 50 mL.        259  C.
Confirm that the peak area of amlexanox obtained from 10            Mobile phase: Dissolve 17.9 g of disodium hydrogen
Supplement I, JP XV                                                                       Official Monographs            1833

phosphate dodecahydrate in water to make 1000 mL. Adjust          about 30 mg of Amlexanox Reference Standard, previously
the pH of this solution to 8.0 by adding a solution prepared      dried at 1059 for 2 hours, and dissolve in the mobile phase
                                                                                C
by dissolving 7.8 g of sodium dihydrogen phosphate dihy-          to make exactly 50 mL. Pipet 25 mL of this solution, add ex-
drate in 1000 mL of water. To 760 mL of this solution add         actly 10 mL of the internal standard solution, add the mo-
240 mL of acetonitrile.                                           bile phase to make 100 mL, and use this solution as the stan-
   Flow rate: Adjust the flow rate so that the retention time     dard solution. Then, proceed as directed in the Assay under
of amlexanox is about 10 minutes.                                 Amlexanox.
System suitability—
                                                                          Amount (mg) of amlexanox (C16H14N2O4)
   System performance: When the procedure is run with 10
                                                                           =WS×(QT/QS)×(V/200)
mL of the standard solution according to the above condi-
tions, amlexanox and the internal standard are eluted in this       WS: Amount (mg) of Amlexanox Reference Standard
order with the resolution between these peaks being not less
                                                                  Internal standard solution—A solution of 3-nitroaniline in
than 2.0.
                                                                  the mobile phase (1 in 500).
   System repeatability: When the test is repeated 6 times
with 10 mL of the standard solution under the above condi-        Dissolution <6.10> When the test is performed at 50 revolu-
tions, the relative standard deviation of the ratio of the peak   tions per minute according to the Paddle method, using 900
area of amlexanox to that of the internal standard is not         mL of 2nd fluid for dissolution test as the dissolution medi-
more than 1.0z.                                                   um, the dissolution rate in 45 minutes of Amlexanox Tablets
                                                                  is not less than 80z.
Containers and storage     Containers—Well-closed contain-
                                                                     Start the test with 1 tablet of Amlexanox Tablets,
ers.
                                                                  withdraw not less than 20 mL of the medium at the specified
                                                                  minute after starting the test, and filter through a membrane
                                                                  filter with a pore size not exceeding 0.45 mm. Discard the
Add the following:
                                                                  first 10 mL of the filtrate, pipet V mL of the subsequent
                                                                  filtrate, add the dissolution medium to make exactly V? mL
Amlexanox Tablets                                                 so that each mL contains about 5.6 mg of amlexanox
                                                                  (C16H14N2O4) according to the labeled amount, and use this
アンレキサノクス錠
                                                                  solution as the sample solution. Separately, weigh accurately
                                                                  about 28 mg of Amlexanox Reference Standard, previously
  Amlexanox Tablets contain not less than 93.0z and
                                                                  dried at 1059 for 2 hours, and dissolve in 2 mL of dilute so-
                                                                                C
not more than 107.0z of the labeled amount of am-
                                                                  dium hydroxide TS, add the dissolution medium to make
lexanox (C16H14N2O4: 298.29).
                                                                  exactly 50 mL. Pipet 1 mL of this solution, add the dissolu-
Method of preparation     Prepare as directed under Tablets,      tion medium to make exactly 100 mL, and use this solution
with Amlexanox.                                                   as the standard solution. Determine the absorbances, AT and
                                                                  AS, at 350 nm of the sample solution and standard solution
Identification (1) Take an amount of powdered Amlexa-
                                                                  as directed under Ultraviolet-visible Spectrophotometry
nox Tablets, equivalent to 10 mg of Amlexanox according to
                                                                  <2.24>.
the labeled amount, add 100 mL of ethanol (99.5), shake
vigorously, and filter. Pipet 1 mL of the filtrate, add 25 mL     Dissolution rate (z) with respect to the labeled amount of
of ethanol (99.5), and use this solution as the sample solu-      amlexanox (C16H14N2O4)
tion. Determine the absorption spectrum of the sample solu-         =WS×(AT/AS)×(V?/V)×(1/C)×18
tion as directed under Ultraviolet-visible Spectrophotometry
                                                                    WS: Amount (mg) of Amlexanox Reference Standard
<2.24>: it exhibits absorption maxima between 240 nm and
                                                                    C: Labeled amount (mg) of amlexanox (C16H14N2O4) in 1
244 nm, between 285 nm and 289 nm, and between 341 nm
                                                                       tablet
and 352 nm.
   (2) Observe the sample solution obtained in (1) under          Assay Weigh accurately not less than 20 Amlexanox
ultraviolet light (main wavelength: 365 nm): the solution         Tablets, and powder. Weigh accurately a portion of the
shows a bluish-white fluorescence.                                powder, equivalent to about 15 mg of amlexanox (C16H14N2
                                                                  O4), add exactly 10 mL of the internal standard solution,
Uniformity of dosage units <6.02> Perform the test accord-
                                                                  add 80 mL of the mobile phase, shake vigorously for 5
ing to the following method: it meets the requirement of the
                                                                  minutes, and then add the mobile phase to make 100 mL.
Content uniformity test.
                                                                  Centrifuge this solution, and use the supernatant liquid as
   Take 1 tablet of Amlexanox Tablets, add exactly 0.6 mL
                                                                  the sample solution. Separately, weigh accurately about 30
of the internal standard solution per 1 mg of amlexanox
                                                                  mg of Amlexanox Reference Standard, previously dried at
(C16H14N2O4), add the mobile phase to make exactly V mL
                                                                  1059 for 2 hours, and dissolve in the mobile phase to make
                                                                       C
so there is about 167 mg of amlexanox (C16H14N2O4) per 1
                                                                  exactly 50 mL. Pipet 25 mL of this solution, add exactly 10
mL, disintegrate the tablet, and then shake vigorously for 5
                                                                  mL of the internal standard solution, add the mobile phase
minutes. Centrifuge this solution, and use the supernatant
                                                                  to make 100 mL, and use this solution as the standard solu-
liquid as the sample solution. Separately, weigh accurately
1834       Official Monographs                                                                      Supplement I, JP XV

tion. Then, proceed as directed in the Assay under Amlexa-       um nitrate and 0.1 g of anhydrous sodium carbonate, mix,
nox.                                                             and gradually ignite. After cooling, dissolve the residue in 2
                                                                 mL of dilute hydrochloric acid and 10 mL of water, filter if
         Amount (mg) of amlexanox (C16H14N2O4)
                                                                 necessary, and add barium chloride TS: a white precipitate is
          =WS×(QT/QS)×(1/2)
                                                                 formed.
  WS: Amount (mg) of Amlexanox Reference Standard
                                                                 Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of
Internal standard solution—A solution of 3-nitroaniline in       Amlodipine Besilate according to Method 4, and perform
the mobile phase (1 in 500).                                     the test. Prepare the control solution with 2.5 mL of Stan-
                                                                 dard Lead Solution (not more than 25 ppm).
Containers and storage    Containers—Tight containers.
                                                                    (2) Related substances—Dissolve 0.10 g of Amlodipine
                                                                 Besilate in 50 mL of a mixture of water and acetonitrile
                                                                 (1:1), and use this solution as the sample solution. Pipet 1
Add the following:
                                                                 mL of the sample solution, and add the mixture of water and
                                                                 acetonitrile (1:1) to make exactly 100 mL. Pipet 3 mL of this
Amlodipine Besilate                                              solution, add the mixture of water and acetonitrile (1:1) to
                                                                 make exactly 10 mL, and use this solution as the standard
アムロジピンベシル酸塩
                                                                 solution. Perform the test with exactly 10 mL each of the
                                                                 sample solution and standard solution as directed under
                                                                 Liquid Chromatography <2.01> according to the following
                                                                 conditions, and determine each peak area by the automatic
                                                                 integration method: the area of the peak having the relative
                                                                 retention time of 0.90 with respect to amlodipine, obtained
                                                                 from the sample solution is not larger than the peak area of
                                                                 amlodipine from the standard solution, and the area of the
C20H25ClN2O5.C6H6O3S: 567.05                                     peak other than amlodipine, other than benzenesulfonic acid
3-Ethyl 5-methyl (4RS)-2-[(2-aminoethoxy)methyl]-                having the relative retention time of about 0.15 with respect
4-(2-chlorophenyl)-6-methyl-1,4-dihydropyridine-3,5-             to amlodipine, and other than the peak mentioned above is
dicarboxylate monobenzenesulfonate [111470-99-6]                 not larger than 1/3 times the peak area of amlodipine from
                                                                 the standard solution. Furthermore, total peak area for
  Amlodipine Besilate contains not less than 98.0z               peaks other than amlodipine and benzenesulfonic acid of the
and not more than 102.0z of C20H25ClN2O5.C6H6O3S,                sample solution is not larger than 2.7 times the peak area of
calculated on the anhydrous basis.                               amlodipine from the standard solution.
Description Amlodipine Besilate occurs as a white to yel-        Operating conditions—
lowish white crystalline powder.                                    Detector: An ultraviolet absorption photometer (wave-
  It is freely soluble in methanol, soluble in ethanol (99.5),   length: 237 nm).
and slightly soluble in water.                                      Column: A stainless steel column 4.6 mm in inside di-
  A solution of Amlodipine Besilate in methanol (1 in 100)       ameter and 15 cm in length, packed with octadecylsilanized
shows no optical rotation.                                       silica gel for liquid chromatography (3 mm in particle di-
  Melting point: about 1989 (with decomposition).
                              C                                  ameter).
                                                                    Column temperature: A constant temperature of about
Identification (1) Determine the absorption spectrum of          359  C.
a solution of Amlodipine Besilate in 0.01 mol/L hydrochlor-         Mobile phase A: A mixture of water and trifluoroacetic
ic acid-methanol TS (1 in 40,000) as directed under Ultrav-      acid (5000:1).
iolet-visible Spectrophotometry <2.24>, and compare the             Mobile phase B: A mixture of acetonitrile and trifluoroa-
spectrum with the Reference Spectrum or the spectrum of a        cetic acid (5000:1).
solution of Amlodipine Besilate Reference Standard pre-             Flowing of the mobile phase: Control the gradient by mix-
pared in the same manner as the sample solution: both spec-      ing the mobile phases A and B as directed in the following
tra exhibit similar intensities of absorption at the same        table.
wavelengths.
   (2) Determine the infrared absorption spectrum of               Time after injection     Mobile phase      Mobile phase
Amlodipine Besilate as directed in the potassium bromide             of sample (min)         A (volz)          B (volz)
disk method under Infrared Spectrophotometry <2.25>, and                   0 – 30              80ª 20             20ª 80
compare the spectrum with the Reference Spectrum or the                   30 – 45                20                 80
spectrum of Amlodipine Besilate Reference Standard: both
spectra exhibit similar intensities of absorption at the same      Flow rate: 1.0 mL per minute.
wave numbers.                                                      Time span of measurement: About 3 times as long as the
   (3) To 30 mg of Amlodipine Besilate add 0.1 g of sodi-        retention time of amlodipine, beginning after the solvent
                                                                 peak.
Supplement I, JP XV                                                                        Official Monographs             1835

System suitability—                                               this order with the resolution between these peaks being not
  Test for required detectability: Pipet 1 mL of the standard     less than 3.
solution, and add a mixture of water and acetonitrile (1:1) to       System repeatability: When the test is repeated 6 times
make exactly 10 mL. Confirm that the peak area of amlodi-         with 20 mL of the standard solution under the above operat-
pine obtained with 10 mL of this solution is equivalent to 7 to   ing conditions, the relative standard deviation of the ratio of
13z of that with 10 mL of the standard solution.                  the peak area of amlodipine to that of the internal standard
  System performance: When the procedure is run with 10           is not more than 1.0z.
mL of the standard solution under the above operating con-
                                                                  Containers and storage Containers—Well-closed contain-
ditions, the number of theoretical plates and the symmetry
                                                                  ers.
factor of the peak of amlodipine are not less than 70,000 and
                                                                    Storage—Light-resistant.
not more than 1.5, respectively.
  System repeatability: When the test is repeated 6 times
with 10 mL of the standard solution under the above operat-
                                                                  Add the following:
ing conditions, the relative standard deviation of the peak
area of amlodipine is not more than 2.0z.
  (3) Residual solvent—Being specified separately.                Amosulalol Hydrochloride
Water <2.48> Not more than 0.5z (1 g, volumetric titra-           アモスラロール塩酸塩
tion, direct titration).

Residue on ignition <2.44>    Not more than 0.2z (1 g).

Assay Weigh accurately about 35 mg each of Amlodipine
Besilate and Amlodipine Besilate Reference Standard
(separately determine the water <2.48> using the same man-
ner as Amlodipine Besilate), dissolve them separately in the      C18H24N2O5S.HCl: 416.92
mobile phase to make exactly 250 mL. Pipet 5 mL each of           5-((1RS)-1-Hydroxy-2-{[2-(2-
these solutions, add exactly 5 mL each of the internal stan-      methoxyphenoxy)ethyl]amino}ethyl)-
dard solution, add the mobile phase to make 25 mL, and use        2-methylbenzenesulfonamide monohydrochloride
these solutions as the sample solution and standard solution.     [70958-86-0]
Perform the test with 20 mL of the sample solution and
standard solution as directed under Liquid Chromatography           Amosulalol Hydrochloride contains not less than
<2.01> according to the following conditions, and calculate       98.5z and not more than 101.0z of C18H24N2O5S.
the ratios, QT and QS, of the peak area of amlodipine to that     HCl, calculated on the anhydrous basis.
of the internal standard.
                                                                  Description Amosulalol Hydrochloride occurs as white
          Amount (mg) ofC20H25ClN2O5.C6H6O3S                      crystals or a white crystalline powder. It has a bitter taste.
           = W S × ( Q T/ Q S )                                     It is very soluble in formic acid, freely soluble in
                                                                  methanol, and sparingly soluble in water and in ethanol
  WS: Amount (mg) of Amlodipine Besilate Reference Stan-
                                                                  (99.5).
      dard, calculated on the anhydrous basis
                                                                    It is hygroscopic.
Internal standard solution—A solution of isobutyl para-             A solution of Amosulalol Hydrochloride in methanol (1 in
hydroxybenzoate in the mobile phase (3 in 20,000).                100) shows no optical rotation.
Operating conditions—
                                                                  Identification (1) Determine the absorption spectrum of
   Detector: An ultraviolet absorption photometer (wave-
                                                                  a solution of Amosulalol Hydrochloride (1 in 20,000) as
length: 237 nm).
                                                                  directed under Ultraviolet-visible Spectrophotometry <2.24>,
   Column: A stainless steel column 4.6 mm in inside di-
                                                                  and compare the spectrum with the Reference Spectrum:
ameter and 15 cm in length, packed with octadecylsilanized
                                                                  both spectra exhibit similar intensities of absorption at the
silica gel for liquid chromatography (5 mm in particle di-
                                                                  same wavelengths.
ameter).
                                                                    (2) Determine the infrared absorption spectrum of
   Column temperature: A constant temperature of about
                                                                  Amosulalol Hydrochloride as directed in the potassium
259  C.
                                                                  chloride disk method under Infrared Spectrophotometry
   Mobile phase: A mixture of methanol and a solution of
                                                                  <2.25>, and compare the spectrum with the Reference Spec-
potassium dihydrogen phosphate (41 in 10,000) (13:7).
                                                                  trum: both spectra exhibit similar intensities of absorption at
   Flow rate: Adjust the flow rate so that the retention time
                                                                  the same wave numbers.
of amlodipine is about 8 minutes.
                                                                    (3) A solution of Amosulalol Hydrochloride (1 in 100)
System suitability—
                                                                  responds to the Qualitative Tests <1.09> for chloride.
   System performance: When the procedure is run with 20
mL of the standard solution under the above operating con-
ditions, amlodipine and the internal standard are eluted in
1836       Official Monographs                                                                     Supplement I, JP XV

Melting point <2.60>            C
                       158 – 1629                                  System repeatability: When the test is repeated 6 times
                                                                 with 10 mL of the standard solution under the above operat-
Purity (1) Heavy metals < 1.07 > —Place 1.0 g of
                                                                 ing conditions, the relative standard deviation of the peak
Amosulalol Hydrochloride in a porcelain crucible, add 1.5
                                                                 area of amosulalol is not more than 1.0z.
mL of sulfuric acid, cover loosely, and heat gently to car-
bonize. After cooling, add 2 mL of nitric acid, heat carefully   Water <2.48> Not more than 4.0z (1 g, volumetric titra-
until white fumes no longer are evolved, and then heat inten-    tion, direct titration).
sely to 500 – 6009 to incinerate. After cooling, add 2 mL of
                  C
                                                                 Residue on ignition <2.44>   Not more than 0.1z (1 g).
hydrochloric acid, proceed according to Method 2, and
perform the test. The control solution, processed in the same    Assay Weigh accurately about 0.6 g of Amosulalol
manner as the test solution using the same amounts of            Hydrochloride, dissolve in 3 mL of formic acid, add 80 mL
reagents, is prepared by combining 2.0 mL of Standard            of a mixture of acetic acid (100) and acetic anhydride (3:2),
Lead Solution and water to make 50 mL (not more than 20          and titrate <2.50> within 5 minutes with 0.1 mol/L perchloric
ppm).                                                            acid VS (potentiometric titration). Perform a blank determi-
   (2) Related substances—Dissolve 0.10 g of Amosulalol          nation using the same procedure, and make any necessary
Hydrochloride in 20 mL of the mobile phase, and use this         correction.
solution as the sample solution. Pipet 1 mL of the sample
                                                                         Each mL of 0.1 mol/L perchloric acid VS
solution, add the mobile phase to make exactly 200 mL, and
                                                                           =41.69 mg of C18H24N2O5S.HCl
use this solution as the standard solution. Perform the test
with exactly 10 mL each of the sample solution and standard      Containers and storage    Containers—Tight containers.
solution as directed under Liquid Chromatography <2.01>
according to the following conditions, and determine each
peak area of both solutions by the automatic integration         Add the following:
method: the total area of the peaks other than amosulalol
obtained from the sample solution is not larger than 2/5         Amosulalol Hydrochloride Tablets
times the peak area of amosulalol from the standard solu-
tion.                                                            アモスラロール塩酸塩錠
Operating conditions—
   Detector: An ultraviolet absorption photometer (wave-           Amosulalol Hydrochloride Tablets contain not less
length: 272 nm).                                                 than 95.0z and not more than 105.0z of the labeled
   Column: A stainless steel column 4.6 mm in inside di-         amount of amosulalol hydrochloride (C18H24N2O5S.
ameter and 15 cm in length, packed with octadecylsilanized       HCl: 416.92).
silica gel for liquid chromatography (5 mm in particle di-
                                                                 Method of preparation Prepare as directed under Tablets,
ameter).
                                                                 with Amosulalol Hydrochloride.
   Column temperature: A constant temperature of about
309  C.                                                          Identification To a quantity of powdered Amosulalol
   Mobile phase: Dissolve 1.36 g of potassium dihydrogen         Hydrochloride Tablets, equivalent to 50 mg of Amosulalol
phosphate in water to make 1000 mL, and adjust to pH 5.7         Hydrochloride according to the labeled amount, add 25 mL
by adding a solution prepared by dissolving 3.58 g of disodi-    of 0.1 mol/L hydrochloric acid TS, shake well, and then
um hydrogen phosphate dodecahydrate in water to make             centrifuge. To 2.5 mL of the supernatant liquid add water to
1000 mL. To 670 mL of this solution add 330 mL of acetoni-       make 100 mL. Determine the absorption spectrum of this
trile.                                                           solution as directed under Ultraviolet-visible Spectrophoto-
   Flow rate: Adjust the flow rate so that the retention time    metry <2.24>: it exhibits a maximum between 270 nm and
of amosulalol is about 7 minutes.                                274 nm, and a shoulder between 275 nm and 281 nm.
   Time span of measurement: About 2 times as long as the
                                                                 Uniformity of dosage units <6.02> Perform the test accord-
retention time of amosulalol, beginning after the solvent
                                                                 ing to the following method: it meets the requirement of the
peak.
                                                                 Content uniformity test.
System suitability—
                                                                    Take 1 tablet of Amosulalol Hydrochloride Tablets, disin-
   Test for required detectability: Pipet 1 mL of the standard
                                                                 tegrate by adding 2 mL of 0.1 mol/L hydrochloric acid TS,
solution, and add the mobile phase to make exactly 10 mL.
                                                                 add 15 mL of methanol, and shake well. Add methanol to
Confirm that the peak area of amosulalol obtained with 10
                                                                 make exactly V mL so that each mL contains about 0.4 mg
mL of this solution is equivalent to 7 to 13z of that with 10
                                                                 of amosulalol hydrochloride (C18H24N2O5S.HCl), and cen-
mL of the standard solution.
                                                                 trifuge. Pipet 5 mL of the supernatant liquid, add exactly 2
   System performance: When the procedure is run with 10
                                                                 mL of the internal standard solution and the mobile phase to
mL of the standard solution under the above operating
                                                                 make 20 mL, and use this solution as the sample solution.
conditions, the number of theoretical plates and the symmet-
                                                                 Separately, weigh accurately about 20 mg of amosulalol
ry factor of the peak of amosulalol are not less than 4000
                                                                 hydrochloride for assay (separately determine the water
and not more than 1.7, respectively.
                                                                 <2.48> in the same manner as Amosulalol Hydrochloride),
Supplement I, JP XV                                                                      Official Monographs            1837

and dissolve in methanol to make exactly 50 mL. Pipet 5 mL       by adding a soluion prepared by dissolving 3.58 g of disodi-
of this solution, add exactly 2 mL of the internal standard      um hydrogen phosphate dodecahydrate in water to make
solution, add the mobile phase to make 20 mL, and use this       1000 mL. To 670 mL of this solution add 330 mL of acetoni-
solution as the standard solution. Then, proceed as directed     trile.
in the Assay.                                                       Flow rate: Adjust the flow rate so that the retention time
                                                                 of amosulalol is about 5 minutes.
        Amount (mg) of amosulalol hydrochloride
                                                                 System suitability—
        (C18H24N2O5S.HCl)
                                                                    System performance: When the procedure is run with 50
          =WS×(QT/QS)×(V/50)
                                                                 mL of the standard solution under the above operating con-
  WS: Amount (mg) of amosulalol hydrochloride for assay,         ditions, the number of theoretical plates and the symmetry
      calculated on the anhydrous basis                          factor of the peak of amosulalol are not less than 4000 and
                                                                 not more than 1.7, respectively.
Internal standard solution—A solution of ethyl parahydrox-
                                                                    System repeatability: When the test is repeated 6 times
ybenzoate in methanol (1 in 6250).
                                                                 with 50 mL of the standard solution under the above operat-
Dissolution <6.10> When the test is performed at 50 revolu-      ing conditions, the relative standard deviation of the peak
tions per minute according to the Paddle method, using 900       area of amosulalol is not more than 1.0z.
mL of water as the dissolution medium, the dissolution rate
                                                                 Assay Take 10 Amosulalol Hydrochloride Tablets, add 20
in 30 minutes of Amosulalol Hydrochloride Tablets is not
                                                                 mL of 0.1 mol/L hydrochloric acid TS, and shake well to
less than 75z.
                                                                 disintegrate. Add 120 mL of methanol, again shake well,
   Start the test with 1 tablet of Amosulalol Hydrochloride
                                                                 add methanol to make exactly 200 mL, and then centrifuge.
Tablets, withdraw not less than 20 mL of the medium at the
                                                                 Pipet a volume of supernatant liquid corresponding to about
specified minute after starting the test, and filter through a
                                                                 5 mg of amosulalol hydrochloride (C18H24N2O5S.HCl), add
membrane filter with a pore size not exceeding 0.5 mm. Dis-
                                                                 exactly 5 mL of the internal standard solution, add the
card the first 10 mL of the filtrate, pipet V mL of the subse-
                                                                 mobile phase to make 50 mL, and use this solution as the
quent filtrate, add water to make exactly V? mL so that each
                                                                 sample solution. Separately, weigh accurately about 25 mg
mL contains about 5.5 mg of amosulalol hydrochloride
                                                                 of amosulalol hydrochloride for assay (separately determine
(C18H24N2O5S.HCl) according to the labeled amount, and
                                                                 the water <2.48> in the same manner as Amosulalol
use this solution as the sample solution. Separately, weigh
                                                                 Hydrochloride), and dissolve in methanol to make exactly
accurately about 22 mg of amosulalol hydrochloride for
                                                                 25 mL. Pipet 5 mL of this solution, add exactly 5 mL of the
assay (separately determine the water <2.48> in the same
                                                                 internal standard solution, add the mobile phase to make 50
manner as Amosulalol Hydrochloride), and dissolve in
                                                                 mL, and use this solution as the standard solution. Perform
water to make exactly 200 mL. Pipet 5 mL of this solution,
                                                                 the test with 10 mL each of the sample solution and standard
add water to make exactly 100 mL, and use this solution as
                                                                 solution as directed under Liquid Chromatography <2.01>
the standard solution. Perform the test with exactly 50 mL
                                                                 according to the following conditions, and calculate the
each of the sample solution and standard solution as direct-
                                                                 ratios, QT and QS, of the peak area of amosulalol to that of
ed under Liquid Chromatography <2.01> according to the
                                                                 the internal standard.
following conditions, and determine the amosulalol peak
areas, AT and AS, of both solutions.                                     Amount (mg) of amosulalol hydrochloride
                                                                         (C18H24N2O5S.HCl)
Dissolution rate (z) with respect to the labeled amount of
                                                                           =WS×(QT/QS)×(1/5)
amosulalol hydrochloride (C18H24N2O5S.HCl)
  =WS×(AT/AS)×(V?/V)×(1/C)×(45/2)                                  WS: Amount (mg) of amosulalol hydrochloride for assay,
                                                                       calculated on the anhydrous basis
  WS: Amount (mg) of amosulalol hydrochloride for assay,
      calculated on the anhydrous basis                          Internal standard solution—A solution of ethyl parahydrox-
  C: Labeled amount (mg) of amosulalol hydrochloride             ybenzoate in methanol (1 in 6250).
     (C18H24N2O5S.HCl) in 1 tablet                               Operating conditions—
                                                                    Detector: An ultraviolet absorption photometer (wave-
Operating conditions—
                                                                 length: 272 nm).
   Detector: An ultraviolet absorption photometer (wave-
                                                                    Column: A stainless steel column 4.6 mm in inside di-
length: 272 nm).
                                                                 ameter and 15 cm in length, packed with octadecylsilanized
   Column: A stainless steel column 4.6 mm in inside di-
                                                                 silica gel for liquid chromatography (5 mm in particle di-
ameter and 15 cm in length, packed with octadecylsilanized
                                                                 ameter).
silica gel for liquid chromatography (5 mm in particle di-
                                                                    Column temperature: A constant temperature of about
ameter).
                                                                 259  C.
   Column temperature: A constant temperature of about
                                                                    Mobile phase: A mixture of diluted acetic acid (100) (1 in
309  C.
                                                                 25), acetonitrile and a solution of ammonium acetate (1 in
   Mobile phase: Dissolve 1.36 g of potassium dihydrogen
                                                                 250) (5:3:2).
phosphate in water to make 1000 mL, and adjust to pH 5.7
                                                                    Flow rate: Adjust the flow rate so that the retention time
1838       Official Monographs                                                                        Supplement I, JP XV

of amosulalol is about 4 minutes.                                  Insoluble particulate matter <6.07>     It meets the require-
System suitability—                                                ment.
   System performance: When the procedure is run with 10
                                                                   Sterility <4.06> Perform the test according to the Mem-
mL of the standard solution under the above operating con-
                                                                   brane filtration method: it meets the requirement.
ditions, amosulalol and the internal standard are eluted in
this order with the resolution between these peaks being not       Assay Weigh accurately the mass of the contents of not
less than 7.                                                       less than 10 containers of Ampicillin Sodium for Injection.
   System repeatability: When the test is repeated 6 times         Weigh accurately an amount of a portion of the contents,
with 10 mL of the standard solution under the above operat-        equivalent to about 50 mg (potency) of Ampicillin Sodium,
ing conditions, the relative standard deviation of the ratio of    add exactly 5 mL of the internal standard solution and dis-
the peak area of amosulalol to that of the internal standard       solve. Then add the mobile phase to make 50 mL, and use
is not more than 1.0z.                                             this solution as the sample solution. Separately, weigh ac-
                                                                   curately an amount of Ampicillin Reference Standard,
Containers and storage     Containers—Tight containers.
                                                                   equivalent to about 50 mg (potency), add exactly 5 mL of
                                                                   the internal standard solution and dissolve. Then add the
                                                                   mobile phase to make 50 mL, and use this solution as the
Add the following:
                                                                   standard solution. Perform the test with 10 mL each of the
                                                                   sample solution and standard solution as directed under Liq-
Ampicillin Sodium for Injection                                    uid Chromatography <2.01> according to the following con-
                                                                   ditions, and calculate the ratios, QT and QS, of the peak area
注射用アンピシリンナトリウム
                                                                   of ampicillin to that of the internal standard.
  Ampicillin Sodium for Injection is a preparation for                Amount [mg (potency)] of ampicillin (C16H19N3O4S)
injection which is dissolved before use.                               = W S ×( Q T / Q S )
  It contains not less than 90.0z and not more than
                                                                     WS: Amount [mg (potency)] of Ampicillin Reference
110.0z of the labeled amount of ampicillin
                                                                         Standard
(C16H19N3O4S: 349.40).
                                                                   Internal standard solution—A solution of guaifenesin in the
Method of preparation Prepare as directed under Injec-
                                                                   mobile phase (1 in 200).
tions, with Ampicillin Sodium.
                                                                   Operating conditions—
Description Ampicillin Sodium for Injection occurs as                 Detector: An ultraviolet absorption photometer (wave-
white to light yellowish white crystals or crystalline powder.     length: 230 nm).
                                                                      Column: A stainless steel column 4.6 mm in inside di-
Identification Proceed as directed in the Identification (1)
                                                                   ameter and 15 cm in length, packed with octadecylsilanized
under Ampicillin Sodium.
                                                                   silica gel for liquid chromatography (5 mm in particle di-
Osmotic pressure ratio    Being specified separately.              ameter).
                                                                      Column temperature: A constant temperature of about
pH <2.54> The pH of a solution prepared by dissolving an
                                                                   259  C.
amount of Ampicillin Sodium for Injection, equivalent to
                                                                      Mobile phase: Dissolve 5.94 mg of diammonium hydro-
1.0 g (potency) of Ampicillin Sodium according to the
                                                                   gen phosphate in 850 mL of water, add 100 mL of
labeled amount, in 10 mL of water is 8.0 to 10.0.
                                                                   acetonitril, add phosphoric acid to adjust the pH to 5.0, then
Purity Clarity and color of solution—Dissolve an amount            add water to make exactly 1000 mL.
of Ampicillin Sodium for Injection, equivalent to 0.25 g              Flow rate: Adjust the flow rate so that the retention time
(potency) of Ampicillin Sodium according to the labeled            of ampicillin is about 6 minutes.
amount, in 0.75 mL of water: the solution is clear. Perform        System suitability—
the test with the solution as directed under Ultraviolet-visible      System performance: When the procedure is run with 10
Spectrophotometry <2.24>: the absorbance at 400 nm is not          mL of the standard solution under the above operating con-
more than 0.40.                                                    ditions, ampicillin and the internal standard are eluted in
                                                                   this order with the resolution between these peaks being not
Water <2.48> Not more than 3.0z (0.2 g, volumetric titra-
                                                                   less than 26.
tion, direct titration).
                                                                      System repeatability: When the test is repeated 6 times
Bacterial endotoxins <4.01>     Less than 0.075 EU/mg (po-         with 10 mL of the standard solution under the above operat-
tency).                                                            ing conditions, the relative standard deviation of the ratio of
                                                                   the peak area of ampicillin to that of the internal standard is
Uniformity of dosage units <6.02>     It meets the requirement
                                                                   not more than1.0z.
of the Mass variation test.
                                                                   Containers and storage    Containers—Hermetic containers.
Foreign insoluble matter <6.06> Perform the test according
to Method 2: it meets the requirement.
Supplement I, JP XV                                                                    Official Monographs            1839

                                                                Azelastine Hydrochloride, when dried, contains not
L-Arginine        Hydrochloride Injection                     less than 99.0z and not more than 101.0z of
                                                              C22H24ClN3O.HCl.
L-アルギニン塩酸塩注射液
                                                              Description Azelastine Hydrochloride occurs as a white
                                                              crystalline powder.
Delete the Pyrogen and add the following next
                                                                It is freely soluble in formic acid, and slightly soluble in
to pH:
                                                              water and in ethanol (99.5).
Bacterial endotoxins <4.01>   Less than 0.50 EU/mL.             Melting point: about 2259 (with decomposition).
                                                                                            C
                                                                A solution of Azelastine Hydrochloride (1 in 200) shows
Add the following next to Extractable volume:                 no optical rotation.
Foreign insoluble matter <6.06> Perform the test according    Identification (1) Determine the absorption spectrum of
to Method 1: it meets the requirement.                        a solution of Azelastine Hydrochloride (3 in 100,000) as
                                                              directed under Ultraviolet-visible Spectrophotometry <2.24>,
Insoluble particulate matter <6.07>   It meets the require-
                                                              and compare the spectrum with the Reference Spectrum:
ment.
                                                              both spectra exhibit similar intensities of absorption at the
Sterility <4.06> Perform the test according to the Mem-       same wavelengths.
brane filtration method: it meets the requirement.               (2) Determine the infrared absorption spectrum of
                                                              Azelastine Hydrochloride as directed in the potassium chlo-
                                                              ride disk method under Infrared Spectrophotometry <2.25>:
Ascorbic Acid Injection                                       both spectra exhibit similar intensities of absorption at the
                                                              same wave numbers.
アスコルビン酸注射液                                                       (3) To 10 mL of a saturated solution of Azelastine
                                                              Hydrochloride add 1 mL of dilute nitric acid, and filter to
Add the following next to pH:                                 separate formed crystals: the filtrate responds to the
Bacterial endotoxins <4.01>   Less than 0.15 EU/mg.           Qualitative Tests <1.09> (2) for chloride.

                                                              Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of
Add the following next to Extractable volume:                 Azelastine Hydrochloride according to Method 2, and per-
Foreign insoluble matter <6.06> Perform the test according    form the test. Prepare the control solution with 2.0 mL of
to Method 1: it meets the requirement.                        Standard Lead Solution (not more than 20 ppm).
                                                                 (2) Arsenic <1.11>—Prepare the test solution with 1.0 g
Insoluble particulate matter <6.07>   It meets the require-   of Azelastine Hydrochloride according to Method 3, and
ment.                                                         perform the test (not more than 2 ppm).
Sterility <4.06> Perform the test according to the Mem-          (3) Related substances—Dissolve 50 mg of Azelastine
brane filtration method: it meets the requirement.            Hydrochloride in 100 mL of the mobile phase, and use this
                                                              solution as the sample solution. Pipet 1 mL of the sample
                                                              solution, add the mobile phase to make exactly 100 mL, and
                                                              use this solution as the standard solution. Perform the test
Add the following:
                                                              with exactly 20 mL each of the sample solution and standard
                                                              solution as directed under Liquid Chromatography <2.01>
Azelastine Hydrochloride                                      according to the following conditions. Determine each peak
アゼラスチン塩酸塩                                                     area of these solutions by the automatic integration method:
                                                              each peak area other than azelastine obtained from the sam-
                                                              ple solution is not larger than 1/10 times the peak area of
                                                              azelastine from the standard solution, and the total area of
                                                              the peaks other than the peak of azelastine from the sample
                                                              solution is not larger than 1/2 times the peak area of
                                                              azelastine from the standard solution.
                                                              Operating conditions—
                                                                 Detector: An ultraviolet absorption photometer (wave-
                                                              length: 240 nm).
C22H24ClN3O.HCl: 418.36
                                                                 Column: A stainless steel column 4.6 mm in inside di-
4-[(4-Chlorophenyl)methyl]-2-[(4RS)-
                                                              ameter and 15 cm in length, packed with octadecylsilanized
(1-methylazepan-4-yl)]phthalazin-1(2H)-one
                                                              silica gel for liquid chromatography (5 mm in particle di-
monohydrochloride [79307-93-0]
                                                              ameter).
                                                                 Column temperature: A constant temperature of about
                                                              359  C.
1840       Official Monographs                                                                       Supplement I, JP XV

  Mobile phase: A mixture of water, acetonitrile and per-        cording to the labeled amount, in 1 mL of hydroxylammoni-
chloric acid (660:340:1).                                        um chloride-ethanol TS, allow to stand for 3 minutes, add 1
  Flow rate: Adjust the flow rate so that the retention time     mL of acidic ammonium iron (III) sulfate TS, and mix: a
of azelastine is about 10 minutes.                               red-brown color develops.
  Time span of measurement: About 2 times as long as the           (2) Dissolve an amount of Aztreonam for Injection,
retention time of azelastine, beginning after the solvent        equivalent to 3 mg (potency) of Aztreonam according to the
peak.                                                            labeled amount, in 100 mL of water, and determine the
System suitability—                                              absorption spectrum of the solution as directed under
  Test for required detectability: Pipet 5 mL of the standard    Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a
solution, and add the mobile phase to make exactly 50 mL.        maximum between 289 nm and 293 nm.
Confirm that the peak area of azelastine obtained from 20
                                                                 pH <2.54> The pH of a solution prepared by dissolving an
mL of this solution is equivalent to 7 to 13z of that from 20
                                                                 amount of Aztreonam for Injection, equivalent to 1.0 g
mL of the standard solution.
                                                                 (potency) of Aztreonam according to the labeled amount, in
  System performance: When the procedure is run with 20
                                                                 10 mL of water is 4.5 to 7.0.
mL of the standard solution under the above operating con-
ditions, the number of theoretical plates and the symmetry       Purity Clarity and color of solution—Dissolve an amount
factor of the peak of azelastine is not less than 5000 and not   of Aztreonam for Injection, equivalent to 1.0 g (potency) of
more than 1.5, respectively.                                     Aztreonam according to the labeled amount, in 10 mL of
  System repeatability: When the test is repeated 6 times        water: the solution is clear, and its absorbance <2.24> at 450
with 20 mL of the standard solution under the above operat-      nm is not more than 0.06.
ing conditions, the relative standard deviation of the peak
                                                                 Water <2.48> Not more than 2.0z (0.5 g, volumetric titra-
area of azelastine is not more than 1.0z.
                                                                 tion, direct titration).
Loss on drying <2.41>                                C,
                         Not more than 1.0z (1 g, 1059
                                                                 Bacterial endotoxins <4.01>   Less than 0.10 EU/mg (poten-
2 hours).
                                                                 cy).
Residue on ignition <2.44>   Not more than 0.1z (1 g).
                                                                 Uniformity of dosage units <6.02>    It meets the requirement
Assay Weigh accurately about 0.6 g of previously dried           of the Mass variation test.
Azelastine Hydrochloride, dissolve in 5 mL of formic acid,
                                                                 Foreign insoluble matter <6.06> Perform the test according
add 70 mL of acetic anhydride, and titrate <2.50> with
                                                                 to Method 2: it meets the requirement.
0.1 mol/L perchloric acid VS (potentiometric titration). Per-
form a blank determination in the same manner, and make          Insoluble particulate matter <6.07>    It meets the require-
any necessary correction.                                        ment.

        Each mL of 0.1 mol/L perchloric acid VS                  Sterility <4.06> Perform the test according to the Mem-
          =41.84 mg of C22H24ClN3O.HCl                           brane filtration method: it meets the requirement.

Containers and storage Containers—Tight containers.              Assay Take an amount of Aztreonam for Injection,
 Storage—Light-resistant.                                        equivalent to about 5 g (potency) of Aztreonam, dissolve the
                                                                 contents with a suitable amount of water, and transfer to a
                                                                 100-mL volumetric flask. Wash each container with water,
Add the following:                                               combine the washings and the solution, and add water to
                                                                 make exactly 100 mL. Pipet 10 mL of this solution, and add
Aztreonam for Injection                                          water to make exactly 50 mL. Pipet 2 mL of this solution,
                                                                 add exactly 10 mL of the internal standard solution and
注射用アズトレオナム                                                       water to make 100 mL, and use this solution as the sample
                                                                 solution. Separately, weigh accurately an amount of Aztreo-
   Aztreonam for Injection is a preparation for injec-           nam Reference Standard, equivalent to about 20 mg (poten-
tion which is dissolved before use.                              cy), dissolve in a suitable amount of water, add exactly 10
   It contains not less than 93.0z and not more than             mL of the internal standard solution and water to make 100
107.0z of the labeled amount of aztreonam                        mL, and use this solution as the standard solution. Then,
(C13H17N5O8S2: 435.43).                                          proceed as directed in the Assay under Aztreonam.
Method of preparation Prepare           as   directed   under      Amount [mg (potency)] of aztreonam (C13H17N5O8S2)
Injections, with Aztreonam.                                         =WS×(QT/QS)×250
Description Aztreonam for Injection is white to yellowish          WS: Amount [mg (potency)] of Aztreonam Reference
white masses or powder.                                                Standard
Identification (1) Dissolve an amount of Aztreonam for           Internal standard solution—A solution of 4-aminobenzoic
Injection, equivalent to 6 mg (potency) of Aztreonam ac-         acid (1 in 6250).
Supplement I, JP XV                                                                       Official Monographs             1841

Containers and storage Containers—Hermetic containers.           Method of preparation Prepare as directed under Injec-
 Storage—Light-resistant.                                        tions, with Benzylpenicillin Potassium.

                                                                 Description Benzylpenicillin Potassium for Injection oc-
                                                                 curs as white crystals or crystalline powder.
Baclofen Tablets
                                                                 Identification Proceed as directed in the Identification (2)
バクロフェン錠                                                          under Benzylpenicillin Potassium.

                                                                 Osmotic pressure ratio    Being specified separately.
Add the following next to Identification:
                                                                 pH <2.54> The pH of a solution prepared by dissolving an
Uniformity of dosage units <6.02> Perform the test accord-
                                                                 amount of Benzylpenicillin Potassium for Injection, equiva-
ing to the following method: it meets the requirement of the
                                                                 lent to 1.0×105 Units of Benzylpenicillin Potassium accord-
Content uniformity test.
                                                                 ing to the labeled amount, in 10 mL of water is 5.0 to 7.5.
   To 1 tablet of Baclofen Tablets add 5 mL of 0.1 mol/L
hydrochloric acid TS, disperse the tablet into small particles   Purity Clarity and color of solution—A solution prepared
with the aid of ultrasonic waves, then shake for 10 minutes,     by dissolving an amount of Benzylpenicillin Potassium for
and add 0.1 mol/L hydrochloric acid TS to make exactly V         Injection, equivalent to 1.0×106 Units of Benzylpenicillin
mL so that each mL contains about 0.5 mg of baclofen             Potassium according to the labeled amount, in 10 mL of
(C10H12ClNO2). Centrifuge, pipet 5 mL of the supernatant         water is clear. Perform the test with this solution as directed
liquid, add 2 drops of phenolphthalein TS, neutralize with       under Ultraviolet-visible Spectrophotometry <2.24>: the
dilute sodium hydroxide TS, then add water to make exactly       absorbance at 400 nm is not more than 0.10.
50 mL, and use this solution as the sample solution.
                                                                 Loss on drying <2.41> Not more than 1.2z (3 g, in vacu-
Separately, weigh accurately about 25 mg of Baclofen Refer-
                                                                 um, below 0.67 kPa, 609 3 hours).
                                                                                       C,
ence Standard (separately determine the water <2.48> in the
same manner as Baclofen), and dissolve in 0.1 mol/L              Bacterial endotoxins <4.01>      Less than 1.25×10-4 EU/
hydrochloric acid TS to make exactly 50 mL. Pipet 5 mL of        Unit.
this solution, add 2 drops of phenolphthalein TS, neutralize
                                                                 Uniformity of dosage units <6.02>    It meets the requirement
with dilute sodium hydroxide TS, then add water to make
                                                                 of the Mass variation test.
exactly 50 mL, and use this solution as the standard solu-
tion. To exactly 2 mL each of the sample solution and stan-      Foreign insoluble matter <6.06> Perform the test according
dard solution add 4 mL of ninhydrin-tin (II) chloride TS,        to Method 2: it meets the requirement.
mix, heat on a water bath for 20 minutes, then immediately
                                                                 Insoluble particulate matter <6.07>     It meets the require-
shake vigorously for 2 minutes. After cooling, add a mixture
                                                                 ment.
of water and 1-propanol (1:1) to make them exactly 25 mL,
and determine the absorbances, AT and AS, of them at 570         Sterility <4.06> Perform the test according to the Mem-
nm as directed under Ultraviolet-visible Spectrophotometry       brane filtration method: it meets the requirement.
<2.24>, using a solution obtained with 2 mL of water by the
                                                                 Assay Weigh accurately the mass of the contents of not
same procedure as above as the blank.
                                                                 less than 10 containers of Benzylpenicillin Potassium for In-
         Amount (mg) of baclofen (C10H12ClNO2)                   jection. Weigh accurately an amount of a portion of the con-
          =WS×(AT/AS)×(V/50)                                     tents, equivalent to about 6×104 Units of Benzylpenicillin
                                                                 Potassium, dissolve in water to make exactly 20 mL, and use
  WS: Amount (mg) of Baclofen Reference Standard, cal-
                                                                 this solution as the sample solution. Separately, weigh ac-
      culated on the anhydrous basis
                                                                 curately an amount of Benzylpenicillin Potassium Reference
                                                                 Standard, equivalent to about 6×104 Units, dissolve in
                                                                 water to make exactly 20 mL, and use this solution as the
Add the following:
                                                                 standard solution. Perform the test with exactly 5 mL each of
                                                                 the sample solution and standard solution as directed under
Benzylpenicillin Potassium for                                   Liquid Chromatography <2.01> according to the following
Injection                                                        conditions. Determine the peak area of benzylpenicillin, AT
                                                                 and AS, from each solution.
注射用ベンジルペニシリンカリウム
                                                                        Amount (unit) of Benzylpenicillin Potassium
  Benzylpenicillin Potassium for Injection is a prepa-                   (C16H17KN2O4S)
ration for injection which is dissolved before use.                        =WS×(AT/AS)
  It contains not less than 93.0z and not more than                WS: Amount (unit) of Benzylpenicillin Potassium Refer-
107.0z of the labeled amount of benzylpenicillin                       ence Standard
potassium (C16H17KN2O4S: 372.48).
1842       Official Monographs                                                                        Supplement I, JP XV

Operating conditions—                                             Identification Determine the infrared absorption spectrum
   Detector: An ultraviolet absorption photometer (wave-          of Biotin as directed in the potassium bromide disc method
length: 254 nm).                                                  under Infrared Spectrophotometry <2.25>, and compare the
   Column: A stainless steel column 4.6 mm in inside di-          spectrum with the Reference Spectrum: both spectra exhibit
ameter and 25 cm in length, packed with octadecylsilanized        similar intensities of absorption at the same wave numbers.
silica gel for liquid chromatography (7 mm in particle di-
                                                                  Optical rotation <2.49> [a]20: +89 – +939(after drying,
                                                                                              D
ameter).
                                                                  0.4 g, dilute sodium hydroxide TS, 20 mL, 100 mm).
   Column temperature: A constant temperature of about
259  C.                                                           Purity (1) Clarity and color of solution—Dissolve 1.0 g
   Mobile phase: To a mixture of diammonium hydrogen              of Biotin in 10 mL of 0.5 mol/L sodium hydroxide TS: the
phosphate solution (33 in 5000) and acetonitril (19:6), add       solution is clear and colorless.
phosphoric acid to adjust the pH of this solution to 8.0.            (2) Heavy metals <1.07>—Proceed with 2.0 g of Biotin
   Flow rate: Adjust the flow rate so that the retention time     according to Method 2, and perform the test. Prepare the
of benzylpenicillin is about 7.5 minutes.                         control solution with 2.0 mL of Standard Lead Solution (not
System suitability—                                               more than 10 ppm).
   System performance: When the procedure is run with 5 mL           (3) Arsenic <1.11>—Place 0.7 g of Biotin in a Kjeldahl
of the standard solution under the above operating condi-         flask, add 5 mL of nitric acid and 2 mL of sulfuric acid,
tions, the number of theoretical plates and the symmetry          place a small funnel on the mouth of the flask, and carefully
factor of the peak of benzylpenicillin are not less than 6000     heat until white fumes are evolved. After cooling, add 2 mL
and not more than 2.0, respectively.                              of nitric acid twice, heat, add 2 mL of hydrogen peroxide
   System repeatability: When the test is repeated 6 times        (30) several times, and heat until the color of the solution
with 5 mL of the standard solution under the above operat-        becomes colorless or pale yellow. After cooling, add 2 mL of
ing conditions, the relative standard deviation of the peak       saturated ammonium oxalate solution, and heat to concen-
area of benzylpenicillin is not more than 1.0z.                   trate until white fumes are evolved again. After cooling, add
                                                                  water to make 5 mL, and perform the test using this solution
Containers and storage    Containers—Hermetic containers.
                                                                  as the test solution (not more than 2.8 ppm).
                                                                     (4) Related substances—Dissolve 0.10 g of Biotin in 10
                                                                  mL of diluted ammonia solution (28) (7 in 100), and use this
Betahistine Mesilate                                              solution as the sample solution. Pipet 1 mL of this solution,
                                                                  and add diluted ammonia solution (28) (7 in 100) to make
ベタヒスチンメシル酸塩
                                                                  exactly 100 mL. Pipet 10 mL of this solution, add diluted
                                                                  ammonia solution (28) (7 in 100) to make exactly 50 mL, and
Change the Identification (3) to read:
                                                                  use this solution as the standard solution. Perform the test
Identification (3) A 30 mg portion of Betahistine Mesi-           with these solutions as directed under Thin-layer Chro-
late responds to the Qualitative Tests <1.09> (2) for mesilate.   matography <2.03>. Spot 5 mL each of the sample solution
                                                                  and standard solution on a plate of silica gel for thin-layer
                                                                  chromatography. Develop the plate with a mixture of 1-
Add the following:                                                butanol, water, and acetic acid (100) (5:2:1) to a distance of
                                                                  about 10 cm, air-dry the plate, and then dry for 30 minutes
Biotin                                                            at 1059 Spray the plate evenly with a mixture of a solution
                                                                          C.
                                                                  of 4-dimethylaminocinnamaldehyde in ethanol (99.5) (1 in
ビオチン                                                              500) and a solution of sulfuric acid in ethanol (99.5) (1 in 50)
                                                                  (1:1): the spots other than the principal spot obtained from
                                                                  the sample solution are not more intense than the spot from
                                                                  the standard solution.

                                                                  Loss on drying <2.41>    Not more than 0.5z (0.5 g, 1059C,
C10H16N2O3S: 244.31                                               4 hours).
5-[(3aS,4S,6aR)-2-Oxohexahydro-1H-
                                                                  Residue on ignition <2.44>    Not more than 0.1z (1 g).
thieno[3,4-d]imidazol-4-yl]pentanoic acid     [58-85-5]
                                                                  Assay Weigh accurately about 0.25 g of Biotin, previously
  Biotin, when dried, contains not less than 98.5z                dried, dissolve by adding exactly 20 mL of 0.1 mol/L sodi-
and not more than 101.0z of C10H16N2O3S.                          um hydroxide VS, and titrate <2.50> the excess sodium
                                                                  hydroxide with 0.1 mol/L hydrochloric acid VS (indicator: 2
Description Biotin occurs as white crystals or a white crys-
                                                                  drops of phenolphthalein TS). Perform a blank determina-
talline powder.
                                                                  tion in the same manner.
   It is very slightly soluble in water and in ethanol (99.5).
   It dissolves in dilute sodium hydroxide TS.                           Each mL of 0.1 mol/L sodium hydroxide VS
   Melting point: about 2319 (with decomposition).
                                C                                          =24.43 mg of C10H16N2O3S
Supplement I, JP XV                                                                     Official Monographs             1843

Containers and storage   Containers—Tight containers.          <2.24>, and compare the spectrum with the Reference Spec-
                                                               trum: both spectra exhibit similar intensities of absorption at
                                                               the same wavelengths.
Bisacodyl Suppositories                                          (2) Determine the infrared absorption spectrum of
                                                               Bisoprolol Fumarate as directed in the potassium bromide
ビサコジル坐剤                                                        disc method under Infrared Spectrophotometry <2.25>, and
                                                               compare the spectrum with the Reference Spectrum: both
Add the following next to Identification:                      spectra exhibit similar intensities of absorption at the same
                                                               wave numbers.
Uniformity of dosage units <6.02> Perform the test accord-
ing to the following method: it meets the requirement of the   Melting point <2.60>             C
                                                                                       101 – 1059
Content uniformity test.
                                                               Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of
  To 1 suppository of Bisacodyl Suppositories add tetra-
                                                               Bisoprolol Fumarate according to Method 2, and perform
hydrofuran to make a solution containing about 0.2 mg of
                                                               the test. Prepare the control solution with 2.0 mL of Stan-
bisacodyl (C22H19NO4) in each mL, warm to 409 and    C,
                                                               dard Lead Solution (not more than 10 ppm).
shake to dissolve. After cooling, add tetrahydrofuran to
                                                                  (2) Related substances—Dissolve 50 mg of Bisoprolol
make exactly V mL so that each mL contains about 10 mg of
                                                               Fumarate in 100 mL of a mixture of water and acetonitrile
bisacodyl (C22H19NO4). Pipet 5 mL of this solution, and pro-
                                                               (4:1), and use this solution as the sample solution. Pipet 1
ceed as directed in the Assay.
                                                               mL of this solution, add the mixture of water and acetoni-
         Amount (mg) of bisacodyl (C22H19NO4)                  trile (4:1) to make exactly 100 mL, and use this solution as
          =WS×(QT/QS)×(V/50)                                   the standard solution. Perform the test with exactly 20 mL
                                                               each of the sample solution and standard solution as direct-
  WS: Amount (mg) of Bisacodyl Reference Standard
                                                               ed under Liquid Chromatography <2.01> according to the
Internal standard solution—A solution of ethyl parahydrox-     following conditions. Determine each peak area of both
ybenzoate in acetonitrile (3 in 100,000).                      solutions by the automatic integration method: the area of
                                                               the peaks other than bisoprolol obtained from the sample
                                                               solution is not larger than 1/2 times the peak area of bi-
Add the following:                                             soprolol from the standard solution. Furthermore, the total
                                                               of the areas of all peaks other than bisoprolol from the sam-
Bisoprolol Fumarate                                            ple solution is not larger than the peak area of bisoprolol
                                                               from the standard solution.
ビソプロロールフマル酸塩                                                   Operating conditions—
                                                                  Detector: An ultraviolet absorption photometer (wave-
                                                               length: 225 nm).
                                                                  Column: A stainless steel column 4.6 mm in inside di-
                                                               ameter and 15 cm in length, packed with octylsilanized silica
                                                               gel for liquid chromatography (5 mm in particle diameter).
                                                                  Column temperature: A constant temperature of about
                                                               409  C.
(C18H31NO4)2.C4H4O4: 766.96                                       Mobile phase: Dissolve 4.08 g of potassium dihydrogen
(2RS)-1-(4- [2-(1-
          {                                                    phosphate in 1000 mL of water, and adjust to pH 2.5 with
Methylethoxy)ethoxy]methyl}phenoxy)-                           phosphoric acid. To 800 mL of this solution add 200 mL of
3-[(1-methylethyl)amino]propan-2-ol hemifumarate               acetonitrile.
[104344-23-2]                                                     Flow rate: Adjust the flow rate so that the retention time
                                                               of bisoprolol is about 8 minutes.
  Bisoprolol Fumarate, when dried, contains not less              Time span of measurement: About 2 times as long as the
than 98.5z and not more than 101.0z of                         retention time of bisoprolol, beginning after the fumaric
(C18H31NO4)2.C4H4O4.                                           acid peak.
Description Bisoprolol Fumarate occurs as white crystals       System suitability—
or a white crystalline powder.                                    Test for required detectability: Pipet 2 mL of the standard
  It is very soluble in water and in methanol, and freely      solution, and add a mixture of water and acetonitrile (4:1) to
soluble in ethanol (99.5) and in acetic acid (100).            make exactly 20 mL. Confirm that the peak area of bi-
  A solution of Bisoprolol Fumarate (1 in 10) shows no opti-   soprolol obtained from 20 mL of this solution is equivalent
cal rotation.                                                  to 7 to 13z of that from 20 mL of the standard solution.
                                                                  System performance: When the procedure is run with 20
Identification (1) Determine the absorption spectrum of        mL of the standard solution under the above operating con-
a solution of Bisoprolol Fumarate in methanol (1 in 10,000)    ditions, the number of theoretical plates and the symmetry
as directed under Ultraviolet-visible Spectrophotometry        factor of the peak of bisoprolol are not less than 5000 and
1844       Official Monographs                                                                      Supplement I, JP XV

not more than 1.5, respectively.                                 oxide as a dessicant, dissolve in water to make exactly 200
  System repeatability: When the test is repeated 6 times        mL, and use as the standard solution. Determine the absor-
with 20 mL of the standard solution under the above operat-      bances, AT and AS, of the sample solution and standard so-
ing conditions, the relative standard deviation of the peak      lution at 271.5 nm as directed under Ultraviolet-visible Spec-
area of bisoprolol is not more than 1.5z.                        trophotometry <2.24>.

Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-          Amount (mg) of bisoprolol fumarate [(C18H31NO4)2.C4H4O4]
um, phosphorus (V) oxide, 809 5 hours).
                            C,                                    =WS×(AT/AS)×(V?/V)×(1/20)

Residue on ignition <2.44>   Not more than 0.1z (1 g).             WS: Amount (mg) of bisoprolol fumarate for assay
Assay Weigh accurately about 0.6 g of Bisoprolol                 Dissolution <6.10>—When the test is performed at 50 revolu-
Fumarate, previously dried, dissolve in 70 mL of acetic acid     tions per minute according to the Paddle method, using 900
(100), and titrate <2.50> with 0.1 mol/L perchloric acid VS      mL of 2nd fluid for dissolution test as the dissolution medi-
(indicator: 2 drops of crystal violet TS). The endpoint of       um, the dissolution rate in 30 minutes of Bisoprolol
titration is when the purple color of the solution turns blue    Fumarate Tablets is not less than 85z.
and then blue-green. Perform a blank determination in the           Start the test with 1 tablet of Bisoprolol Fumarate Tablets,
same manner, and make any necessary correction.                  withdraw not less than 20 mL of the medium at the specified
                                                                 minute after starting the test, and filter through a membrane
        Each mL of 0.1 mol/L perchloric acid VS
                                                                 filter with a pore size not exceeding 0.45 mm. Discard the
          =38.35 mg of (C18H31NO4)2.C4H4O4
                                                                 first 10 mL of the filtrate, pipet V mL of the subsequent
Containers and storage    Containers—Tight containers.           filtrate, add the dissolution medium to make exactly V? mL
                                                                 so that each mL contains about 2.8 mg of bisoprolol
                                                                 fumarate [(C18H31NO4)2.C4H4O4] according to the labeled
Add the following:                                               amount, and use this solution as the sample solution.
                                                                 Separately, weigh accurately about 14 mg of bisoprolol
Bisoprolol Fumarate Tablets                                      fumarate for assay, previously dried in vacuum at 809 for 5
                                                                                                                         C
                                                                 hours using phosphorus (V) oxide as a dessicant, and dis-
ビソプロロールフマル酸塩錠                                                    solve in the dissolution medium to make exactly 100 mL.
                                                                 Pipet 2 mL of this solution, add the dissolution medium to
  Bisoprolol Fumarate Tablets contain not less than              make exactly 100 mL, and use this solution as the standard
95.0z and not more than 105.0z of the labeled am-                solution. Perform the test with exactly 50 mL each of the
ount of bisoprolol fumarate [(C18H31NO4)2.C4H4O4:                sample solution and standard solution as directed under Liq-
766.96].                                                         uid Chromatography <2.01> according to the following con-
                                                                 ditions, and determine the bisoprolol peak areas, AT and AS,
Method of preparation Prepare as directed under Tablets,
                                                                 of both solutions.
with Bisoprolol Fumarate.
                                                                 Dissolution rate (z) with respect to the labeled amount of
Identification To a quantity of powdered Bisoprolol
                                                                 bisoprolol fumarate [(C18H31NO4)2.C4H4O4]
Fumarate Tablets, equivalent to 10 mg of Bisoprolol
                                                                   =WS×(AT/AS)×(V?/V)×(1/C)×18
Fumarate according to the labeled amount, add 60 mL of
methanol, shake vigorously for 10 minutes, add methanol to         WS: Amount (mg) of bisoprolol fumarate for assay
make 100 mL, and filter through a membrane filter with a           C: Labeled amount (mg) of bisoprolol fumarate
pore size not exceeding 0.45 mm. Determine the absorption             [(C18H31NO4)2.C4H4O4] in 1 tablet
spectrum of the filtrate as directed under Ultraviolet-visible
                                                                 Operating conditions—
Spectrophotometry <2.24>: it exhibits a maximum between
                                                                   Detector, column, column temperature, and flow rate:
271 nm and 275 nm.
                                                                 Proceed as directed in the operating conditions in the Assay.
Uniformity of dosage units <6.02>—Perform the test accord-         Mobile phase: Dissolve 4.08 g of potassium dihydrogen
ing to the following method: it meets the requirement of the     phosphate in 1000 mL of water, and adjust to pH 2.5 with
Content uniformity test.                                         phosphoric acid. To 750 mL of this solution add 250 mL of
   Take 1 tablet of Bisoprolol Fumarate Tablets, disintegrate    acetonitrile.
by adding 8 mL of water, and add water to make exactly 10        System suitability—
mL, and then filter through a membrane filter with a pore          System performance: When the procedure is run with 50
size not exceeding 0.45 mm. Discard the first 3 mL of the        mL of the standard solution under the above operating con-
filtrate, pipet V mL of the subsequent filtrate, add water to    ditions, the number of theoretical plates and the symmetry
make exactly V? mL so that each mL contains about 0.1 mg         factor of the peak of bisoprolol are not less than 3000 and
of bisoprolol fumarate [(C18H31NO4)2.C4H4O4], and use as         not more than 2.0, respectively.
the sample solution. Separately, weigh accurately about 20         System repeatability: When the test is repeated 6 times
mg of bisoprolol fumarate for assay, previously dried under      with 50 mL of the standard solution under the above operat-
reduced pressure at 809 for 5 hours, using phosphorus (V)
                        C                                        ing conditions, the relative standard deviation of the peak
Supplement I, JP XV                                                                         Official Monographs             1845

area of bisoprolol is not more than 2.0z.                          Add the following:
Assay Weigh accurately not less than 20 Bisoprolol
Fumarate Tablets and powder. Weigh accurately a portion            Bucillamine Tablets
of the powder, equivalent to about 20 mg of bisoprolol
                                                                   ブシラミン錠
fumarate [(C18H31NO4)2.C4H4O4], add exactly 70 mL of a
mixture of water and acetonitrile (3:1) and exactly 10 mL of
                                                                     Bucillamine Tablets contain not less than 95.0z and
the internal standard solution, shake vigorously for 10
                                                                   not more than 105.0z of the labeled amount of bucil-
minutes, and add the mixture of water and acetonitrile (3:1)
                                                                   lamine (C7H13NO3S2: 223.31).
to make 100 mL. Filter this solution through a membrane
filter with a pore size not exceeding 0.45 mm, discard the first   Method of preparation     Prepare as directed under Tablets,
3 mL of the filtrate, and use the subsequent filtrate as the       with Bucillamine.
sample solution. Separately, weigh accurately about 20 mg
                                                                   Identification (1) To a quantity of powdered Bucillamine
of bisoprolol fumarate for assay, previously dried in vacuum
                                                                   Tablets, equivalent to 0.1 g of Bucillamine according to the
at 809 for 5 hours using phosphorus (V) oxide as the des-
       C
                                                                   labeled amount, add 0.1 g of sodium hydrogen carbonate
sicant, add exactly 10 mL of the internal standard solution,
                                                                   and 10 mL of water, shake well, filter, and add 1 or 2 drops
dissolve in the mixture of water and acetonitrile (3:1) to
                                                                   of ninhydrin TS to the filtrate: it exhibits a red-brown color.
make 100 mL, and use this solution as the standard solution.
                                                                     (2) To a quantity of powdered Bucillamine Tablets,
Perform the test with 20 mL each of the sample solution and
                                                                   equivalent to 0.1 g of Bucillamine according to the labeled
standard solution as directed under Liquid Chromatography
                                                                   amount, add 25 mL of water, shake well, and filter. To 5
<2.01> according to the following conditions, and calculate
                                                                   mL of the filtrate, add 2 mL of dilute sodium hydroxide TS
the ratios, QT and QS, of the peak area of bisoprolol to that
                                                                   and 1 or 2 drops of sodium pentacyanonitrosylferrate (III)
of the internal standard.
                                                                   TS: it exhibits a red-purple color.
Amount (mg) of bisoprolol fumarate [(C18H31NO4)2.C4H4O4]
                                                                   Uniformity of dosage units <6.02>—Perform the test accord-
 = W S ×( Q T / Q S )
                                                                   ing to the following method: it meets the requirement of the
  WS: Amount (mg) of bisoprolol fumarate for assay                 Content uniformity test.
                                                                      Store the sample solution and standard solution in a cold
Internal standard solution—A solution of isopropyl para-
                                                                   place until performing the measurements. Take 1 tablet of
hydroxybenzoate in the mixture of water and acetonitrile
                                                                   Bucillamine Tablets, add exactly 1 mL of the internal stan-
(3:1) (1 in 250).
                                                                   dard solution per 0.1 g of bucillamine (C7H13NO3S2), then
Operating conditions—
                                                                   add 3 mL of water and 6 mL of methanol per 0.1 g of bucil-
  Detector: An ultraviolet absorption photometer (wave-
                                                                   lamine (C7H13NO3S2), and stir well until the tablet complete-
length: 225 nm).
                                                                   ly disintegrated. To 1 mL of this solution add the mobile
  Column: A stainless steel column 4.6 mm in inside di-
                                                                   phase to make 25 mL, filter through a membrane filter with
ameter and 15 cm in length, packed with octylsilanized silica
                                                                   a pore size not exceeding 0.45 mm, and use the filtrate as the
gel for liquid chromatography (5 mm in particle diameter).
                                                                   sample solution. Then, proceed as directed in the Assay.
  Column temperature: A constant temperature of about
409 C.                                                                     Amount (mg) of bucillamine (C7H13NO3S2)
  Mobile phase: Dissolve 4.08 g of potassium dihydrogen                     =WS×(QT/QS)×C×(1/200)
phosphate in 1000 mL of water, and adjust to pH 2.5 with
                                                                     WS: Amount (mg) of bucillamine for assay
phosphoric acid. To 800 mL of this solution add 200 mL of
                                                                     C: Labeled amount (mg) of bucillamine (C7H13NO3S2) in
acetonitrile.
                                                                        1 tablet
  Flow rate: Adjust the flow rate so that the retention time
of bisoprolol is about 8 minutes.                                  Internal standard solution—A solution of 4-fluorobenzoic
System suitability—                                                acid in methanol (1 in 100).
  System performance: When the procedure is run with 20
                                                                   Dissolution <6.10>—When the test is performed at 50 revolu-
mL of the standard solution under the above operating con-
                                                                   tions per minute according to the Paddle method, using 900
ditions, fumaric acid, bisoprolol and the internal standard
                                                                   mL of water as the dissolution medium, the dissolution rate
are eluted in this order with the resolution between the peaks
                                                                   in 30 minutes of Bucillamine Tablets is not less than 80z.
of bisoprolol and the internal standard being not less than
                                                                      Store the sample solution and standard solution in a cold
12.
                                                                   place until performing the measurements. Start the test with
  System repeatability: When the test is repeated 6 times
                                                                   1 tablet of Bucillamine Tablets, withdraw not less than 20
with 20 mL of the standard solution under the above operat-
                                                                   mL of the medium at the specified minute after starting the
ing conditions, the relative standard deviation of the ratio of
                                                                   test, and filter through a membrane filter with a pore size
the peak area of bisoprolol to that of the internal standard is
                                                                   not exceeding 0.45 mm. Discard the first 10 mL of the
not more than 1.0z.
                                                                   filtrate, and use the subsequent filtrate as the sample solu-
Containers and storage     Containers—Tight containers.            tion. Separately, weigh accurately an amount of bucillamine
1846       Official Monographs                                                                      Supplement I, JP XV

for assay equivalent to the labeled amount of the tablet,
                                                                         Amount (mg) of bucillamine (C7H13NO3S2)
previously dried in vacuum at 609 for 6 hours using phos-
                                  C
                                                                          =WS×(QT/QS)×C×(1/200)
phorus (V) oxide as a dessicant, and dissolve in methanol to
make exactly 10 mL. Pipet 1 mL of this solution, add water         WS: Amount (mg) of bucillamine for assay
to make exactly 100 mL, and use this solution as the stan-         C: Labeled amount (mg) of bucillamine (C7H13NO3S2) in
dard solution. Perform the test with exactly 20 mL each of            1 tablet
the sample solution and standard solution as directed under
                                                                 Internal standard solution—A solution of 4-fluorobenzoic
Liquid Chromatography <2.01> according to the following
                                                                 acid in methanol (1 in 100).
conditions, and determine the bucillamine peak areas, AT
                                                                 Operating conditions—
and AS, of both solutions.
                                                                    Detector: An ultraviolet absorption photometer (wave-
Dissolution rate (z) with respect to the labeled amount of       length: 254 nm).
bucillamine (C7H13NO3S2)                                            Column: A stainless steel column 4.6 mm in inside di-
  =WS×(AT/AS)×(1/C)×90                                           ameter and 15 cm in length, packed with octadecylsilanized
                                                                 silica gel for liquid chromatography (5 mm in particle di-
  WS: Amount (mg) of bucillamine for assay
                                                                 ameter).
  C: Labeled amount (mg) of bucillamine (C7H13NO3S2) in
                                                                    Column temperature: A constant temperature of about
     1 tablet
                                                                 409  C.
Operating conditions—                                               Mobile phase: A mixture of diluted phosphoric acid (1 in
  Detector, column, and column temperature: Proceed as           1000) and methanol (3:2).
directed in the operating conditions in the Assay.                  Flow rate: Adjust the flow rate so that the retention time
  Mobile phase: A mixture of diluted phosphoric acid (1 in       of bucillamine is about 5 minutes.
1000) and methanol (11:9).                                       System suitability—
  Flow rate: Adjust the flow rate so that the retention time        System performance: When the procedure is run with 10
of bucillamine is about 4 minutes.                               mL of the standard solution under the above operating con-
System suitability—                                              ditions, bucillamine and the internal standard are eluted in
  System performance: When the procedure is run with 20          this order with the resolution between these peaks being not
mL of the standard solution under the above operating con-       less than 3.
ditions, the number of theoretical plates and the symmetry          System repeatability: When the test is repeated 6 times
factor of the peak of bucillamine are not less than 3000 and     with 10 mL of the standard solution under the above operat-
not more than 1.5, respectively.                                 ing conditions, the relative standard deviation of the ratio of
  System repeatability: When the test is repeated 6 times        the peak area of bucillamine to that of the internal standard
with 20 mL of the standard solution under the above operat-      is not more than 1.0z.
ing conditions, the relative standard deviation of the peak
                                                                 Containers and storage     Containers—Tight containers.
area of bucillamine is not more than 2.0z.

Assay Store the sample solution and standard solution in a
cold place until performing the measurements. Take 10            Add the following:
tablets of Bucillamine Tablets, add exactly 1 mL of the inter-
nal standard solution per 0.1 g of bucillamine (C7H13NO3S2),     Buformin Hydrochloride
add 3 mL of water and 6 mL of methanol, and stir well until
the tablets completely disintegrated. To 1 mL of this solu-      ブホルミン塩酸塩
tion add the mobile phase to make 25 mL, filter through a
membrane filter with a pore size not exceeding 0.45 mm, and
use this solution as the sample solution. Separately, weigh
accurately about 0.2 g of bucillamine for assay, previously
                                                                 C6H15N5.HCl: 193.68
dried in vacuum for 6 hours at 609 using phosphorus (V)
                                     C
                                                                 1-Butylbiguanide hydrochloride     [1190-53-0]
oxide as a dessicant, add exactly 2 mL of the internal stan-
dard solution, and add 6 mL of water and 12 mL of
                                                                   Buformin Hydrochloride, when dried, contains not
methanol. To 1 mL of this solution add 25 mL of the mobile
                                                                 less than 98.5z and not more than 101.0z of
phase, filter through a membrane filter with a pore size not
                                                                 C6H15N5.HCl.
exceeding 0.45 mm, and use this solution as the standard so-
lution. Perform the test with 10 mL each of the sample solu-     Description Buformin Hydrochloride occurs as a white
tion and standard solution as directed under Liquid Chro-        crystalline powder.
matography <2.01> according to the following conditions,           It is freely soluble in water and in ethanol (99.5).
and calculate the ratios, QT and QS, of the peak area of
                                                                 Identification (1) To 5 mL of a solution of Buformin
bucillamine to that of the internal standard.
                                                                 Hydrochloride (1 in 2000) add 1 mL of dilute sodium pen-
                                                                 tacyanonitrosylferrate (III)-potassium hexacyanoferrate
Supplement I, JP XV                                                                        Official Monographs           1847

(III) TS: a red-brown color develops.                             solution, and add the mobile phase to make exactly 10 mL.
   (2) Determine the absorption spectrum of a solution of         Confirm that the peak area of buformin obtained from 10
Buformin Hydrochloride (1 in 125,000) as directed under           mL of this solution is equivalent to 7 to 13z of that from 10
Ultraviolet-visible Spectrophotometry <2.24>, and compare         mL of the standard solution.
the spectrum with the Reference Spectrum: both spectra              System performance: When the procedure is run with 10
exhibit similar intensities of absorption at the same             mL of the standard solution under the above operating con-
wavelengths.                                                      ditions, the number of theoretical plates and the symmetry
   (3) Determin the infrared absorption spectrum of Bufor-        factor of the peak of buformin are not less than 5000 and
min Hydrochloride as directed in the potassium chloride           not more than 2.0, respectively.
disk method under Infrared Spectrophotometry <2.25>, and            System repeatability: When the test is repeated 6 times
compare the spectrum with the Reference Spectrum: both            with 10 mL of the standard solution under the above operat-
spectra exhibit similar intensities of absorption at the same     ing conditions, the relative standard deviation of the peak
wave numbers.                                                     area of buformin is not more than 1.0z.
   (4) A solution of Buformin Hydrochloride (1 in 20)
                                                                  Loss on drying <2.41>    Not more than 0.5z (1 g, 1059 3
                                                                                                                       C,
responds to the Qualitative Tests <1.09> for chlorides.
                                                                  hours).
Melting point <2.60>    175 – 1809
                                 C
                                                                  Residue on ignition <2.44>   Not more than 0.1z (1 g).
Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of
                                                                  Assay Weigh accurately about 0.15 g of Buformin
Buformin Hydrochloride according to Method 1, and per-
                                                                  Hydrochloride, previously dried, dissolve in 50 mL of a mix-
form the test. Prepare the control solution with 2.0 mL of
                                                                  ture of acetic anhydride and acetic acid (100) (7:3), and im-
Standard Lead Solution (not more than 20 ppm).
                                                                  mediately titrate <2.50> with 0.1 mol/L perchloric acid VS
   (2) Arsenic <1.11>—Prepare the test solution with 1.0 g
                                                                  (potentiometric titration). Perform a blank determination in
of Buformin Hydrochloride according to Method 1, and
                                                                  the same manner, and make any necessary correction.
perform the test (not more than 2 ppm).
   (3) Related substances—Dissolve 0.10 g of Buformin                     Each mL of 0.1 mol/L perchloric acid VS
Hydrochloride in 200 mL of the mobile phase, and use this                   =9.684 mg of C6H15N5.HCl
solution as the sample solution. Pipet 1 mL of the sample
                                                                  Containers and storage    Containers—Tight containers.
solution, add the mobile phase to make exactly 100 mL, and
use this solution as the standard solution. Perform the test
with exactly 10 mL each of the sample solution and standard
                                                                  Add the following:
solution as directed under Liquid Chromatography <2.01>
according to the following conditions, and determine each
peak area of both solutions by the automatic integration          Buformin Hydrochloride
method: the area of the peak other than buformin obtained         Enteric-coated Tablets
from the sample solution is not larger than 1/5 times the
peak area of buformin from the standard solution. Further-        ブホルミン塩酸塩腸溶錠
more, the total of the areas of all peaks other than the bufor-
min peak from the sample solution is not larger than 1/2            Buformin Hydrochloride Enteric-coated Tablets
times the peak area of buformin from the standard solution.       contain not less than 93.0z and not more than 107.0
Operating conditions—                                             z of the labeled amount of buformin hydrochloride
   Detector: An ultraviolet absorption photometer (wave-          (C6H15N5.HCl: 193.68).
length: 230 nm).                                                  Method of preparation Prepare as directed under Tablets,
   Column: A stainless steel column 4.6 mm in inside di-          with Buformin Hydrochloride.
ameter and 15 cm in length, packed with octadecylsilanized
silica gel for liquid chromatography (5 mm in particle di-        Identification To a quantity of powdered Buformin
ameter).                                                          Hydrochloride Enteric-coated Tablets, equivalent to 0.1 g of
   Column temperature: A constant temperature of about            Buformin Hydrochloride according to the labeled amount,
359  C.                                                           add 10 mL of water, shake well, and then filter. To 4 mL of
   Mobile phase: A mixture of a solution of sodium                the filtrate add 1 mL of a mixture of hydrogen peroxide TS,
perchlorate monohydrate in diluted phosphoric acid (1 in          sodium pentacyanonitrosylferrate (III) TS and a solution of
1000) (7 in 250) and acetonitrile (7:1).                          sodium hydroxide (1 in 10) (2:1:1): the solution exhibits a
   Flow rate: Adjust the flow rate so that the retention time     red to red-purple color.
of buformin is about 6 minutes.                                   Uniformity of dosage units <6.02> Perform the test accord-
   Time span of measurement: About 2 times as long as the         ing to the following method: it meets the requirement of the
retention time of buformin, beginning after the solvent           Content uniformity test.
peak.                                                               To 1 tablet of Buformin Hydrochloride Enteric-coated
System suitability—                                               Tablets add 5 mL of a mixture of ethanol (99.5) and acetone
   Test for required detectability: Pipet 1 mL of the standard
1848       Official Monographs                                                                        Supplement I, JP XV

(1:1), disperse the pellicle to smaller using ultrasonic waves,      Column: A stainless steel column 4.6 mm in inside di-
add exactly 10 mL of the internal standard solution per 50        ameter and 15 cm in length, packed with octadecylsilanized
mg of buformin hydrochloride (C6H15N5.HCl), and then add          silica gel for liquid chromatography (5 mm in particle di-
diluted acetonitrile (1 in 2) to make 13V/20 mL. Disintegrate     ameter).
the tablet using ultrasonic waves, then shake for 20 minutes,        Column temperature: A constant temperature of about
and add diluted acetonitrile (1 in 2) to make a solution,         359  C.
volume V mL, containing about 0.5 mg of buformin                     Mobile phase: A mixture of a solution of sodium
hydrochloride (C6H15N5.HCl) per mL. Centrifuge this               perchlorate in diluted phosphoric acid (1 in 1000) (7 in 500)
solution, pipet 1 mL of the supernatant liquid, and add the       and acetonitrile (7:1).
mobile phase to make 50 mL. If necessary, filter this solu-          Flow rate: Adjust the flow rate so that the retention time
tion through a membrane filter with a pore size not exceed-       of buformin is about 6 minutes.
ing 0.5 mm, and use the filtrate as the sample solution. Then,    System suitability—
proceed as directed in the Assay.                                    System performance: When the procedure is run with 20
                                                                  mL of the standard solution under the above operating con-
 Amount (mg) of buformin hydrochloride (C6H15N5.HCl)
                                                                  ditions, the number of theoretical plates and the symmetry
  =WS×(QT/QS)×(V/50)
                                                                  factor of the peak of buformin are not less than 3000 and
  WS: Amount (mg) of buformin hydrochloride for assay             not more than 2.0, respectively.
                                                                     System repeatability: When the test is repeated 6 times
Internal standard solution—A solution of p-acetanisidide in
                                                                  with 20 mL of the standard solution under the above operat-
diluted acetonitrile (1 in 2) (1 in 150).
                                                                  ing conditions, the relative standard deviation of the peak
Dissolution <6.10> When the tests are performed at 50             area of buformin is not more than 2.0z.
revolutions per minute according to the Paddle method,
                                                                  Assay Add 20 mL of a mixture of ethanol (99.5) and
using 900 mL each of 1st fluid for dissolution test and 2nd
                                                                  acetone (1:1) to an amount of Buformin Hydrochloride
fluid for dissolution test as the dissolution medium, the dis-
                                                                  Enteric-coated Tablets equivalent to 0.5 g of buformin
solution rate in 120 minutes of Buformin Hydrochloride En-
                                                                  hydrochloride (C6H15N5.HCl), disperse the pellicles to
teric-coated Tablets using the 1st fluid is not more than 5z,
                                                                  smaller using ultrasonic waves, and then add 100 mL of
and that in 90 minutes of Buformin Hydrochloride Enteric-
                                                                  diluted acetonitrile (1 in 2). Disintegrate the tablets with the
coated Tablets using the 2nd fluid is not less than 80z.
                                                                  aid of ultrasonic waves, shake for 20 minutes, and then add
   Start the test with 1 tablet of Buformin Hydrochloride En-
                                                                  diluted acetonitrile (1 in 2) to make exactly 200 mL. Cen-
teric-coated Tablets, withdraw not less than 20 mL of the
                                                                  trifuge this solution, pipet 10 mL of the supernatant liquid,
medium at the specified minute after starting the test, and
                                                                  add exactly 5 mL of the internal standard solution, and then
filter through a membrane filter with a pore size not exceed-
                                                                  add diluted acetonitrile (1 in 2) to make 50 mL. Pipet 1 mL
ing 0.5 mm. Discard the first 10 mL of the filtrate, pipet V
                                                                  of this solution, and add the mobile phase to make 50 mL. If
mL of the subsequent filtrate, add the relevant dissolution
                                                                  necessary, filter this solution through a membrane filter with
medium to make exactly V? mL so that each mL contains
                                                                  a pore size not exceeding 0.5 mm, and use the filtrate as the
about 56 mg of buformin hydrochloride (C6H15N5.HCl) ac-
                                                                  sample solution. Separately, weigh accurately about 25 mL
cording to the labeled amount, and use this solution as the
                                                                  of buformin hydrochloride for assay, previously dried at
sample solution. Separately, weigh accurately about 28 mg
                                                                  1059 for 3 hours, dissolve in an adequate amount of dilut-
                                                                       C
of buformin hydrochloride for assay, previously dried at
                                                                  ed acetonitrile (1 in 2), add exactly 5 mL of the internal stan-
1059 for 3 hours, and dissolve in the relevant dissolution
      C
                                                                  dard solution, and then add diluted acetonitrile (1 in 2) to
medium to make exactly 100 mL. Pipet 4 mL of this solu-
                                                                  make 50 mL. To 1 mL of this solution add the mobile phase
tion, add the relevant dissolution medium to make exactly 20
                                                                  to make 50 mL, and use this solution as the standard solu-
mL, and use this solution as the standard solution. Perform
                                                                  tion. Perform the test with 10 mL each of the sample solution
the test with exactly 20 mL each of the sample solution and
                                                                  and standard solution as directed under Liquid Chro-
standard solution as directed under Liquid Chromatography
                                                                  matography <2.01> according to the following conditions,
<2.01> according to the following conditions, and determine
                                                                  and calculate the ratios, QT and QS, of the peak area of
the buformin peak areas, AT and AS, of both solutions.
                                                                  buformin to that of the internal standard.
Dissolution rate (z) with respect to the labeled amount of
                                                                   Amount (mg) of buformin hydrochloride (C6H15N5.HCl)
buformin hydrochloride (C6H15N5.HCl)
                                                                    =WS×(QT/QS)×20
  =WS×(AT/AS)×(V?/V)×(1/C)×180
                                                                    WS: Amount (mg) of buformin hydrochloride for assay
  WS: Amount (mg) of buformin hydrochloride for assay
  C: Labeled amount (mg) of buformin hydrochloride                Internal standard solution—A solution of p-acetanisidide in
     (C6H15N5.HCl) in 1 tablet                                    diluted acetonitrile (1 in 2) (1 in 150).
                                                                  Operating conditions—
Operating conditions—
                                                                     Detector: An ultraviolet absorption photometer (wave-
  Detector: An ultraviolet absorption photometer (wave-
                                                                  length: 233 nm).
length: 230 nm).
                                                                     Column: A stainless steel column 4.6 mm in inside di-
Supplement I, JP XV                                                                        Official Monographs            1849

ameter and 15 cm in length, packed with octadecylsilanized        100 mL, and use this solution as the standard solution. De-
silica gel for liquid chromatography (5 mm in particle di-        termine the absorbances, AT and AS, of the sample solution
ameter).                                                          and standard solution at 233 nm as directed under Ultrav-
   Column temperature: A constant temperature of about            iolet-visible Spectrophotometry <2.24>.
359  C.
                                                                   Amount (mg) of buformin hydrochloride (C6H15N5.HCl)
   Mobile phase: A mixture of a solution of sodium
                                                                    =WS×(AT/AS)×(2/V)
perchlorate (7 in 250) and acetonitrile (7:1).
   Flow rate: Adjust the flow rate so that the retention time       WS: Amount (mg) of buformin hydrochloride for assay
of buformin is about 7 minutes.
                                                                  Dissolution <6.10> When the test is performed at 50 revolu-
System suitability—
                                                                  tions per minute according to the Paddle method, using 900
   System performance: When the procedure is run with 10
                                                                  mL of water as the dissolution medium, the dissolution rate
mL of the standard solution under the above operating con-
                                                                  in 15 minutes of Buformin Hydrochloride Tablets is not less
ditions, buformin and the internal standard are eluted in this
                                                                  than 80z.
order with the resolution between these peaks being not less
                                                                     Start the test with 1 tablet of Buformin Hydrochloride
than 5.
                                                                  Tablets, withdraw not less than 20 mL of the medium at the
   System repeatability: When the test is repeated 6 times
                                                                  specified minute after starting the test, and filter through a
with 10 mL of the standard solution under the above operat-
                                                                  membrane filter with a pore size not exceeding 0.5 mm. Dis-
ing conditions, the relative standard deviation of the ratio of
                                                                  card the first 10 mL of the filtrate, pipet V mL of the subse-
the peak area of buformin to that of the internal standard is
                                                                  quent filtrate, and add water to make exactly V? mL so that
not more than 1.0z.
                                                                  each mL contains about 5.6 mg of buformin hydrochloride
Containers and storage     Containers—Well-closed contain-        (C6H15N5.HCl) according to the labeled amount, and use
ers.                                                              this solution as the sample solution. Separately, weigh ac-
                                                                  curately about 28 mg of buformin hydrochloride for assay,
                                                                  previously dried at 1059 for 3 hours, and dissolve in water
                                                                                           C
Add the following:                                                to make exactly 100 mL. Pipet 2 mL of this solution, add
                                                                  water to make exactly 100 mL, and use this solution as the
Buformin Hydrochloride Tablets                                    standard solution. Perform the test with the sample solution
                                                                  and standard solution as directed under Ultraviolet-visible
ブホルミン塩酸塩錠                                                         Spectrophotometry <2.24>, and determine the absorbances,
                                                                  AT and AS, at 233 nm.
  Buformin Hydrochloride Tablets contain not less
                                                                  Dissolution rate (z) with respect to the labeled amount of
than 95.0z and not more than 105.0z of the labeled
                                                                  buformin hydrochloride (C6H15N5.HCl)
amount of buformin hydrochloride (C6H15N5.HCl:
                                                                    =WS×(AT/AS)×(V?/V)×(1/C)×18
193.68).
                                                                    WS: Amount (mg) of buformin hydrochloride for assay
Method of preparation Prepare as directed under Tablets,
                                                                    C: Labeled amount (mg) of buformin hydrochloride
with Buformin Hydrochloride.
                                                                       (C6H15N5.HCl) in 1 tablet
Identification To a quantity of powdered Buformin
                                                                  Assay Weigh accurately not less than 20 Buformin
Hydrochloride Tablets, equivalent to 1 g of Buformin
                                                                  Hydrochloride Tablets, and powder. Weigh accurately a
Hydrochloride according to the labeled amount, add 100
                                                                  portion of the powder, equivalent to about 60 mg of bufor-
mL of water, shake well, and then filter. To 4 mL of the
                                                                  min hydrochloride (C6H15N5.HCl), add water to make
filtrate add 1 mL of dilute sodium pentacyanonitrosylferrate
                                                                  exactly 200 mL, and treat with ultrasonic waves for 5
(III)-potassium hexacyanoferrate (III) TS: the solution ex-
                                                                  minutes. Take 40 mL of this solution, centrifuge, pipet 2 mL
hibits a red-brown color.
                                                                  of the supernatant liquid, add water to make exactly 100
Uniformity of dosage units <6.02> Perform the test accord-        mL, and use this solution as the sample solution. Separately,
ing to the following method: it meets the requirement of the      weigh accurately about 60 mg of buformin hydrochloride
Content uniformity test.                                          for assay, previously dried at 1059 for 3 hours, and dis-
                                                                                                      C
   Take 1 tablet of Buformin Hydrochloride Tablets, add           solve in water to make exactly 200 mL. Pipet 2 mL of this
water to make exactly 200 mL, and then treat with ultrasonic      solution, add water to make exactly 100 mL, and use this so-
waves for 5 minutes. Take 40 mL of this solution and cen-         lution as the standard solution. Perform the test with the
trifuge. Pipet V mL of the supernatant liquid equivalent to       sample solution and standard solution as directed under
about 0.5 mg of buformin hydrochloride (C6H15N5.HCl),             Ultraviolet-visible Spectrophotometry <2.24>, and determine
add water to make exactly 100 mL, and use this solution as        the absorbances, AT and AS, at 233 nm.
the sample solution. Separately, weigh accurately about 50
                                                                   Amount (mg) of buformin hydrochloride (C6H15N5.HCl)
mg of buformin hydrochloride for assay, previously dried at
                                                                    = W S × ( A T/ A S )
1059 for 3 hours, and dissolve in water to make exactly 200
     C
mL. Pipet 2 mL of this solution, add water to make exactly          WS: Amount (mg) of buformin hydrochloride for assay
1850       Official Monographs                                                                       Supplement I, JP XV

Containers and storage     Containers—Well-closed contain-        phine Hydrochloride in 20 mL of the mobile phase, and use
ers.                                                              this solution as the sample solution. Pipet 1 mL of the sam-
                                                                  ple solution, add the mobile phase to make exactly 100 mL,
                                                                  and use this solution as the standard solution. Perform the
Add the following:                                                test with exactly 20 mL each of the sample solution and stan-
                                                                  dard solution as directed under Liquid Chromatography
Buprenorphine Hydrochloride                                       <2.01> according to the following conditions. Determine
                                                                  each peak area of both solutions by the automatic integra-
ブプレノルフィン塩酸塩                                                       tion method: the area of each peak other than buprenor-
                                                                  phine obtained from the sample solution is not larger than
                                                                  1/4 times the peak area of buprenorphine from the standard
                                                                  solution. Furthermore, the total area of the peaks other than
                                                                  buprenorphine from the sample solution is not larger than
                                                                  13/20 times the peak area of buprenorphine from the stan-
                                                                  dard solution.
                                                                  Operating conditions—
C29H41NO4.HCl: 504.10
                                                                     Detector: An ultraviolet absorption photometer (wave-
(2S)-2-[(5R,6R,7R,14S)-17-(Cyclopropylmethyl)-4,5-
                                                                  length: 288 nm).
epoxy-3-hydroxy-6-methoxy-6,14-ethanomorphinan-7-yl]-
                                                                     Column: A stainless steel column 4.6 mm in inside di-
3,3-dimethylbutan-2-ol monohydrochloride [53152-21-9]
                                                                  ameter and 25 cm in length, packed with octadecylsilanized
                                                                  silica gel for liquid chromatography (5 mm in particle di-
  Buprenorphine Hydrochloride, when dried, con-
                                                                  ameter).
tains not less than 98.5z and not more than 101.0z
                                                                     Column temperature: A constant temperature of about
of C29H41NO4.HCl.
                                                                  409  C.
Description Buprenorphine Hydrochloride occurs as white              Mobile phase: A mixture of methanol, ammonium acetate
to yellowish white, crystals or a crystalline powder.             solution (1 in 100), and acetic acid (100) (6000:1000:1).
  It is freely soluble in methanol and in acetic acid (100),         Flow rate: Adjust the flow rate so that the retention time
and sparingly soluble in water and in ethanol (99.5).             of buprenorphine is about 17 minutes.
  Melting point: about 2689 (with decomposition).
                              C                                      Time span of measurement: About 2.5 times as long as the
                                                                  retention time of buprenorphine, beginning after the solvent
Identification (1) Determine the absorption spectrum of
                                                                  peak.
a solution of Buprenorphine Hydrochloride (1 in 5000) as
directed under Ultraviolet-visible Spectrophotometry <2.24>,
                                                                  System suitability—
                                                                     Test for required detectability: Pipet 5 mL of the standard
and compare the spectrum with the Reference Spectrum:
                                                                  solution, and add the mobile phase to make exactly 50 mL.
both spectra exhibit similar intensities of absorption at the
                                                                  Confirm that the peak area of buprenorphine obtained from
same wavelengths.
                                                                  20 mL of this solution is equivalent to 7 to 13z of that from
  (2) Determine the infrared absorption spectrum of
                                                                  20 mL of the standard solution.
Buprenorphine Hydrochloride as directed in the potassium
                                                                     System performance: When the procedure is run with 20
chloride disk method under Infrared Spectrophotometry
<2.25>, and compare the spectrum with the Reference Spec-         mL of the standard solution under the above operating
                                                                  conditions, the number of theoretical plates and the symmet-
trum: both spectra exhibit similar intensities of absorption at
                                                                  ry factor of the peak of buprenorphine are not less than 6500
the same wave numbers.
                                                                  and not more than 1.2, respectively.
  (3) A solution of Buprenorphine Hydrochloride (1 in
                                                                     System repeatability: When the test is repeated 6 times
100) responds to the Qualitative Tests <1.09> for chloride.
                                                                  with 20 mL of the standard solution under the above operat-
Optical rotation <2.49> [a]20: -92 – -989(after drying,
                           D                                      ing conditions, the relative standard deviation of the peak
0.4 g, methanol, 20 mL, 100 mm).                                  area of buprenorphine is not more than 2.0z.
pH <2.54> The pH of a solution prepared by dissolving 1.0         Loss on drying <2.41>   Not more than 1.0z (1 g, 1159 3
                                                                                                                      C,
g of Buprenorphine Hydrochloride in 200 mL of water is            hours).
between 4.0 and 6.0.
                                                                  Residue on ignition <2.44>   Not more than 0.1z (1 g).
Purity (1) Clarity and color of solution—A solution ob-
                                                                  Assay Weigh accurately about 0.5 g of Buprenorphine
tained by dissolving 0.1 g of Buprenorphine Hydrochloride
                                                                  Hydrochloride, previously dried, dissolve in 5 mL of acetic
in 10 mL of water is clear and colorless.
                                                                  acid (100), add 50 mL of acetic anhydride, and titrate <2.50>
   (2) Heavy metals <1.07>—Proceed with 1.0 g of
                                                                  with 0.1 mol/L perchloric acid VS (potentiometric titra-
Buprenorphine Hydrochloride according to Method 4, and
                                                                  tion). Perform a blank determination in the same manner,
perform the test. Prepare the control solution with 1.0 mL
                                                                  and make any necessary correction.
of Standard Lead Solution (not more than 10 ppm).
   (3) Related substances—Dissolve 0.10 g of Buprenor-
Supplement I, JP XV                                                                         Official Monographs            1851

         Each mL of 0.1 mol/L perchloric acid VS                   Change the Purity to read:
           =50.41 mg of C29H41NO4.HCl
                                                                   Purity (1) Clarity and color of solution—To 1.25 g of
Containers and storage     Containers—Well-closed contain-         Calcium Folinate add 50 mL of freshly boiled and cooled
ers.                                                               water, and warm to 409 if necessary, to dissolve: the solu-
                                                                                            C,
                                                                   tion is clear, and the absorbance at 420 nm of it, determined
                                                                   as directed under Ultraviolet-visible Spectrophotometry
Calcium Chloride Injection                                         <2.24>, is not more than 0.25.
                                                                      (2) Heavy metals <1.07>—Proceed with 0.40 g of Calci-
塩化カルシウム注射液                                                         um Folinate according to Method 2, and perform the test.
                                                                   Prepare the control solution with 2.0 mL of Standard Lead
Add the following next to Extractable volume:                      Solution (not more than 50 ppm).
                                                                      (3) Related substances—Dissolve 10 mg of Calcium
Foreign insoluble matter <6.06> Perform the test according
                                                                   Folinate in 25 mL of water, and use this solution as the
to Method 1: it meets the requirement.
                                                                   sample solution. Pipet 2 mL of the sample solution, add
Insoluble particulate matter <6.07>     It meets the require-      water to make exactly 200 mL, and use this solution as the
ment.                                                              standard solution. Perform the test with exactly 20 mL each
                                                                   of the sample solution and standard solution as directed
Sterility <4.06> Perform the test according to the Mem-
                                                                   under Liquid Chromatography <2.01> according to the
brane filtration method: it meets the requirement.
                                                                   following conditions, and determine each peak area by the
                                                                   automatic integration method: the area of the peak other
                                                                   than folinate obtained from the sample solution is not larger
Calcium Folinate                                                   than the peak area of folinate from the standard solution,
ホリナートカルシウム                                                         and the total area of the peaks other than the peak of
                                                                   folinate from the sample solution is not larger than 5 times
                                                                   the peak area of folinate from the standard solution.
Change the Description and the Identification to
read:                                                              Operating conditions—
                                                                      Detector, column, column temperature, mobile phase,
Description Calcium Folinate occurs as a white to light yel-       and flow rate: Proceed as directed in the operating condi-
low, crystalline powder.                                           tions in the Assay.
  It is sparingly soluble in water, and practically insoluble in      Time span of measurement: About 2.5 times as long as the
methanol and in ethanol (99.5).                                    retention time of folinate, beginning after the solvent peak.
Identification (1) Determine the absorption spectrum of            System suitability—
a solution of Calcium Folinate (1 in 100,000) as directed             Test for required detectability: Pipet 5 mL of the standard
under Ultraviolet-visible Spectrophotometry <2.24>, and            solution, and add water to make exactly 50 mL. Confirm
compare the spectrum with the Reference Spectrum or the            that the peak area of folinate obtained from 20 mL of this so-
spectrum of a solution of Calcium Folinate Reference Stan-         lution is equivalent to 7 to 13z of that from 20 mL of the
dard prepared in the same manner as the sample solution:           standard solution.
both spectra exhibit similar intensities of absorption at the         System performance: Proceed as directed in the system
same wavelengths.                                                  suitability in the Assay.
   (2) Determine the infrared absorption spectrum of                  System repeatability: When the test is repeated 6 times
Calcium Folinate as directed in the potassium bromide disk         with 20 mL of the standard solution under the above operat-
method under Infrared Spectrophotometry <2.25>, and com-           ing conditions, the relative standard deviation of the peak
pare the spectrum with the Reference Spectrum: both spec-          area of folinate is not more than 2.0z.
tra exhibit similar intensities of absorption at the same wave
numbers.                                                           Change the Water to read:
   (3) A solution of Calcium Folinate (1 in 100) responds to       Water <2.48> Not less than 7.0z and not more than 17.0z
the Qualitative Tests <1.09> (2) and (3) for calcium salt.         (0.2 g, volumetric titration, direct titration).

Add the following next to Identification:                          Change the Assay to read:
Optical rotation <2.49>   [a]20:
                             +14 – +199
                             D            (0.1 g calculated        Assay Weigh accurately about 10 mg each of Calcium
on the anhydrous basis, water, 10 mL, 100 mm).                     Folinate and Calcium Folinate Reference Standard
pH <2.54> To 1.25 g of Calcium Folinate add 50 mL of               (separately determine the water <2.48> in the same manner as
freshly boiled and cooled water, and warm to 409 if neces-
                                                  C,               Calcium Folinate), dissolve in water to make them exactly 25
sary, to dissolve: the pH of this solution is between 6.8 and      mL. Pipet 5 mL each of these solutions, add the mobile
8.0.                                                               phase to make exactly 25 mL, and use these solutions as the
                                                                   sample solution and the standard solution, respectively. Per-
1852       Official Monographs                                                                        Supplement I, JP XV

form the test with exactly 20 mL each of the sample solution     Add the following:
and standard solution as directed under Liquid Chro-
matography <2.01> according to the following conditions,         Cefadroxil Capsules
and determine the peak areas, AT and AS, of folinate of both
solutions.                                                       セファドロキシルカプセル

              Amount (mg) of C20H21CaN7O7
                                                                   Cefadroxil Capsules contain not less than 95.0z
               = W S × ( A T/ A S )
                                                                 and not more than 105.0z of the labeled amount of
  WS: Amount (mg) of Calcium Folinate Reference Stan-            cefadroxil (C16H17N3O5S: 363.39).
      dard, calculated on the anhydrous basis
                                                                 Method of preparation       Prepare as directed under Cap-
Operating conditions—                                            sules, with Cefadroxil.
   Detector: An ultraviolet absorption photometer (wave-
                                                                 Identification Dissolve the contents of Cefadroxil Cap-
length: 254 nm).
                                                                 sules, equivalent to 10 mg (potency) of Cefadroxil according
   Column: A stainless steel column 4.6 mm in inside di-
                                                                 to the labeled amount, in 500 mL of water, and filter. Deter-
ameter and 15 cm in length, packed with octadecylsilanized
                                                                 mine the absorption spectrum of the filtrate as directed un-
silica gel for liquid chromatography (5 mm in particle di-
                                                                 der Ultraviolet-visible Spectrophotometry <2.24>: it exhibits
ameter).
                                                                 maxima between 228 nm and 232 nm, and between 261 nm
   Column temperature: A constant temperature of about
                                                                 and 265 nm.
459  C.
   Mobile phase: A mixture of disodium hydrogen phos-            Water <2.48> Not more than 7.0z (0.15 g, volumetric
phate dodecahydrate solution (287 in 100,000), methanol          titration, direct titration).
and tetrabutylammonium hydroxide TS (385:110:4), adjust-
                                                                 Uniformity of dosage units <6.02> Perform the test accord-
ed to pH7.5 with phosphoric acid.
                                                                 ing to the following method: it meets the requirement of the
   Flow rate: Adjust the flow rate so that the retention time
                                                                 Content uniformity test.
of folinate is about 10 minutes.
                                                                    Place 1 capsule of Cefadroxil Capsules in 300 mL of
System suitability—
                                                                 water, disperse with the aid of ultrasonic waves, shake for 30
   System performance: Dissolve 10 mg each of Calcium
                                                                 minutes, and add water to make exactly 500 mL. Pipet 5 mL
Folinate and folic acid in 100 mL of the mobile phase. When
                                                                 of this solution, and add water to make exactly V mL so that
the procedure is run with 20 mL of this solution under the
                                                                 each mL contains about 0.1 mg (potency) of Cefadroxil.
above operating conditions, folinate and folic acid are eluted
                                                                 Filter the solution, discard the first 10 mL of the filtrate, and
in this order with the resolution between these peaks being
                                                                 use the subsequent filtrate as the sample solution. Separate-
not less than 10.
                                                                 ly, weigh accurately about 20 mg (potency) of Cefadroxil
   System repeatability: When the test is repeated 6 times
                                                                 Reference Standard, dissolve in water to make exactly 200
with 20 mL of the standard solution under the above operat-
                                                                 mL, and use this solution as the standard solution. Then,
ing conditions, the relative standard deviation of the peak
                                                                 proceed as directed in the Assay under Cefadroxil.
area of folinate is not more than 1.0z.
                                                                    Amount [mg (potency)] of cefadroxil (C16H17N3O5S)
                                                                     =WS×(AT/AS)×(V/2)
Camostat Mesilate                                                  WS: Amount [mg (potency)] of Cefadroxil Reference
                                                                       Standard
カモスタットメシル酸塩
                                                                 Dissolution <6.10> When the test is performed at 50 revolu-
Change the Identification (3) to read:                           tions per minute according to the Paddle method, using 900
                                                                 mL of 0.05 mol/L acetic acid-sodium acetate buffer solu-
Identification (3) A 0.1 g portion of Camostat Mesilate
                                                                 tion, pH 4.0 as the dissolution medium, the dissolution rate
responds to the Qualitative Tests <1.09> (1) for mesilate.
                                                                 in 90 minutes of Cefadroxil Capsules is not less than 80z.
                                                                    Start the test with 1 capsule of Cefadroxil Capsules,
                                                                 withdraw not less than 20 mL of the medium at the specified
                                                                 minute after starting the test, and filter through a membrane
                                                                 filter with a pore size not exceeding 0.45 mm. Discard the
                                                                 first 10 mL of the filtrate, pipet V mL of the subsequent
                                                                 filtrate, add water to make exactly V? mL so that each mL
                                                                 contains about 22 mg (potency) of Cefadroxil according to
                                                                 the labeled amount, and use this solution as the sample solu-
                                                                 tion. Separately, weigh accurately about 22 mg (potency) of
                                                                 Cefadroxil Reference Standard, and add water to make ex-
                                                                 actly 100 mL. Pipet 5 mL of this solution, add water to
Supplement I, JP XV                                                                       Official Monographs            1853

make exactly 50 mL, and use this solution as the standard        Uniformity of dosage units <6.02> The syrup in single-unit
solution. Determine the absorbances, AT and AS, at 263 nm        container meets the requirement of the Mass variation test.
of the sample solution and standard solution as directed un-
                                                                 Dissolution <6.10> When the test is performed at 50 revolu-
der Ultraviolet-visible Spectrophotometry <2.24>, using
                                                                 tions per minute according to the Paddle method (put the
water as the blank.
                                                                 sample in the dissolution medium so that it disperses), using
Dissolution rate (z) with respect to the labeled amount of       900 mL of water as the dissolution medium, the dissolution
cefadroxil (C16H17N3O5S)                                         rate in 15 minutes of Cefadroxil for Syrup is not less than 85
  =WS×(AT/AS)×(V?/V)×(1/C)×90                                    z.
                                                                    Start the test with accurately weighed amount of
  WS: Amount [mg (potency)] of Cefadroxil Reference
                                                                 Cefadroxil for Syrup, equivalent to about 0.1 g (potency) of
      Standard
                                                                 Cefadroxil according to the labeled amount, withdraw not
  C: Labeled amount [mg (potency)] of cefadroxil in 1 cap-
                                                                 less than 20 mL of the medium at the specified minute after
     sule
                                                                 starting the test, and filter through a membrane filter with a
Assay Take out the contents of 20 Cefadroxil Capsules,           pore size not exceeding 0.45 mm. Discard the first 10 mL of
and combine. Weigh accurately the mass of the combined           the filtrate, pipet 4 mL of the subsequent filtrate, add water
contents, and powder. Weigh accurately a portion of the          to make exactly 20 mL, and use this solution as the sample
powder, equivalent to about 50 mg (potency) of Cefadroxil,       solution. Separately, weigh accurately about 22 mg (poten-
add 300 mL of water, shake for 30 minutes, then add water        cy) of Cefadroxil Reference Standard, and dissolve in water
to make exactly 500 mL, and filter. Discard the first 10 mL      to make exactly 100 mL. Pipet 5 mL of this solution, add
of the filtrate, and use the subsequent filtrate as the sample   water to make exactly 50 mL, and use this solution as the
solution. Separately, weigh accurately an amount of              standard solution. Determine the absorbances, AT and AS,
Cefadroxil Reference Standard, equivalent to about 20 mg         at 263 nm of the sample solution and standard solution as
(potency), dissolve in water to make exactly 200 mL, and use     directed under Ultraviolet-visible Spectrophotometry <2.24>.
this solution as the standard solution. Then, proceed as
                                                                 Dissolution rate (z) with respect to the labeled amount of
directed in the Assay under Cefadroxil.
                                                                 cefadroxil (C16H17N3O5S)
   Amount [mg (potency)] of cefadroxil (C16H17N3O5S)               =(WS/WT)×(AT/AS)×(1/C)×450
    =WS×(AT/AS)×(5/2)
                                                                   WS: Amount [mg (potency)] of Cefadroxil Reference
  WS: Amount [mg (potency)] of Cefadroxil Reference                    Standard
      Standard                                                     WT: Amount (g) of sample
                                                                   C: Labeled amount [mg (potency)] of cefadroxil in 1 g
Containers and storage    Containers—Tight containers.
                                                                 Assay Weigh accurately an amount of powdered
                                                                 Cefadroxil for Syrup, equivalent to about 50 mg (potency)
Add the following:                                               of Cefadroxil, dissolve in water to make exactly 500 mL, and
                                                                 use this solution as the sample solution. Separately, weigh
Cefadroxil for Syrup                                             accurately an amount of Cefadroxil Reference Standard,
                                                                 equivalent to about 20 mg (potency), dissolve in water to
シロップ用セファドロキシル                                                    make exactly 200 mL, and use this solution as the standard
                                                                 solution. Then, proceed as directed in the Assay under
  Cefadroxil for Syrup is a preparation for syrup                Cefadroxil.
which is suspended before use.
                                                                    Amount [mg (potency)] of cefadroxil (C16H17N3O5S)
  It contains not less than 95.0z and not more than
                                                                     =WS×(AT/AS)×(5/2)
110.0z of the labeled amount of cefadroxil
(C16H17N3O5S: 363.39).                                             WS: Amount [mg (potency)] of Cefadroxil Reference
                                                                       Standard
Method of preparation     Prepare as directed under Syrups,
with Cefadroxil.                                                 Containers and storage    Containers—Tight containers.
Identification Dissolve an amount of Cefadroxil for
Syrup, equivalent to 10 mg (potency) of Cefadroxil accord-
ing to the labeled amount, in 500 mL of water, and deter-
mine the absorption spectrum of the solution as directed un-
der Ultraviolet-visible Spectrophotometry <2.24>: it exhibits
maxima between 228 nm and 232 nm, and between 261 nm
and 265 nm.

Water <2.48> Not more than 3.0z (0.5 g, volumetric titra-
tion, direct titration).
1854       Official Monographs                                                                      Supplement I, JP XV

                                                                Purity (1) Clarity and color of solution—Conduct this
Cefalotin Sodium                                                procedure within 10 minutes after the preparation of the
                                                                solutions. A solution prepared by dissolving an amount of
セファロチンナトリウム                                                     Cefazolin Sodium for Injection, equivalent to 1.0 g (poten-
                                                                cy) of Cefazolin Sodium according to the labeled amount, in
Change the origin/limits of content to read:                    10 mL of water is clear, and the absorbance of this solution
                                                                at 400 nm, determined as directed under Ultraviolet-visible
  Cefalotin Sodium contains not less than 920 mg                Spectrophotometry <2.24>, is not more than 0.35.
(potency) and not more than 980 mg (potency) per mg,               (2) Related substances—Dissolve an amount of Cefazo-
calculated on the anhydrous basis. The potency of               lin Sodium for Injection, equivalent to 0.10 g (potency) of
Cefalotin Sodium is expressed as mass (potency) of              Cefazolin Sodium according to the labeled amount, in 20
cefalotin (C16H16N2O6S2: 396.44).                               mL of 0.1 mol/L phosphate buffer solution, pH 7.0, and
                                                                use this solution as the sample solution. Prepare the sample
                                                                solution before use. Perform the test with 5 mL of the sample
Cefatrizine Propylene Glycolate                                 solution as directed under Liquid Chromatography <2.01>
                                                                according to the following conditions, and determine each
セファトリジンプロピレングリコール                                               peak area by the automatic integration method. Calculate
                                                                the amount of each peak by the area percentage method:
Change the origin/limits of content to read:                    each area of the peaks other than cefazolin is not more than
                                                                1.5z. Furthermore the total area of the peaks other than
  Cefatrizine Propylene Glycolate contains not less             cefazolin is not more than 2.5z. For these calculations, use
than 816 mg (potency) and not more than 876 mg                  the area of the peak, having the relative retention time of
(potency) per mg, calculated on the anhydrous basis.            about 0.2 with respect to cefazolin, after multiplying by the
The potency of Cefatrizine Propylene Glycolate is               relative response factor, 1.43.
expressed as mass (potency) of cefatrizine                      Operating conditions—
(C18H18N6O5S2: 462.50).                                            Detector, column, column temperature, mobile phase,
                                                                and flow rate: Proceed as directed in the operating condi-
                                                                tions in the Assay under Cefazolin Sodium.
Add the following:                                                 Time span of measurement: About 3 times as long as the
                                                                retention time of cefazolin, beginning after the solvent peak.
Cefazolin Sodium for Injection                                  System suitability—
                                                                   Test for required detectability: Pipet 8 mL of the sample
注射用セファゾリンナトリウム                                                  solution, add 0.1 mol/L phosphate buffer solution, pH 7.0
                                                                to make exactly 50 mL, and use this solution as the solution
  Cefazolin Sodium for Injection is a preparation for           for system suitability test. Pipet 1 mL of the solution for sys-
injection which is dissolved before use.                        tem suitability test, add 0.1 mol/L phosphate buffer solu-
  It contains not less than 90.0z and not more                  tion, pH 7.0 to make exactly 20 mL. Confirm that the peak
than 110.0z of the labeled amount of cefazolin                  area of cefazolin obtained from 5 mL of this solution is
(C14H14N8O4S3: 454.51).                                         equivalent to 3 to 7z of that from 5 mL of the solution for
Method of preparation Prepare as directed under Injec-          system suitability test.
tions, with Cefazolin Sodium.                                      System performance: Proceed as directed in the system
                                                                suitability in the Assay under Cefazolin Sodium.
Description Cefazolin Sodium for Injection occurs as               System repeatability: When the test is repeated 6 times
white to light yellowish white crystals or crystalline powder   with 5 mL of the solution for system suitability test under the
or masses.                                                      above operating conditions, the relative standard deviation
Identification (1) Determine the absorption spectrum of         of the peak area of cefazolin is not more than 1.0z.
a solution of Cefazolin Sodium for Injection (1 in 50,000) as   Water <2.48> Not more than 3.0z (0.5 g, volumetric titra-
directed under Ultraviolet-visible Spectrophotometry <2.24>:    tion, direct titration). Use a mixture of formamide for Karl
it exhibits a maximum between 270 nm and 274 nm.                Fischer method and methanol for Karl Fischer method (2:1)
   (2) Cefazolin Sodium for Injection responds to the           instead of methanol for Karl Fischer method.
Qualitative Tests <1.09> (1) for chloride.
                                                                Bacterial endotoxins <4.01>    Less than 0.05 EU/mg (poten-
Osmotic pressure ratio   Being specified separately.            cy).
pH <2.54> The pH of a solution prepared by dissolving an        Uniformity of dosage units <6.02>     It meets the requirement
amount of Cefazolin Sodium for Injection, equivalent to 1.0     of the Mass variation test.
g (potency) of Cefazolin Sodium according to the labeled
amount, in 10 mL of water is 4.5 to 6.5.                        Foreign insoluble matter <6.06> Perform the test according
                                                                to Method 2: it meets the requirement.
Supplement I, JP XV                                                                        Official Monographs             1855

Insoluble particulate matter <6.07>     It meets the require-     pH <2.54> Take an amount of Cefmetazole Sodium for In-
ment.                                                             jection equivalent to 1.0 g (potency) of Cefmetazole Sodium
                                                                  according to the labeled amount, and dissolve in 10 mL of
Sterility <4.06> Perform the test according to the Mem-
                                                                  water: the pH of the solution is 4.2 to 6.2.
brane filtration method: it meets the requirement.
                                                                  Purity (1) Clarity and color of solution—Dissolve an
Assay Weigh accurately the mass of the contents of not
                                                                  amount of Cefmetazole Sodium for Injection, equivalent to
less than 10 containers of Cefazolin Sodium for Injection.
                                                                  1.0 g (potency) of Cefmetazole Sodium according to the
Weigh accurately an amount of the contents, equivalent to
                                                                  labeled amount, in 10 mL of water: the solution is clear and
about 50 mg (potency) of Cefazolin Sodium, dissolve in the
                                                                  the color is not darker than the following control solution.
internal standard solution to make exactly 50 mL, and use
                                                                     Control solution: Pipet 5 mL of Iron (III) Chloride
this solution as the sample solution. Separately, weigh ac-
                                                                  Colorimetric Stock Solution and 0.5 mL of Cobalt (II) Chlo-
curately an amount of Cefazolin Reference Standard,
                                                                  ride Colorimetric Stock Solution, and add water to make
equivalent to about 50 mg (potency), dissolve in the internal
                                                                  exactly 50 mL. Pipet 15 mL of this solution, and add water
standard solution to make exactly 50 mL, and use this solu-
                                                                  to make exactly 20 mL.
tion as the standard solution. Then, proceed as directed in
                                                                     (2) Related substances—Proceed as directed in the Puri-
the Assay under Cefazolin Sodium.
                                                                  ty (4) under Cefmetazole Sodium.
   Amount [mg (potency)] of cefazolin (C14H14N8O4S3)
                                                                  Bacterial endotoxins <4.01>    Less than 0.06 EU/mg (poten-
    = W S × ( Q T/ Q S )
                                                                  cy).
  WS: Amount [mg (potency)] of Cefazolin Reference Stan-
                                                                  Uniformity of dosage units <6.02>    It meets the requirement
      dard
                                                                  of the Mass variation test.
Internal standard solution—A solution of p-acetanisidide in
                                                                  Foreign particulate matter <6.06> Perform the test accord-
0.1 mol/L phosphate buffer solution, pH 7.0 (11 in 20,000).
                                                                  ing to Method 2: it meets the requirement.
Containers and storage Containers—hermetic containers.
                                                                  Insoluble particulate matter <6.07>     It meets the require-
Plastic containers for aqueous injections may be used.
                                                                  ment.

                                                                  Sterility <4.06> Perform the test according to the Mem-
Add the following:                                                brane filtration method: it meets the requirement.

                                                                  Assay Take 10 containers of Cefmetazole Sodium for In-
Cefmetazole Sodium for Injection                                  jection, dissolve the contents of each in the mobile phase,
                                                                  rinse each of the containers with the mobile phase, combine
注射用セフメタゾールナトリウム
                                                                  the rinse with the respective previous solution, and add the
                                                                  mobile phase to make exactly 500 mL. Take exactly a
  Cefmetazole Sodium for Injection is a preparation
                                                                  volume of this solution equivalent to about 0.2 g (potency)
for injection which is dissolved before use.
                                                                  of Cefmetazole Sodium, and add the mobile phase to make
  It contains not less than 90.0z and not more than
                                                                  exactly 100 mL. Pipet 1 mL of this solution, add exactly 10
110.0z of the labeled amount of cefmetazole
                                                                  mL of the internal standard solution, and use this solution as
(C15H17N7O5S3: 471.53).
                                                                  the sample solution. Separately, weigh accurately an amount
Method of preparation Prepare as directed under Injec-            of Cefmetazole Reference Standard, equivalent to about 50
tions, with Cefmetazole Sodium.                                   mg (potency), and dissolve in the mobile phase to make ex-
                                                                  actly 25 mL. Pipet 1 mL of this solution, add exactly 10 mL
Description Cefmetazole Sodium for Injection is a white to
                                                                  of the internal standard solution, and use this solution as the
light yellow powder or masses.
                                                                  standard solution. Then, proceed as directed in the Assay
   It is hygroscopic.
                                                                  under Cefmetazole Sodium.
Identification (1) Determine the absorption spectrum of
                                                                    Amount [mg (potency)] of cefmetazole (C15H17N7O5S3)
a solution of Cefmetazole Sodium for Injection (1 in 40,000)
                                                                     =WS×(QT/QS)×4
as directed under Ultraviolet-visible Spectrophotometry
<2.24>, and compare the spectrum with the Reference Spec-           WS: Amount [mg (potency)] of Cefmetazole Reference
trum: both spectra exhibit similar intensities of absorption at         Standard
the same wavelengths.
                                                                  Internal standard solution—A solution of methyl para-
   (2) Determine the infrared absorption spectrum of Cef-
                                                                  hydroxybenzoate in the mobile phase (1 in 10,000).
metazole Sodium for Injection as directed in the potassium
bromide disk method under Infrared Spectrophotometry              Containers and storage Containers—Hermetic containers.
<2.25>, and compare the spectrum with the Reference Spec-         Plastic containers for aqueous injections may be used.
trum: both spectra exhibit similar intensities of absorption at
the same wave numbers.
1856       Official Monographs                                                                     Supplement I, JP XV

Add the following:                                              buffer solution, pH 7.0, to make 50 mL, and use this solu-
                                                                tion as the sample solution. Separately, weigh accurately an
Ceftazidime for Injection                                       amount of Ceftazidime Reference Standard, equivalent to
                                                                about 25 mg (potency), and dissolve in 0.05 mol/L phos-
注射用セフタジジム                                                       phate buffer solution, pH 7.0, to make exactly 25 mL. Pipet
                                                                10 mL of this solution, add exactly 5 mL of the internal stan-
   Ceftazidime for Injection is a preparation for injec-        dard solution, then add 0.05 mol/L phosphate buffer solu-
tion which is dissolved before use.                             tion, pH 7.0, to make 50 mL, and use this solution as the
   It contains not less than 93.0z and not more than            standard solution. Then, proceed as directed in the Assay
107.0z of the labeled amount of ceftazidime                     under Ceftazidime Hydrate.
(C22H22N6O7S2: 546.58).
                                                                  Amount [mg (potency)] of ceftazidime (C22H22N6O7S2)
Method of preparation Prepare as directed under Injec-             =WS×(QT/QS)×10
tions, with Ceftazidime Hydrate.
                                                                  WS: Amount [mg(potency)] of Ceftazidime Reference
Description Ceftazidime for Injection is a white to pale              Standard
yellowish white powder.
                                                                Internal standard solution—A solution of dimedon in 0.05
Identification Determine the absorption spectrum of a so-       mol/L phosphate buffer solution, pH 7.0 (11 in 10,000).
lution of Ceftazidime for Injection (1 in 100,000) in phos-
                                                                Containers and storage Containers—Hermetic containers.
phate buffer solution, pH 6.0, as directed under Ultraviolet-
                                                                 Storage—Light-resistant.
visible Spectrophotometry <2.24>: it exhibits a maximum be-
tween 255 nm and 259 nm.

pH <2.54> Dissolve an amount of Ceftazidime for Injec-          Add the following:
tion, equivalent to 1.0 g (potency) of Ceftazidime Hydrate
according to the labeled amount, in 10 mL of water: the pH      Cetirizine Hydrochloride
of this solution is 5.8 to 7.8.
                                                                セチリジン塩酸塩
Purity Clarity and color of solution—Dissolve 5 g of diso-
dium hydrogen phosphate and 1 g of potassium dihydrogen
phosphate in water to make 100 mL. In 10 mL of this solu-
tion dissolve an amount of Ceftazidime for Injection,
equivalent to 1.0 g (potency) of Ceftazidime Hydrate ac-
cording to the labeled amount: the solution is clear and
colorless. Also, determine the absorption spectra of this so-   C21H25ClN2O3.2HCl: 461.81
lution as directed under Ultraviolet-visible Spectrophoto-      2-(2-{4-[(RS)-(4-Chlorophenyl)phenylmethyl]piperazin-
metry <2.24>: the absorbance at 420 nm is not more than 0.3.    1-yl}ethoxy)acetic acid dihydrochloride [83881-52-1]
Loss on drying <2.41> Not more than 14.0z (0.1 g, in
vacuum not exceeding 0.67 kPa, 609 3 hours).
                                 C,                               Cetirizine Hydrochloride, when dried, contains not
                                                                less than 99.0z and not more than 101.0z of
Bacterial endotoxins <4.01>   Less than 0.067 EU/mg (po-        C21H25ClN2O3.2HCl.
tency).
                                                                Description Cetirizine Hydrochloride occurs as a white
Uniformity of dosage units <6.02>   It meets the requirement    crystalline powder.
of the Mass variation test.                                       It is very soluble in water, and slightly soluble in ethanol
Foreign insoluble matter <6.06> Perform the test according      (99.5).
to Method 2: it meets the requirement.                            It dissolves in 0.1 mol/L hydrochloric acid TS.
                                                                  A solution of Cetirizine Hydrochloride (1 in 10) shows no
Insoluble particulate matter <6.07>   It meets the require-     optical rotation.
ment.
                                                                Identification (1) Determine the absorption spectrum of
Sterility <4.06> Perform the test according to the Mem-         a solution of Cetirizine Hydrochloride in 0.1 mol/L
brane filter method: it meets the requirement.                  hydrochloric acid TS (1 in 50,000) as directed under Ultrav-
Assay Weigh accurately the mass of the contents of not          iolet-visible Spectrophotometry <2.24>, and compare the
less than 10 containers of Ceftazidime for Injection. Weigh     spectrum with the Reference Spectrum: both spectra exhibit
accurately an amount of Ceftazidime Hydrate, equivalent to      similar intensities of absorption at the same wavelengths.
about 0.25 g (potency), and dissolve in 0.05 mol/L phos-           (2) Determine the infrared absorption spectrum of
phate buffer solution, pH 7.0, to make exactly 250 mL.          Cetirizine Hydrochloride as directed in the potassium chlo-
Pipet 10 mL of this solution, add exactly 5 mL of the inter-    ride disk method under Infrared Spectrophotometry <2.25>,
nal standard solution, add more 0.05 mol/L phosphate            and compare the spectrum with the Reference Spectrum:
Supplement I, JP XV                                                                        Official Monographs           1857

both spectra exhibit similar intensities of absorption at the           C,
                                                                  um, 609 3 hours).
same wave numbers.
                                                                  Residue on ignition <2.44>   Not more than 0.2z (1 g).
  (3) A solution of Cetirizine Hydrochloride (1 in 100)
responds to the Qualitative Tests <1.09> for chloride.            Assay Weigh accurately about 0.1 g of Cetirizine
                                                                  Hydrochloride, previously dried, dissolve in 70 mL of a
Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of
                                                                  mixture of acetone and water (7:3), and titrate <2.50> to the
Cetirizine Hydrochloride according to Method 2, and per-
                                                                  second equivalence point with 0.1 mol/L sodium hydroxide
form the test. Prepare the control solution with 2.0 mL of
                                                                  VS (potentiometric titration). Perform a blank determina-
Standard Lead Solution (not more than 10 ppm).
                                                                  tion in the same manner, and make any necessary correc-
   (2) Related substances—Dissolve 0.10 g of Cetirizine
                                                                  tion.
Hydrochloride in 50 mL of the mobile phase, and use this
solution as the sample solution. Pipet 2 mL of the sample so-            Each mL of 0.1 mol/L sodium hydroxide VS
lution, add the mobile phase to make exactly 50 mL. Pipet 5                =15.39 mg of C21H25ClN2O3.2HCl
mL of this solution, add the mobile phase to make exactly
                                                                  Containers and storage    Containers—Well-closed contain-
100 mL, and use this solution as the standard solution. Per-
                                                                  ers.
form the test with exactly 10 mL each of the sample solution
and standard solution as directed under Liquid Chro-
matography <2.01> according to the following conditions,
                                                                  Add the following:
and determine each peak area of each solution by the auto-
matic integration method: the area of each peak other than
cetirizine obtained from the sample solution is not larger        Cetirizine Hydrochloride Tablets
than the peak area of cetirizine from the standard solution.
                                                                  セチリジン塩酸塩錠
Furthermore, the total area of the peaks other than cetirizine
from the sample solution is not larger than 2.5 times the
                                                                    Cetirizine Hydrochloride Tablets contain not less
peak area of cetirizine from the standard solution.
                                                                  than 95.0z and not more than 105.0z of the labeled
Operating conditions—
                                                                  amount of cetirizine hydrochloride (C21H25ClN2O3.
   Detector: An ultraviolet absorption photometer (wave-
                                                                  2HCl: 461.81).
length: 230 nm).
   Column: A stainless steel column 4.0 mm in inside di-          Method of preparation Prepare as directed under Tablets,
ameter and 25 cm in length, packed with silica gel for liquid     with Cetirizine Hydrochloride.
chromatography (5 mm in particle diameter).
                                                                  Identification To a quantity of powdered Cetirizine
   Column temperature: A constant temperature of about
                                                                  Hydrochloride Tablets, equivalent to 10 mg of Cetirizine
259 C.
                                                                  Hydrochloride according to the labeled amount, add about
   Mobile phase: A mixture of acetonitrile and diluted 0.5
                                                                  70 mL of 0.1 mol/L hydrochloric acid TS, shake, add 0.1
mol/L sulfuric acid TS (2 in 25) (47:3).
                                                                  mol/L hydrochloric acid TS to make 100 mL, and filter. To
   Flow rate: Adjust the flow rate so that the retention time
                                                                  4 mL of the filtrate add 0.1 mol/L hydrochloric acid TS to
of cetirizine is about 9 minutes.
                                                                  make 25 mL, and determine the absorption spectrum of this
   Time span of measurement: About 3 times as long as the
                                                                  solution as directed under Ultraviolet-visible Spectrophoto-
retention time of cetirizine, beginning after the solvent peak.
                                                                  metry <2.24>: it exhibits a maximum between 230 nm and
System suitability—
                                                                  234 nm.
   Test for required detectability: Pipet 5 mL of the standard
solution, and add the mobile phase to make exactly 10 mL.         Uniformity of dosage units <6.02> Perform the test accord-
Confirm that the peak area of cetirizine obtained from 10 mL      ing to the following method: it meets the requirement of the
of this solution is equivalent to 35 to 65z of that from 10 mL    Content uniformity test.
of the standard solution.                                            Take 1 tablet of Cetirizine Hydrochloride Tablets, add
   System performance: Dissolve 20 mg of Cetirizine               4V/5 mL of sodium 1-heptanesulfonate solution (1 in 5000)
Hydrochloride in the mobile phase to make 100 mL. To 5            adjusted to pH 3.0 with 0.5 mol/L sulfuric acid TS, treat
mL of this solution, add 3 mL of a solution of aminopyrine        with ultrasonic waves for 20 minutes, adjust the volume to
in the mobile phase (1 in 2500), and add the mobile phase to      exactly V mL, by adding sodium 1-heptanesulfonate solu-
make 20 mL. When the procedure is run with 10 mL of this          tion (1 in 5000) adjusted to pH 3.0 with 0.5 mol/L sulfuric
solution under the above operating conditions, cetirizine and     acid TS, so that each mL contains about 0.2 mg of cetirizine
aminopyrine are eluted in this order with the resolution be-      hydrochloride (C21H25ClN2O3.2HCl), and filter through a
tween these peaks being not less than 7.                          membrane filter with a pore size not exceeding 0.45 mm. Dis-
   System repeatability: When the test is repeated 6 times        card the first 3 mL of the filtrate, pipet 5 mL of the subse-
with 10 mL of the standard solution under the above operat-       quent filtrate, add exactly 2 mL of the internal standard so-
ing conditions, the relative standard deviation of the peak       lution, add acetonitrile to make 10 mL, and use this solution
area of cetirizine is not more than 2.0z.                         as the sample solution. Then, proceed as directed in the As-
                                                                  say.
Loss on drying <2.41>    Not more than 0.5z (1 g, in vacu-
1858       Official Monographs                                                                          Supplement I, JP XV

                                                                   ditions, cetirizine and the internal standard are eluted in this
         Amount (mg) of cetirizine hydrochloride
                                                                   order with the resolution between these peaks being not less
         (C21H25ClN2O3.2HCl)
                                                                   than 7.
           =WS×(QT/QS)×(V/100)
                                                                     System repeatability: When the test is repeated 6 times
  WS: Amount (mg) of cetirizine hydrochloride for assay            with 20 mL of the standard solution under the above operat-
                                                                   ing conditions, the relative standard deviation of the ratio of
Internal standard solution—A solution of propyl para-
                                                                   the peak area of cetirizine to that of the internal standard is
hydroxybenzoate in the mobile phase (1 in 1000).
                                                                   not more than 1.0z.
Assay Weigh accurately not less than 20 Cetirizine
                                                                   Containers and storage      Containers—Well-closed contain-
Hydrochloride Tablets, and powder. Weigh accurately a
                                                                   ers.
portion of the powder, equivalent to about 10 mg of cetiri-
zine hydrochloride (C21H25ClN2O3.2HCl), add 40 mL of so-
dium 1-heptanesulfonate solution (1 in 5000) adjusted to pH
3.0 with 0.5 mol/L sulfuric acid TS, treat with ultrasonic         Chlordiazepoxide Tablets
waves for 20 minutes, add sodium 1-heptanesulfonate solu-
                                                                   クロルジアゼポキシド錠
tion (1 in 5000), adjusted to pH 3.0 with 0.5 mol/L sulfuric
acid TS, to make exactly 50 mL, and filter through a mem-
                                                                   Add the following next to Purity:
brane with a pore size not exceeding 0.45 mm. Discard the
first 3 mL of the filtrate, pipet 5 mL of the subsequent           Uniformity of dosage units <6.02> Perform the test accord-
filtrate, add exactly 2 mL of the internal standard solution,      ing to the following method: it meets the requirement of the
add acetonitrile to make exactly 10 mL, and use this solution      Content uniformity test.
as the sample solution. Separately, weigh accurately about            Conduct this procedures using light-resistant vessels. To 1
20 mg of cetirizine hydrochloride for assay, previously dried      tablet of Chlordiazepoxide Tablets add 1 mL of water, shake
in vacuum at 609 for 3 hours, and add sodium 1-hep-
                    C                                              to disintegrate the tablet, then add 20 mL of methanol,
tanesulfonate solution (1 in 5000), adjusted to pH 3.0 with        shake, add methanol to make exactly 25 mL, and filter
0.5 mol/L sulfuric acid TS, to make exactly 100 mL. Pipet 5        through a membrane filter with a pore size not exceeding 0.5
mL of this solution, add exactly 2 mL of the internal stan-        mm. Discard the first 5 mL of the filtrate, take exactly V mL
dard solution, add acetonitrile to make 10 mL, and use this        of the subsequent filtrate equivalent to about 2 mg of chlor-
solution as the standard solution. Perform the test with 20        diazepoxide (C16H14ClN3O), add exactly 1 mL of the internal
mL each of the sample solution and standard solution as            standard solution, then add methanol to make 20 mL, and
directed under Liquid Chromatography <2.01> according to           use this solution as the sample solution. Then, proceed as
the following conditions, and calculate the ratios, QT and         directed in the Assay.
QS, of the peak area of cetirizine to that of the internal stan-
                                                                        Amount (mg) of chlordiazepoxide (C16H14ClN3O)
dard.
                                                                         =WS×(QT/QS)×(5/V)
         Amount (mg) of cetirizine hydrochloride
                                                                     WS: Amount (mg) of Chlordiazepoxide Reference Stan-
         (C21H25ClN2O3.2HCl)
                                                                         dard
           =WS×(QT/QS)×(1/2)
                                                                   Internal standard solution—A solution of isobutyl salicylate
  WS: Amount (mg) of cetirizine hydrochloride for assay
                                                                   in methanol (1 in 20).
Internal standard solution—A solution of propyl para-
hydroxybenzoate in the mobile phase (1 in 1000).
Operating conditions—                                              Chlorphenesin Carbamate
  Detector: An ultraviolet absorption photometer (wave-
length: 230 nm).                                                   クロルフェネシンカルバミン酸エステル
  Column: A stainless steel column 4.0 mm in inside di-
ameter and 25 cm in length, packed with octylsilanized silica      Change the Description to read:
gel for liquid chromatography (5 mm in particle diameter).         Description Chlorphenesin Carbamate occurs as white
  Column temperature: A constant temperature of about              crystals or a crystalline powder.
259 C.                                                                It is freely soluble in methanol, in ethanol (95) and in pyri-
  Mobile phase: A mixture of a solution of sodium 1-hep-           dine, and slightly soluble in water.
tansulfonate (1 in 2900) and acetonitrile (29:21), adjusted to        A solution of Chlorphenesin Carbamate in ethanol (95)
pH 3.0 with 0.5 mol/L sulfuric acid TS.                            (1 in 20) shows no optical rotation.
  Flow rate: Adjust the flow rate so that the retention time
of cetirizine is about 5 minutes.                                  Change the Identification (2) to read:
System suitability—
  System performance: When the procedure is run with 20            Identification
mL of the standard solution under the above operating con-           (2) Determine the infrared absorption spectrum of
Supplement I, JP XV                                                                       Official Monographs            1859

Chlorphenesin Carbamate, as directed in the potassium             with 10 mL of the solution for system suitability test under
bromide disk method under Infrared Spectrophotometry              the above operating conditions, the relative standard devia-
<2.25>, and compare the spectrum with the Reference Spec-         tion of the peak areas of chlorphenesin carbamate is not
trum: both spectra exhibit similar intensities of absorption at   more than 2.0z.
the same wave numbers.
                                                                  Add the following next to Purity (3) :
Change the Purity (3) to read:
                                                                     (4) Related substances—Dissolve 0.10 g of Chlorphene-
Purity                                                            sin Carbamate in 10 mL of ethanol (95), and use this solu-
   (3) Chlorphenesin-2-carbamate—Dissolve 0.10 g of               tion as the sample solution. Pipet 1 mL of the sample solu-
Chlorphenesin Carbamate in 20 mL of a mixture of hexane           tion, add ethanol (95) to make exactly 20 mL. Pipet 2 mL of
for liquid chromatography and 2-propanol (7:3), and use           this solution, add ethanol (95) to make exactly 20 mL, and
this solution as the sample solution. Perform the test with 10    use this solution as the standard solution. Perform the test
mL of the sample solution as directed under Liquid Chro-          with these solutions as directed under Thin-layer Chro-
matography <2.01> according to the following conditions.          matography <2.03>. Spot 50 mL each of the sample solution
Determine the peak area, Aa, of chlorphenesin carbamate           and standard solution on a plate of silica gel for thin-layer
and the peak area, Ab, of chlorphenesin-2-carbamate by the        chromatography. Develop the plate with a mixture of ethyl
automatic integration method: the ratio, Ab/(Aa+Ab), is           acetate, methanol and ammonia solution (28) (17:2:1) to a
not more than 0.007.                                              distance of about 10 cm, and air-dry the plate. Allow the
Operating conditions—                                             plate to stand in iodine vapor for 20 minutes: the spot other
   Detector: An ultraviolet absorption photometer (wave-          than the principal spot from the sample solution is not more
length: 280 nm).                                                  than one, and it is not more intense than the spot from the
   Column: A stainless steel column 4 mm in inside diameter       standard solution.
and 30 cm in length, packed with silica gel for liquid chro-
matography (5 mm in particle diameter).
   Column temperature: A constant temperature of about            Add the following:
409 C.
   Mobile phase: A mixture of hexane for liquid chro-             Chlorphenesin Carbamate Tablets
matography, 2-propanol and acetic acid (100) (700:300:1).
   Flow rate: Adjust the flow rate so that the retention time     クロルフェネシンカルバミン酸エステル錠
of chlorphenesin carbamate is about 9 minutes.
System suitability—                                                 Chlorphenesin Carbamate Tablets contain not less
   Test for required detection: Pipet 1 mL of the sample          than 93.0z and not more than 107.0z of the labeled
solution, add a mixture of hexane for liquid chro-                amount of chlorphenesin carbamate (C10H12ClNO4:
matography and 2-propanol (7:3) to make exactly 100 mL,           245.66).
and use this solution as the solution for system suitability
                                                                  Method of preparation Prepare as directed under Tablets,
test. To exactly 5 mL of the solution for system suitability
                                                                  with Chlorphenesin Carbamate.
test add the mixture of hexane for liquid chromatography
and 2-propanol (7:3) to make exactly 10 mL. Confirm that          Identification To a quantity of powdered Chlorphenesin
the peak area of chlorphenesin carbamate obtained from 10         Carbamate Tablets, equivalent to 0.15 g of Chlorphenesin
mL of this solution is equivalent to 40 to 60z of that of         Carbamate according to the labeled amount, add 60 mL of
chlorphenesin carbamate obtained from 10 mL of the solu-          ethanol (95), treat with ultrasonic waves, and add ethanol
tion for system suitability test.                                 (95) to make 100 mL. Centrifuge 20 mL of this solution, add
   System performance: Dissolve 0.1 g of Chlorphenesin            ethanol (95) to 1 mL of the supernatant liquid to make 100
Carbamate in 50 mL of methanol. To 25 mL of this solution         mL, and determine the absorption spectrum of this solution
add 25 mL of dilute sodium hydroxide TS, and warm at              as directed under Ultraviolet-visible Spectrophotometry
609 for 20 minutes. To 20 mL of this solution add 5 mL of
    C                                                             <2.24>: it exhibits maxima between 226 nm and 230 nm, be-
1 mol/L hydrochloric acid TS, shake well with 20 mL of            tween 279 nm and 283 nm, and between 286 nm and 290 nm.
ethyl acetate, and allow to stand to separate the upper layer.
                                                                  Uniformity of dosage units <6.02> Perform the test accord-
When the procedure is run with 10 mL of this layer under the
                                                                  ing to the following method: it meets the requirement of the
above operating conditions, chlorphenesin, chlorphenesin
                                                                  Content uniformity test.
carbamate and chlorphenesin-2-carbamate are eluted in this
                                                                     To 1 tablet of Chlorphenesin Carbamate Tablets add 10
order, with the relative retention times of chlorphenesin and
                                                                  mL of water to disintegrate the tablet, add 70 mL of a mix-
chlorphenesin-2-carbamate with respect to chlorphenesin
                                                                  ture of water and methanol (1:1), treat with ultrasonic waves
carbamate being about 0.7 and about 1.2, respectively, and
                                                                  for 15 minutes with occasional stirring, then add the mixture
with the resolution between the peaks of chlorphenesin and
                                                                  of water and methanol (1:1) to make exactly 100 mL. Cen-
chlorphenesin carbamate being not less than 2.0.
                                                                  trifuge this solution, pipet V mL of the supernatant liquid
   System repeatability: When the test is repeated 6 times
                                                                  equivalent to about 2.5 mg of chlorphenesin carbamate
1860       Official Monographs                                                                        Supplement I, JP XV

(C10H12ClNO4), add the mixture of water and methanol (1:1)         ly dried in a desiccator (in vacuum, silica gel) for 4 hours,
to make exactly 25 mL, and use this solution as the sample         and dissolve in ethyl acetate to make exactly 50 mL. Pipet 5
solution. Separately, weigh accurately about 50 mg of chlor-       mL of this solution, add exactly 2 mL of the internal stan-
phenesin carbamate for assay, previously dried in a desicca-       dard solution, then add ethyl acetate to make 20 mL, and
tor (in vacuum, silica gel) for 4 hours, and dissolve in the       use this solution as the standard solution. Perform the test
mixture of water and methanol (1:1) to make exactly 50 mL.         with 10 mL each of the sample solution and standard solu-
Pipet 2 mL of this solution, add the mixture of water and          tion as directed under Liquid Chromatography <2.01> ac-
methanol (1:1) to make exactly 20 mL, and use this solution        cording to the following conditions, and calculate the ratios,
as the standard solution. Determine the absorbances at 280         QT and QS, of the peak area of chlorphenesin carbamate to
nm, AT and AS, of the sample solution and standard solution        that of the internal standard.
as directed under Ultraviolet-visible Spectrophotometry
                                                                   Amount (mg) of chlorphenesin carbamate (C10H12ClNO4)
<2.24>.
                                                                    =WS×(QT/QS)×(5/2)
         Amount (mg) of chlorphenesin carbamate
                                                                     WS: Amount (mg) of chlorphenesin carbamate for assay
         (C10H12ClNO4)
           =WS×(AT/AS)×(1/V)×5                                     Internal standard solution—A solution of ethenzamide in
                                                                   ethyl acetate (1 in 400).
  WS: Amount (mg) of chlorphenesin carbamate for assay
                                                                   Operating conditions—
Dissolution <6.10> When the test is performed at 50 revolu-           Detector: An ultraviolet absorption photometer (wave-
tions per minute according to the Paddle method, using 900         length: 280 nm).
mL of water as the dissolution medium, the dissolution rate           Column: A stainless steel column 4 mm in inside diameter
in 15 minutes of Chlorphenesin Carbamate Tablets is not            and 30 cm in length, packed with silica gel for liquid chro-
less than 85z.                                                     matography (5 mm in particle diameter).
   Start the test with 1 tablet of Chlorphenesin Carbamate            Column temperature: A constant temperature of about
Tablets, withdraw not less than 20 mL of the medium at the         409 C.
specified minute after starting the test, and filter through a        Mobile phase: A mixture of hexane for liquid chro-
membrane filter with a pore size not exceeding 0.45 mm. Dis-       matography, 2-propanol and acetic acid (100) (700:300:1).
card the first 10 mL of the filtrate, pipet V mL of the subse-        Flow rate: Adjust the flow rate so that the retention time
quent filtrate, add water to make exactly V? mL so that each       of chlorphenesin carbamate is about 9 minutes.
mL contains about 0.14 mg of chlorphenesin carbamate               System suitability—
(C10H12ClNO4) according to the labeled amount, and use this           System performance: Proceed as directed in the Purity (3)
solution as the sample solution. Separately, weigh accurately      under Chlorphenesin Carbamate.
about 28 mg of chlorphenesin carbamate for assay, previ-              System repeatability: When the test is repeated 6 times
ously dried in a desiccator (in vacuum, silica gel) for 4 hours,   with 10 mL of the standard solution under the above operat-
dissolve in 1 mL of methanol, and add water to make exactly        ing conditions, the relative standard deviation of the ratio of
50 mL. Pipet 5 mL of this solution, add water to make ex-          the peak area of chlorphenesin carbamate to that of the in-
actly 20 mL, and use this solution as the standard solution.       ternal standard is not more than 1.5z.
Determine the absorbances, AT and AS, at 278 nm of the
                                                                   Containers and storage     Containers—Well-closed contain-
sample solution and standard solution as directed under
                                                                   ers.
Ultraviolet-visible Spectrophotometry <2.24>.

Dissolution rate (z) with respect to the labeled amount of
chlorphenesin carbamate (C10H12ClNO4)                              Chlorpromazine Hydrochloride
  =WS×(AT/AS)×(V?/V)×(1/C)×450
                                                                   Injection
  WS: Amount (mg) of chlorphenesin carbamate for assay
  C: Labeled amount (mg) of chlorphenesin carbamate                クロルプロマジン塩酸塩注射液
     (C10H12ClNO4) in 1 tablet
                                                                   Add the following next to Extractable volume:
Assay Weigh accurately the mass of not less than 20 Chlor-
phenesin Carbamate Tablets, and powder them in an agate            Foreign insoluble matter <6.06> Perform the test according
mortar. Weigh accurately a portion of the powder, equiva-          to Method 1: it meets the requirement.
lent to about 0.25 g of chlorphenesin carbamate                    Insoluble particulate matter <6.07>     It meets the require-
(C10H12ClNO4), add 30 mL of ethyl acetate, disperse using          ment.
ultrasonic waves, then add ethyl acetate to make exactly 50
mL. Centrifuge 20 mL of this solution, pipet 2 mL of the su-       Sterility <4.06> Perform the test according to the Mem-
pernatant liquid, add exactly 2 mL of the internal standard        brane filtration method: it meets the requirement.
solution, add ethyl acetate to make 20 mL, and use this solu-
tion as the sample solution. Separately, weigh accurately
about 0.1 g of chlorphenesin carbamate for assay, previous-
Supplement I, JP XV                                                                      Official Monographs            1861

                                                                     Amount (mg) of chlorpropamide (C10H13ClN2O3S)
Chlorpromazine Hydrochloride                                          =WS×(AT/AS)×(V/20)
Tablets                                                            WS: Amount (mg) of chlorpropamide for assay
クロルプロマジン塩酸塩錠

Add the following next to Identification:                        Add the following:

Uniformity of dosage units <6.02> Perform the test accord-       Cibenzoline Succinate
ing to the following method: it meets the requirement of the
Content uniformity test.                                         シベンゾリンコハク酸塩
   Conduct this procedures using light-resistant vessels. To 1
tablet of Chlorpromazine Hydrochloride Tablets add an
amount of a mixture of diluted phosphoric acid (1 in 500)
and ethanol (99.5) (1:1) so that each mL contains about 0.83
mg of chlorpromazine hydrochloride (C17H19ClN2S.HCl),
treat with the ultrasonic waves for 5 minutes, then shake
vigorously for 20 minutes, and add the mixture of diluted        C18H18N2.C4H6O4: 380.44
phosphoric acid (1 in 500) and ethanol (99.5) (1:1) to make      2-[(1RS)-2,2-Diphenylcyclopropan-1-yl]-4,5-dihydro-1H-
exactly V mL so that each mL contains about 0.5 mg of            imidazole monosuccinate [100678-32-8]
chlorpromazine hydrochloride (C17H19ClN2S.HCl). Filter
through a membrane filter with a pore size not exceeding            Cibenzoline Succinate, when dried, contains not less
0.45 mm. Discard the first 3 mL of the filtrate, pipet 2.5 mL    than 98.5z and not more than 101.0z of C18H18N2.
of the subsequent filtrate, add exactly 5 mL of the internal     C 4 H 6 O4 .
standard solution, then add the mixture of diluted phos-
                                                                 Description Cibenzoline Succinate occurs as a white crys-
phoric acid (1 in 500) and ethanol (99.5) (1:1) to make 25
                                                                 talline powder.
mL, and use this solution as the sample solution. Then, pro-
                                                                    It is freely soluble in methanol and in acetic acid (100),
ceed as directed in the Assay.
                                                                 and sparingly soluble in water and in ethanol (99.5).
      Amount (mg) of chlorpromazine hydrochloride                   A solution of Cibenzoline Succinate in methanol (1 in 10)
      (C17H19ClN2S.HCl)                                          shows no optical rotation.
        =WS×(QT/QS)×(V/50)
                                                                 Identification (1) Determine the absorption spectrum of
  WS: Amount (mg) of chlorpromazine hydrochloride for            a solution of Cibenzoline Succinate (1 in 50,000) as directed
      assay                                                      under Ultraviolet-visible Spectrophotometry <2.24>, and
                                                                 compare the spectrum with the Reference Spectrum: both
Internal standard solution—A solution of ethyl parahydrox-
                                                                 spectra exhibit similar intensities of absorption at the same
ybenzoate in the mixture of diluted phosphoric acid (1 in
                                                                 wavelengths.
500) and ethanol (99.5) (1:1) (1 in 4500).
                                                                    (2) Determine the infrared absorption spectrum of
                                                                 Cibenzoline Succinate as directed in the paste method under
                                                                 Infrared Spectrophotometry <2.25>, and compare the spec-
Chlorpropamide Tablets                                           trum with the Reference Spectrum: both spectra exhibit
                                                                 similar intensities of absorption at the same wave numbers.
クロルプロパミド錠
                                                                    (3) Shake 0.4 g of Cibenzoline Succinate with 2.5 mL of
                                                                 sodium hydroxide TS and 5 mL of ethyl acetate, allow to
Add the following next to Identification:
                                                                 stand, and to 1 mL of the water layer so obtained add 0.5
Uniformity of dosage units <6.02> Perform the test accord-       mL of 1 mol/L hydrochloric acid TS and 0.5 mL of iron
ing to the following method: it meets the requirement of the     (III) chloride TS: a blown precipitate is formed.
Content uniformity test.
                                                                 Melting point <2.60>   163 – 1679
                                                                                                 C
  To 1 tablet of Chlorpropamide Tablets add 75 mL of the
mobile phase, treat with the ultrasonic waves for 20 minutes     pH <2.54> Dissolve 0.20 g of Cibenzoline Succinate in 10
with occasional strong shaking, then add the mobile phase to     mL of water: the pH of this solution is between 4.0 and 6.0.
make exactly V mL so that each mL contains about 2.5 mg
                                                                 Purity (1) Clarity and color of solution—A solution ob-
of Chlorpropamide. Centrifuge the solution, pipet 2 mL of
                                                                 tained by dissolving 0.20 g of Cibenzoline Succinate in 10
the supernatant liquid, add the mobile phase to make exactly
                                                                 mL of water is clear and colorless.
100 mL, and use this solution as the sample solution. Then,
                                                                    (2) Heavy metals <1.07>—Proceed with 1.0 g of Ciben-
proceed as directed in the Assay.
                                                                 zoline Succinate according to Method 2, and perform the
                                                                 test. Prepare the control solution with 2.0 mL of Standard
1862       Official Monographs                                                                       Supplement I, JP XV

Lead Solution (not more than 20 ppm).                             Identification To a quantity of powdered Cibenzoline Suc-
   (3) Arsenic <1.11>—Prepare the test solution with 1.0 g        cinate Tablets, equivalent to 50 mg of Cibenzoline Succinate
of Cibenzoline Succinate according to Method 3, using a           according to the labeled amount, add 100 mL of water,
solution of magnesium nitrate hexahydrate in ethanol (95)         shake for 10 minutes, and centrifuge. To 2 mL of the super-
(1 in 25), and perform the test (not more than 2 ppm).            natant liquid add water to make 50 mL, and determine the
   (4) Related substances—Dissolve 0.10 g of Cibenzoline          absorption spectrum of this solution as directed under
Succinate in 10 mL of methanol, and use this solution as the      Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a
sample solution. Pipet 1 mL of the sample solution, and add       maximum between 221 nm and 225 nm.
methanol to make exactly 100 mL. Pipet 5 mL and 2 mL of
                                                                  Uniformity of dosage units <6.02> Perform the test accord-
this solution, add methanol to make them exactly 10 mL,
                                                                  ing to the following method: it meets the requirement of the
and use these solutions as the standard solution (1) and the
                                                                  Content uniformity test.
standard solution (2). Perform the test with these solutions
                                                                     To 1 tablet of Cibenzoline Succinate Tablets add a suita-
as directed under Thin-layer Chromatography <2.03>. Spot
                                                                  ble amount of water so that each mL contains about 10 mg
10 mL each of the sample solution and standard solutions (1)
                                                                  of cibenzoline succinate (C18H18N2.C4H6O4), and allow
and (2) on a plate of silica gel with fluorescent indicator for
                                                                  standing for 10 minutes while occasional shaking. To this so-
thin-layer chromatography. Develop the plate with a mix-
                                                                  lution add methanol so that each mL contains about 2 mg of
ture of ethyl acetate, methanol and ammonia solution (28)
                                                                  cibenzoline succinate (C18H18N2.C4H6O4), add exactly 1 mL
(20:3:2) to a distance of about 10 cm, air-dry the plate, and
                                                                  of the internal standard solution per 10 mg of cibenzoline
dry more at 809 for 30 minutes. After cooling, examine un-
                 C
                                                                  succinate (C18H18N2.C4H6O4), then add methanol so that
der ultraviolet light (main wavelength: 254 nm): the spot
                                                                  each mL contains about 1 mg of cibenzoline succinate
other than the principal spot obtained with the sample solu-
                                                                  (C18H18N2.C4H6O4). Centrifuge the solution, and use the su-
tion is not more intense than the spot with the standard solu-
                                                                  pernatant liquid as the sample solution. Then, proceed as
tion (1). Allow the plate to stand for 30 minutes in iodine
                                                                  directed in the Assay.
vapor: the spot other than the principal spot obtained with
the sample solution is not more intense than the spot with        Amount (mg) of cibenzoline succinate (C18H18N2.C4H6O4)
the standard solution (1), and the spot, which is more intense     =WS×(QT/QS)×(C/100)
than the spot with the standard solution (2), is not more than
                                                                    WS: Amount (mg) of cibenzoline succinate for assay
two.
                                                                    C: Labeled amount (mg) of cibenzoline succinate
Loss on drying <2.41>    Not more than 0.3z (1 g, 1059 2
                                                      C,          (C18H18N2.C4H6O4) in 1 tablet
hours).
                                                                  Internal standard solution—Dissolve 0.1 g of 2-ethylhexyl
Residue on ignition <2.44>    Not more than 0.1z (1 g).           parahydroxybenzoate in methanol to make 100 mL.

Assay Weigh accurately about 0.4 g of Cibenzoline Suc-            Dissolution <6.10> When the test is performed at 50 revolu-
cinate, previously dried, dissolve in 50 mL of acetic acid        tions per minute according to the Paddle method, using 900
(100), and titrate <2.50> with 0.1 mol/L perchloric acid VS       mL of water as the dissolution medium, the dissolution rate
until the color of the solution changes from violet to blue-      in 15 minutes of Cibenzoline Succinate Tablets is not less
green through blue (indicator: 2 drops of crystal violet TS).     than 80z.
Perform a blank determination in the same manner, and                Start the test with 1 tablet of Cibenzoline Succinate
make any necessary correction.                                    Tablets, withdraw not less than 20 mL of the medium at the
                                                                  specified minute after starting the test, and filter through a
         Each mL of 0.1 mol/L perchloric acid VS
                                                                  membrane filter with a pore size not exceeding 0.45 mm. Dis-
           =38.04 mg of C18H18N2.C4H6O4
                                                                  card the first 10 mL of the filtrate, pipet V mL of the subse-
Containers and storage     Containers—Tight containers.           quent filtrate, add water to make exactly V? mL so that each
                                                                  mL contains about 11 mg of cibenzoline succinate (C18H18N2.
                                                                  C4H6O4) according to the labeled amount, and use this solu-
Add the following:                                                tion as the sample solution. Separately, weigh accurately
                                                                  about 28 mg of cibenzoline succinate for assay, previously
Cibenzoline Succinate Tablets                                     dried at 1059 for 2 hours, and dissolve in water to make
                                                                                 C
                                                                  exactly 100 mL. Pipet 2 mL of this solution, add water to
シベンゾリンコハク酸塩錠                                                      make exactly 50 mL, and use this solution as the standard
                                                                  solution. Determine the absorbances, AT and AS, of the
  Cibenzoline Succinate Tablets contain not less than             sample solution and standard solution at 222 nm as directed
95.0z and not more than 105.0z of the labeled                     under Ultraviolet-visible Spectrophotometry <2.24>.
amount of cibenzoline succinate (C18H18N2.C4H6O4:
                                                                  Dissolution rate (z) with respect to the labeled amount of
380.44).
                                                                  cibenzoline succinate (C18H18N2.C4H6O4)
Method of preparation Prepare as directed under Tablets,             =WS×(AT/AS)×(V?/V)×(1/C)×36
with Cibenzoline Succinate.
Supplement I, JP XV                                                                        Official Monographs             1863

  WS: Amount (mg) of cibenzoline succinate for assay              Add the following:
  C: Labeled amount (mg) of cibenzoline succinate
     (C18H18N2.C4H6O4) in 1 tablet                                Cilazapril Hydrate
Assay Weigh accurately not less than 20 Cibenzoline Suc-
                                                                  シラザプリル水和物
cinate Tablets, and powder. Weigh accurately a portion of
the powder, equivalent to about 0.1 g of cibenzoline suc-
cinate (C18H18N2.C4H6O4), add 10 mL of water, shake, and
add 40 mL of methanol and exactly 10 mL of the internal
standard solution. Shake for 20 minutes, add methanol to
make 100 mL, centrifuge, and use the supernatant liquid as
                                                                  C22H31N3O5.H2O: 435.51
the sample solution. Separately, weigh accurately about
                                                                  (1S,9S)-9-[(1S)-(1-Ethoxycarbonyl-
0.1 g of cibenzoline succinate for assay, previously dried at
                                                                  3-phenylpropyl)amino]-10-oxooctahydro-
1059 for 2 hours, add 10 mL of water and 40 mL of
     C
                                                                  6H-pyridazino[1,2-a][1,2]diazepine-1-carboxylic acid
methanol to dissolve, then add exactly 10 mL of the internal
                                                                  monohydrate [92077-78-6]
standard solution and methanol to make 100 mL, and use
this solution as the standard solution. Perform the test with
                                                                    Cilazapril Hydrate contains not less than 98.5z and
5 mL each of the sample solution and standard solution as
                                                                  not more than 101.0z of cilazapril (C22H31N3O5:
directed under Liquid Chromatography <2.01> according to
                                                                  417.50), calculated on the anhydrous basis.
the following conditions, and calculate the ratios, QT and
QS, of the peak area of cibenzoline to that of the internal       Description Cilazapril Hydrate occurs as white to yellow-
standard.                                                         ish white crystals or crystalline powder.
                                                                     It is very soluble in methanol, freely soluble in ethanol
Amount (mg) of cibenzoline succinate (C18H18N2.C4H6O4)
                                                                  (99.5) and in acetic acid (100), and slightly soluble in water.
 = W S ×( Q T / Q S )
                                                                     It gradually turns yellow on exposure to light.
  WS: Amount (mg) of cibenzoline succinate for assay                 Melting point: about 1019 (with decomposition).
                                                                                                 C

Internal standard solution—Dissolve 0.1 g of 2-ethylhexyl         Identification (1) To 4 mL of a solution of Cilazapril
parahydroxybenzoate in methanol to make 100 mL.                   Hydrate (1 in 1000) add 2 mL of Dragendorff's TS: an
Operating conditions—                                             orange precipitate is produced.
   Detector: An ultraviolet absorption photometer (wave-             (2) Determine the infrared absorption spectrum of
length: 254 nm).                                                  Cilazapril Hydrate as directed in the potassium bromide disk
   Column: A stainless steel column 4.6 mm in inside di-          method under Infrared Spectrophotometry <2.25>, and com-
ameter and 5 cm in length, packed with octadecylsilanized         pare the spectrum with the Reference Spectrum: both spec-
silica gel for liquid chromatography (3 mm in particle di-        tra exhibit similar intensities of absorption at the same wave
ameter).                                                          numbers.
   Column temperature: A constant temperature of about
                                                                  Optical rotation <2.49> [a]20: -53 – -589
                                                                                             D             (0.2 g calculated
259  C.
                                                                  on the anhydrous basis, methanol, 20 mL, 100 mm).
   Mobile phase: Dissolve 2.67 g of sodium di-2-ethylhexyl
sulfosuccinate in 2000 mL of a mixture of water, acetonitrile     Purity (1) Chloride <1.03>—Perform the test using 1.0 g
and diluted phosphoric acid (1 in 10) (1000:1000:1).              of Cilazapril Hydrate. Prepare the control solution with 0.25
   Flow rate: Adjust the flow rate so that the retention time     mL of 0.01 mol/L hydrochloric acid VS (not more than
of cibenzoline is about 3 minutes.                                0.009z).
System suitability—                                                  (2) Sulfate <1.14>—Dissolve 1.0 g of Cilazapril Hydrate
   System performance: When the procedure is run with 5 mL        in 40 mL of water and 1.5 mL of dilute hydrochloric acid,
of the standard solution under the above operating condi-         and add water to make 50 mL. Perform the test using this
tions, cibenzoline and the internal standard are eluted in this   solution as the test solution. Prepare the control solution
order with the resolution between these peaks being not less      with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more
than 6.                                                           than 0.019z).
   System repeatability: When the test is repeated 6 times           (3) Heavy metals <1.07>—Proceed with 1.0 g of
with 5 mL of the standard solution under the above operat-        Cilazapril Hydrate according to Method 4, and perform the
ing conditions, the relative standard deviation of the ratio of   test. However, use 10 mL of a solution of magnesium nitrate
the peak area of cibenzoline to that of the internal standard     hexahydrate in ethanol (95) (1 in 8). Prepare the control so-
is not more than 1.0z.                                            lution with 2.0 mL of Standard Lead Solution (not more
                                                                  than 20 ppm).
Containers and storage     Containers—Tight containers.
                                                                     (4) Related substances—Dissolve 0.10 g of Cilazapril
                                                                  Hydrate in 20 mL of methanol, and use this solution as the
                                                                  sample solution. Pipet 1 mL of the sample solution, add
                                                                  methanol to make exactly 100 mL, and use this solution as
1864       Official Monographs                                                                        Supplement I, JP XV

the standard solution (1). Pipet 3 mL of the standard solu-       mg of cilazapril in 5 mL of the mixture of acetonitrile and
tion (1), add methanol to make exactly 10 mL, and use this        ethyl acetate (3:1), and use this solution as the standard solu-
solution as the standard solution (2). Separately, pipet 2 mL     tion. Perform the test with these solutions as directed under
of the standard solution (1), add methanol to make exactly        Thin-layer Chromatography <2.03>. Spot 20 mL each of the
10 mL, and use this solution as the standard solution (3).        sample solution and standard solution on a plate of silica gel
Perform the test with these solutions as directed under Thin-     with fluorescent indicator for thin-layer chromatography.
layer Chromatography <2.03>. Spot 20 mL each of the sam-          Develop the plate with a mixture of ethyl acetate, methanol,
ple solution and three standard solutions on a plate of silica    acetic acid (100), hexane and water (62:15:10:10:3) to a dis-
gel with fluorescent indicator for thin-layer chro-               tance of about 15 cm, and air-dry the plate. Place the plate
matography. Develop the plate with a mixture of ethyl             in iodine vapor for 2 hours, and immediately examine under
acetate, methanol, acetic acid (100), hexane, and water           ultraviolet light (main wavelength: 254 nm): the spots ob-
(62:15:10:10:3) to a distance of about 15 cm, and air-dry the     tained from the sample and standard solutions are dark
plate. Leave the plate in iodine vapor for 2 hours, and exa-      brown and they show the same Rf value.
mine the plate under ultraviolet light (main wavelength: 254
                                                                  Uniformity of dosage units <6.02> Perform the test accord-
nm): of the spots other than the principal spot with an Rf
                                                                  ing to the following method: it meets the requirement of the
value close to 0.40 obtained from the sample solution, the
                                                                  Content uniformity test.
spot in the vicinity of Rf value 0.17 is not more intense than
                                                                     To 1 tablet of Cilazapril Tablets add 5 mL of a mixture of
the spot from the standard solution (1), and the spot in the
                                                                  water and acetonitrile (7:3), shake well until disintegration,
vicinity of Rf value 0.44 is not more intense than the spot
                                                                  add the mixture of water and acetonitrile (7:3) to make
from the standard solution (2). The number of all other spot
                                                                  exactly V mL so that each mL contains about 25 mg of
does not exceed 3, and of these spots, no more than one is
                                                                  cilazapril (C22H31N3O5), and centrifuge. Pipet 4 mL of the
more intense than the spot from the standard solution (3)
                                                                  supernatant liquid, add exactly 1 mL of the internal stan-
and none are more intense than the spot from the standard
                                                                  dard solution, add the mixture of water and acetonitrile (7:3)
solution (2).
                                                                  to make 10 mL, and use this solution as the sample solution.
Water <2.48>   3.5 – 5.0z (0.3 g, volumetric titration, direct    Separately, weigh accurately about 26 mg of cilazapril for
titration).                                                       assay (separately determine the water <2.48> in the same
                                                                  manner as Cilazapril Hydrate), and dissolve in the mixture
Residue on ignition <2.44>    Not more than 0.1z (0.5 g).
                                                                  of water and acetonitrile (7:3) to make exactly 50 mL. Pipet
Assay Weigh accurately about 0.2 g of Cilazapril Hydrate,         2 mL of this solution, add exactly 10 mL of the internal stan-
dissolve in 50 mL of acetic acid (100), and titrate <2.50> with   dard solution, add the mixture of water and acetonitrile (7:3)
0.02 mol/L perchloric acid VS (potentiometric titration).         to make 100 mL, and use this solution as the standard solu-
Perform a blank determination in the same manner and              tion. Perform the test with 100 mL each of the sample solu-
make any necessary correction.                                    tion and standard solution as directed under Liquid Chro-
                                                                  matography <2.01> according to the following conditions,
        Each mL of 0.02 mol/L perchloric acid VS
                                                                  and calculate the ratios, QT and QS, of the peak area of
          =8.350 mg of C22H31N3O5
                                                                  cilazapril to that of the internal standard.
Containers and storage Containers—Tight containers.
                                                                            Amount (mg) of cilazapril (C22H31N3O5)
 Storage—Light-resistant.
                                                                             =WS×(QT/QS)×(V/1000)

                                                                    WS: Amount (mg) of cilazapril for assay, calculated on
Add the following:
                                                                        the anhydrous basis

Cilazapril Tablets                                                Internal standard solution—A solution of dimethyl phtha-
                                                                  late in a mixture of water and acetonitrile (7:3) (1 in 12,500).
シラザプリル錠                                                           Operating conditions—
                                                                     Proceed as directed in the operating conditions in the
   Cilazapril Tablets contain not less than 93.0z and             Assay.
not more than 107.0z of the labeled amount of                     System suitability—
cilazapril (C22H31N3O5: 417.50).                                     System performance: When the procedure is run with 100
                                                                  mL of the standard solution under the above conditions,
Method of preparation Prepare as directed under Tablets,
                                                                  cilazapril and the internal standard are eluted in this order
with Cilazapril Hydrate.
                                                                  with the resolution between these peaks being not less than
Identification To a quantity of powdered Cilazapril               6.
Tablets, equivalent to 2 mg of cilazapril (C22H31N3O5) ac-           System repeatability: When the test is repeated 6 times
cording to the labeled amount, add 2 mL of a mixture of           with 100 mL of the standard solution under the above condi-
acetonitrile and ethyl acetate (3:1), shake, treat with ultra-    tions, the relative standard deviation of the ratio of the peak
sonic waves for 30 seconds, centrifuge, and use the super-        area of cilazapril to that of the internal standard is not more
natant liquid as the sample solution. Separately, dissolve 5      than 2.0z.
Supplement I, JP XV                                                                        Official Monographs              1865

Dissolution <6.10> When the test is performed at 50 revolu-       tions, the relative standard deviation of the peak area of
tions per minute according to the Paddle method, using 900        cilazapril is not more than 2.0z.
mL of water as the dissolution medium, the dissolution rate
                                                                  Assay Weigh acurately 20 Cilazapril Tablets, and powder.
in 15 minutes of Cilazapril Tablets is not less than 85z.
                                                                  Weigh accurately a portion of the powder, equivalent to
   Start the test with 1 tablet of Cilazapril Tablets, withdraw
                                                                  about 1 mg of cilazapril (C22H31N3O5), add 30 mL of a mix-
not less than 20 mL of the medium at the specified minute
                                                                  ture of water and acetonitrile (7:3), and treat with ultrasonic
after starting the test, and filter through a membrane filter
                                                                  waves for 5 minutes. Next, add exactly 5 mL of the internal
with a pore size not exceeding 0.45 mm. Discard the first 10
                                                                  standard solution, add the mixture of water and acetonitrile
mL of the filtrate, pipet V mL of the subsequent filtrate, and
                                                                  (7:3) to make 50 mL, and centrifuge. Filter the supernatant
add water to make exactly V? mL so that each mL contains
                                                                  liquid through a membrane filter with a pore size not exceed-
about 0.28 mg of cilazapril (C22H31N3O5) according to the la-
                                                                  ing 0.5 mm, and use the filtrate as the sample solution.
beled amount. Pipet 10 mL of the solution, add exactly 5
                                                                  Separately, weigh accurately about 26 mg of cilazapril for
mL of acetonitrile, and use this solution as the sample solu-
                                                                  assay (separately determine the water <2.48> in the same
tion. Separately, weigh accurately about 29 mg of cilazapril
                                                                  manner as Cilazapril Hydrate), and dissolve in the mixture
for assay (separately determine the water <2.48> in the same
                                                                  of water and acetonitrile (7:3) to make exactly 50 mL. Pipet
manner as Cilazapril Hydrate), and dissolve in water to
                                                                  2 mL of this solution, add exactly 5 mL of the internal stan-
make exactly 100 mL. Pipet 5 mL of the solution, add water
                                                                  dard solution, add the mixture of water and acetonitrile (7:3)
to make exactly 100 mL. Then, pipet 2 mL of this solution,
                                                                  to make 50 mL, and use this solution as the standard solu-
add water to make exactly 100 mL. Pipet 10 mL of this solu-
                                                                  tion. Perform the test with 50 mL each of the sample solution
tion, add exactly 5 mL of acetonitrile, and use this solution
                                                                  and standard solution as directed under Liquid Chro-
as the standard solution. Perform the test with exactly 100
                                                                  matography <2.01> according to the following conditions,
mL each of the sample solution and standard solution as
                                                                  and calculate the ratios, QT and QS, of the peak area of
directed under Liquid Chromatography <2.01> according to
                                                                  cilazapril to that of the internal standard.
the following conditions, and determine the peak areas of
cilazapril, AT and AS, of both solutions.                                   Amount (mg) of cilazapril (C22H31N3O5)
                                                                             =WS×(QT/QS)×(1/25)
Dissolution rate (z) with respect to the labeled amount of
cilazapril (C22H31N3O5)                                           WS: Amount (mg) of cilazapril for assay, calculated on the
   =WS×(AT/AS)×(V?/V)×(1/C)×(9/10)                                    anhydrous basis

  WS: Amount (mg) of cilazapril for assay, calculated on          Internal standard solution—A solution of dimethyl phtha-
       the anhydrous basis                                        late in a mixture of water and acetonitrile (7:3) (1 in 12,500).
  C: Labeled amount (mg) of cilazapril (C22H31N3O5) in 1          Operating conditions—
     tablet                                                          Detector: An ultraviolet absorption photometer (wave-
                                                                  length: 210 nm).
Operating conditions—
                                                                     Column: A stainless steel column 6 mm in inside diameter
   Detector: An ultraviolet absorption photometer (wave-
                                                                  and 15 cm in length, packed with octadecylsilanized silica gel
length: 210 nm).
                                                                  for liquid chromatography (5 mm in particle diameter).
   Column: A stainless steel column 4.6 mm in inside di-
                                                                     Column temperature: A constant temperature of about
ameter and 15 cm in length, packed with octadecylsilanized
                                                                  239  C.
silica gel for liquid chromatography (5 mm in particle di-
                                                                     Mobile phase: To a solution consisting of 180 mL of tetra-
ameter).
                                                                  hydrofuran for liquid chromatography, 120 mL of acetoni-
   Column temperature: A constant temperature of about
                                                                  trile for liquid chromatography and 3 mL of triethylamine
259  C.
                                                                  add water to make 1000 mL, and adjust the pH to 2.5 with
   Mobile phase: To a solution consisting of 180 mL of tetra-
                                                                  phosphoric acid.
hydrofuran for liquid chromatography, 120 mL of acetoni-
                                                                     Flow rate: Adjust the flow rate so that the retention time
trile for liquid chromatography and 3 mL of triethylamine
                                                                  of cilazapril is about 10 minutes.
add water to make 1000 mL, and adjust the pH to 2.5 with
                                                                  System suitability—
phosphoric acid.
                                                                     System performance: When the procedure is run with 50
   Flow rate: Adjust the flow rate so that the retention time
                                                                  mL of the standard solution under the above conditions,
of cilazapril is about 10 minutes.
                                                                  cilazapril and the internal standard are eluted in this order
System suitability—
                                                                  with the resolution between these peaks being not less than
   System performance: When the procedure is run with 100
                                                                  6.
mL of the standard solution under the above conditions, the
                                                                     System repeatability: When the test is repeated 6 times
number of theoretical plates and the symmetry factor of the
                                                                  with 50 mL of the standard solution under the above condi-
peak of cilazapril are not less than 3000 and not more than
                                                                  tions, the relative standard deviation of the ratio of the peak
2.0, respectively.
                                                                  area of cilazapril to that of the internal standard is not more
   System repeatability: When the test is repeated 6 times
                                                                  than 1.0z.
with 100 mL of the standard solution under the above condi-
1866       Official Monographs                                                                       Supplement I, JP XV

Containers and storage     Containers—Tight containers.           Method of preparation Prepare as            directed    under
                                                                  Injections, with Clindamycin Phosphate.

                                                                  Description Clindamycin Phosphate Injection is a clear,
Cilostazol Tablets                                                colorless or light yellow liquid.
シロスタゾール錠                                                          Identification To a volume of Clindamycin Phosphate In-
                                                                  jection, equivalent to 0.15 g (potency) of Clindamycin Phos-
Change the Assay to read:                                         phate according to the labeled amount, add 4 mL of water, 2
                                                                  mL of 8 mol/L sodium hydroxide TS and 0.1 mL of sodium
Assay Weigh accurately not less than 20 Cilostazol
                                                                  pentacyanonitrosylferrate (III) TS, mix, heat in a water bath
Tablets, and powder. Weigh accurately a portion of the pow-
                                                                  for 10 minutes, and add 2 mL of hydrochloric acid: a blue-
der, equivalent to about 50 mg of cilostazol (C20H27N5O2),
                                                                  green color develops.
add exactly 5 mL of the internal standard solution and
methanol to make 50 mL, and shake well for 10 minutes. To         Osmotic pressure ratio    Being specified separately.
1 mL of this solution add methanol to make 10 mL, filter
                                                                  pH <2.54>   6.0 – 7.0
through a membrane filter with a pore size not exceeding 0.5
mm, and use the filtrate as the sample solution. Separately,      Bacterial endotoxins <4.01>    Less than 0.1 EU/mg (poten-
weigh accurately about 50 mg of Cilostazol Reference Stan-        cy).
dard, previously dried at 1059 for 2 hours, dissolve in a
                                C
                                                                  Extractable volume <6.05>     It meets the requirement.
suitable amount of methanol, and add exactly 5 mL of the
internal standard solution, and add methanol to make 50           Foreign insoluble matter <6.06> Perform the test according
mL. To 1 mL of this solution add methanol to make 10 mL,          to Method 1: it meets the requirement.
and use this solution as the standard solution. Perform the
                                                                  Insoluble particulate matter <6.07>    It meets the require-
test with 10 mL each of the sample solution and standard so-
                                                                  ment.
lution as directed under Liquid Chromatography <2.01> ac-
cording to the following conditions, and calculate the ratios,    Sterility <4.06> Perform the test according to the Mem-
QT and QS, of the peak area of cilostazol to that of the inter-   brane filtration method: it meets the requirement.
nal standard.
                                                                  Assay Measure exactly a volume of Clindamycin Phos-
 Amount (mg) of cilostazol (C20H27N5O2)=WS×(QT/QS)                phate Injection, equivalent to about 0.3 g (potency) of Clin-
                                                                  damycin Phosphate, and add the mobile phase to make ex-
  WS: Amount (mg) of Cilostazol Reference Standard
                                                                  actly 100 mL. Pipet 7 mL of this solution, add exactly 25 mL
Internal standard solution—A solution of benzophenone in          of the internal standard solution and the mobile phase to
methanol (1 in 250).                                              make 100 mL, and use this solution as the sample solution.
Operating conditions—                                             Separately, weigh accurately an amount of Clindamycin
  Proceed as directed in the operating conditions in the As-      Phosphate Reference Standard, equivalent to about 20 mg
say under Cilostazol.                                             (potency), dissolve in exactly 25 mL of the internal standard
System suitability—                                               solution, add the mobile phase to make 100 mL, and use this
  System performance: Proceed as directed in the system           solution as the standard solution. Then, proceed as directed
suitability in the Assay under Cilostazol.                        in the Assay under Clindamycin Phosphate.
  System repeatability: When the test is repeated 6 times
                                                                      Amount [mg (potency)] of clindamycin phosphate
with 10 mL of the standard solution under the above operat-
                                                                      (C18H34ClN2O8PS)
ing conditions, the relative standard deviation of the ratio of
                                                                        =WS×(QT/QS)×(100/7)
the peak area of cilostazol to that of the internal standard is
not more than 1.5z.                                                 WS: Amount [mg (potency)] of Clindamycin Phosphate
                                                                        Reference Standard

                                                                  Internal standard solution—A solution of methyl para-
Add the following:
                                                                  hydroxybenzoate in the mobile phase (3 in 50,000).

Clindamycin Phosphate Injection                                   Containers and storage    Containers—Hermetic containers.

クリンダマイシンリン酸エステル注射液

  Clindamycin Phosphate Injection is an aqueous in-
jection.
   It contains not less than 90.0z and not more than
110.0z of the labeled amount of clindamycin phos-
phate (C18H34ClN2O8PS: 504.96).
Supplement I, JP XV                                                                       Official Monographs            1867

Add the following:                                               Operating conditions—
                                                                    Detector, column, column temperature, mobile phase,
Clobetasol Propionate                                            and flow rate: Proceed as directed in the operating condi-
                                                                 tions in the Assay.
クロベタゾールプロピオン酸エステル                                                   Time span of measurement: About 2.5 times as long as the
                                                                 retention time of clobetasol propionate, beginning after the
                                                                 solvent peak.
                                                                 System suitability—
                                                                    Test for required detectability: Pipet 2 mL of the standard
                                                                 solution, and add the mobile phase to make exactly 50 mL.
                                                                 Confirm that the peak area of clobetasol propionate ob-
                                                                 tained from 10 mL of this solution is equivalent to 2.8 to 5.2
C25H32ClFO5: 466.97                                              z of that from 10 mL of the standard solution.
21-Chloro-9-fluoro-11b,17-dihydroxy-                                System performance: Dissolve 20 mg of Clobetasol
16b-methylpregna-1,4-diene-3,20-dione 17-propanoate              Propionate in 20 mL of methanol. To 5 mL of this solution
[25122-46-7]                                                     add 10 mL of a solution of beclometasone dipropionate in
                                                                 methanol (1 in 1000), and then add the mobile phase to make
  Clobetasol Propionate, when dried, contains not                50 mL. When the procedure is run with 10 mL of this solu-
less than 97.0z and not more than 102.0z of                      tion under the above conditions, clobetasol propionate and
C25H32ClFO5.                                                     beclometasone dipropionate are eluted in this order with the
                                                                 resolution between these peaks being not less than 8.
Description Clobetasol Propionate occurs as a white to
                                                                    System repeatability: When the test is repeated 6 times
pale yellowish white crystalline powder.
                                                                 with 10 mL of the standard solution under the above condi-
  It is soluble in methanol and in ethanol (99.5), and practi-
                                                                 tions, the relative standard deviation of the peak area of
cally insoluble in water.
                                                                 clobetasol propionate is not more than 2.0z.
  It gradually turns yellow by light.
  Melting point: about 1969 (with decomposition).
                              C                                  Loss on drying <2.41>   Not more than 0.5z (1 g, 1059 3
                                                                                                                     C,
                                                                 hours).
Identification Determine the infrared absorption spectra
of Clobetasol Propionate as directed in the paste method         Residue on ignition <2.44>   Not more than 0.1z (1 g, plati-
under Infrared Spectrophotometry <2.25>, and compare the         num crucible).
spectrum with the Reference Spectrum or the spectrum of
                                                                 Assay Weigh accurately about 10 mg each of Clobetasol
Clobetasol Propionate Reference Standard: both spectra
                                                                 Propionate and Clobetasol Propionate Reference Standard,
exhibit similar intensities of absorbance at the same wave
                                                                 both previously dried, dissolve each in the mobile phase, add
numbers.
                                                                 exactly 100 mL of the internal standard solution, add the
Optical rotation <2.49> [a]20: +109 – +1159
                           D               (after drying,        mobile phase to make 250 mL, and use these solutions as the
0.1 g, methanol, 10 mL, 100 mm).                                 sample solution and standard solution. Perform the test with
                                                                 10 mL each of the sample solution and standard solution as
Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of
                                                                 directed under Liquid Chromatography <2.01> according to
Clobetasol Propionate according to Method 2, and perform
                                                                 the following conditions, and calculate the ratios, QT and
the test. Prepare the control solution with 2.0 mL of Stan-
                                                                 QS, of the peak area of clobetasol propionate to that of the
dard Lead Solution (not more than 20 ppm).
                                                                 internal standard.
   (2) Related substances—Dissolve 10 mg of Clobetasol
Propionate in 100 mL of the mobile phase, and use this solu-                    Amount (mg) of C25H32ClFO5
tion as the sample solution. Pipet 5 mL of the sample solu-                      =WS×(QT/QS)
tion, add the mobile phase to make exactly 200 mL, and use
                                                                   WS: Amount (mg) of Clobetasol Propionate Reference
this solution as the standard solution. Perform the test with
                                                                       Standard
exactly 10 mL each of the sample solution and standard solu-
tion as directed under Liquid Chromatography <2.01> ac-          Internal standard solution—A solution of beclometasone
cording to the following conditions, and determine each          dipropionate in the mobile phase (1 in 5000).
peak area of these solutions by the automatic integration        Operating conditions—
method: the area of the peak other than clobetasol                  Detector: An ultraviolet absorption photometer (wave-
propionate obtained from the sample solution is not larger       length: 240 nm).
than 2/5 times the peak area of clobetasol propionate from          Column: A stainless steel column 4.6 mm in inside di-
the standard solution. Furthermore, the total area of the        ameter and 15 cm in length, packed with octadecylsilanized
peaks other than clobetasol propionate from the sample so-       silica gel for liquid chromatography (5 mm in particle di-
lution is not larger than the peak area of clobetasol            ameter).
propionate from the standard solution.                              Column temperature: A constant temperature of about
                                                                 259  C.
1868       Official Monographs                                                                       Supplement I, JP XV

   Mobile phase: Dissolve 7.80 g of sodium dihydrogen                (2) Determine the absorption spectrum of a solution of
phosphate dihydrate in 900 mL of water, adjust the pH to          Clorazepate Dipotassium (1 in 200,000) as directed under
2.5 with phosphoric acid, and then add water to make 1000         Ultraviolet-visible Spectrophotometry <2.24>, and compare
mL. To 425 mL of this solution add 475 mL of acetonitrile         the spectrum with the Reference Spectrum: both spectra
and 100 mL of methanol.                                           exhibit similar intensities of absorption at the same wave-
   Flow rate: Adjust the flow rate so that the retention time     lengths.
of clobetasol propionate is about 10 minutes.                       (3) Determine the infrared absorption spectrum of
System suitability—                                               Clorazepate Dipotassium as directed in the potassium
   System performance: When the procedure is run with 10          bromide disk method under Infrared Spectrophotometry
mL of the standard solution under the above conditions,           <2.25>, and compare the spectrum with the Reference Spec-
clobetasol propionate and the internal standard are eluted in     trum: both spectra exhibit similar intensities of absorption at
this order with the resolution between these peaks being not      the same wave numbers.
less than 8.                                                         (4) Clorazepate Dipotassium responds to the Qualitative
   System repeatability: When the test is repeated 6 times        Tests <1.09> (1) for potassium salt.
with 10 mL of the standard solution under the above condi-
                                                                  Purity (1) Chloride <1.03>—Dissolve 1.0 g of Cloraze-
tions, the relative standard deviation of the ratio of the peak
                                                                  pate Dipotassium in 20 mL of water, add 20 mL of acetone,
area of clobetasol propionate to that of the internal standard
                                                                  6 mL of dilute nitric acid and water to make 50 mL. Perform
is not more than 1.0z.
                                                                  the test with this solution as the test solution. Prepare the
Containers and storage Containers—Tight containers.               control solution as follows: To 0.40 mL of 0.01 mol/L
 Storage—Light-resistant.                                         hydrochloric acid VS add 20 mL of acetone, 6 mL of dilute
                                                                  nitric acid and water to make 50 mL (not more than 0.014
                                                                  z).
Add the following:                                                   (2) Heavy metals <1.07>—Proceed with 1.0 g of Cloraze-
                                                                  pate Dipotassium according to Method 2, and perform the
Clorazepate Dipotassium                                           test. Prepare the control solution with 2.0 mL of Standard
                                                                  Lead Solution (not more than 20 ppm).
クロラゼプ酸二カリウム                                                          (3) Arsenic <1.11>—Prepare the test solution with 1.0 g
                                                                  of Clorazepate Dipotassium according to Method 3, and
                                                                  perform the test (not more than 2 ppm).
                                                                     (4) Related substances—Dissolve 15 mg of Clorazepate
                                                                  Dipotassium in 25 mL of a mixture of water, potassium car-
                                                                  bonate solution (97 in 1000) and acetonitrile (3:1:1), and use
                                                                  this solution as the sample solution. Pipet 1 mL of the sam-
                                                                  ple solution, add the mixture of water, potassium carbonate
C16H10ClKN2O3.KOH: 408.92                                         solution (97 in 1000) and acetonitrile (3:1:1) to make exactly
Monopotassium 7-chloro-2-oxo-5-phenyl-2,3-dihydro-                200 mL, and use this solution as the standard solution. Pre-
1H-1,4-benzodiazepine-3-carboxylate                               pare these solutions quickly and perform the test within 3
mono (potassium hydroxide) [57109-90-7]                           minutes. Perform the test with exactly 5 mL each of the sam-
                                                                  ple solution and standard solution as directed under Liquid
  Clorazepate Dipotassium, when dried, contains not               Chromatography <2.01> according to the following condi-
less than 98.5z and not more than 101.0z of                       tions, and determine each peak area by the automatic in-
C16H10ClKN2O3.KOH.                                                tegration method: the peak area of nordiazepam, having the
Description Clorazepate Dipotassium occurs as white to            relative retention time of about 3.0 with respect to clorazepic
light yellow, crystals or crystalline powder.                     acid, obtained from the sample solution is not larger than
   It is freely soluble in water, and very slightly soluble in    the peak area of clorazepic acid from the standard solution,
ethanol (99.5).                                                   the area of the peak other than clorazepic acid and nordia-
   It dissolves in acetic acid (100).                             zepam is not larger than 1/5 times the peak area of clorazep-
   The pH of a solution obtained by dissolving 1 g of             ic acid from the standard solution, and the total area of the
Clorazepate Dipotassium in 100 mL of water is between 11.5        peaks other than clorazepic acid from the sample solution is
and 12.5.                                                         not larger than 2 times the peak area of clorazepic acid from
   It gradually turns yellow on exposure to light.                the standard solution. For this comparison, use the peak
                                                                  area of nordiazepam, having the relative retention time of
Identification (1) Carefully and gradually ignite to red-         about 3.0 with respect to clorazepic acid, after multiplying
ness 30 mg of Clorazepate Dipotassium with 50 mg of sodi-         by the relative response factor, 0.64.
um. After cooling, add 3 drops of ethanol (99.5) and 5 mL         Operating conditions—
of water, mix well, and filter: the filtrate responds to the         Detector: An ultraviolet absorption photometer (wave-
Qualitative Tests <1.09> for chloride.                            length: 232 nm).
Supplement I, JP XV                                                                       Official Monographs              1869

   Column: A stainless steel column 4.6 mm in inside di-         Method of preparation Prepare as directed under Cap-
ameter and 15 cm in length, packed with octadecylsilanized       sules, with Clorazepate Dipotassium.
silica gel for liquid chromatography (5 mm in particle di-
                                                                 Identification To 10 mL of the sample solution obtained in
ameter).
                                                                 the Assay add water to make 20 mL. Determine the absorp-
   Column temperature: A constant temperature of about
                                                                 tion spectrum of this solution as directed under Ultraviolet-
259  C.
                                                                 visible Spectrophotometry <2.24>: it exhibits a maximum be-
   Mobile phase: Dissolve 13.8 g of sodium dihydrogen
                                                                 tween 228 nm and 232 nm.
phosphate dihydrate in 500 mL of water, and adjust to pH
8.0 with sodium hydroxide TS. To 100 mL of this solution         Purity Related substances—Take out the contents of
add 400 mL of acetonitrile and 300 mL of water.                  Clorazepate Dipotassium Capsules, and powder. To a por-
   Flow rate: Adjust the flow rate so that the retention time    tion of the powder, equivalent to 15 mg of Clorazepate
of clorazepic acid is about 1.3 minutes.                         Dipotassium according to the labeled amount, add a mixture
   Time span of measurement: About 10 times as long as the       of water, potassium carbonate solution (97 in 1000) and
retention time of clorazepic acid, beginning after the solvent   acetonitrile (3:1:1) to make 25 mL, and mix for 10 minutes.
peak.                                                            Filter the solution through a membrane filter with a pore size
System suitability—                                              not exceeding 0.45 mm, discard the first 5 mL of the filtrate,
   Test for required detectability: To exactly 5 mL of the       and use the subsequent filtrate as the sample solution. Pipet
standard solution add the mixture of water, potassium car-       1 mL of the sample solution, add a mixture of water, potas-
bonate solution (97 in 1000) and acetonitrile (3:1:1) to make    sium carbonate solution (97 in 1000) and acetonitrile (3:1:1)
exactly 25 mL. Confirm that the peak area of clorazepic acid     to make exactly 200 mL, and use this solution as the stan-
obtained from 5 mL of this solution is equivalent to 15 to 25    dard solution. Then, proceed as directed in the Purity (4) un-
z of that from 5 mL of the standard solution.                    der Clorazepate Dipotassium: the peak area of nordiazep-
   System performance: When the procedure is run with 5 mL       am, having the relative retention time of about 3.0 with
of the standard solution under the above operating condi-        respect to clorazepic acid, obtained from the sample solution
tions, the number of theoretical plates and the symmetry         is not larger than 3 times the peak area of clorazepic acid
factor of the peak of clorazepic acid are not less than 3000     from the standard solution, and the total area of the peaks
and not more than 1.5, respectively.                             other than clorazepic acid and nordiazepam from the sample
   System repeatability: When the test is repeated 6 times       solution is not larger than the peak area of clorazepic acid
with 5 mL of the standard solution under the above operat-       from the standard solution. For this comparison, use the
ing conditions, the relative standard deviation of the peak      peak area of nordiazepam after multiplying by the relative
area of clorazepic acid is not more than 1.5z.                   response factor, 0.64.

Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-          Uniformity of dosage units <6.02> Perform the test accord-
um, phosphorus (V) oxide, 609 5 hours).
                            C,                                   ing to the following method: it meets the requirement of the
                                                                 Content uniformity test.
Assay Weigh accurately about 0.15 g of Clorazepate
                                                                   To 1 capsule of Clorazepate Dipotassium Capsules add 70
Dipotassium, previously dried, dissolve in 100 mL of acetic
                                                                 mL of water, shake for 15 minutes, and add water to make
acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid
                                                                 exactly 100 mL. Centrifuge the solution, pipet V mL of the
VS until the color of solution changes from violet to blue-
                                                                 supernatant liquid, add water to make exactly V? mL so that
green through blue (indicator: 3 drops of crystal violet TS).
                                                                 each mL contains about 12 mg of clorazepate dipotassium
Perform a blank determination in the same manner, and
                                                                 (C16H10ClKN2O3.KOH), and use this solution as the sample
make any necessary correction.
                                                                 solution. Then, proceed as directed in the Assay.
        Each mL of 0.1 mol/L perchloric acid VS
                                                                          Amount (mg) of clorazepate dipotassium
          =13.63 mg of C16H10ClKN2O3.KOH
                                                                          (C16H10ClKN2O3.KOH)
Containers and storage Containers—Tight containers.                         =WS×(AT/AS)×(V?/V)×(2/25)
 Storage—Light-resistant.
                                                                   WS: Amount (mg) of clorazepate dipotassium for assay
                                                                 Dissolution <6.10> When the test is performed at 50 revolu-
Add the following:                                               tions per minute according to the Paddle method using the
                                                                 sinker, using 900 mL of water as the dissolution medium,
Clorazepate Dipotassium Capsules                                 the dissolution rate in 30 minutes of Clorazepate Dipotassi-
                                                                 um Capsules is not less than 80z.
クロラゼプ酸二カリウムカプセル                                                     Start the test with 1 capsule of Clorazepate Dipotassium
                                                                 Capsules, withdraw not less than 20 mL of the medium at
  Clorazepate Dipotassium Capsules contain not less              the specified minute after starting the test, and filter through
than 93.0z and not more than 107.0z of the labeled               a membrane filter with a pore size not exceeding 0.45 mm.
amount of clorazepate dipotassium (C16H10ClKN2O3.                Discard the first 10 mL of the filtrate, pipet V mL of the
KOH: 408.92).                                                    subsequent filtrate, add water to make exactly V? mL so that
1870      Official Monographs                                                                     Supplement I, JP XV

each mL contains about 8.3 mg of clorazepate dipotassium
(C16H10ClKN2O3.KOH) according to the labeled amount,           Cyanocobalamin
and use this solution as the sample solution. Separately,
weigh accurately about 21 mg of clorazepate dipotassium        シアノコバラミン
for assay, previously dried in vacuum over phosphorus (V)
oxide at 609 for 5 hours, and dissolve in water to make ex-
            C                                                  Change the Identification (2) and (3) to read:
actly 100 mL. Pipet 4 mL of this solution, add water to
                                                               Identification
make exactly 100 mL, and use this solution as the standard
                                                                  (2) Mix 1 mg of Cyanocobalamin with 50 mg of potassi-
solution. Determine the absorbances, AT and AS, at 252 nm
                                                               um hydrogen sulfate, and fuse by igniting. Cool, break up
of the sample solution and standard solution as directed un-
                                                               the mass with a glass rod, add 3 mL of water, and dissolve
der Ultraviolet-visible Spectrophotometry <2.24>.
                                                               by boiling. Add 1 drop of phenolphthalein TS, then add
Dissolution rate (z) with respect to the labeled amount of     dropwise sodium hydroxide TS until a light red color just
clorazepate dipotassium (C16H10ClKN2O3.KOH)                    develops. Add 0.5 g of sodium acetate trihydrate, 0.5 mL of
  =WS×(AT/AS)×(V?/V)×(1/C)×36                                  dilute acetic acid and 0.5 mL of a solution of disodium 1-
                                                               nitroso-2-naphthol-3,6-disulfonate (1 in 500): a red to oran-
  WS: Amount (mg) of clorazepate dipotassium for assay
                                                               ge-red color is immediately produced. Then add 0.5 mL of
  C: Labeled amount (mg) of clorazepate dipotassium
                                                               hydrochloric acid, and boil for 1 minute: the red color does
     (C16H10ClKN2O3.KOH) in 1 capsule
                                                               not disappear.
Assay Carefully take out the contents of not less than 20         (3) Transfer 5 mg of Cyanocobalamin to a 50-mL distil-
Clorazepate Dipotassium Capsules, weigh accurately the         ling flask, dissolve in 5 mL of water, and add 2.5 mL of
mass of the contents, and powder. Weigh accurately a por-      hypophosphorous acid. Connect the flask with a short con-
tion of the powder, equivalent to about 15 mg of clorazepate   denser, and dips its tip into a test tube containing 1 mL of a
dipotassium (C16H10ClKN2O3.KOH), add 70 mL of water,           solution of sodium hydroxide (1 in 50). Heat gently for 10
shake for 15 minutes, and add water to make exactly 100        minutes, then distil 1 mL into a test tube. To the test tube
mL. Centrifuge the solution, pipet 4 mL of the supernatant     add 4 drops of a saturated solution of ammonium iron (II)
liquid, add water to make exactly 50 mL, and use this solu-    sulfate hexahydrate, shake gently, then add about 30 mg of
tion as the sample solution. Separately, weigh accurately      sodium fluoride, and heat the contents to boil. Immediately
about 15 mg of clorazepate dipotassium for assay, previous-    add dropwise diluted sulfuric acid (1 in 7) until a clear solu-
ly dried in vacuum over phosphorus (V) oxide at 609 for 5
                                                     C         tion results, then add 3 to 5 drops more of diluted sulfuric
hours, and dissolve in water to make exactly 100 mL. Pipet     acid (1 in 7): a blue to blue-green color develops.
4 mL of this solution, add water to make exactly 50 mL, and
use this solution as the standard solution. Perform the test   Change the Purity (2) to read:
with the sample solution and standard solution as directed
                                                               Purity
under Ultraviolet-visible Spectrophotometry <2.24>, and
                                                                  (2) Related substances—Conduct this procedure using
determine the absorbances, AT and AS, at 252 nm.
                                                               light-resistant vessels. Dissolve 10 mg of Cyanocobalamin in
         Amount (mg) of clorazepate dipotassium                10 mL of the mobile phase, and use this solution as the sam-
         (C16H10ClKN2O3.KOH)                                   ple solution. Pipet 3 mL of the sample solution, add the mo-
           =WS×(AT/AS)                                         bile phase to make exactly 100 mL, and use this solution as
                                                               the standard solution. Perform the test with exactly 20 mL
  WS: Amount (mg) of clorazepate dipotassium for assay
                                                               each of the sample solution and standard solution as direct-
Containers and storage   Containers—Tight containers.          ed under Liquid Chromatography <2.01> according to the
                                                               following conditions, and determine each peak area by the
                                                               automatic integration method: the total area of the peak
Creosote                                                       other than cyanocobalamin obtained from the sample solu-
                                                               tion is not larger than the peak area of cyanocobalamin from
                                                               the standard solution.
Change the title of the monograph as follows:
                                                               Operating conditions—
                                                                  Detector: An ultraviolet absorption photometer (wave-
Wood Creosote                                                  length: 361 nm).
木クレオソート                                                           Column: A stainless steel column 4.6 mm in inside di-
                                                               ameter and 25 cm in length, packed with octylsilanized silica
                                                               gel for liquid chromatography (5 mm in particle diameter).
                                                                  Column temperature: A constant temperature of about
                                                               309 C.
                                                                  Mobile phase: Dissolve 10 g of anhydrous disodium
                                                               hydrogen phosphate in 1000 mL of water, and adjust to pH
                                                               3.5 with phosphoric acid. To 147 mL of this solution add 53
Supplement I, JP XV                                                                       Official Monographs            1871

mL of methanol.                                                  Add the following:
   Flow rate: Adjust the flow rate so that the retention time
of cyanocobalamin is about 7 minutes.                            L-Cysteine
   Time span of measurement: About 4 times as long as the
retention time of cyanocobalamin, beginning after the sol-       L-システイン
vent peak.
System suitability—
   Test for required detectability: Pipet 1 mL of the sample
solution, add the mobile phase to make exactly 100 mL, and       C3H7NO2S: 121.16
use this solution as the solution for system suitability test.   (2R)-2-Amino-3-sulfanylpropanoic acid      [52-90-4]
Pipet 1 mL of the solution for system suitability test, and
add the mobile phase to make exactly 10 mL. Confirm that           L-Cysteine contains not less than 98.5z and not
the peak area of cyanocobalamin obtained with 20 mL of this      more than 101.0z of C3H7NO2S, calculated on the
solution is equivalent to 7 to 13z of that with 20 mL of the     dried basis.
solution for system suitability test.
                                                                 Description L-Cysteine occurs as white crystals or a white
   System performance: Perform this procedure quickly
                                                                 crystalline powder. It has a characteristic odor and a pun-
after the solution is prepared. To 25 mg of cyanocobalamin
                                                                 gent taste.
add 10 mL of water, and warm, if necessary, to dissolve.
                                                                   It is freely soluble in water, and practically insoluble in
After cooling, add 0.5 mL of sodium toluenesulfon-
                                                                 ethanol (99.5).
chloramide TS, 0.5 mL of 0.05 mol/L hydrochloric acid TS
                                                                   It dissolves in 1 mol/L hydrochloric acid TS.
and water to make 25 mL, mix, and allow the solution to
stand for 5 minutes. To 1 mL of the solution add the mobile      Identification Determine the infrared absorption spectrum
phase to make 10 mL. When the procedure is run with 20 mL        of L-Cysteine as directed in the potassium bromide disk
of the solution under the above operating conditions, two        method under Infrared Spectrophotometry <2.25>, and com-
principal peaks appear with the resolution between these         pare the spectrum with the Reference Spectrum: both spec-
peaks being not less than 2.5.                                   tra exhibit similar intensities of absorption at the same wave
   System repeatability: When the test is repeated 6 times       numbers.
with 20 mL of the solution for system suitability test under
                                                                 Optical rotation <2.49> [a]20: +8.0 – +10.09(2 g calculat-
                                                                                              D
the above operating conditions, the relative standard devia-
                                                                 ed on the dried basis, 1 mol/L hydrochloric acid TS, 25 mL,
tion of the peak area of cyanocobalamin is not more than
                                                                 100 mm).
3.0z.
                                                                 pH <2.54> The pH of a solution prepared by dissolving
                                                                 1.25 g of L-Cysteine in 50 mL of water is 4.7 to 5.7.
Cyanocobalamin Injection                                         Purity (1) Clarity and color of solution—Dissolve 1.0 g
シアノコバラミン注射液                                                      of L-Cysteine in 20 mL of water: the solution is clear and
                                                                 colorless.
Change the Description to read:                                     (2) Chloride <1.03>—Dissolve 0.30 g of L-Cysteine in 10
                                                                 mL of diluted nitric acid (1 in 4), add 10 mL of hydrogen
Description Cyanocobalamin Injection is a clear, light red       peroxide (30), heat for 20 minutes in a boiling water bath,
to red liquid.                                                   cool, and then add water to make 50 mL. Perform the test
                                                                 using this solution as the test solution. Prepare the control
Add the following next to Identification:                        solution with 0.35 mL of 0.01 mol/L hydrochloric acid VS
Bacterial endotoxins <4.01>   Less than 0.30 EU/mg.              (not more than 0.041z).
                                                                    (3) Sulfate <1.14>—Dissolve 0.6 g of L-Cysteine in 30
Add the following next to Extractable volume:                    mL of water and 3 mL of dilute hydrochloric acid, and add
                                                                 water to make 50 mL. Perform the test using this solution as
Foreign insoluble matter <6.06> Perform the test according       the test solution. Prepare the control solution as follows: To
to Method 1: it meets the requirement.                           0.35 mL of 0.005 mol/L sulfuric acid VS add 3 mL of dilute
Insoluble particulate matter <6.07>    It meets the require-     hydrochloric acid and water to make 50 mL. Prepare the test
ment.                                                            solution and the control solution with 4 mL of barium chlo-
                                                                 ride TS, respectively (not more than 0.028z).
Sterility <4.06> Perform the test according to the Mem-             (4) Ammonium <1.02>—Perform the test with 0.25 g of
brane filtration method: it meets the requirement.               L-Cysteine, using the distillation under reduced pressure.
                                                                 Prepare the control solution with 5.0 mL of Standard Am-
                                                                 monium Solution (not more than 0.02z).
                                                                    (5) Heavy metals <1.07>—Proceed with 1.0 g of
                                                                 L-Cysteine according to Method 4, and perform the test.
1872       Official Monographs                                                                       Supplement I, JP XV

Prepare the control solution with 1.0 mL of Standard Lead         Add the following:
Solution (not more than 10 ppm).
   (6) Iron <1.10>—Prepare the test solution with 1.0 g of        L-Cysteine         Hydrochloride Hydrate
L-Cysteine according to Method 1, and perform the test us-
ing Method A. Prepare the control solution with 1.0 mL of         L-システイン塩酸塩水和物
Standard Iron Solution (not more than 10 ppm).
   (7) Related substances—Dissolve 0.10 g of L-Cysteine in
N-ethylmaleimide solution (1 in 50) to make 10 mL, leave
for 30 minutes, and use this solution as the sample solution.     C3H7NO2S.HCl.H2O: 175.63
Pipet 1 mL of the sample solution, add water to make exact-       (2R)-2-Amino-3-sulfanylpropanoic acid monohydrochloride
ly 10 mL, pipet 1 mL of this solution, add water to make          monohydrate [7048-04-6]
exactly 50 mL, and use this solution as the standard solution
(1). Separately, dissolve 0.10 g of L-cystine in 0.5 mol/L          L-Cysteine Hydrochloride Hydrate contains not less
hydrochloric acid TS to make 20 mL. Pipet 1 mL of this            than 98.5z and not more than 101.0z of L-cysteine
solution, add water to make 100 mL, and use this solution as      hydrochloride (C3H7NO2S.HCl: 157.62), calculated
the standard solution (2). Perform the test with these solu-      on the dried basis.
tions as directed under Thin-layer Chromatography <2.03>.
                                                                  Description L-Cysteine Hydrochloride Hydrate occurs as
Spot 10 mL each of the sample solution and standard solu-
                                                                  white crystals or crystalline powder. It has a characteristic
tions (1) and (2) on a plate of silica gel for thin-layer chro-
                                                                  odor and a strong acid taste.
matography. Develop the plate with a mixture of 1-butanol,
                                                                    It is very soluble in water, and soluble in ethanol (99.5).
water, and acetic acid (100) (3:1:1) to a distance of about 10
                                                                    It dissolves in 6 mol/L hydrochloric acid TS.
cm, and dry the plate for 30 minutes at 809 Spray the plate
                                             C.
evenly with a solution of ninhydrin in a mixture of methanol      Identification (1) Determine the infrared absorption
and acetic acid (100) (97:3) (1 in 100), and then heat at 809C    spectrum of L-Cysteine Hydrochloride Hydrate as directed
for 10 minutes: the spot obtained from the sample solution        in the potassium chloride disk method under Infrared Spec-
corresponding to the spot obtained from the standard solu-        trophotometry <2.25>, and compare the spectrum with the
tion (2) is not more intense than the spot from the standard      Reference Spectrum: both spectra exhibit similar intensities
solution (2). Also, the spots other than the principal spot       of absorption at the same wave numbers.
and the spots mentioned above from the sample solution are           (2) To 10 mL of a solution of L-Cysteine Hydrochloride
not more intense than the spot from the standard solution         Hydrate (1 in 50) add 1 mL of hydrogen peroxide (30), heat
(1).                                                              on a water bath for 20 minutes, and cool: the solution
                                                                  responds to the Qualitative Tests <1.09> (2) for chloride.
Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-
um, phosphorus (V) oxide, 3 hours).                               Optical rotation <2.49> [a]20: +6.0 – +7.59(2 g, calculat-
                                                                                               D
                                                                  ed on the dried basis, 6 mol/L hydrochloric acid TS, 25 mL,
Residue on ignition <2.44>    Not more than 0.1z (1 g).
                                                                  100 mm).
Assay Weigh accurately about 0.2 g of L-Cysteine, place it
                                                                  pH <2.54> The pH of a solution prepared by dissolving
in a stoppered flask, and dissolve in 20 mL of water. Dis-
                                                                  1.0 g of L-Cysteine Hydrochloride Hydrate in 100 mL of
solve 4 g of potassium iodide in this solution, immediately
                                                                  water is between 1.3 and 2.3.
place in ice cold water, add 5 mL of dilute hydrochloric acid
and exactly 25 mL of 0.05 mol/L iodine VS, leave in a dark        Purity (1) Clarity and color of solution—A solution
place for 20 minutes, and then titrate <2.50> an excess           obtained by dissolving 1.0 g of L-Cysteine Hydrochloride
amount of iodine with 0.1 mol/L sodium thiosulfate VS (in-        Hydrate in 10 mL of water is clear and colorless.
dicator: starch TS). Perform a blank determination using            (2) Sulfate < 1.14 > —Dissolve 0.8 g of L-Cysteine
the same method.                                                  Hydrochloride Hydrate in 30 mL of water and 3 mL of di-
                                                                  lute hydrochloric acid, and add water to make 50 mL. Per-
Each mL of 0.05 mol/L iodine VS=12.12 mg of C3H7NO2S
                                                                  form the test using this solution as the test solution. Prepare
Containers and storage     Containers—Tight containers.           the control solution as follows: To 0.35 mL of 0.005 mol/L
                                                                  sulfuric acid VS add 3 mL of dilute hydrochloric acid and
                                                                  water to make 50 mL. To both of the test solution and the
                                                                  control solution add 4 mL of barium chloride TS (not more
                                                                  than 0.021z).
                                                                    (3) Ammonium <1.02>—Perform the test with 0.25 g of
                                                                  L-Cysteine Hydrochloride Hydrate using the distillation
                                                                  under reduced pressure. Prepare the control solution with
                                                                  5.0 mL of Standard Ammonium Solution (not more than
                                                                  0.02z).
                                                                    (4) Heavy metals <1.07>—Proceed with 1.0 g of
Supplement I, JP XV                                                                     Official Monographs           1873

L-Cysteine Hydrochloride Hydrate according to Method 4,
and perform the test. Prepare the control solution with 1.0      Dehydrocholic Acid Injection
mL of Standard Lead Solution (not more than 10 ppm).
  (5) Iron <1.10>—Prepare the test solution with 1.0 g of        デヒドロコール酸注射液
L-Cysteine Hydrochloride Hydrate according to Method 1,
and perform the test according to Method A. Prepare the          Add the following next to Extractable volume:
control solution with 1.0 mL of Standard Iron Solution (not
                                                                 Foreign insoluble matter <6.06> Perform the test according
more than 10 ppm).
                                                                 to Method 1: it meets the requirement.
  (6) Related substances—Dissolve 0.10 g of L-Cysteine
Hydrochloride Hydrate in N-ethylmaleimide solution (1 in         Insoluble particulate matter <6.07>   It meets the require-
50) to make 10 mL, allow to stand for 30 minutes, and use        ment.
this solution as the sample solution. Pipet 1 mL of the sam-
                                                                 Sterility <4.06> Perform the test according to the Mem-
ple solution, add water to make exactly 10 mL. Pipet 1 mL
                                                                 brane filtration method: it meets the requirement.
of this solution, add water to make exactly 50 mL, and use
this solution as the standard solution. Perform the test with
these solutions as directed under Thin-Layer Chro-
matography <2.03>. Spot 5 mL each of the sample solution
                                                                 Deslanoside Injection
and standard solution on a plate of silica gel for thin-layer    デスラノシド注射液
chromatography. Then develop with a mixture of 1-butanol,
water and acetic acid (100) (3:1:1) to a distance of about 10    Add the following next to Identification:
cm, and dry the plate at 809 for 30 minutes. Spray evenly a
                            C
solution of ninhydrin in a mixture of methanol and acetic        Bacterial endotoxins <4.01>   Less than 500 EU/mg.
acid (100) (97:3) (1 in 100) on the plate, and then heat at
809 for 10 minutes: the spot other than the principal spot
    C                                                            Add the following next to Extractable volume:
obtained with the sample solution is not more intense than       Foreign insoluble matter <6.06> Perform the test according
the spot with the standard solution.                             to Method 1: it meets the requirement.
Loss on drying <2.41> 8.5 – 12.0z (1 g, in vacuum, phos-         Insoluble particulate matter <6.07>   It meets the require-
phorus (V) oxide, 20 hours).                                     ment.
Residue on ignition <2.44>   Not more than 0.1z (1 g).           Sterility <4.06> Perform the test according to the Mem-
Assay Weigh accurately about 0.25 g of L-Cysteine                brane filtration method: it meets the requirement.
Hydrochloride Hydrate, place in a glass-stoppered flask,
and dissolve in 20 mL of water. Dissolve 4 g of potassium
iodide in this solution, soak immediately in ice cold water,     Dextran 40
add 5 mL of dilute hydrochloric acid and exactly 25 mL of
0.05 mol/L iodine VS, allow to stand for 20 minutes in a         デキストラン 40
dark place, titrate <2.50> the excess of iodine with 0.1 mol/L
sodium thiosulfate VS (indicator: starch TS). Perform a          Delete the Pyrogen and add the following next
blank determination in the same manner, and make any             to Residue on ignition:
necessary correction.                                            Bacterial endotoxins <4.01>   Less than 2.5 EU/g.
            Each mL of 0.05 mol/L iodine VS
              =15.76 mg of C3H7NO2S.HCl

Containers and storage    Containers—Tight containers.



Deferoxamine Mesilate
デフェロキサミンメシル酸塩

Change the Identification (2) to read:
Identification
  (2) A 50 mg portion of Deferoxamine Mesilate responds
to the Qualitative Tests <1.09> (1) for mesilate.
1874       Official Monographs                                                                        Supplement I, JP XV

Change to read:                                                   um Phosphate with 5 mL of freshly boiled and cooled water,
                                                                  and add immediately 2 mL of hydrochloric acid: no efferves-
Anhydrous Dibasic Calcium                                         cence occurs.
                                                                     (5)  Heavy metals <1.07>—Dissolve 0.65 g of Anhy-
Phosphate                                                         drous Dibasic Calcium Phosphate in a mixture of 5 mL of
無水リン酸水素カルシウム                                                      water and 5 mL of dilute hydrochloric acid by warming,
                                                                  cool, and add ammonia TS until precipitates begin to form
                                                                  in the solution. Dissolve the precipitates by adding a small
CaHPO4: 136.06
                                                                  amount of dilute hydrochloric acid dropwise, add 10 mL of
[7757-93-9]
                                                                  hydrochloric acid-ammonium acetate buffer solution, pH
  This monograph is harmonized with the European Phar-            3.5, and water to make 50 mL, and perform the test using
macopoeia and the U.S. Pharmacopeia. The parts of the text        this solution as the test solution. Prepare the control solu-
that are not harmonized are marked with symbols ( ).            tion as follows: to 10 mL of hydrochloric acid-ammonium
                                                                  acetate buffer solution, pH 3.5, add 2.0 mL of Standard
  Anhydrous Dibasic Calcium Phosphate contains                    Lead Solution and water to make 50 mL (not more than 31
not less than 98.0z and not more than 103.0z of                   ppm).
CaHPO4.                                                              (6) Barium—Heat 0.5 g of Anhydrous Dibasic Calcium
                                                                  Phosphate with 10 mL of water, add 1 mL of hydrochloric
Description     Anhydrous Dibasic Calcium Phosphate oc-
                                                                  acid dropwise with stirring, and filter after cooling, if neces-
curs as white, crystalline powder or granules.
                                                                  sary. Add 2 mL of potassium sulfate TS to this solution, and
  It is practically insoluble in water and in ethanol (99.5).
                                                                  allow to stand for 10 minutes: no turbidity forms.
  It dissolves in dilute hydrochloric acid and in dilute nitric      
                                                                       (7) Arsenic <1.11>—Dissolve 1.0 g of Anhydrous Di-
acid.
                                                                  basic Calcium Phosphate in 5 mL of dilute hydrochloric
Identification (1) Dissolve 0.1 g of Anhydrous Dibasic            acid, and perform the test with this solution as the test solu-
Calcium Phosphate in 10 mL of 2 mol/L hydrochloric acid           tion (not more than 2 ppm).
TS by warming, add 2.5 mL of ammonia TS dropwise with
                                                                  Loss on ignition <2.43> Not less than 6.6z and not more
shaking, and add 5 mL of ammonium oxalate TS: a white
                                                                  than 8.5z (1 g, 800 – 8259 constant mass).
                                                                                           C,
precipitate is produced.
  (2) Dissolve 0.1 g of Anhydrous Dibasic Calcium Phos-           Assay Weigh accurately about 0.4 g of Anhydrous Dibasic
phate in 5 mL of dilute nitric acid, and add 2 mL of hexaam-      Calcium Phosphate, dissolve in 12 mL of dilute hydrochlor-
monium heptamolybdate TS after warming at 709 for 1 to
                                                   C              ic acid, and add water to make exactly 200 mL. Pipet 20 mL
2 minutes: a yellow precipitate is produced.                      of this solution, add exactly 25 mL of 0.02 mol/L disodium
                                                                  dihydrogen ethylenediamine tetraacetate VS, 50 mL of
Purity (1) Acid-insoluble substances—Dissolve 5.0 g of
                                                                  water and 5 mL of ammonia-ammonium chloride buffer
Anhydrous Dibasic Calcium Phosphate in 40 mL of water
                                                                  solution, pH 10.7, and titrate <2.50> the excess of disodium
and 10 mL of hydrochloric acid, and boil gently for 5
                                                                  dihydrogen ethylenediamine tetraacetate with 0.02 mol/L
minutes. After cooling, collect the insoluble substance using
                                                                  zinc sulfate VS (indicator: 25 mg of eriochrome black T-so-
a filter paper for assay. Wash with water until no more
                                                                  dium chloride indicator). Perform a blank determination in
turbidity of the washings is produced when silver nitrate TS
                                                                  the same manner.
is added. Ignite to incinerate the residue and the filter paper
at 600±509 the mass is not more than 10 mg (not more
             C:                                                   Each mL of 0.02 mol/L disodium dihydrogen ethylenedia-
than 0.2z).                                                       mine tetraacetate VS
   (2) Chloride <1.03>—Dissolve 0.20 g of Anhydrous Di-             =2.721 mg of CaHPO4
basic Calcium Phosphate in 20 mL of water and 13 mL of            Containers    and storage     Containers—Well-closed con-
dilute nitric acid, add water to make 100 mL, and filter, if
                                                                  tainers.
necessary. Perform the test using a 50-mL portion of this so-
lution as the test solution. Prepare the control solution with
0.70 mL of 0.01 mol/L hydrochloric acid VS (not more than
0.248z).
   (3) Sulfate <1.14>—Dissolve 0.5 g of Anhydrous Dibasic
Calcium Phosphate in 5 mL of water and 5 mL of dilute
hydrochloric acid, add water to make 100 mL, and filter, if
necessary. Take 20 mL of this solution, add 1 mL of dilute
hydrochloric acid, and add water to make 50 mL. Perform
the test using this solution as the test solution. Prepare the
control solution with 1.0 mL of 0.005 mol/L sulfuric acid
VS (not more than 0.48z).
   (4) Carbonate—Mix 1.0 g of Anhydrous Dibasic Calci-
Supplement I, JP XV                                                                        Official Monographs              1875

Change to read:                                                   and add immediately 2 mL of hydrochloric acid: no efferves-
                                                                  cence occurs.
Dibasic Calcium Phosphate                                            (5)  Heavy metals <1.07>—Dissolve 0.65 g of Dibasic
                                                                  Calcium Phosphate Hydrate in a mixture of 5 mL of water
Hydrate                                                           and 5 mL of dilute hydrochloric acid by warming, cool, and
リン酸水素カルシウム水和物                                                     add ammonia TS until precipitates begin to form in the solu-
                                                                  tion. Dissolve the precipitates by adding a small amount of
                                                                  dilute hydrochloric acid dropwise, add 10 mL of
CaHPO4.2H2O: 172.09                                               hydrochloric acid-ammonium acetate buffer solution, pH
[7789-77-7]                                                       3.5, and water to make 50 mL, and perform the test using
  This monograph is harmonized with the European Phar-            this solution as the test solution. Prepare the control solu-
macopoeia and the U.S. Pharmacopeia. The parts of the text        tion as follows: to 10 mL of hydrochloric acid-ammonium
that are not harmonized are marked with symbols ( ).            acetate buffer solution, pH 3.5, add 2.0 mL of Standard
                                                                  Lead Solution and water to make 50 mL (not more than 31
  Dibasic Calcium Phosphate Hydrate contains not                  ppm).
less than 98.0z and not more than 105.0z of                          (6) Barium—Heat 0.5 g of Dibasic Calcium Phosphate
CaHPO4.2H2O.                                                      Hydrate with 10 mL of water, add 1 mL of hydrochloric
                                                                  acid dropwise with stirring, and filter after cooling, if neces-

 Description Dibasic Calcium Phosphate Hydrate occurs             sary. Add 2 mL of potassium sulfate TS to this solution, and
as a white, crystalline powder.                                   allow to stand for 10 minutes: no turbidity forms.
  It is practically insoluble in water and in ethanol (99.5).        (7)  Arsenic <1.11>—Dissolve 1.0 g of Dibasic Calcium
  It dissolves in dilute hydrochloric acid and in dilute nitric   Phosphate Hydrate in 5 mL of dilute hydrochloric acid, and
acid.                                                            perform the test with this solution as the test solution (not
Identification (1) Dissolve 0.1 g of Dibasic Calcium              more than 2 ppm).
Phosphate Hydrate in 10 mL of 2 mol/L hydrochloric acid           Loss on ignition <2.43> Not less than 24.5z and not more
TS by warming, add 2.5 mL of ammonia TS dropwise with             than 26.5z (1 g, 800 – 8259 constant mass).
                                                                                             C,
shaking, and add 5 mL of ammonium oxalate TS: a white
precipitate is produced.                                          Assay Weigh accurately about 0.4 g of Dibasic Calcium
  (2) Dissolve 0.1 g of Dibasic Calcium Phosphate Hy-             Phosphate Hydrate, dissolve in 12 mL of dilute hydrochloric
drate in 5 mL of dilute nitric acid, and add 2 mL of hexaam-      acid, and add water to make exactly 200 mL. Pipet 20 mL of
monium heptamolybdate TS after warming at 709 for 1 to
                                                   C              this solution, add exactly 25 mL of 0.02 mol/L disodium di-
2 minutes: a yellow precipitate is produced.                      hydrogen ethylenediamine tetraacetate VS, 50 mL of water
                                                                  and 5 mL of ammonia-ammonium chloride buffer solution,
Purity (1) Acid-insoluble substance—Dissolve 5.0 g of             pH 10.7, and titrate <2.50> the excess of disodium dihydro-
Dibasic Calcium Phosphate Hydrate in 40 mL of water and           gen ethylenediamine tetraacetate with 0.02 mol/L zinc sul-
10 mL of hydrochloric acid, and boil gently for 5 minutes.        fate VS (indicator: 25 mg of eriochrome black T-sodium
After cooling, collect the insoluble substance using a filter     chloride indicator). Perform a blank determination in the
paper for assay. Wash with water until no more turbidity of       same manner.
the washing is produced when silver nitrate TS is added. Ig-
nite to incinerate the residue and filter paper at 600±509  C:    Each mL of 0.02 mol/L disodium dihydrogen ethylenedia-
the mass is not more than 10 mg (not more than 0.2z).             mine tetraacetate VS
  (2) Chloride <1.03>—Dissolve 0.20 g of Dibasic Calcium            =3.442 mg of CaHPO4.2H2O
Phosphate Hydrate in 20 mL of water and 13 mL of dilute           Containers   and storage Containers—Well-closed contain-
nitric acid, add water to make 100 mL, and filter, if necessa-    ers.
ry. Perform the test using a 50-mL portion of this solution as
the test solution. Prepare the control solution with 0.70 mL
of 0.01 mol/L hydrochloric acid VS (not more than
0.248z).
  (3) Sulfate <1.14>—Dissolve 0.5 g of Dibasic Calcium
Phosphate Hydrate in 5 mL of water and 5 mL of dilute
hydrochloric acid, add water to make 100 mL, and filter, if
necessary. Take 20 mL of this solution, add 1 mL of dilute
hydrochloric acid, and add water to make 50 mL. Perform
the test using this solution as the test solution. Prepare the
control solution with 1.0 mL of 0.005 mol/L sulfuric acid
VS (not more than 0.48z).
  (4) Carbonate—Mix 1.0 g of Dibasic Calcium Phos-
phate Hydrate with 5 mL of freshly boiled and cooled water,
1876       Official Monographs                                                                        Supplement I, JP XV

Add the following:                                                 total area of the peaks other than domperidone from the
                                                                   sample solution is not larger than the peak area of domperi-
Domperidone                                                        done from the standard solution.
                                                                   Operating conditions—
ドンペリドン                                                                Detector: An ultraviolet absorption photometer (wave-
                                                                   length: 287 nm).
                                                                      Column: A stainless steel column 4.6 mm in inside di-
                                                                   ameter and 25 cm in length, packed with octylsilanized silica
                                                                   gel for liquid chromatography (5 mm in particle diameter).
                                                                     Column temperature: A constant temperature of about
                                                                   359 C.
                                                                      Mobile phase: Dissolve 2.72 g of dipotassium hydrogen
                                                                   phosphate in water to make 1000 mL, and adjust the pH to
                                                                   3.5 of this solution with a solution prepared by dissolving
C22H24ClN5O2: 425.91                                               2.31 g of phosphoric acid in water to make 1000 mL. To 500
5-Chloro-1-{1-[3-(2-oxo-2,3-dihydro-1H-benzoimidazol-              mL of this solution add 500 mL of methanol.
1-yl)propyl]piperidin-4-yl}-1,3-dihydro-2H-benzoimidazol-             Flow rate: Adjust the flow rate so that the retention time
2-one [57808-66-9]                                                 of domperidone is about 9 minutes.
                                                                      Time span of measurement: About 4 times as long as the
  Domperidone, when dried, contains not less than                  retention time of domperidone, beginning after the solvent
99.0z and not more than 101.0z of C22H24ClN5O2.                    peak.
                                                                   System suitability—
Description Domperidone occurs as a white to pale yellow,
                                                                      Test for required detectability: Pipet 2 mL of the standard
crystalline powder or powder.
                                                                   solution, and add methanol to make exactly 5 mL. Confirm
  It is freely soluble in acetic acid (100), slightly soluble in
                                                                   that the peak area of domperidone obtained from 10 mL of
methanol and in ethanol (99.5), very slightly soluble in 2-
                                                                   this solution is equivalent to 30 to 50z of that from 10 mL of
propanol, and practically insoluble in water.
                                                                   the standard solution.
  Melting point: about 2439 (with decomposition).
                               C
                                                                     System performance: Dissolve 10 mg of Domperidone
Identification (1) Determine the absorption spectrum of            and 20 mg of ethyl parahydroxybenzoate in 100 mL of
a solution of Domperidone in a mixture of 2-propanol and           methanol. When the procedure is run with 10 mL of this
0.1 mol/L hydrochloric acid TS (9:1) (1 in 50,000) as direct-      solution under the above operating conditions, domperidone
ed under Ultraviolet-visible Spectrophotometry <2.24>, and         and ethyl parahydroxybenzoate are eluted in this order with
compare the spectrum with the Reference Spectrum: both             the resolution between these peaks being not less than 1.5.
spectra exhibit similar intensities of absorption at the same        System repeatability: When the test is repeated 6 times
wavelengths.                                                       with 10 mL of the standard solution under the above operat-
   (2) Determine the infrared absorption spectrum of               ing conditions, the relative standard deviation of the peak
Domperidone as directed in the potassium bromide disk              area of domperidone is not more than 3.0z.
method under Infrared Spectrophotometry <2.25>, and com-
                                                                   Loss on drying <2.41>                                C,
                                                                                            Not more than 0.5z (1 g, 1059
pare the spectrum with the Reference Spectrum: both spec-
                                                                   4 hours).
tra exhibit similar intensities of absorption at the same wave
numbers.                                                           Residue on ignition <2.44>   Not more than 0.1z (1 g).
Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of               Assay Weigh accurately about 0.5 g of Domperidone,
Domperidone according to Method 2, and perform the test.           previously dried, dissolve in 50 mL of acetic acid (100), and
Prepare the control solution with 2.0 mL of Standard Lead          titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio-
Solution (not more than 10 ppm).                                   metric titration). Perform a blank determination in the same
  (2) Related substances—Dissolve 30 mg of Domperi-                manner, and make any necessary correction.
done in 100 mL of methanol, and use this solution as the
                                                                           Each mL of 0.1 mol/L perchloric acid VS
sample solution. Pipet 1 mL of the sample solution, add
                                                                             =42.59 mg of C22H24ClN5O2
methanol to make exactly 200 mL, and use this solution as
the standard solution. Perform the test with exactly 10 mL         Containers and storage Containers—Well-closed contain-
each of the sample solution and standard solution as direct-       ers.
ed under Liquid Chromatography <2.01> according to the               Storage—Light-resistant.
following conditions, and determine each peak area of each
solution by the automatic integration method: the area of
each peak other than domperidone obtained from the sam-
ple solution is not larger than 1/2 times the peak area of
domperidone from the standard solution. Furthermore, the
Supplement I, JP XV                                                                      Official Monographs            1877

                                                                 Bacterial endotoxins <4.01>   Less than 2.50 EU/mg (poten-
Dopamine Hydrochloride Injection                                 cy).

                                                                 Uniformity of dosage units <6.02>   It meets the requirement
ドパミン塩酸塩注射液
                                                                 of the Mass variation test.
Add the following next to Extractable volume:                    Foreign insoluble matter <6.06> Perform the test according
                                                                 to Method 2: it meets the requirement.
Foreign insoluble matter <6.06> Perform the test according
to Method 1: it meets the requirement.                           Insoluble particulate matter <6.07>   It meets the require-
                                                                 ment.
Insoluble particulate matter <6.07>    It meets the require-
ment.                                                            Sterility <4.06> Perform the test according to the Mem-
                                                                 brane filtration method: it meets the requirement.
Sterility <4.06> Perform the test according to the Mem-
brane filtration method: it meets the requirement.               Assay Weigh accurately the mass of the contents of not
                                                                 less than 10 containers of Doxorubicin Hydrochloride for
                                                                 Injection. Weigh accurately an amount of the contents, e-
Add the following:                                               quivalent to about 10 mg (potency) of Doxorubicin
                                                                 Hydrochloride, add exactly 5 mL of the internal standard
Doxorubicin Hydrochloride for                                    solution and the mobile phase to make 100 mL, and use the
                                                                 solution as the sample solution. Separately, weigh accurately
Injection                                                        an amount of Doxorubicin Hydrochloride Reference Stan-
注射用ドキソルビシン塩酸塩                                                    dard, equivalent to 10 mg (potency), add exactly 5 mL of the
                                                                 internal standard solution and the mobile phase to make 100
  Doxorubicin Hydrochloride for Injection is a prepa-            mL, and use this solution as the standard solution. Perform
ration for injection, which is dissolved before use.             the test with 10 mL each of the sample solution and standard
  It contains not less than 90.0z and not more than              solution as directed under Liquid Chromatography <2.01>,
110.0z of the labeled amount of doxorubicin                      and calculate the ratios, QT and QS, of the peak area of dox-
hydrochloride (C27H29NO11.HCl: 579.98).                          orubicin to that of the internal standard.

Method of preparation Prepare as directed under Injec-             Amount [mg (potency)] of doxorubicin hydrochloride
tions, with Doxorubicin Hydrochloride.                             (C27H29NO11.HCl)
                                                                     =WS×(QT/QS)
Description Doxorubicin Hydrochloride for Injection oc-
curs as red-orange, powder or masses.                              WS: Amount [mg (potency)] of Doxorubicin Hydrochlo-
                                                                       ride Reference Standard
Identification Dissolve an amount of Doxorubicin
Hydrochloride for Injection, equivalent to 10 mg (potency)       Internal standard solution—A solution of butyl parahydrox-
of Doxorubicin Hydrochloride according to the labeled            ybenzoate in the mobile phase (1 in 1000).
amount, in methanol to make 100 mL. To 5 mL of this solu-        Operating conditions—
tion add methanol to make 50 mL, and determine the ab-              Detector: An ultraviolet absorption photometer (wave-
sorption spectrum of the solution as directed under Ultrav-      length: 254 nm).
iolet-visible Spectrophotometry <2.24>: it exhibits maxima          Column: A stainless steel column 4.6 mm in inside di-
between 231 nm and 235 nm, between 250 nm and 254 nm,            ameter and 25 cm in length, packed with octadecylsilanized
between 477 nm and 481 nm, and between 493 nm and 497            silica gel for liquid chromatography (5 mm in particle di-
nm, and exhibits a shoulder between 528 nm and 538 nm.           ameter).
                                                                    Column temperature: A constant temperature of about
pH <2.54> The pH of a solution, prepared by dissolving an        259  C.
amount of Doxorubicin Hydrochloride for Injection equiva-           Mobile phase: Dissolve 3 g of sodium lauryl sulfate in
lent to 10 mg (potency) of Doxorubicin Hydrochloride ac-         1000 mL of diluted phosphoric acid (7 in 5000). To this solu-
cording to the labeled amount in 2 mL of water, is 5.0 to 6.0.   tion add 1000 mL of acetonitrile.
Purity Clarity and color of solution—Dissolve an amount             Flow rate: Adjust the flow rate so that the retention time
of Doxorubicin Hydrochloride for Injection, equivalent to        of doxorubicin is about 8 minutes.
50 mg (potency) of Doxorubicin Hydrochloride according to        System suitability—
the labeled amount, in 10 mL of water: the solution is clear        System performance: When the procedure is run with 10
and red.                                                         mL of the standard solution under the above operating con-
                                                                 ditions, doxorubicin and the internal standard are eluted in
Water <2.48> Not more than 4.0z (0.25 g, volumetric              this order with the resolution between these peaks being not
titration, direct titration).                                    less than 5, and the symmetry factor of the peak of dox-
                                                                 orubicin is between 0.8 and 1.2.
                                                                    System repeatability: When the test is repeated 6 times
1878       Official Monographs                                                                       Supplement I, JP XV

with 10 mL of the standard solution under the above operat-          (3) Determine the infrared absorption spectrum of
ing conditions, the relative standard deviation of the ratio of   Emorfazone as directed in the potassium bromide disk
the peak area of doxorubicin to that of the internal standard     method under Infrared Spectrophotometry <2.25>, and com-
is not more than 1.0z.                                            pare the spectrum with the Reference Spectrum: both spec-
                                                                  tra exhibit similar intensities of absorption at the same wave
Containers and storage    Containers—Hermetic containers.
                                                                  numbers.

                                                                  Melting point <2.60>   89 – 929 (after drying).
                                                                                                 C
Edrophonium Chloride Injection                                    Purity (1) Chloride <1.03>—Perform the test with 1.0 g
                                                                  of Emorfazone. Prepare the control solution with 0.50 mL
エドロホニウム塩化物注射液
                                                                  of 0.01 mol/L hydrochloric acid VS (not more than 0.018
                                                                  z).
Add the following next to pH:
                                                                     (2) Heavy metals <1.07>—Proceed with 2.0 g of Emorfa-
Bacterial endotoxins <4.01>    Less than 15 EU/mg.                zone according to Method 2, and perform the test. Prepare
                                                                  the control solution with 2.0 mL of Standard Lead Solution
Add the following next to Extractable volume:                     (not more than 10 ppm).
                                                                     (3) Arsenic <1.11>—Prepare the test solution with 2.0 g
Foreign insoluble matter <6.06> Perform the test according
                                                                  of Emorfazone according to Method 3, and perform the test
to Method 1: it meets the requirement.
                                                                  (not more than 1 ppm).
Insoluble particulate matter <6.07>     It meets the require-        (4) Related substances—Conduct this procedure using
ment.                                                             light-resistant vessels. Dissolve 0.5 g of Emorfazone in 50
                                                                  mL of the mobile phase, and use this solution as the sample
Sterility <4.06> Perform the test according to the Mem-
                                                                  solution. Pipet 1 mL of the sample solution, add the mobile
brane filtration method: it meets the requirement.
                                                                  phase to make exactly 100 mL, and use this solution as the
                                                                  standard solution. Perform the test with exactly 20 mL each
                                                                  of the sample solution and standard solution as directed un-
Add the following:
                                                                  der Liquid Chromatography <2.01> according to the follow-
                                                                  ing conditions, and determine each peak area by the auto-
Emorfazone                                                        matic integration method: each peak area other than emor-
エモルファゾン                                                           fazone obtained from the sample solution is not larger than
                                                                  1/10 times the peak area of emorfazone from the standard
                                                                  solution, and the total area of the peaks other than the peak
                                                                  of emorfazone from the sample solution is not larger than
                                                                  1/2 times the peak area of emorfazone from the standard so-
                                                                  lution.
C11H17N3O3: 239.27                                                Operating conditions—
4-Ethoxy-2-methyl-5-(morpholin-4-yl)pyridazin-                       Detector: An ultraviolet absorption photometer (wave-
3(2H)-one [38957-41-4]                                            length: 254 nm).
                                                                     Column: A stainless steel column 4.6 mm in inside di-
 Emorfazone, when dried, contains not less than 98.5              ameter and 15 cm in length, packed with octadecylsilanized
z and not more than 101.0z of C11H17N3O3.                         silica gel for liquid chromatography (5 mm in particle di-
                                                                  ameter).
Description Emorfazone occurs as colorless crystals or a
                                                                     Column temperature: A constant temperature of about
white to light yellow crystalline powder.
                                                                  259  C.
   It is very soluble in ethanol (99.5), and freely soluble in
                                                                     Mobile phase: A mixture of water and methanol (11:10).
water and in acetic anhydride.
                                                                     Flow rate: Adjust the flow rate so that the retention time
   It dissolves in 1 mol/L hydrochloric acid TS.
                                                                  of emorfazone is about 5 minutes.
   It gradually turns yellow and decomposes on exposure to
                                                                     Time span of measurement: About 2.5 times as long as the
light.
                                                                  retention time of emorfazone, beginning after the solvent
Identification (1) Dissolve 20 mg of Emorfazone in 2 mL           peak.
of 1 mol/L hydrochloric acid TS, and add 5 drops of               System suitability—
Reinecke's TS: light red floating matters are formed.                Test for required detectability: Pipet 1 mL of the standard
  (2) Determine the absorption spectrum of a solution of          solution, add the mobile phase to make exactly 20 mL. Con-
Emorfazone (1 in 100,000) as directed under Ultraviolet-visi-     firm that the peak area of emorfazone obtained with 20 mL
ble Spectrophotometry <2.24>, and compare the spectrum            of this solution is equivalent to 3.5 to 6.5z of that with 20
with the Reference Spectrum: both spectra exhibit similar         mL of the standard solution.
intensities of absorption at the same wavelengths.                   System performance: Dissolve 16 mg of Emorfazone and
                                                                  30 mg of 2,4-dinitrophenylhydrazine in 100 mL of
Supplement I, JP XV                                                                        Official Monographs            1879

methanol. When the procedure is run with 20 mL of this so-        hydrochloric acid TS, shake, add 5 mL of diethyl ether, and
lution under the above operating conditions, emorfazone           shake for 5 minutes. Take 3 mL of the upper layer, distil off
and 2,4-dinitrophenylhydrazine are eluted in this order with      the diethyl ether on a water bath, add 5 mL of water to the
the resolution between these peaks being not less than 2.5.       residue with shaking, and add 1 drop of potassium perman-
  System repeatability: When the test is repeated 6 times         ganate TS: the red color of the test solution immediately dis-
with 20 mL of the standard solution under the above operat-       appears.
ing conditions, the relative standard deviation of the peak
                                                                  Optical rotation <2.49> [a]20: -41.0 – -43.59 (after
                                                                                                D
area of emorfazone is not more than 1.0z.
                                                                  drying, 0.25 g, methanol, 25 mL, 100 mm).
Loss on drying <2.41>    Not more than 0.5z (1 g, in vacu-
                                                                  Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of
um, 609 4 hours).
       C,
                                                                  Enalapril Maleate according to Method 2, and perform the
Residue on ignition <2.44>    Not more than 0.1z (1 g).           test. Prepare the control solution with 2.0 mL of Standard
                                                                  Lead Solution (not more than 10 ppm).
Assay Weigh accurately about 0.2 g of Emorfazone, previ-
                                                                     (2) Related substances—Dissolve 30 mg of Enalapril
ously dried, dissolve in 60 mL of acetic anhydride, and
                                                                  Maleate in 100 mL of a mixture of sodium dihydrogen phos-
titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio-
                                                                  phate TS, pH 2.5 and acetonitrile (19:1), and use this solu-
metric titration). Perform a blank determination in the same
                                                                  tion as the sample solution. Pipet 1 mL of the sample solu-
manner, and make any necessary correction.
                                                                  tion, add the mixture of sodium dihydrogen phosphate TS,
         Each mL of 0.1 mol/L perchloric acid VS                  pH 2.5 and acetonitrile (19:1) to make exactly 100 mL, and
           =23.93 mg of C11H17N3O3                                use this solution as the standard solution. Perform the test
                                                                  with exactly 50 mL each of the sample solution and standard
Containers and storage Containers—Tight containers.
                                                                  solution as directed under Liquid Chromatography <2.01>
 Storage—Light-resistant.
                                                                  acccording to the following conditions, and determine each
                                                                  peak area of these solutions by the automatic integration
                                                                  method: the area of the peak other than maleic acid and
Add the following:
                                                                  enalapril obtained from the sample solution is not larger
                                                                  than the peak area of enalapril from the standard solution.
Enalapril Maleate                                                 Furthermore, the total area of the peaks other than maleic
                                                                  acid and enalapril from the sample solution is not larger
エナラプリルマレイン酸塩
                                                                  than twice the peak area of enalapril from the standard solu-
                                                                  tion.
                                                                  Operating conditions—
                                                                     Detector, column, column temperature, mobile phases,
                                                                  mobile phase flow, and flow rate: Proceed as directed in the
                                                                  operating conditions in the Assay.
                                                                     Time span of measurement: About 2 times as long as the
C20H28N2O5.C4H4O4: 492.52
                                                                  retention time of enalapril, beginning after the peak of
      {
(2S)-1- (2S)-2-[(1S)-1-Ethoxycarbonyl-
                                                                  maleic acid.
3-phenylpropylamino]propanoyl}pyrrolidine-2-carboxylic
                                                                  System suitability—
acid monomaleate [76095-16-4]
                                                                     Test for required detectability: Pipet 1 mL of the standard
                                                                  solution, and add a mixture of sodium dihydrogen phos-
  Enalapril Maleate, when dried, contains not less                phate TS, pH 2.5 and acetonitrile (19:1) to make exactly 10
than 98.0z and not more than 102.0z of                            mL. Confirm that the peak area of enalapril obtained from
C20H28N2O5.C4H4O4.                                                50 mL of this solution is equivalent to 7 to 13z of that from
Description Enalapril Maleate occurs as white crystals or a       50 mL of the standard solution.
white crystalline powder.                                            System performance: When the procedure is run with 50
  It is freely soluble in methanol, sparingly soluble in water    mL of the standard solution under the above conditions, the
and in ethanol (99.5), and slightly soluble in acetonitrile.      number of theoretical plates and the symmetry factor of the
  Melting point: about 1459 (with decomposition).
                              C                                   peak of enalapril are not less than 3000 and not more than
                                                                  2.0, respectively.
Identification (1) Determine the infrared absorption
                                                                     System repeatability: When the test is repeated 6 times
spectra of Enalapril Maleate as directed in the potassium
                                                                  with 50 mL of the standard solution under the above condi-
bromide disc method under Infrared Spectrophotometry
                                                                  tions, the relative standard deviation of the peak area of
<2.25>, and compare the spectrum with the Reference Spec-
                                                                  enalapril is not more than 2.0z.
trum or the spectrum of Enalapril Maleate Reference Stan-
dard: both spectra exhibit similar intensities of absorption at   Loss on drying <2.41>    Not more than 1.0z (1 g, in vacu-
the same wave numbers.                                            um, 609 2 hours).
                                                                         C,
  (2) To 20 mg of Enalapril Maleate add 5 mL of 1 mol/L
                                                                  Residue on ignition <2.44>   Not more than 0.2z (1 g).
1880       Official Monographs                                                                     Supplement I, JP XV

                                                                Add the following:
Assay Weigh accurately about 30 mg each of Enalapril
Maleate and Enalapril Maleate Reference Standard, both
previously dried, and dissolve in a mixture of sodium di-       Enalapril Maleate Tablets
hydrogen phosphate TS, pH 2.5 and acetonitrile (19:1) to
                                                                エナラプリルマレイン酸塩錠
make exactly 100 mL, and use these solutions as the sample
solution and standard solution. Perform the test with exactly
                                                                  Enalapril Maleate Tablets contain not less than 93.0
50 mL each of the sample solution and standard solution as
                                                                z and not more than 107.0z of the labeled amount
directed under Liquid Chromatography <2.01> according to
                                                                of enalapril maleate (C20H28N2O5.C4H4O4: 492.52).
the following conditions, and determine the peak areas of
enalapril, AT and AS, of both solutions.                        Method of preparation Prepare as directed under Tablets,
                                                                with Enalapril Maleate.
           Amount (mg) of C20H28N2O5.C4H4O4
            =WS×(AT/AS)                                         Identification To a quantity of powdered Enalapril Male-
                                                                ate Tablets equivalent to 50 mg of Enalapril Maleate accord-
  WS: Amount (mg) of Enalapril Maleate Reference Stan-
                                                                ing to the labeled amount, add 20 mL of methanol, shake,
      dard
                                                                centrifuge, and then use the supernatant liquid as the sample
Operating conditions—                                           solution. Separately, dissolve 25 mg of enalapril maleate in
   Detector: An ultraviolet absorption photometer (wave-        10 mL of methanol, and use this solution as the standard
length: 215 nm).                                                solution. Perform the test with these solutions as directed
   Column: A stainless steel column 4.1 mm in inside di-        under Thin-Layer Chromatography <2.03>. Spot 20 mL each
ameter and 15 cm in length, packed with porous styrene-         of the sample solution and standard solution on a plate of
divinylbenzene copolymer for liquid chromatography (5 mm        silica gel with fluorescent indicator for thin-layer chro-
in particle diameter).                                          matography. Develop the plate with a mixture of water, ace-
   Column temperature: A constant temperature of about          tone, 1-butanol, acetic acid (100) and toluene (1:1:1:1:1) to a
709 C.                                                          distance of about 10 cm, and air-dry the plate. Examine
   Mobile phase A: Dissolve 3.1 g of sodium dihydrogen          under ultraviolet light (main wavelength: 254 nm): the Rf
phosphate dihydrate in 900 mL of water, adjust the pH to        values of the 2 spots obtained from the sample solution and
6.8 with a solution of sodium hydroxide (1 in 4), and add       the 2 spots obtained from the standard solution are equiva-
water to make 1000 mL. To 950 mL of this solution, add 50       lent.
mL of acetonitrile for liquid chromatography.
                                                                Purity Enalaprilat and enalapril diketopiperazine—Use
   Mobile phase B: Dissolve 3.1 g of sodium dihydrogen
                                                                the sample solution obtained in the Assay as the sample solu-
phosphate dihydrate in 900 mL of water, adjust the pH to
                                                                tion. Pipet 1 mL of the sample solution, add sodium di-
6.8 with a solution of sodium hydroxide (1 in 4), and add
                                                                hydrogen phosphate TS, pH 2.2 to make exactly 100 mL,
water to make 1000 mL. To 340 mL of this solution, add 660
                                                                and use this solution as the standard solution. Perform the
mL of acetonitrile for liquid chromatography.
                                                                test with exactly 50 mL each of the sample solution and stan-
   Mobile phase flow: Control the concentration gradient by
                                                                dard solution as directed under Liquid Chromatography
changing the ratio of the mobile phases A and B as follows.
                                                                <2.01> according to the following conditions, and determine
  Time after injection     Mobile phase      Mobile phase       each peak area of both solutions by the automatic integra-
    of sample (min)         A (volz)          B (volz)          tion method: the peak area of enalaprilat, having the relative
                                                                retention time of about 0.5 with respect to enalapril obtained
           0                    95                 5
                                                                from the sample solution, is not larger than 2 times the peak
         0 – 20              95 → 40            5 → 60
                                                                area of enalapril from the standard solution. Also, the peak
        20 – 25                 40                60
                                                                area of enalapril diketopiperazine, having the relative reten-
   Flow rate: 1.4 mL per minute.                                tion time of about 1.5 is not larger than the peak area of
System suitability—                                             enalapril from the standard solution.
   System performance: When the procedure is run with 50        Operating conditions—
mL of the standard solution under the above conditions, the        Proceed as directed in the operating conditions in the As-
number of theoretical plates and the symmetry factor of the     say.
peak of enalapril are not less than 3000 and not more than      System suitability—
2.0, respectively.                                                 Test for required detectability: Pipet 1 mL of the standard
   System repeatability: When the test is repeated 6 times      solution, and add sodium dihydrogen phosphate TS, pH 2.2
with 50 mL of the standard solution under the above condi-      to make exactly 10 mL. Confirm that the peak area of
tions, the relative standard deviation of the peak area of      enalapril obtained from 50 mL of this solution is equivalent
enalapril is not more than 1.0z.                                to 7 to 13z of that from 50 mL of the standard solution.
                                                                   System performance: Proceed as directed in the system
Containers and storage    Containers—Well-closed contain-
                                                                suitability in the Assay.
ers.
                                                                   System repeatability: When the test is repeated 6 times
Supplement I, JP XV                                                                      Official Monographs             1881

with 50 mL of the standard solution under the above condi-         Mobile phase: Dissolve 1.88 g of sodium dihydrogen
tions, the relative standard deviation of the peak area of      phosphate dihydrate in 900 mL of water, adjust the pH to
enalapril is not more than 2.0z.                                2.2 with phosphoric acid, and add water to make 1000 mL.
                                                                To 750 mL of this solution add 250 mL of acetonitrile.
Uniformity of dosage units <6.02> Perform the test accord-
                                                                System suitability—
ing to the following method: it meets the requirement of the
                                                                   System performance: When the procedure is run with 50
Content uniformity test.
                                                                mL of the standard solution under the above conditions, the
  Take 1 tablet of Enalapril Maleate Tablets, add V/2 mL
                                                                number of theoretical plates and the symmetry factor of the
of sodium dihydrogen phosphate TS, pH 2.2, treat with
                                                                peak of enalapril are not less than 300 and not more than
ultrasonic waves for 15 minutes, shake for 30 minutes, and
                                                                2.0, respectively.
add sodium dihydrogen phosphate TS, pH 2.2 to make
                                                                   System repeatability: When the test is repeated 6 times
exactly V mL so that 1 mL of the solution contains about 0.1
                                                                with 50 mL of the standard solution under the above condi-
mg of enalapril maleate (C20H28N2O5.C4H4O4). Treat this so-
                                                                tions, the relative standard deviation of the peak area of
lution with ultrasonic waves for 15 minutes, filter through a
                                                                enalapril is not more than 2.0z.
membrane filter with a pore size not exceeding 0.45 mm, and
use the filtrate as the sample solution. Then, proceed as       Assay Weigh accurately not less than 20 Enalapril Maleate
directed in the Assay.                                          Tablets, and powder. Weigh accurately a portion of the
                                                                powder equivalent to about 10 mg of enalapril maleate
 Amount (mg) of enalapril maleate (C20H28N2O5.C4H4O4)
                                                                (C20H28N2O5.C4H4O4), add 50 mL of sodium dihydrogen
  =WS×(AT/AS)×(V/200)
                                                                phosphate TS, pH 2.2, treat with ultrasonic waves for 15
   WS: Amount (mg) of Enalapril Maleate Reference Stan-         minutes, shake for 30 minutes, and then add sodium di-
       dard                                                     hydrogen phosphate TS, pH 2.2 to make exactly 100 mL.
                                                                Treat this solution with ultrasonic waves for 15 minutes,
Dissolution <6.10> When the test is performed at 50 revolu-
                                                                filter through a membrane filter with a pore size not exceed-
tions per minute according to the Paddle method, using 900
                                                                ing 0.45 mm, and use this filtrate as the sample solution.
mL of water as the dissolution medium, the dissolution rates
                                                                Separately, weigh accurately about 20 mg of Enalapril Male-
in 15 minutes of a 2.5- and 5-mg tablet of Enalapril Maleate
                                                                ate Reference Standard, previously dried in vacuum at 609  C
Tablets and in 30 minutes of a 10-mg tablet of Enalapril
                                                                for 2 hours, dissolve in sodium dihydrogen phosphate TS,
Maleate Tablets are not less than 85z, respectively.
                                                                pH 2.2 to make exactly 200 mL, and use this solution as the
   Start the test with 1 tablet of Enalapril Maleate Tablets,
                                                                standard solution. Perform the test with exactly 50 mL each
withdraw not less than 20 mL of the medium at the specified
                                                                of the sample solution and standard solution as directed un-
minute after starting the test, and filter through a membrane
                                                                der Liquid Chromatography <2.01> according to the follow-
filter with a pore size not exceeding 0.45 mm. Discard the
                                                                ing conditions, and determine the enalapril peak areas, AT
first 10 mL of the filtrate, pipet V mL of the subsequent
                                                                and AS, of both solutions.
filtrate, add water to make exactly V? mL so that each mL
contains about 2.8 mg of enalapril maleate (C20H28N2O5.          Amount (mg) of enalapril maleate (C20H28N2O5.C4H4O4)
C4H4O4) according to the labeled amount, and use this solu-       =WS×(AT/AS)×(1/2)
tion as the sample solution. Separately, weigh accurately
                                                                  WS: Amount (mg) of Enalapril Maleate Reference Stan-
about 14 mg of Enalapril Maleate Reference Standard,
                                                                      dard
previously dried in vacuum at 609 for 2 hours, and dissolve
                                   C
in water to make exactly 500 mL. Pipet 5 mL of this solu-       Operating conditions—
tion, add water to make exactly 50 mL, and use this solution       Detector: An ultraviolet absorption photometer (wave-
as the standard solution. Perform the test with exactly 50 mL   length: 215 nm).
each of the sample solution and standard solution as direct-       Column: A stainless steel column 4.6 mm in inside di-
ed under Liquid Chromatography <2.01> according to the          ameter and 25 cm in length, packed with octylsilanized silica
following conditions, and determine the enalapril peak          gel for liquid chromatography (5 mm in particle diameter).
areas, AT and AS, of both solutions.                              Column temperature: A constant temperature of about
                                                                509 C.
Dissolution rate (z) with respect to the labeled amount of
                                                                   Mobile phase: A mixture of sodium dihydrogen phos-
enalapril maleate (C20H28N2O5.C4H4O4)
                                                                phate TS, pH 2.2 and acetonitrile (3:1).
  =WS×(AT/AS)×(V?/V)×(1/C)×18
                                                                   Flow rate: Adjust the flow rate so that the retention time
  WS: Amount (mg) of Enalapril Maleate Reference Stan-          of enalapril is about 5 minutes.
      dard                                                      System suitability—
  C: Labeled amount (mg) of enalapril maleate                      System performance: Heat to fusion about 20 mg of
     (C20H28N2O5.C4H4O4) in 1 tablet                            enalapril maleate. After cooling, add 50 mL of acetonitrile,
                                                                and treat with ultrasonic waves to dissolve. To 1 mL of this
Operating conditions—
                                                                solution, add the standard solution to make 50 mL, and use
  Detector, column, column temperature, and flow rate:
                                                                this solution as the solution for system suitability test. When
Proceed as directed in the operating conditions in the Assay.
                                                                the procedure is run with 50 mL of the solution for system
1882       Official Monographs                                                                          Supplement I, JP XV

suitability test under the above conditions, enalapril and          Add the following:
enalapril diketopiperazine, which has a relative retention
time of 1.5 to enalapril, are eluted in this order with the reso-   Erythromycin Enteric-Coated
lution between these peaks being not less than 2.0.
  System repeatability: When the test is repeated 6 times
                                                                    Tablets
with 50 mL of the solution for system suitability test under        エリスロマイシン腸溶錠
the above conditions, the relative standard deviation of the
peak area of enalapril is not more than 1.0z.
                                                                      Erythromycin Enteric-Coated Tablets contain not
Containers and storage      Containers—Well-closed contain-         less than 90.0z and not more than 110.0z of the la-
ers.                                                                beled amount of erythromycin (C37H67NO13: 733.93).
                                                                    Method of preparation     Prepare as directed under Tablets,
                                                                    with Erythromycin.
Ephedrine Hydrochloride Injection
                                                                    Identification To a quantity of powdered Erythromycin
エフェドリン塩酸塩注射液                                                        Enteric-Coated Tablets, equivalent to 10 mg (potency) of
                                                                    Erythromycin according to the labeled amount, add 1 mL of
Add the following next to Extractable volume:                       methanol, shake well, filter, and use the filtrate as the sam-
                                                                    ple solution. Separately, dissolve 10 mg of Erythromycin
Foreign insoluble matter <6.06> Perform the test according
                                                                    Reference Standard in 1 mL of methanol, and use this solu-
to Method 1: it meets the requirement.
                                                                    tion as the standard solution. Then, proceed as directed in
Insoluble particulate matter <6.07>      It meets the require-      the Identification (2) under Erythromycin.
ment.
                                                                    Loss on drying <2.41> Not more than 10.0z (0.2 g, in
Sterility <4.06> Perform the test according to the Mem-             vacuum not exceeding 0.67 kPa, 609 3 hours).
                                                                                                     C,
brane filtration method: it meets the requirement.
                                                                    Uniformity of dosage units <6.02>    It meets the requirement
                                                                    of the Mass variation test.

Ephedrine Hydrochloride Tablets                                     Disintegration <6.09>   It meets the requirement.

                                                                    Assay Perform the test according to the Cylinder-plate
エフェドリン塩酸塩錠
                                                                    method as directed under Microbial Assay for Antibiotics
                                                                    <4.02> according to the following conditions.
Add the following next to Identification:
                                                                       (i) Test organism, culture medium, and standard solu-
Uniformity of dosage units <6.02> Perform the test accord-          tions—
ing to the following method: it meets the requirement of the        Proceed as directed in the Assay under Erythromycin.
Content uniformity test.                                               (ii) Sample solutions—Weigh accurately the mass of not
  To 1 tablet of Ephedrine Hydrochloride Tablets add V              less than 20 Erythromycin Enteric-Coated Tablets, and pul-
mL of water so that each mL contains 0.25 mg of ephedrine           verize into a powder. Weigh accurately a portion of the pow-
hydrochloride (C10H15NO.HCl), then add exactly V/4 mL of            der, equivalent to about 25 mg (potency) of Erythromycin,
the internal standard solution, disperse the tablet into small      add 25 mL of methanol, shake vigorously, add 0.1 mol/L
particles using ultrasonic waves, then stir for a further 10        phosphate buffer solution, pH 8.0, to make exactly 100 mL,
minutes in the same way. Shake this solution for 10 minutes,        and filter. Take exactly an appropriate volume of the
centrifuge, and use the supernatant liquid as the sample so-        filtrate, add 0.1 mol/L phosphate buffer solution, pH 8.0,
lution. Separately, weigh accurately about 25 mg of ephe-           to prepare solutions containing 20 mg (potency) and 5 mg
drine hydrochloride for assay, previously dried at 1059 for
                                                         C          (potency) per mL, and use these solutions as the high and
3 hours, dissolve in water to make exactly 100 mL. Pipet 20         low concentration sample solutions, respectively.
mL of this solution, add exactly 5 mL of the internal stan-
                                                                    Containers and storage    Containers—Well-closed contain-
dard solution, and use this solution as the standard solution.
                                                                    ers.
Then, proceed as directed in the Assay.

Amount (mg) of ephedrine hydrochloride (C10H15NO.HCl)
 =WS×(QT/QS)×(V/100)

  WS: Amount (mg) of ephedrine hydrochloride for assay
Internal standard solution—A           solution   of   etilefrine
hydrochloride (1 in 2000).
Supplement I, JP XV                                                                        Official Monographs           1883

Add the following:                                               Dissolution rate (z) with respect to the labeled amount of
                                                                 etizolam (C17H15ClN4S)
Etizolam Fine Granules                                              =(WS/WT)×(AT/AS)×(1/C)×(18/5)

                                                                   WS: Amount (mg) of etizolam for assay
エチゾラム細粒
                                                                   WT: Amount (g) of sample
                                                                   C: Labeled amount (mg) of etizolam (C17H15ClN4S) in 1 g
  Etizolam Fine Granules contain not less than 93.0z
and not more than 107.0z of the labeled amount of                Operating conditions—
etizolam (C17H15ClN4S: 342.85).                                     Detector: An ultraviolet absorption photometer (wave-
                                                                 length: 243 nm).
Method of preparation Prepare fine particles as directed
                                                                    Column: A stainless steel column 4.6 mm in inside di-
under Powders, with Etizolam.
                                                                 ameter and 15 cm in length, packed with octadecylsilanized
Identification (1) To a quantity of powdered Etizolam            silica gel for liquid chromatography (5 mm in particle di-
Fine Granules, equivalent to 5 mg of Etizolam according to       ameter).
the labeled amount, add 10 mL of methanol, shake, and               Column temperature: A constant temperature of about
filter through a membrane filter with a pore size not exceed-    309  C.
ing 0.45 mm. Evaporate the filtrate to dryness on a water           Mobile phase: A mixture of water and acetonitrile (1:1).
bath, cool, and then dissolve the residue in 2 mL of sulfuric       Flow rate: Adjust the flow rate so that the retention time
acid. The solution gives off a light yellow-green fluorescent    of etizolam is about 7 minutes.
when exposed to ultraviolet light (main wavelength: 365          System suitability—
nm).                                                                System performance: When the procedure is run with 50
   (2) To a quantity of powdered Etizolam Fine Granules,         mL of the standard solution under the above operating con-
equivalent to 1 mg of Etizolam according to the labeled          ditions, the number of theoretical plates and the symmetry
amount, add 80 mL of 0.1 mol/L hydrochloric acid TS,             factor of the peak of etizolam are not less than 3000 and not
shake, and then filter. Determine the absorption spectrum of     more than 2.0, respectively.
the filtrate as directed under Ultraviolet-visible Spec-            System repeatability: When the test is repeated 6 times
trophotometry <2.24>: it exhibits absorption maxima be-          with 50 mL of the standard solution under the above operat-
tween 249 nm and 253 nm, and between 292 nm and 296 nm,          ing conditions, the relative standard deviation of the peak
when perform the measurement within 10 minutes.                  area of etizolam is not more than 2.0z.
Uniformity of dosage units <6.02> The granules in single-u-      Particle size <6.03>   It meets the requirement.
nit container meet the requirement of the Mass variation
                                                                 Assay Weigh accurately an amount of Etizolam Fine Gran-
test.
                                                                 ules, equivalent to about 4 mg of etizolam (C17H15ClN4S),
Dissolution <6.10> When the test is performed at 50 revolu-      add 30 mL of water, and stir. Add 60 mL of methanol, stir
tions per minute according to the Paddle method, using 900       for 20 minutes, add methanol to make exactly 100 mL, and
mL of water as the dissolution medium, the dissolution rate      centrifuge. Pipet 5 mL of the supernatant liquid, add exactly
in 30 minutes of Etizolam Fine Granules is not less than         10 mL of the internal standard solution, add diluted
75z.                                                             methanol (7 in 10) to make 25 mL, and use this solution as
   Start the test with an accurately weighed amount of           the sample solution. Separately, weigh accurately about 0.1
Etizolam Fine Granules, equivalent to about 1 mg of etizol-      g of etizolam for assay, previously dried at 1059 for 3
                                                                                                                      C
am (C17H15ClN4S) according to the labeled amount,                hours, and dissolve in diluted methanol (7 in 10) to make
withdraw not less than 20 mL of the medium at the specified      exactly 100 mL. Pipet 2 mL of this solution, and add diluted
minute after starting the test, and filter through a membrane    methanol (7 in 10) to make exactly 100 mL. Pipet 10 mL of
filter with a pore size not exceeding 0.45 mm. Discard the       this solution, add exactly 10 mL of the internal standard
first 10 mL of filtrate, pipet the subsequent 2 mL, add exact-   solution, add diluted methanol (7 in 10) to make 25 mL, and
ly 2 mL of acetonitrile, and use this solution as the sample     use this solution as the standard solution. Perform the test
solution. Separately, weigh accurately about 28 mg of etizol-    with 10 mL each of the sample solution and standard solu-
am for assay, previously dried at 1059 for 3 hours, and dis-
                                         C                       tion as directed under Liquid Chromatography <2.01> ac-
solve in methanol to make exactly 50 mL. Pipet 5 mL of this      cording to the following conditions, and calculate the ratios,
solution, and add water to make exactly 100 mL. Pipet 4 mL       QT and QS, of the peak area of etizolam to that of the inter-
of this solution, and add water to make exactly 100 mL.          nal standard.
Pipet 2 mL of this solution, add exactly 2 mL of acetonitrile,
                                                                           Amount (mg) of etizolam (C17H15ClN4S)
and use this solution as the standard solution. Perform the
                                                                            =WS×(QT/QS)×(1/25)
test with exactly 50 mL each of the sample solution and stan-
dard solution as directed under Liquid Chromatography              WS: Amount (mg) of etizolam for assay
<2.01> according to the following conditions, and determine
                                                                 Internal standard solution—A solution of ethyl parahydrox-
the enalapril peak areas, AT and AS, of both solutions.
                                                                 ybenzoate in diluted methanol (7 in 10) (1 in 50,000).
1884       Official Monographs                                                                       Supplement I, JP XV

Operating conditions—                                             when perform the measurement within 10 minutes.
   Detector: An ultraviolet absorption photometer (wave-
                                                                  Uniformity of dosage units <6.02> Perform the test accord-
length: 240 nm).
                                                                  ing to the following method: it meets the requirement of the
   Column: A stainless steel column 4.6 mm in inside di-
                                                                  Content uniformity test.
ameter and 15 cm in length, packed with octadecylsilanized
                                                                    Take 1 tablet of Etizolam Tablets, add 2.5 mL of water,
silica gel for liquid chromatography (5 mm in particle di-
                                                                  and stir until it disintegrates. Add 20 mL of methanol, stir
ameter).
                                                                  for 20 minutes, add methanol to make exactly 25 mL, and
   Column temperature: A constant temperature of about
                                                                  centrifuge. Pipet V mL of the supernatant liquid, add exact-
359  C.
                                                                  ly 10 mL of the internal standard solution, add diluted
   Mobile phase: Dissolve 1.36 g of potassium dihydrogen
                                                                  methanol (9 in 10) to make 25 mL so that each mL contains
phosphate in water to make 1000 mL, and adjust the pH to
                                                                  about 8 mg of etizolam (C17H15ClN4S), and use this solution
3.5 with diluted phosphoric acid (1 in 10). To 550 mL of this
                                                                  as the sample solution. Then, proceed as directed in the As-
solution add 450 mL of acetonitrile.
                                                                  say.
   Flow rate: Adjust the flow rate so that the retention time
of etizolam is about 6 minutes.                                            Amount (mg) of etizolam (C17H15ClN4S)
System suitability—                                                         =WS×(QT/QS)×(1/V)×(1/20)
   System performance: When the procedure is run with 10
                                                                    WS: Amount (mg) of etizolam for assay
mL of the standard solution under the above operating con-
ditions, the internal standard and etizolam are eluted in this    Internal standard solution—A solution of ethyl parahydrox-
order with the resolution between these peaks being not less      ybenzoate in diluted methanol (9 in 10) (1 in 50,000).
than 3.
                                                                  Dissolution <6.10> When the test is performed at 50 revolu-
   System repeatability: When the test is repeated 6 times
                                                                  tions per minute according to the Paddle method, using 900
with 10 mL of the standard solution under the above operat-
                                                                  mL of water as the dissolution medium, the dissolution rate
ing conditions, the relative standard deviation of the ratio of
                                                                  in 30 minutes of Etizolam Tablets is not less than 70z.
the peak area of etizolam to that of the internal standard is
                                                                     Start the test with 1 tablet of Etizolam Tablets, withdraw
not more than 1.0z.
                                                                  not less than 20 mL of the medium at the specified minute
Containers and storage Containers—Tight containers.               after starting the test, and filter through a membrane filter
 Storage—Light-resistant.                                         with a pore size not exceeding 0.45 mm. Discard the first 10
                                                                  mL of the filtrate, pipet V mL of the subsequent filtrate, add
                                                                  water to make exactly V? mL so that each mL contains about
Add the following:                                                0.56 mg of etizolam (C17H15ClN4S) according to the labeled
                                                                  amount. Pipet 2 mL of the solution, add exactly 2 mL of
Etizolam Tablets                                                  acetonitrile, and use this solution as the sample solution.
                                                                  Separately, weigh accurately about 28 mg of etizolam for
エチゾラム錠                                                            assay, previously dried at 1059 for 3 hours, dissolve in 50
                                                                                                    C
                                                                  mL of methanol, and add water to make exactly 100 mL.
  Etizolam Tablets contain not less than 93.0z and                Pipet 5 mL of this solution, add water to make exactly 100
not more than 107.0z of the labeled amount of etizol-             mL. Pipet 4 mL of this solution, add water to make exactly
am (C17H15ClN4S: 342.85).                                         100 mL. Pipet 2 mL of this solution, add exactly 2 mL of
                                                                  acetonitrile, and use this solution as the standard solution.
Method of preparation     Prepare as directed under Tablets,
                                                                  Perform the test with exactly 50 mL each of the sample solu-
with Etizolam.
                                                                  tion and standard solution as directed under Liquid Chro-
Identification (1) To a quantity of powdered Etizolam             matography <2.01> according to the following conditions,
Tablets, equivalent to 5 mg of Etizolam according to the          and determine the peak areas of etizolam, AT and AS, of
labeled amount, add 10 mL of methanol, shake, and filter.         both solutions.
Evaporate the filtrate to dryness on a water bath, cool, and
                                                                  Dissolution rate (z) with respect to the labeled amount
then dissolve the residue in 2 mL of sulfuric acid. The solu-
                                                                  of etizolam (C17H15ClN4S)
tion gives off a light yellow-green fluorescence when exposed
                                                                    =WS×(AT/AS)×(V?/V)×(1/C)×(9/5)
to ultraviolet light (main wavelength: 365 nm).
   (2) To a quantity of powdered Etizolam Tablets, equiva-          WS: Amount (mg) of etizolam for assay
lent to 1 mg of Etizolam according to the labeled amount,           C: Labeled amount (mg) of etizolam (C17H15ClN4S) in 1
add 80 mL of 0.1 mol/L hydrochloric acid TS, shake, and                tablet
then filter through a membrane filter with a pore size not
                                                                  Operating conditions—
exceeding 0.45 mm. Determine the absorption spectrum of
                                                                     Detector: An ultraviolet absorption photometer (wave-
this filtrate as directed under Ultraviolet-visible Spec-
                                                                  length: 243 nm).
trophotometry <2.24>: it exhibits absorption maxima be-
                                                                     Column: A stainless steel column 4.6 mm in inside di-
tween 249 nm and 253 nm, and between 292 nm and 296 nm
                                                                  ameter and 15 cm in length, packed with octadecylsilanized
Supplement I, JP XV                                                                       Official Monographs             1885

silica gel for liquid chromatography (5 mm in particle di-       of etizolam is about 6 minutes.
ameter).                                                         System suitability—
   Column temperature: A constant temperature of about             System performance: When the procedure is run with 10
309  C.                                                          mL of the standard solution under the above operating con-
   Mobile phase: A mixture of water and acetonitrile (1:1).      ditions, the internal standard and etizolam are eluted in this
   Flow rate: Adjust the flow rate so that the retention time    order with the resolution between these peaks being not less
of etizolam is about 7 minutes.                                  than 3.
System suitability—                                                System repeatability: When the test is repeated 6 times
   System performance: When the procedure is run with 50         with 10 mL of the standard solution under the above operat-
mL of the standard solution under the above operating con-       ing conditions, the relative standard deviation of the ratio of
ditions, the number of theoretical plates and the symmetry       the peak area of etizolam to that of the internal standard is
factor of the peak of etizolam are not less than 3000 and not    not more than 1.0z.
more than 2.0, respectively.
                                                                 Containers and storage Containers—Tight containers.
   System repeatability: When the test is repeated 6 times
                                                                   Storage—Light-resistant.
with 50 mL of the standard solution under the above operat-
ing conditions, the relative standard deviation of the peak
area of etizolam is not more than 2.0z.
                                                                 Famotidine for Injection
Assay To 20 Etizolam Tablets add 50 mL of water, and stir
until they disintegrate. Add 400 mL of methanol, stir for 20     注射用ファモチジン
minutes, add methanol to make exactly 500 mL, and cen-
trifuge. Pipet an amount of the supernatant liquid, equiva-      Add the following next to Bacterial endotoxins:
lent to about 0.2 mg of etizolam (C17H15ClN4S), add exactly
                                                                 Uniformity of dosage units <6.02>    It meets the requirement
10 mL of the internal standard solution, add diluted
                                                                 of the Mass variation test.
methanol (9 in 10) to make 25 mL, and use this solution as
the sample solution. Separately, weigh accurately about 100      Foreign insoluble matter <6.06> Perform the test according
mg of etizolam for assay, previously dried at 1059 for 3
                                                     C           to Method 2: it meets the requirement.
hours, and dissolve in diluted methanol (9 in 10) to make
                                                                 Insoluble particulate matter <6.07>     It meets the require-
exactly 100 mL. Pipet 2 mL of this solution, and add diluted
                                                                 ment.
methanol (9 in 10) to make exactly 100 mL. Pipet 10 mL of
this solution, add exactly 10 mL of the internal standard        Sterility <4.06> Perform the test according to the Mem-
solution, add diluted methanol (9 in 10) to make 25 mL, and      brane filtration method: it meets the requirement.
use this solution as the standard solution. Perform the test
with 10 mL each of the sample solution and standard solu-
tion as directed under Liquid Chromatography <2.01> ac-          Faropenem Sodium Hydrate
cording to the following conditions, and calculate the ratios,
QT and QS, of the peak area of etizolam to that of the inter-    ファロペネムナトリウム水和物
nal standard.
                                                                 Change the Purity to read:
         Amount (mg) of etizolam (C17H15ClN4S)
          =WS×(QT/QS)×(1/500)                                    Purity
                                                                    (1) Heavy metals <1.07>—Proceed with 2.0 g of
  WS: Amount (mg) of etizolam for assay                          Faropenem Sodium Hydrate according to Method 4, and
Internal standard solution—A solution of ethyl parahydrox-       perform the test. Prepare the control solution with 2.0 mL
ybenzoate in diluted methanol (9 in 10) (1 in 50,000).           of Standard Lead Solution (not more than 10 ppm).
Operating conditions—                                               (2) Related substances—Dissolve a quantity of Faropen-
   Detector: An ultraviolet absorption photometer (wave-         em Sodium Hydrate equivalent to 0.10 g (potency) in 200
length: 240 nm).                                                 mL of water, and use this solution as the sample solution.
   Column: A stainless steel column 4.6 mm in inside di-         Pipet 2 mL of the sample solution, add water to make exact-
ameter and 15 cm in length, packed with octadecylsilanized       ly 200 mL, and use this solution as the standard solution.
silica gel for liquid chromatography (5 mm particle di-          Perform the test with exactly 20 mL each of the sample solu-
ameter).                                                         tion and standard solution as directed under Liquid Chro-
   Column temperature: A constant temperature of about           matography <2.01> according to the following conditions,
359  C.                                                          and determine each peak area by the automatic integration
   Mobile phase: Dissolve 1.36 g of potassium dihydrogen         method: the peak area of the epimer, having the relative
phosphate in water to make 1000 mL, and adjust the pH to         retention time of about 1.1 with respect to faropenem, ob-
3.5 with diluted phosphoric acid (1 in 10). To 550 mL of this    tained from the sample solution is not larger than 3/10 times
solution add 450 mL of acetonitrile.                             the peak area of faropenem from the standard solution, and
   Flow rate: Adjust the flow rate so that the retention time    the total area of the peaks other than the peak of faropenem
1886       Official Monographs                                                                          Supplement I, JP XV

from the sample solution is not larger than 1/2 times the           Operating conditions—
peak area of faropenem from the standard solution.                     Detector: An ultraviolet absorption photometer (wave-
Operating conditions—                                               length: 240 nm).
   Column, column temperature, mobile phase, and flow                  Column: A stainless steel column 4 mm in inside diameter
rate: Proceed as directed in the operating conditions in the        and 25 cm in length, packed with octadecylsilanized silica gel
Assay.                                                              for liquid chromatography (5 mm in particle diameter).
   Detector: An ultraviolet absorption photometer (wave-               Column temperature: A constant temperature of about
length: 240 nm).                                                    409 C.
   Time span of measurement: About 6 times as long as the              Mobile phase A: Dissolve 6.12 g of potassium dihydrogen
retention time of faropenem, beginning after the solvent            phosphate, 1.79 g of disodium hydrogen phosphate dodeca-
peak.                                                               hydrate and 1.61 g of tetra n-butylammonium bromide in
System suitability—                                                 water to make 1000 mL.
   Test for required detectability: To exactly 2 mL of the            Mobile phase B: A mixture of the mobile phase A and
standard solution add water to make exactly 20 mL. Con-             acetonitrile (1:1).
firm that the peak area of faropenem obtained with 20 mL of           Flowing of mobile phase: Control the gradient by mixing
this solution is equivalent to 7 to 13z of that with 20 mL of       the mobile phases A and B as directed in the following table.
the standard solution.
   System performance: When the procedure is run with 20              Time after injection      Mobile phase       Mobile phase
                                                                        of sample (min)          A (volz)           B (volz)
mL of the standard solution obtained in the Assay under the
above operating conditions, m-hydroxyacetophenone and                        0 – 54                84ª 30             16ª 70
faropenem are eluted in this order with the resolution be-
                                                                       Flow rate: About 1.5 mL per minute
tween these peaks being not less than 1.5.
                                                                       Time span of measurement: About 2.5 times as long as the
   System repeatability: When the test is repeated 6 times
                                                                    retention time of faropenem, beginning after the solvent
with 20 mL of the standard solution under the above operat-
                                                                    peak.
ing conditions, the relative standard deviation of the peak
                                                                    System suitability—
area of faropenem is not more than 2.0z.
                                                                       Test for required detectability: To exactly 2 mL of the
                                                                    standard solution add water to make exactly 20 mL. Con-
                                                                    firm that the peak area of faropenem obtained with 20 mL of
Faropenem Sodium for Syrup                                          this solution is equivalent to 7 to 13z of that with 20 mL of
                                                                    the standard solution.
シロップ用ファロペネムナトリウム
                                                                       System performance: When the procedure is run with 20
                                                                    mL of the standard solution obtained in the Assay under the
Add the following next to Identification:
                                                                    above operating conditions, m-hydroxyacetophenone and
Purity Related substances—Powder Faropenem Sodium                   faropenem are eluted in this order with the resolution be-
for Syrup, if necessary. To a part of the powder, equivalent        tween these peaks being not less than 11.
to about 25 mg (potency) of Faropenem Sodium Hydrate                   System repeatability: When the test is repeated 6 times
according to the labeled amount, add about 10 mL of water,          with 20 mL of the standard solution under the above operat-
shake well, then add water to make exactly 50 mL, and               ing conditions, the relative standard deviation of the peak
filter. Discard the first 10 mL of the filtrate, and use the sub-   area of faropenem is not more than 3.0z.
sequent filtrate as the sample solution. Pipet 2 mL of the
sample solution, add water to make exactly 200 mL, and use
this solution as the standard solution. Perform the test with       Faropenem Sodium Tablets
exactly 20 mL each of the sample solution and standard solu-
tion as directed under Liquid Chromatography <2.01> ac-             ファロペネムナトリウム錠
cording to the following conditions, and determine each
peak area of both solutions by the automatic integration            Add the following next to Identification:
method: the area of the peak of cleaved derivative, having
                                                                    Purity Related substances—Powder not less than 5
the relative retention time of about 0.71 with respect to
                                                                    Faropenem Sodium Tablets. To a part of the powder,
faropenem, obtained from the sample solution is not larger
                                                                    equivalent to about 25 mg (potency) of Faropenem Sodium
than 1.5 times the peak area of faropenem from the standard
                                                                    Hydrate according to the labeled amount, add about 10 mL
solution, and the total area of the peaks other than the peak
                                                                    of water, shake well, then add water to make exactly 50 mL,
of faropenem from the sample solution is not larger than 2
                                                                    and filter. Discard the first 10 mL of the filtrate, and use the
times the peak area of faropenem from the standard solu-
                                                                    subsequent filtrate as the sample solution. Pipet 2 mL of the
tion. For these calculations use the area of the peak of
                                                                    sample solution, add water to make exactly 200 mL, and use
cleaved derivative, having the relative retention time of 0.71,
                                                                    this solution as the standard solution. Perform the test with
after multiplying by the relative response factor 0.37.
                                                                    exactly 20 mL each of the sample solution and standard solu-
                                                                    tion as directed under Liquid Chromatography <2.01>
Supplement I, JP XV                                                                          Official Monographs          1887

according to the following conditions, and determine each         Add the following:
peak area of both solutions by the automatic integration
method: the area of the peak of cleaved derivative, having        Felbinac
the relative retention time of about 0.71 with respect to
faropenem, obtained from the sample solution is not larger        フェルビナク
than 1.5 times the peak area of faropenem from the standard
solution, and the total area of the peaks other than the peak
of faropenem from the sample solution is not larger than 2.5
times the peak area of faropenem from the standard solu-
tion. For these calculation, use the area of the peak of
                                                                  C14H12O2: 212.24
cleaved derivative, having the relative retention time of 0.71,
                                                                  Biphenyl-4-ylacetic acid    [5728-52-9]
after multiplying by the relative response factor 0.37.
Operating conditions—
                                                                    Felbinac, when dried, contains not less than 98.5z
   Detector: An ultraviolet absorption photometer (wave-
                                                                  and not more than 101.0z of C14H12O2.
length: 240 nm).
   Column: A stainless steel column 4 mm in inside diameter       Description Felbinac occurs as white to pale yellowish
and 25 cm in length, packed with octadecylsilanized silica gel    white crystals or crystalline powder.
for liquid chromatography (5 mm in particle diameter).               It is soluble in methanol and in acetone, sparingly soluble
   Column temperature: A constant temperature of about            in ethanol (95), and practically insoluble in water.
409 C.
                                                                  Identification (1) Determine the absorption spectrum of
   Mobile phase A: Dissolve 6.12 g of potassium dihydrogen
                                                                  a solution of Felbinac in methanol (95) (1 in 200,000) as
phosphate, 1.79 g of disodium hydrogen phosphate dodeca-
                                                                  directed under Ultraviolet-visible Spectrophotometry <2.24>,
hydrate and 1.61 g of tetra n-butylammonium bromide in
                                                                  and compare the spectrum with the Reference Spectrum:
water to make 1000 mL.
                                                                  both spectra exhibit similar intensities of absorption at the
   Mobile phase B: A mixture of the mobile phase A and
                                                                  same wavelengths.
acetonitrile (1:1).
                                                                    (2) Determine the infrared absorption spectrum of Fel-
   Flowing of mobile phase: Control the gradient by mixing
                                                                  binac as directed in the potassium bromide disc method un-
the mobile phases A and B as directed in the following table.
                                                                  der Infrared Spectrophotometry <2.25>, and compare the
  Time after injection      Mobile phase      Mobile phase        spectrum with the Reference Spectrum: both spectra exhibit
    of sample (min)          A (volz)          B (volz)           similar intensities of absorption at the same wave numbers.

         0 – 54                84ª 30            16ª 70           Melting point <2.60>   163 – 1669
                                                                                                  C

   Flow rate: About 1.5 mL per minute                             Purity (1) Chloride <1.03>—Dissolve 1.0 g of Felbinac in
   Time span of measurement: About 2.5 times as long as the       40 mL of acetone, add 6 mL of dilute nitric acid and water
retention time of faropenem, beginning after the solvent          to make 50 mL. Perform the test using this solution as the
peak.                                                             test solution. Prepare the control solution by combining 0.3
System suitability—                                               mL of 0.01 mol/L hydrochloric acid VS, 40 mL of acetone
   Test for required detectability: To exactly 2 mL of the        and 6 mL of dilute nitric acid, and add water to make 50 mL
standard solution add water to make exactly 20 mL. Con-           (not more than 0.011z).
firm that the peak area of faropenem obtained with 20 mL of          (2) Heavy metals <1.07>—Proceed with 1.0 g of Felbinac
this solution is equivalent to 7 to 13z of that with 20 mL of     according to Method 2, and perform the test. Prepare the
the standard solution.                                            control solution with 1.0 mL of Standard Lead Solution (not
   System performance: When the procedure is run with 20          more than 10 ppm).
mL of the standard solution obtained in the Assay under the          (3) Related substances—Dissolve 0.10 g of Felbinac in
above operating conditions, m-hydroxyacetophenone and             10 mL of acetone, and use this solution as the sample solu-
faropenem are eluted in this order with the resolution be-        tion. Pipet 2 mL of this solution, and add acetone to make
tween these peaks being not less than 11.                         exactly 100 mL. Pipet 5 mL of this solution, add acetone to
   System repeatability: When the test is repeated 6 times        make exactly 50 mL, and use this solution as the standard
with 20 mL of the standard solution under the above operat-       solution. Perform the test with these solutions as directed
ing conditions, the relative standard deviation of the peak       under Thin-layer Chromatography <2.03>. Spot 10 mL each
area of faropenem is not more than 3.0z.                          of the sample solution and standard solution on a plate of
                                                                  silica gel with fluorescent indicator for thin-layer chro-
                                                                  matography. Develop the plate with a mixture of heptane,
Add the following next to Uniformity of dosage                    acetone, and acetic acid (100) (50:25:1) to a distance of
units:                                                            about 12 cm, and air-dry the plate. Examine the plate under
                                                                  ultraviolet light (main wavelength: 254 nm): spots other than
Disintegration <6.09>    It meets the requirement.
                                                                  the principal spot from the sample solution are not more in-
1888       Official Monographs                                                                       Supplement I, JP XV

tense than the spot from the standard solution.                   hydrochloric acid and water to make exactly 100 mL. Pipet
                                                                  60 mL of this solution, add 0.5 g of zinc powder, shake fre-
Loss on drying <2.41>    Not more than 0.3z (1 g, 1059 3
                                                      C,
                                                                  quently, allow to stand for 20 minutes, and filter the solu-
hours).
                                                                  tion through a dried filter paper. Discard the first 10 mL of
Residue on ignition <2.44>    Not more than 0.1z (1 g).           the filtrate, pipet 10 mL of the subsequent filtrate, add water
                                                                  to make exactly 100 mL, and use this solution as the stan-
Assay Weigh accurately about 0.5 g of Felbinac, previous-
                                                                  dard solution. Pipet 4 mL each of the sample solution and
ly dried, dissolve in 50 mL of methanol, add 15 mL of water,
                                                                  standard solution, add 1 mL of water, 1 mL of dilute
and titrate <2.50> with 0.1 mol/L sodium hydroxide VS
                                                                  hydrochloric acid and 1 mL of sodium nitrite solution (1 in
(potentiometric titration). Perform a blank determination in
                                                                  1000) to them, mix, and allow to stand for 2 minutes. To
the same manner, and make any necessary correction.
                                                                  these solutions add 1 mL of a solution of ammonium
       Each mL of 0.1 mol/L sodium hydroxide VS                   amidosulfate (1 in 200), shake, and allow them to stand for 2
         =21.22 mg of C14H12O2                                    minutes. To these solutions add 1 mL of a solution of N,N-
                                                                  diethyl-N'-1-naphthylethylenediamine oxalate (1 in 1000),
Containers and storage     Containers—Tight containers.
                                                                  shake, allow to stand for 10 minutes, and add water to make
                                                                  exactly 20 mL. Separately, pipet 30 mL of the sample stock
                                                                  solution, add 20 mL of dilute hydrochloric acid, and add
Folic Acid Injection                                              water to make exactly 100 mL. Pipet V mL of this solution,
                                                                  and add water to make exactly V? mL so that each mL con-
葉酸注射液
                                                                  tains about 15 mg of folic acid (C19H19N7O6). With exactly 4
                                                                  mL of this solution perform the same procedure described
Add the following next to Extractable volume:
                                                                  above for obtaining the sample solution, and use the solu-
Foreign insoluble matter <6.06> Perform the test according        tion so obtained as the blank solution. Determine the absor-
to Method 1: it meets the requirement.                            bances at 550 nm, AT, AS and AC, of the solutions obtained
                                                                  from the sample solution and standard solution, and the
Insoluble particulate matter <6.07>     It meets the require-
                                                                  blank solution as directed under Ultraviolet-visible Spec-
ment.
                                                                  trophotometry <2.24>, using a control solution obtained with
Sterility <4.06> Perform the test according to the Mem-           4 mL of water in the same manner as described above.
brane filtration method: it meets the requirement.
                                                                            Amount (mg) of folic acid (C19H19N7O6)
                                                                             =WS×{(AT-AC)/AS}×(V?/V)×(1/10)

Folic Acid Tablets                                                  WS: Amount (mg) of Folic Acid Reference Standard, cal-
                                                                        culated on the anhydrous basis
葉酸錠

Add the following next to Identification:                         Delete the following two Monographs:
Uniformity of dosage units <6.02> Perform the test accord-
ing to the following method: it meets the requirement of the      Fosfestrol
Content uniformity test.                                          ホスフェストロール
   To 1 tablet of Folic Acid Tablets add 50 mL of dilute sodi-
um hydroxide TS, shake frequently, and filter. Wash the           Fosfestrol Tablets
residue with dilute sodium hydroxide TS, combine the
                                                                  ホスフェストロール錠
filtrate and the washings, then add dilute sodium hydroxide
TS to make exactly 100 mL, and use this solution as the sam-
ple stock solution. Pipet 30 mL of this solution, add 20 mL
of dilute hydrochloric acid and water to make exactly 100         Fructose Injection
mL. Pipet 60 mL of this solution, add 0.5 g of zinc powder,
                                                                  果糖注射液
shake frequently, allow to stand for 20 minutes, and filter
the solution through a dried filter paper. Discard the first 10
                                                                  Delete the Pyrogen and add the following next
mL of the filtrate, pipet V mL of the subsequent filtrate, add
                                                                  to Residue on ignition:
water to make exactly V? mL so that each mL contains about
15 mg of folic acid (C19H19N7O6), and use this solution as the    Bacterial endotoxins <4.01>    Less than 0.5 EU/mL.
sample solution. Separately, weigh accurately about 50 mg
of Folic Acid Reference Standard (separately determine the
water <2.48> in the same manner as Folic Acid), and dissolve      Add the following next to Extractable volume:
in dilute sodium hydroxide TS to make exactly 100 mL.
                                                                  Foreign insoluble matter <6.06> Perform the test according
Pipet 30 mL of this solutions, add 20 mL of dilute
                                                                  to Method 1: it meets the requirement.
Supplement I, JP XV                                                                       Official Monographs             1889

Insoluble particulate matter <6.07>    It meets the require-                                  C
                                                                 mL of water, warm to 409 to dissolve, and after cooling,
ment.                                                            add water to make exactly 50 mL. Determine the optical ro-
                                                                 tation of this solution in a 100-mm cell, within 60 minutes.
Sterility <4.06> Perform the test according to the Mem-
brane filtration method: it meets the requirement.               pH <2.54> The pH of a solution prepared by dissolving
                                                                 1.0 g of L-Glutamine in 50 mL of water is between 4.5 and
                                                                 6.0.
Gabexate Mesilate                                                Purity (1) Clarity and color of solution—A solution ob-
                                                                 tained by dissolving 0.5 g of L-Glutamine in 20 mL of water
ガベキサートメシル酸塩
                                                                 is clear and colorless.
                                                                    (2) Chloride <1.03>—Perform the test with 0.5 g of
Change the Identification (4) to read:
                                                                 L-Glutamine. Prepare the control solution with 0.30 mL of
Identification (4) A 0.1 g portion of Gabexate Mesilate          0.01 mol/L hydrochloric acid VS (not more than 0.021z).
responds to the Qualitative Tests <1.09> (1) for mesilate.          (3) Sulfate <1.14>—Perform the test with 0.6 g of
                                                                 L-Glutamine. Prepare the control solution with 0.35 mL of
                                                                 0.005 mol/L sulfuric acid VS (not more than 0.028z).
Glucose Injection                                                   (4) Ammonium <1.02>—Perform the test with 0.10 g of
                                                                 L-Glutamine, using the distillation under reduced pressure.
ブドウ糖注射液                                                          Prepare the control solution with 10.0 mL of Standard Am-
                                                                 monium Solution. The temperature of the water bath is
Add the following next to Extractable volume:                    459 (not more than 0.1z).
                                                                     C
Foreign insoluble matter <6.06> Perform the test according          (5) Heavy metals <1.07>—Proceed with 1.0 g of L-Gluta-
to Method 1: it meets the requirement.                           mine according to Method 1, and perform the test. Prepare
                                                                 the control solution with 1.0 mL of Standard Lead Solution
Insoluble particulate matter <6.07>    It meets the require-     (not more than 10 ppm).
ment.                                                               (6) Iron <1.10>—Prepare the test solution with 1.0 g of
                                                                 L-Glutamine according to Method 1, and perform the test
Sterility <4.06> Perform the test according to the Mem-
brane filtration method: it meets the requirement.               according to Method A. Prepare the control solution with
                                                                 1.0 mL of Standard Iron Solution (not more than 10 ppm).
                                                                    (7) Related substances—Dissolve 0.10 g of L-Glutamine
                                                                 in 10 mL of water, and use this solution as the sample solu-
Add the following:
                                                                 tion. Pipet 1 mL of this solution, add water to make exactly
                                                                 10 mL. Pipet 1 mL of this solution, add water to make ex-
L-Glutamine
                                                                 actly 50 mL, and use this solution as the standard solution.
L-グルタミン                                                          Perform the test with these solutions as directed under Thin-
                                                                 layer Chromatography <2.03>. Spot 5 mL each of the sample
                                                                 solution and standard solution on a plate of silica gel for
                                                                 thin-layer chromatography. Then develop with a mixture of
                                                                 1-butanol, water and acetic acid (100) (3:1:1) to a distance of
                                                                 about 10 cm, and dry the plate at 809 for 30 minutes. Spray
                                                                                                       C
C5H10N2O3: 146.14
                                                                 evenly a solution of ninhydrin in a mixture of methanol and
(2S)-2,5-Diamino-5-oxopentanoic acid      [56-85-9]
                                                                 acetic acid (100) (97:3) (1 in 100) on the plate, and heat at
                                                                 809 for 10 minutes: the spot other than the principal spot
                                                                     C
  L-Glutamine, when dried, contains not less than
                                                                 obtained with the sample solution is not more intense than
99.0z and not more than 101.0z of C5H10N2O3.
                                                                 the spot with the standard solution.
Description L-Glutamine occurs as white crystals or a crys-
                                                                 Loss on drying <2.41>    Not more than 0.3z (1 g, 1059 3
                                                                                                                      C,
talline powder. It has a slight characteristic taste.
                                                                 hours).
   It is freely soluble in formic acid, soluble in water, and
practically insoluble in ethanol (99.5).                         Residue on Ignition <2.44>    Not more than 0.1z (1 g).
Identification Determine the infrared absorption spectrum        Assay Weigh accurately about 0.15 g of L-Glutamine,
of L-Glutamine as directed in the potassium bromide disk         previously dried, dissolve in 3 mL of formic acid, add 50 mL
method under Infrared Spectrophotometry <2.25>, and com-         of acetic acid (100), and titrate <2.50> with 0.1 mol/L per-
pare the spectrum with the Reference Spectrum: both spec-        chloric acid VS (potentiometric titration). Perform a blank
tra exhibit similar intensities of absorption at the same wave   determination in the same manner, and make any necessary
numbers.                                                         correction.
Optical rotation <2.49> [a]20: +6.3 – +7.39 Weigh ac-
                             D                                            Each mL of 0.1 mol/L perchloric acid VS
curately about 2 g of L-Glutamine, previously dried, add 45                 =14.61 mg of C5H10N2O3
1890       Official Monographs                                                                       Supplement I, JP XV

Containers and storage     Containers—Tight containers.          filter through a membrane filter with a pore size not exceed-
                                                                 ing 0.5 mm, discard the first 5 mL of the filtrate, and use the
                                                                 subsequent filtrate as the sample solution. Separately, weigh
Add the following:                                               accurately an amount of Griseofulvin Reference Standard,
                                                                 equivalent to about 40 mg (potency), and dissolve in N,N-
Griseofulvin Tablets                                             dimethylformamide to make exactly 20 mL. Pipet 5 mL of
                                                                 this solution, add exactly 20 mL of the internal standard so-
グリセオフルビン錠                                                        lution, add water to make 100 mL, and use this solution as
                                                                 the standard solution. Perform the test with 10 mL each of
  Griseofulvin Tablets contain not less than 95.0z               the sample solution and standard solution as directed under
and not more than 105.0z of the labeled amount of                Liquid Chromatography <2.01> according to the following
griseofulvin (C17H17ClO6: 352.77).                               conditions, and calculate the ratios, QT and QS, of the peak
                                                                 area of griseofulvin to that of the internal standard.
Method of preparation     Prepare as directed under Tablets,
with Griseofulvin.                                                  Amount [mg (potency)] of griseofulvin (C17H17ClO6)
                                                                     =WS×(QT/QS)×(25/2)
Identification To a quantity of powdered Griseofulvin
Tablets, equivalent to 15 mg (potency) of Griseofulvin ac-         WS: Amount [mg (potency)] of Griseofulvin Reference
cording to the labeled amount, add 100 mL of ethanol (95),             Standard
shake vigorously, and filter. To 1 mL of the filtrate add
                                                                 Internal standard solution—A solution of butyl parahydrox-
ethanol (95) to make 10 mL, and determine the absorption
                                                                 ybenzoate in acetonitrile (1 in 2000).
spectrum of this solution as directed under Ultraviolet-visi-
                                                                 Operating conditions—
ble Spectrophotometry <2.24>: it exhibits maxima between
                                                                    Proceed as directed in the operating conditions in the
234 nm and 238 nm, between 290 nm and 294 nm, and be-
                                                                 Assay under Griseofulvin.
tween 323 nm and 328 nm.
                                                                 System suitability—
Uniformity of dosage units <6.02>—Perform the test accord-          System performance: When the procedure is run with 10
ing to the following method: it meets the requirement of the     mL of the standard solution under the above operating con-
Content uniformity test.                                         ditions, griseofulvin and the internal standard are eluted in
   Take 1 tablet of Griseofulvin Tablets, add V/5 mL of          this order with the resolution between these peaks being not
water, treat with ultrasonic waves to disintegrate the tablet,   less than 4.
add N,N-dimethylformamide to make 5V/8 mL, shake                    System repeatability: When the test is repeated 6 times
vigorously for 20 minutes, add N,N-dimethylformamide to          with 10 mL of the standard solution under the above operat-
make exactly V mL so that each mL contains 1.25 mg               ing conditions, the relative standard deviation of the ratio of
(potency) of Griseofulvin, and centrifuge. Pipet 8 mL of the     the peak area of griseofulvin to that of the internal standard
supernatant liquid, add exactly 20 mL of the internal stan-      is not more than 1.0z.
dard solution, add water to make 100 mL, filter through a
                                                                 Containers and storage     Containers—Tight containers.
membrane filter with a pore size not exceeding 0.5 mm, dis-
card the first 5 mL of the filtrate, and use the subsequent
filtrate as the sample solution. Then, proceed as directed un-
der the Assay.                                                   Hydralazine Hydrochloride Tablets
   Amount [mg (potency)] of griseofulvin (C17H17ClO6)            ヒドララジン塩酸塩錠
    =WS×(QT/QS)×(V/32)
                                                                 Add the following next to Identification:
  WS: Amount [mg (potency)] of Griseofulvin Reference
      Standard                                                   Uniformity of dosage units <6.02> Perform the test accord-
                                                                 ing to the following method: it meets the requirement of the
Internal standard solution—A solution of butyl parahydrox-
                                                                 Content uniformity test.
ybenzoate in acetonitrile (1 in 2000).
                                                                   To 1 tablet of Hydralazine Hydrochloride Tablets add 25
Disintegration <6.09>    It meets the requirement.               mL of 0.1 mol/L hydrochloric acid TS, disperse the tablet
                                                                 into a small particles using ultrasonic waves, then shake
Assay Weigh accurately not less than 20 Griseofulvin
                                                                 well, add 0.1 mol/L hydrochloric acid TS to make exactly 50
Tablets, and pulverize into a powder. Weigh accurately a
                                                                 mL, and centrifuge. Pipet V mL of the supernatant liquid,
portion of the powder, equivalent to about 0.5 g (potency)
                                                                 add 0.1 mol/L hydrochloric acid TS to make exactly V? mL
of Griseofulvin, add 50 mL of water, and treat with ultra-
                                                                 so that each mL contains about 10 mg of hydralazine
sonic waves. Add 100 mL of N,N-dimethylformamide,
                                                                 hydrochloride (C8H8N4.HCl), and use this solution as the
shake vigorously for 20 minutes, and add N,N-dimethylfor-
                                                                 sample solution. Separately, weigh accurately about 25 mg
mamide to make exactly 250 mL. Centrifuge this solution,
                                                                 of hydralazine hydrochloride for assay, previously dried at
pipet 5 mL of the supernatant liquid, add exactly 20 mL of
                                                                 1059 for 3 hours, dissolve in 0.1 mol/L hydrochloric acid
                                                                      C
the internal standard solution, add water to make 100 mL,
Supplement I, JP XV                                                                        Official Monographs            1891

TS to make exactly 50 mL. Pipet 2 mL of this solution, add                           C
                                                                  ously dried at 1059 for 1 hour, add 90 g of a mixture of
0.1 mol/L hydrochloric acid TS to make exactly 100 mL,            methanol and dichloromethane in equal mass ratio, and stir
and use this solution as the standard solution. Determine the     to dissolve. Determine the viscosity at 20±0.19 as directed
                                                                                                                C
absorbances at 260 nm, AT1 and AS1, and at 350 nm, AT2 and        in Method 1 under Viscosity Determination: the viscosity is
AS2, of the sample solution and standard solution as directed     not less than 80z and not more than 120z of the labeled
under Ultraviolet-visible Spectrophotometry <2.24>.               unit.

Amount (mg) of hydralazine hydrochloride (C8H8N4.HCl)             Purity (1) Chloride <1.03>—Dissolve 1.0 g of Hypromel-
 =WS×{(AT1-AT2)/(AS1—AS2)}×(V?/V)×(1/50)                          lose Phthalate in 40 mL of 0.2 mol/L sodium hydroxide VS,
                                                                  add 1 drop of phenolphthalein TS, and add dilute nitric acid
    WS: Amount (mg) of hydralazine hydrochloride for assay
                                                                  dropwise with vigorous stirring until the red color is dis-
                                                                  charged. Further add 20 mL of dilute nitric acid with stir-
                                                                  ring. Heat on a water bath with stirring until the gelatinous
Change to read:
                                                                  precipitate formed turns to granular particles. After cooling,
                                                                  centrifuge, and take off the supernatant liquid. Wash the
Hypromellose Phthalate                                            precipitate with three 20-mL portions of water by centrifug-
                                                                  ing each time, combine the supernatant liquid and the wash-
ヒプロメロースフタル酸エステル
                                                                  ings, add water to make 200 mL, and filter. Perform the test
                                                                  with 50 mL of the filtrate. Control solution: To 0.50 mL of
[9050-31-1]                                                       0.01 mol/L hydrochloric acid VS add 10 mL of 0.2 mol/L
  This monograph is harmonized with the European Phar-            sodium hydroxide VS and 7 mL of dilute nitric acid, and add
macopoeia and the U.S. Pharmacopeia. The parts of the text        water to make 50 mL (not more than 0.07z).
that are not harmonized are marked with symbols ( ).
                                                                     
                                                                       (2) Heavy metals <1.07>—Proceed with 2.0 g of
                                                                  Hypromellose Phthalate according to Method 2, and per-
  Hypromellose Phthalate is a monophthalic acid                   form the test. Prepare the control solution with 2.0 mL of
ester of hypromellose.                                            Standard Lead Solution (not more than 10 ppm).
   It contains methoxy group (-OCH3: 31.03),                         (3) Phthalic acid—Weigh accurately about 0.2 g of
hydroxypropoxy group (-OCH2CHOHCH3: 75.09),                       Hypromellose Phthalate, add about 50 mL of acetonitrile to
and carboxybenzoyl group ( - COC6H4COOH:                          dissolve partially with the aid of ultrasonic waves, add 10
149.12).                                                          mL of water, and dissolve further with the ultrasonic waves.
   It contains not less than 21.0z and not more than              After cooling, add acetonitrile to make exactly 100 mL, and
35.0z of carboxybenzoyl group, calculated on the                  use this solution as the sample solution. Separately, weigh
anhydrous basis.                                                  accurately about 12.5 mg of phthalic acid, dissolve in about
    Its substitution type and its viscosity in millipascal       125 mL of acetonitrile by mixing, add 25 mL of water, then
second (mPa・s) are shown on the label.                            add acetonitrile to make exactly 250 mL, and use this solu-
                                                                  tion as the standard solution. Perform the test with exactly
        Substitution Carboxybenzoyl group (z)                     10 mL each of the sample solution and standard solution as
         Type            Min.          Max.                       directed under Liquid Chromatography <2.01> according to
        200731           27.0          35.0                       the following conditions, and determine the peak areas of
        220824           21.0          27.0                       phthalic acid, AT and AS, of both solutions: amount of
                                                                 phthalic acid (C8H6O4: 166.13) is not more than 1.0z.

Description    Hypromellose Phthalate occurs as white             Amount (z) of phthalic acid=(WS/WT)×(AT/AS)×40
powder or granules.                                                 WS: Amount (mg) of phthalic acid
  It is practically insoluble in water, in acetonitrile and in      WT: Amount (mg) of sample, calculated on the anhydrous
ethanol (99.5).                                                         basis
  It becomes a viscous liquid when a mixture of methanol
and dichloromethane (1:1) or a mixture of ethanol (99.5)          Operating conditions—
and acetone (1:1) is added.                                          Detector: An ultraviolet absorption photometer (wave-
  It dissolves in sodium hydroxide TS.                           length: 235 nm).
                                                                     Column: A stainless steel column about 4.6 mm in inside

  Identification Determine the infrared absorption spec-          diameter and 25 cm in length, packed with octadecyl-
trum of Hypromellose Phthalate as directed in the potassi-        silanized silica gel for liquid chromatography (3 to 10 mm in
um bromide disk method under Infrared Spectrophotometry           particle diameter).
<2.25>, and compare the spectrum with the Reference Spec-            Column temperature: A constant temperature of about
trum: both spectra exhibit similar intensities of absorption at   209 C.
the same wave numbers.                                              Mobile phase: A mixture of 0.1z trifluoroacetic acid and
Viscosity <2.53>   To 10 g of Hypromellose Phthalate, previ-      acetonitrile (9:1).
                                                                     Flow rate: About 2.0 mL per minute.
1892       Official Monographs                                                                      Supplement I, JP XV

System suitability—                                              compare the spectrum with the Reference Spectrum: both
  System performance: When the procedure is run with 10
                                                                 spectra exhibit similar intensities of absorption at the same
mL of the standard solution under the above operating con-       wavelengths.
ditions, the number of theoretical plates and the symmetry         (2) Determine the infrared absorption spectrum of
factor of the peak of phthalic acid are not less than 2500 and   Ibudilast as directed in the potassium bromide disk method
not more than 1.5, respectively.                                under Infrared Spectrophotometry <2.25>, and compare the
  System repeatability: When repeat the test 6 times with 10     spectrum with the Reference Spectrum: both spectra exhibit
mL of the standard solution under the above operating con-       similar intensities of absorption at the same wave numbers.
ditions, the relative standard deviation of the peak area of
                                                                 Melting point <2.60>   54 – 589C
phthalic acid is not more than 1.0z.
                                                                 Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of
Water <2.48> Not more than 5.0z (1 g, direct titration,
                                                                 Ibudilast according to Method 2, and perform the test. Pre-
using a mixture of ethanol (99.5) and dichloromethane (3:2)
                                                                 pare the control solution with 2.0 mL of Standard Lead
instead of methanol for Karl Fischer method).
                                                                 Solution (not more than 20 ppm).
Residue on ignition <2.44>   Not more than 0.2z (1 g).              (2) Related substances—Dissolve 50 mg of Ibudilast in
                                                                 50 mL of the mobile phase, and use this solution as the
Assay Weigh accurately about 1 g of Hypromellose Phtha-
                                                                 sample solution. Pipet 1 mL of the sample solution, and add
late, dissolve in 50 mL of a mixture of ethanol (95), acetone
                                                                 the mobile phase to make exactly 50 mL. Pipet 1 mL of this
and water (2:2:1), and titrate <2.50> with 0.1 mol/L sodium
                                                                 solution, add the mobile phase to make exactly 20 mL, and
hydroxide VS (indicator: 2 drops of phenolphthalein TS).
                                                                 use this solution as the standard solution. Perform the test
Perform a blank determination in the same manner, and
                                                                 with exactly 10 mL each of the sample solution and standard
make any necessary correction.
                                                                 solution as directed under Liquid Chromatography <2.01>
       Amount (z) of carboxybenzoyl group (C8H5O3)               according to the following conditions, and determine each
        ={(0.01×149.1×V)/W}-{(2×149.1×P)/166.1}                  peak area by the automatic integration method: the peak
                                                                 area other than ibudilast obtained from the sample solution
  P: Amount (z) of phthalic acid obtained in the Purity (3)
                                                                 is not larger than the peak area of ibudilast from the stan-
  V: Amount (mL) of 0.1 mol/L sodium hydroxide VS con-
                                                                 dard solution, and the total area of the peaks other than
     sumed
                                                                 ibudilast from the sample solution is not larger than 3 times
  W: Amount (g) of sample, calculated on the anhydrous
                                                                 the peak area of ibudilast from the standard solution.
      basis
                                                                 Operating conditions—
Containers and storage    Containers—Tight containers.              Detector: An ultraviolet absorption photometer (wave-
                                                                 length: 292 nm).
                                                                    Column: A stainless steel column 2.6 mm in inside di-
Add the following:                                               ameter and 15 cm in length, packed with silica gel for liquid
                                                                 chromatography (5 mm in particle diameter).
Ibudilast                                                           Column temperature: A constant temperature of about
                                                                 259 C.
イブジラスト                                                              Mobile phase: A mixture of hexane and ethyl acetate
                                                                 (50:1)
                                                                    Flow rate: Adjust the flow rate so that the retention time
                                                                 of ibudilast is about 9 minutes.
                                                                    Time span of measurement: About 4 times as long as the
                                                                 retention time of ibudilast, beginning after the solvent peak.
                                                                 System suitability—
C14H18N2O: 230.31                                                   Test for required detectability: To exactly 5 mL of the
1-[2-(1-Methylethyl)pyrazolo[1,5-a]pyridin-3-yl]-                standard solution add the mobile phase to make exactly 10
2-methylpropan-1-one [50847-11-5]                                mL. Confirm that the peak area of ibudilast obtained with
                                                                 10 mL of this solution is equivalent to 40 to 60z of that with
  Ibudilast, when dried, contains not less than 98.5z            10 mL of the standard solution.
and not more than 101.0z of C14H18N2O.                              System performance: To 5 mL of the sample solution add
Description Ibudilast occurs as a white crystalline powder.      the mobile phase to make 50 mL. To 2 mL of this solution
  It is very soluble in methanol, freely soluble in ethanol      add the mobile phase to make 20 mL. When the procedure is
(99.5) and in acetic anhydride, and very slightly soluble in     run with 10 mL of this solution under the above operating
water.                                                           conditions, the number of theoretical plates and the symmet-
                                                                 ry factor of the peak of ibudilast are not less than 3500 and
Identification (1) Determine the absorption spectrum of          not more than 2.0, respectively.
a solution of Ibudilast in methanol (1 in 250,000) as directed      System repeatability: When the test is repeated 6 times
under Ultraviolet-visible Spectrophotometry <2.24>, and          with 10 mL of the standard solution under the above operat-
Supplement I, JP XV                                                                       Official Monographs            1893

ing conditions, the relative standard deviation of the peak        WS: Amount (mg) of Imipramine Hydrochloride Refer-
area of ibudilast is not more than 3.0z.                               ence Standard

Loss on drying <2.41>    Not more than 0.3z (1 g, in vacu-
um, 4 hours).
                                                                 Indometacin Capsules
Residue on ignition <2.44>   Not more than 0.1z (1 g).
                                                                 インドメタシンカプセル
Assay Weigh accurately about 0.2 g of Ibudilast, previous-
ly dried, dissolve in 50 mL of acetic anhydride, and titrate
                                                                 Add the following next to Purity:
<2.50> with 0.1 mol/L perchloric acid VS (potentiometric
titration). Perform a blank determination in the same            Uniformity of dosage units <6.02> Perform the test accord-
manner, and make any necessary correction.                       ing to the following method: it meets the requirement of the
                                                                 Content uniformity test.
        Each mL of 0.1 mol/L perchloric acid VS
                                                                    Take out the content of 1 capsule of Indometacin Cap-
          =23.03 mg of C14H18N2O
                                                                 sules, and dissolve in methanol to make exactly V mL so that
Containers and storage    Containers—Tight containers.           each mL contains about 1 mg of indometacin (C19H16ClNO4).
                                                                 Filter the solution, discard the first 10 mL of the filtrate,
                                                                 pipet 5 mL of the subsequent filtrate, add exactly 3 mL of
Idoxuridine Ophthalmic Solution                                  the internal standard solution, then add the mobile phase to
                                                                 make 100 mL, and use this solution as the sample solution.
イドクスウリジン点眼液                                                      Separately, weigh accurately about 25 mg of Indometacin
                                                                 Reference Standard, previously dried at 1059 for 4 hours,
                                                                                                                C
Add the following next to Purity:                                dissolve in methanol to make exactly 25 mL. Pipet 5 mL of
                                                                 this solution, add exactly 3 mL of the internal standard solu-
Foreign insoluble matter <6.11>    It meets the requirement.
                                                                 tion, then add the mobile phase to make 100 mL, and use
Insoluble particulate matter <6.08>    It meets the require-     this solution as the standard solution. Then, proceed as
ment.                                                            directed in the Assay.

Sterility <4.06> Perform the test according to the Mem-                 Amount (mg) of indometacin (C19H16ClNO4)
brane filtration method: it meets the requirement.                       =WS×(QT/QS)×(V/25)

                                                                   WS: Amount (mg) of Indometacin Reference Standard

Imipramine Hydrochloride Tablets                                 Internal standard solution—A solution of butyl parahydrox-
                                                                 ybenzoate in methanol (1 in 1000).
イミプラミン塩酸塩錠

Add the following next to Identification:                        Isotonic Sodium Chloride Solution
Uniformity of dosage units <6.02> Perform the test accord-
                                                                 生理食塩液
ing to the following method: it meets the requirement of the
Content uniformity test.
                                                                 Add the following next to Extractable volume:
   To 1 tablet of Imipramine Hydrochloride Tablets add ex-
actly 40 mL of 0.01 mol/L hydrochloric acid TS, disperse         Foreign insoluble matter <6.06> Perform the test according
the tablet into a small particles using ultrasonic waves, then   to Method 1: it meets the requirement.
shake well. Centrifuge the solution, pipet V mL of the super-
                                                                 Insoluble particulate matter <6.07>    It meets the require-
natant liquid, add water to make exactly V? mL so that each
                                                                 ment.
mL contains about 20 mg of imipramine hydrochloride
(C19H24N2.HCl), and use this solution as the sample solu-        Sterility <4.06> Perform the test according to the Mem-
tion. Separately, weigh accurately about 25 mg of Imipra-        brane filtration method: it meets the requirement.
mine Hydrochloride Reference Standard, previously dried at
1059 for 2 hours, dissolve in 0.01 mol/L hydrochloric acid
     C
TS to make exactly 100 mL. Pipet 2 mL of this solution, add
water to make exactly 25 mL, and use this solution as the
standard solution. Determine the absorbances at 251 nm,
AT1 and AS1, and at 330 nm, AT2 and AS2, of the sample solu-
tion and standard solution as directed under Ultraviolet-visi-
ble Spectrophotometry <2.24>.

Amount (mg) of imipramine hydrochloride (C19H24N2.HCl)
 =WS×{(AT1-AT2)/(AS1-AS2)}×(V?/V)×(4/125)
1894       Official Monographs                                                                     Supplement I, JP XV

Add the following:                                              solution as directed under Liquid Chromatography <2.01>
                                                                according to the following conditions. Determine each peak
Isoxsuprine Hydrochloride                                       area by the automatic integration method: each peak area
                                                                other than isoxsuprine obtained from the sample solution is
イソクスプリン塩酸塩                                                      not larger than the peak area of isoxsuprine from the stan-
                                                                dard solution, and the total area of the peaks other than the
                                                                peak of isoxsuprine from the sample solution is not larger
                                                                than 2 times the peak area of isoxsuprine from the standard
                                                                solution.
                                                                Operating conditions—
C18H23NO3.HCl: 337.84                                              Detector: An ultraviolet absorption photometer (wave-
(1RS,2SR)-1-(4-Hydroxyphenyl)-2-{[(2SR)-1-                      length: 269 nm).
phenoxypropan-2-yl]amino}propan-1-ol                               Column: A stainless steel column 4.6 mm in inside di-
monohydrochloride [579-56-6]                                    ameter and 25 cm in length, packed with octadecylsilanized
                                                                silica gel for liquid chromatography (5 mm in particle di-
  Isoxsuprine Hydrochloride, when dried, contains               ameter).
not less than 99.0z and not more than 101.0z of                    Column temperature: A constant temperature of about
C18H23NO3.HCl.                                                  409  C.
                                                                   Mobile phase: Dissolve 4.3 g of diammonium hydrogen
Description Isoxsuprine Hydrochloride occurs as a white,
                                                                phosphate and 3.2 g of sodium 1-pentane sulfonate in water
powder or crystalline powder.
                                                                to make 1000 mL, and adjust to pH 2.5 with phosphoric
  It is soluble in formic acid and in methanol, and slightly
                                                                acid. To 770 mL of this solution add 230 mL of acetonitrile.
soluble in water and in ethanol (99.5).
                                                                   Flow rate: Adjust the flow rate so that the retention time
  Melting point: about 2049 (with decomposition).
                              C
                                                                of isoxsuprine is about 18 minutes.
  A solution of Isoxsuprine Hydrochloride in methanol (1 in
                                                                   Time span of measurement: About 3 times as long as the
50) shows no optical rotation.
                                                                retention time of isoxsuprine, beginning after the solvent
Identification (1) Determine the absorption spectrum of         peak.
a solution of Isoxsuprine Hydrochloride (1 in 20,000) as        System suitability—
directed under Ultraviolet-visible Spectrophotometry <2.24>,       Test for required detectability: Pipet 1 mL of the standard
and compare the spectrum with the Reference Spectrum:           solution, and add the mobile phase to make exactly 10 mL.
both spectra exhibit similar intensities of absorption at the   Confirm that the peak area of isoxsuprine obtained with 10
same wavelengths.                                               mL of this solution is equivalent to 7 to 13z of that with 10
  (2) Determine the infrared absorption spectrum of Isox-       mL of the standard solution.
suprine Hydrochloride as directed in the potassium chloride        System performance: To 1 mL of the sample solution add
disk method under Infrared Spectrophotometry <2.25>, and        2.5 mL of a solution of methyl parahydroxybenzoate (1 in
compare the spectrum with the Reference Spectrum: both          25,000) and the mobile phase to make 50 mL. When the
spectra exhibit similar intensities of absorption at the same   procedure is run with 10 mL of this solution under the above
wave numbers.                                                   operating conditions, methyl parahydroxybenzoate and
  (3) Dissolve 0.5 g of Isoxsuprine Hydrochloride in 50         isoxsuprine are eluted in this order with the resolution be-
mL of water by warming, and cool: the solution responds to      tween these peaks being not less than 4.
the Qualitative Tests <1.09> (2) for chloride.                     System repeatability: When the test is repeated 6 times
                                                                with 10 mL of the standard solution under the above operat-
pH <2.54> Dissolve 0.5 g of Isoxsuprine Hydrochloride in
                                                                ing conditions, the relative standard deviation of the peak
50 mL of water by warming, and cool: the pH of the solu-
                                                                area of isoxsuprine is not more than 2.5z.
tion is between 4.5 and 6.0.
                                                                Loss on drying <2.41>                               C,
                                                                                        Not more than 0.5z (1 g, 1059 1
Purity (1) Clarity and color of solution—Dissolve 0.1 g
                                                                hour).
of Isoxsuprine Hydrochloride in 10 mL of water, warm if
necessary, and cool: the solution is clear and colorless.       Residue on ignition <2.44>   Not more than 0.2z (1 g).
  (2) Heavy metals <1.07>—Proceed with 1.0 g of Isox-
                                                                Assay Weigh accurately about 0.3 g of Isoxsuprine
suprine Hydrochloride according to Method 2, and perform
                                                                Hydrochloride, previously dried, dissolve in 5 mL of formic
the test. Prepare the control solution with 2.0 mL of Stan-
                                                                acid, add 50 mL of a mixture of acetic anhydride and acetic
dard Lead Solution (not more than 20 ppm).
                                                                acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric
  (3) Related substances—Dissolve 20 mg of Isoxsuprine
                                                                acid VS (potentiometric titration). Perform a blank determi-
Hydrochloride in 20 mL of the mobile phase, and use this
                                                                nation in the same manner, and make any necessary correc-
solution as the sample solution. Pipet 1 mL of the sample
                                                                tion.
solution, add the mobile phase to make exactly 100 mL, and
use this solution as the standard solution. Perform the test            Each mL of 0.1 mol/L perchloric acid VS
with exactly 10 mL each of the sample solution and standard               =33.78 mg of C18H23NO3.HCl
Supplement I, JP XV                                                                      Official Monographs            1895

Containers and storage    Containers—Well-closed contain-                                     C
                                                                 say, previously dried at 1059 for 1 hour, and dissolve in
ers.                                                             water to make exactly 100 mL. Pipet 4 mL of this solution,
                                                                 add water to make exactly 100 mL, and use this solution as
                                                                 the standard solution. Perform the test with exactly 10 mL
Add the following:                                               each of the sample solution and standard solution as direct-
                                                                 ed under Liquid Chromatography <2.01> according to the
Isoxsuprine Hydrochloride Tablets                                following conditions, and determine the isoxsuprine peak
                                                                 areas, AT and AS, of both solutions.
イソクスプリン塩酸塩錠
                                                                 Dissolution rate (z) with respect to the labeled amount of
                                                                 isoxsuprine hydrochloride (C18H23NO3.HCl)
  Isoxsuprine Hydrochloride Tablets contain not less
                                                                    =WS×(AT/AS)×(V?/V)×(1/C)×36
than 95.0z and not more than 105.0z of the labeled
amount of isoxsuprine hydrochloride (C18H23NO3.                    WS: Amount (mg) of isoxsuprine hydrochloride for assay
HCl: 337.84).                                                      C: Labeled amount (mg) of isoxsuprine hydrochloride
                                                                      (C18H23NO3.HCl) in 1 tablet
Method of preparation Prepare as directed under Tablets,
with Isoxsuprine Hydrochloride.                                  Operating conditions—
                                                                   Proceed as directed in the operating conditions in the
Identification To a quantity of powdered Isoxsuprine
                                                                 Assay.
Hydrochloride Tablets, equivalent to 10 mg of Isoxsuprine
                                                                 System suitability—
Hydrochloride according to the labeled amount, add 150
                                                                   System performance: When the procedure is run with 10
mL of water, shake, and then add water to make 200 mL.
                                                                 mL of the standard solution under the above operating con-
Centrifuge this solution, filter the supernatant liquid
                                                                 ditions, the number of theoretical plates and the symmetry
through a membrane filter with a pore size not exceeding
                                                                 factor of the peak of isoxsuprine are not less than 2000 and
0.45 mm, discard the first 10 mL of filtrate, and determine
                                                                 not more than 2.0, respectively.
the absorption spectrum of the subsequent filtrate as direct-
                                                                   System repeatability: When the test is repeated 6 times
ed under Ultraviolet-visible Spectrophotometry <2.24>: it ex-
                                                                 with 10 mL of the standard solution under the above operat-
hibits maxima between 267 nm and 271 nm, and between
                                                                 ing conditions, the relative standard deviation of the peak
272 nm and 276 nm.
                                                                 area of isoxsuprine is not more than 2.0z.
Uniformity of dosage units <6.02> Perform the test accord-
                                                                 Assay Weigh accurately not less than 20 Isoxsuprine
ing to the following method: it meets the requirement of the
                                                                 Hydrochloride Tablets, and powder. Weigh accurately a
Content uniformity test.
                                                                 portion of the powder equivalent to about 40 mg of isox-
  Add methanol to 1 tablet of Isoxsuprine Hydrochloride
                                                                 suprine hydrochloride (C18H23NO3.HCl), add 60 mL of
Tablets, and shake to disintegrate. Add methanol to make
                                                                 methanol, shake for 20 minutes, and then add methanol to
exactly V mL so that each mL contains about 0.4 mg of isox-
                                                                 make exactly 100 mL. Centrifuge a portion of this solution,
suprine hydrochloride (C18H23NO3.HCl). Centrifuge this so-
                                                                 filter the supernatant liquid through a membrane filter with
lution, and use the supernatant liquid as the sample solution.
                                                                 a pore size not exceeding 0.45 mm, discard the first 10 mL of
Then, proceed as directed in the Assay.
                                                                 filtrate, and use the subsequent filtrate as the sample solu-
        Amount (mg) of isoxsuprine hydrochloride                 tion. Separately, weigh accurately about 40 mg of isox-
        (C18H23NO3.HCl)                                          suprine hydrochloride for assay, previously dried at 1059   C
          =WS×(AT/AS)×V×(1/100)                                  for 1 hour, and dissolve in methanol to make exactly 100
                                                                 mL. Filter through a membrane filter with a pore size not ex-
  WS: Amount (mg) of isoxsuprine hydrochloride for assay
                                                                 ceeding 0.45 mm, discard the first 10 mL of the filtrate, and
Dissolution <6.10> When the test is performed at 50 revolu-      use the subsequent filtrate as the standard solution. Perform
tions per minute according to the Paddle method, using 900       the test with exactly 10 mL each of the sample solution and
mL of water as the dissolution medium, the dissolution rate      standard solution as directed under Liquid Chromatography
in 15 minutes of Isoxsuprine Hydrochloride Tablets is not        <2.01> according to the following conditions, and determine
less than 80z.                                                   the peak areas, AT and AS, of isoxsuprine in each solution.
   Start the test with 1 tablet of Isoxsuprine Hydrochloride             Amount (mg) of isoxsuprine hydrochloride
Tablets, withdraw not less than 20 mL of the medium at the               (C18H23NO3.HCl)
specified minute after starting the test, and filter through a             =WS×(AT/AS)
membrane filter with a pore size not exceeding 0.45 mm. Dis-
card the first 10 mL of the filtrate, pipet V mL of the subse-     WS: Amount (mg) of isoxsuprine hydrochloride for assay
quent filtrate, add water to make exactly V? mL so that each     Operating conditions—
mL contains about 11 mg of isoxsuprine hydrochloride                Detector: An ultraviolet absorption photometer (wave-
(C18H23NO3.HCl) according to the labeled amount, and use         length: 269 nm).
this solution as the sample solution. Separately, weigh ac-         Column: A stainless steel column 4.6 mm in inside di-
curately about 28 mg of isoxsuprine hydrochloride for as-        ameter and 15 cm in length, packed with octadecylsilanized
1896       Official Monographs                                                                     Supplement I, JP XV

silica gel for liquid chromatography (5 mm in particle di-      Description Itraconazole occurs as a white powder.
ameter).                                                           It is soluble in N,N-dimethylformamide, very slightly
   Column temperature: A constant temperature of about          soluble in ethanol (99.5), and practically insoluble in water
409  C.                                                         and in 2-propanol.
   Mobile phase: Dissolve 4.3 g of diammonium hydrogen             A solution of Itraconazole in N,N-dimethylformamide (1
phosphate and 3.2 g of sodium 1-pentane sulfonate in water      in 100) shows no optical rotation.
to make 1000 mL, and adjust to pH 2.5 with phosphoric
                                                                Identification (1) Determine the absorption spectrum of
acid. To 600 mL of this solution add 400 mL of methanol.
                                                                a solution of Itraconazole in 2-propanol (1 in 100,000) as
   Flow rate: Adjust the flow rate so that the retention time
                                                                directed under Ultraviolet-visible Spectrophotometry <2.24>,
of isoxsuprine is about 9 minutes.
                                                                and compare the spectrum with the Reference Spectrum:
System suitability—
                                                                both spectra exhibit similar intensities of absorption at the
   System performance: To exactly 1 mL of the standard
                                                                same wavelengths.
solution add the mobile phase to make exactly 50 mL. When
                                                                   (2) Determine the infrared absorption spectrum of
the procedure is run with 10 mL of this solution under the
                                                                Itraconazole, previously dried, as directed in the potassium
above operating conditions, the number of theoretical plates
                                                                bromide disk method under Infrared Spectrophotometry
and the symmetry factor of the peak of isoxsuprine are not
                                                                <2.25>, and compare the spectrum with the Reference Spec-
less than 2000 and not more than 2.0, respectively.
                                                                trum: both spectra exhibit similar intensities of absorption at
   System repeatability: When the test is repeated 6 times
                                                                the same wave numbers.
with 10 mL of the standard solution under the above operat-
                                                                   (3) Perform the test with Itraconazole as directed under
ing conditions, the relative standard deviation of the peak
                                                                Flame Coloration Test <1.04> (2): a green color appears.
area of isoxsuprine is not more than 1.0z.
                                                                Melting point <2.60>    166 – 1709
                                                                                                 C
Containers and storage    Containers—Well-closed contain-
ers.                                                            Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of
                                                                Itraconazole according to Method 2, and perform the test.
                                                                Prepare the control solution with 2.0 mL of Standard Lead
Add the following:                                              Solution (not more than 20 ppm).
                                                                   (2) Related substances—Dissolve 0.10 g of Itraconazole
Itraconazole                                                    in 10 mL of a mixture of methanol and tetrahydrofuran
                                                                (1:1), and use this solution as the sample solution. Pipet 1
イトラコナゾール                                                        mL of the sample solution, add the mixture of methanol and
                                                                tetrahydrofuran (1:1) to make exactly 100 mL. Pipet 5 mL
                                                                of this solution, add the mixture of methanol and tetra-
                                                                hydrofuran (1:1) to make exactly 10 mL, and use this solu-
                                                                tion as the standard solution. Perform the test with exactly
                                                                10 mL each of the sample solution and standard solution as
                                                                directed under Liquid Chromatography <2.01> according to
                                                                the following conditions. Determine each peak area of each
                                                                solution by the automatic integration method: the area of
                                                                each peak other than itraconazole obtained from the sample
                                                                solution is not larger than the peak area of itraconazole from
                                                                the standard solution. Furthermore, the total area of the
                                                                peaks other than itraconazole from the sample solution is
                                                                not larger than 2.5 times the peak area of itraconazole from
                                                                the standard solution.
C35H38Cl2N8O4: 705.63                                           Operating conditions—
4-(4-{4-[4-({(2RS,4SR)-2-(2,4-Dichlorophenyl)-                     Detector: An ultraviolet absorption photometer (wave-
2-[(1H-1,2,4-triazol-1-yl)methyl]-1,3-dioxolan-                 length: 225 nm).
4-yl}methoxy)phenyl]piperazin-1-yl}phenyl)-2-[(1RS)-               Column: A stainless steel column 4.6 mm in inside di-
1-methylpropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one              ameter and 10 cm in length, packed with octadecylsilanized
4-(4-{4-[4-({(2SR,4RS)-2-(2,4-Dichlorophenyl)-                  silica gel for liquid chromatography (3 mm in particle di-
2-[(1H-1,2,4-triazol-1-yl)methyl]-1,3-dioxolan-                 ameter).
4-yl}methoxy)phenyl]piperazin-1-yl}phenyl)-2-[(1RS)-               Column temperature: A constant temperature of about
1-methylpropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one              309  C.
[84625-61-6]                                                       Mobile phase A: A solution of Tetrabutylammonium
                                                                hydrogensulfate (17 in 625).
  Itraconazole contains not less than 98.5z and not                Mobile phase B: Acetonitrile.
more than 101.0z of C35H38Cl2N8O4, calculated on                   Flowing of the mobile phase: Control the gradient by mix-
the dried basis.
Supplement I, JP XV                                                                       Official Monographs           1897

ing the mobile phases A and B as directed in the following       Identification To a quantity of powdered Josamycin
table.                                                           Tablets, equivalent to 10 mg (potency) of Josamycin accord-
                                                                 ing to the labeled amount, add 100 mL of methanol, shake
  Time after injection       Mobile phase    Mobile phase        vigorously, and centrifuge. To 5 mL of the supernatant liq-
    of sample (min)           A (volz)        B (volz)
                                                                 uid, add methanol to make 50 mL, and determine the ab-
          0 – 20               80ª 50            20ª 50          sorption spectrum of this solution as directed under Ultrav-
         20 – 25                 50                50            iolet-visible Spectrophotometry <2.24>: it exhibits a maxi-
                                                                 mum between 229 nm and 233 nm.
   Flow rate: 1.5 mL per minute.
   Time span of measurement: About 2 times as long as the        Loss on drying <2.41>    Not more than 5.0z (0.5 g, in vacu-
retention time of itraconazole, beginning after the solvent      um, 609 3 hours).
                                                                         C,
peak.                                                            Uniformity of dosage units <6.02>—Perform the test accord-
System suitability—                                              ing to the following method: it meets the requirement of the
   Test for required detectability: To exactly 1 mL of the       Content uniformity test.
standard solution add the mixture of methanol and tetra-            Take 1 tablet of Josamycin Tablets, add 5 mL of water,
hydrofuran (1:1) to make exactly 10 mL. Confirm that the         and shake vigorously to disintegrate the tablet. Add
peak area of itraconazole obtained from 10 mL of this solu-      methanol and then use ultrasonic waves to disperse the parti-
tion is equivalent to 7 to 13z of that from 10 mL of the stan-   cles, add methanol to make exactly V mL so that each mL
dard solution.                                                   contains about 2 mg (potency) of Josamycin, and cen-
   System performance: Dissolve 1 mg of Itraconazole and 1       trifuge. Pipet 3 mL of the supernatant liquid, and add
mg of miconazole nitrate in 20 mL of the mixture of              methanol to make exactly 100 mL. Pipet 10 mL of this solu-
methanol and tetrahydrofuran (1:1). When the procedure is        tion, add methanol to make exactly 50 mL, and use this solu-
run with 10 mL of this solution under the above operating        tion as the sample solution. Separately, accurately weigh
conditions, miconazole and itraconazole are eluted in this       about 50 mg (potency) of Josamycin Reference Standard,
order with the resolution between these peaks being not less     dissolve in 5 mL of water and methanol to make exactly 25
than 2.0.                                                        mL. Pipet 3 mL of this solution, and add methanol to make
   System repeatability: When the test is repeated 6 times       exactly 100 mL. Pipet 10 mL of this solution, add methanol
with 10 mL of the standard solution under the above operat-      to make exactly 50 mL, and use this solution as the standard
ing conditions, the relative standard deviation of the peak      solution. Determine the absorbances, AT and AS, of the
area of itraconazole is not more than 2.0z.                      sample solution and the standard solution at 231 nm as
Loss on drying <2.41>    Not more than 0.5z (1 g, 1059 4
                                                      C,         directed under Ultraviolet-visible Spectrophotometry <2.24>.
                                                                             —
hours).                                                          However, X in the formula for calculation of acceptance
                                                                 value is the result of the assay.
Residue on ignition <2.44>     Not more than 0.1z (1 g).
                                                                     Amount [mg (potency)] of josamycin (C42H69NO15)
Assay Weigh accurately about 0.3 g of Itraconazole, dis-              =WS×(AT/AS)×(V/25)
solve in 70 mL of a mixture of 2-butanone and acetic acid
(100) (7:1), and titrate <2.50> with 0.1 mol/L perchloric acid     WS: Amount [mg (potency)] of Josamycin Reference
VS (potentiometric titration). Perform a blank determina-              Standard
tion in the same manner, and make any necessary correc-          Disintegration <6.09>    It meets the requirement.
tion.
                                                                 Assay Perform the test according to the Cylinder-plate
        Each mL of 0.1 mol/L perchloric acid VS                  method as directed under Microbial Assay for Antibiotics
          =35.28 mg of C35H38Cl2N8O4                             <4.02> according to the following conditions.
Containers and storage    Containers—Tight containers.              (i) Test organism, culture medium, and standard
                                                                 solutions—Proceed as directed in the Assay under Josamy-
                                                                 cin.
Add the following:                                                  (ii) Sample solutions—Weigh accurately the mass of not
                                                                 less than 20 Josamycin Tablets and pulverize into a powder.
Josamycin Tablets                                                Weigh accurately a portion of the powder, equivalent to
                                                                 about 0.3 g (potency) of Josamycin, add 50 mL of
ジョサマイシン錠                                                         methanol, shake vigorously, and add water to make exactly
                                                                 1000 mL. Take exactly an appropriate amount of this solu-
  Josamycin Tablets contain not less than 90.0z and              tion, add water to prepare solutions containing 30 mg (poten-
not more than 110.0z of the labeled amount of                    cy) and 7.5 mg (potency) per mL, and use these solutions as
josamycin (C42H69NO15: 827.99).                                  the high and low concentration sample solutions, respective-
                                                                 ly.
Method of preparation     Prepare as directed under Tablets,
with Josamycin.                                                  Containers and storage     Containers—Tight containers.
1898       Official Monographs                                                                      Supplement I, JP XV

Add the following:                                               Standard Lead Solution (not more than 20 ppm).
                                                                    (2) Related substances—Dissolve 0.8 g of Labetalol
Labetalol Hydrochloride                                          Hydrochloride in 10 mL of methanol, and use this solution
                                                                 as the sample solution. Pipet 1 mL of the sample solution,
ラベタロール塩酸塩                                                        add methanol to make exactly 200 mL, and use this solution
                                                                 as the standard solution. Perform the test with these solu-
                                                                 tions as directed under Thin-layer Chromatography <2.03>.
                                                                 Spot 5 mL each of the sample solution and standard solution
                                                                 on a plate of silica gel for thin-layer chromatography. De-
                                                                 velop the plate with a mixture of ethyl acetate, 2-propanol,
                                                                 water, and ammonia solution (28) (25:15:8:2) to a distance
                                                                 of about 10 cm, and air-dry the plate. Allow the plate to
                                                                 stand in iodine vapor for 30 minutes: the spots other than
                                                                 the principal spot from the sample solution do not exceed 2
                                                                 in number and are not more intense than the spot obtained
                                                                 from the standard solution.

                                                                 Loss on drying <2.41>    Not more than 1.0z (1 g, 1059 3
                                                                                                                      C,
                                                                 hours).

                                                                 Residue on ignition <2.44>    Not more than 0.1z (1 g).
C19H24N2O3.HCl: 364.87
2-Hydroxy-5-{(1RS)-1-hydroxy-2-[(1RS)-1-methyl-                  Isomer ratio Dissolve 5 mg of Labetalol Hydrochloride in
3-phenylpropylamino]ethyl}benzamide monohydrochloride            0.7 mL of a solution of n-butylboronic acid in anhydrous
2-Hydroxy-5-{(1RS)-1-hydroxy-2-[(1SR)-1-methyl-                  pyridine (3 in 250), allow to stand for 20 minutes, and use
3-phenylpropylamino]ethyl}benzamide monohydrochloride            this solution as the sample solution. Perform the test with 2
[32780-64-6]                                                     mL of the sample solution as directed under Gas Chro-
                                                                 matography <2.02> according to the following conditions.
  Labetalol Hydrochloride, when dried, contains not              Determine the areas of two adjacent peaks, Aa and Ab,
less than 98.5z and not more than 101.0z of                      where Aa is the peak area of the shorter retention time and
C19H24N2O3.HCl.                                                  Ab is the peak area of the longer retention time, using the au-
                                                                 tomatic integration method: the ratio Ab/(Aa+Ab) is be-
Description Labetalol Hydrochloride occurs as a white            tween 0.45 and 0.55.
crystalline powder.                                              Operating conditions—
  It is freely soluble in methanol, and sparingly soluble in        Detector: A hydrogen flame-ionization detector.
water and in ethanol (99.5).                                        Column: A fused silica column 0.53 mm in inside di-
  It dissolves in 0.05 mol/L sulfuric acid TS.                   ameter and 25 m in length, coated inside with methyl silicone
  Melting point: about 1819 (with decomposition).
                             C                                   polymer for gas chromatography in 5 mm thickness.
Identification (1) Determine the absorption spectrum of             Column temperature: A constant temperature of about
a solution of Labetalol Hydrochloride in 0.05 mol/L sulfur-      2909 C.
ic acid TS (1 in 20,000) as directed under Ultraviolet-visible      Injection port temperature: A constant temperature of
Spectrophotometry <2.24>, and compare the spectrum with          about 3509  C.
the Reference Spectrum: both spectra exhibit similar intensi-       Detector temperature: A constant temperature of about
ties of absorption at the same wavelengths.                      3509 C.
   (2) Determine the infrared absorption spectrum of                Carrier gas: Helium
Labetalol Hydrochloride as directed in the potassium chlo-          Flow rate: Adjust the flow rate so that the retention time
ride disc method under Infrared Spectrophotometry <2.25>,        of the peak showing earlier elution of the two peaks of
and compare the spectrum with the Reference Spectrum:            labetalol is about 9 minutes.
both spectra exhibit similar intensities of absorption at the    System suitability—
same wave numbers.                                                  System performance: Proceed with 2 mL of the sample so-
   (3) A solution of Labetalol Hydrochloride (1 in 50)           lution under the above conditions: the resolution between
responds to the Qualitative Tests <1.09> for chloride.           the two labetalol peaks is not less than 1.5.
                                                                    System repeatability: Repeat the test 6 times under the
pH <2.54> The pH of a solution prepared by dissolving 0.5 g      above conditions with 2 mL of the sample solution: the rela-
of Labetalol Hydrochloride in 50 mL of water is between 4.0      tive standard deviation of the ratio of the peak area of
and 5.0.                                                         labetalol with the shorter retention time to that of the longer
Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of             retention time is not more than 2.0z.
Labetalol Hydrochloride according to Method 2, and per-          Assay Weigh accurately about 0.3 g of Labetalol
form the test. Prepare the control solution with 2.0 mL of       Hydrochloride, previously dried, dissolve in 100 mL of a
Supplement I, JP XV                                                                         Official Monographs            1899

mixture of acetic anhydride and acetic acid (100) (7:3), and       of labetalol hydrochloride (C19H24N2O3.HCl), and use this
titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio-        solution as the sample solution. Separately, weigh accurately
metric titration). Perform a blank determination in the same       about 20 mg of labetalol hydrochloride for assay, previously
manner and make any necessary correction.                          dried at 1059 for 3 hours, and dissolve in 0.05 mol/L sul-
                                                                                  C
                                                                   furic acid TS to make exactly 50 mL. Pipet 5 mL of this so-
         Each mL of 0.1 mol/L perchloric acid VS
                                                                   lution, add 0.05 mol/L sulfuric acid TS to make exactly 50
           =36.49 mg of C19H24N2O3.HCl
                                                                   mL, and use this solution as the standard solution. Deter-
Containers and storage     Containers—Tight containers.            mine the absorbances, AT and AS, of the sample solution
                                                                   and standard solution at 302 nm as directed under Ultrav-
                                                                   iolet-visible Spectrophotometry <2.24>.
Add the following:
                                                                   Amount (mg) of labetalol hydrochloride (C19H24N2O3.HCl)
                                                                    =WS×(AT/AS)×(V/40)
Labetalol Hydrochloride Tablets
                                                                     WS: Amount (mg) of labetalol hydrochloride for assay
ラベタロール塩酸塩錠
                                                                   Dissolution <6.10> When the test is performed at 50 revolu-
                                                                   tions per minute according to the Paddle method, using 900
                                                                   mL of water as the dissolution medium, the dissolution rate
  Labetalol Hydrochloride Tablets contain not less
                                                                   in 30 minutes of Labetalol Hydrochloride Tablets is not less
than 93.0z and not more than 107.0z of the labeled
                                                                   than 75z.
amount of labetalol hydrochloride (C19H24N2O3.HCl:
                                                                      Start the test with 1 tablet of Labetalol Hydrochloride
364.87).
                                                                   Tablets, withdraw not less than 20 mL of the medium at spe-
Method of preparation Prepare as directed under Tablets,           cified minute after starting the test, and filter through a
with Labetalol Hydrochloride.                                      membrane filter with a pore size not exceeding 0.8 mm. Dis-
                                                                   card the first 10 mL of the filtrate, pipet V mL of the subse-
Identification (1) To a quantity of powdered Labetalol
                                                                   quent filtrate, and add water to make exactly V? mL so that
Hydrochloride Tablets equivalent to 5 mg of Labetalol
                                                                   each mL contains about 50 mg of labetalol hydrochloride
Hydrochloride according to the labeled amount, add 100
                                                                   (C19H24N2O3.HCl) according to the labeled amount, and use
mL of 0.05 mol/L sulfuric acid TS, shake, and filter. Deter-
                                                                   this solution as the sample solution. Separately, weigh ac-
mine the absorption spectrum of the filtrate as directed un-
                                                                   curately about 50 mg of labetalol hydrochloride for assay,
der Ultraviolet-visible Spectrophotometry <2.24>: it exhibits
                                                                   previously dried at 1059 for 3 hours, and dissolve in water
                                                                                            C
a maximum between 300 nm and 304 nm.
                                                                   to make exactly 100 mL. Pipet 10 mL of this solution, add
  (2) To a quantity of powdered Labetalol Hydrochloride
                                                                   water to make exactly 100 mL, and use this solution as the
Tablets equivalent to 0.25 g of Labetalol Hydrochloride ac-
                                                                   standard solution. Perform the test with the sample solution
cording to the labeled amount, add 25 mL of methanol,
                                                                   and standard solution as directed under Ultraviolet-visible
shake vigorously for 30 minutes, filter, and use the filtrate as
                                                                   Spectrophotometry <2.24>, and determine the absorbances,
the sample solution. Separately, dissolve 10 mg of labetalol
                                                                   AT and AS, at 302 nm.
hydrochloride in 1 mL of methanol, and use this solution as
the standard solution. Perform the test using these solutions      Dissolution rate (z) with respect to the labeled amount of
as directed under Thin-layer Chromatography <2.03>. Spot 5         labetalol hydrochloride (C19H24N2O3.HCl)
mL each of the sample solution and standard solution on a            =WS×(AT/AS)×(V?/V)×(1/C)×90
plate of silica gel with fluorescent indicator for thin-layer
                                                                     WS: Amount (mg) of labetalol hydrochloride for assay
chromatography. Develop the plate with a mixture of ethyl
                                                                     C: Labeled amount (mg) of labetalol hydrochloride
acetate, 2-propanol, water, and ammonia solution (28)
                                                                        (C19H24N2O3.HCl) in 1 tablet
(25:15:8:2) to a distance of about 10 cm, and air-dry the
plate. Examine under ultraviolet light (main wavelength: 254       Assay Weigh accurately not less than 20 Labetalol
nm): the principal spot obtained from the sample solution          Hydrochloride Tablets, and powder. Weigh accurately a
and the spot obtained from the standard solution show the          portion of the powder, equivalent to about 1 g of labetalol
same Rf value.                                                     hydrochloride (C19H24N2O3.HCl), add 100 mL of 0.5 mol/L
                                                                   sulfuric acid TS and 600 mL of water, shake vigorously for
Uniformity of dosage units <6.02> Perform the test accord-
                                                                   30 minutes, add water to make exactly 1000 mL, and filter.
ing to the following method: it meets the requirement of the
                                                                   Discard the first 5 mL of the filtrate, pipet 5 mL of the sub-
Content uniformity test.
                                                                   sequent filtrate, and add 0.05 mol/L sulfuric acid TS to
  To 1 tablet of Labetalol Hydrochloride Tablets add 5 mL
                                                                   make exactly 25 mL. Pipet 5 mL of this solution, add 0.05
of 0.5 mol/L sulfuric acid TS and 30 mL of water, shake
                                                                   mol/L sulfuric acid TS to make exactly 25 mL, and use this
vigorously for 30 minutes, add water to make exactly 50 mL,
                                                                   solution as the sample solution. Separately, weigh accurately
and filter. Discard the first 5 mL of the filtrate, pipet 4 mL
                                                                   about 40 mg of labetalol hydrochloride for assay, previously
of the subsequent filtrate, add 0.05 mol/L sulfuric acid TS
                                                                   dried at 1059 for 3 hours, and dissolve in 0.05 mol/L sul-
                                                                                 C
to make exactly V mL so that each mL contains about 40 mg
                                                                   furic acid TS to make exactly 100 mL. Pipet 5 mL of this so-
1900        Official Monographs                                                                   Supplement I, JP XV

lution, add 0.05 mol/L sulfuric acid TS to make exactly 50     to Method 1: it meets the requirement.
mL, and use this solution as the standard solution. Perform
                                                               Insoluble particulate matter <6.07>     It meets the require-
the test with the sample solution and standard solution as
                                                               ment.
directed under Ultraviolet-visible Spectrophotometry <2.24>,
and determine the absorbances, AT and AS, at 302 nm.           Sterility <4.06> Perform the test according to the Mem-
                                                               brane filtration method: it meets the requirement.
Amount (mg) of labetalol hydrochloride (C19H24N2O3.HCl)
 =WS×(AT/AS)×25

    WS: Amount (mg) of labetalol hydrochloride for assay       Add the following:
Containers and storage   Containers—Tight containers.
                                                               Manidipine Hydrochloride
                                                               マニジピン塩酸塩
Anhydrous Lactose
無水乳糖

Change the origin/limits of content to read:

  Anhydrous Lactose is b-lactose or a mixture of b-
lactose and a-lactose.
    The relative quantities of a-lactose and b-lactose
                                                               C35H38N4O6.2HCl: 683.62
in Anhydrous Lactose is labeled as the isomer ratio.
                                                               3-{2-[4-(Diphenymethyl)piperazin-1-yl]ethyl}
                                                               5-methyl (4RS)-2,6-dimethyl-4-(3-nitrophenyl)-
Change the Identification to read:
                                                               1,4-dihydropyridine-3,5-dicarboxylate dihydrochloride

  Identification Determine the infrared absorption spec-       [126229-12-7]
trum of Anhydrous Lactose, previously dried, as directed in
the potassium bromide disk method under Infrared Spec-           Manidipine Hydrochloride, when dried, contains
trophotometry <2.25>, and compare the spectrum with the        not less than 98.5z and not more than 101.0z of
Reference Spectrum or the spectrum of Anhydrous Lactose        C35H38N4O6.2HCl.
Reference Standard: both spectra exhibit similar intensities
                                                               Description Manidipine Hydrochloride occurs as white to
of absorption at the same wave numbers.
                                                               pale yellow crystals or crystalline powder.
                                                                  It is freely soluble in dimethylsulfoxide, sparingly soluble
                                                               in methanol, slightly soluble in ethanol (99.5), and practical-
Levallorphan Tartrate Injection                                ly insoluble in water.
レバロルファン酒石酸塩注射液                                                    A solution of Manidipine Hydrochloride in dimethylsul-
                                                               foxide (1 in 100) shows no optical rotation.
Add the following next to Identification:                         Manidipine Hydrochloride turns slightly brown-yellowish
                                                               white on exposure to light.
Bacterial endotoxins <4.01>   Less than 150 EU/mg.                Melting point: about 2079 (with decomposition).
                                                                                              C

Add the following next to Extractable volume:                  Identification (1) Determine the absorption spectrum of
                                                               a solution of Manidipine Hydrochloride in methanol (1 in
Foreign insoluble matter <6.06> Perform the test according     100,000) as directed under Ultraviolet-visible Spectrophoto-
to Method 1: it meets the requirement.                         metry <2.24>, and compare the spectrum with the Reference
Insoluble particulate matter <6.07>   It meets the require-    Spectrum or the spectrum of a solution of Manidipine
ment.                                                          Hydrochloride Reference Standard prepared in the same
                                                               manner as the sample solution: both spectra exhibit similar
Sterility <4.06> Perform the test according to the Mem-        intensities of absorption at the same wavelengths.
brane filtration method: it meets the requirement.                (2) Determine the infrared absorption spectrum of
                                                               Manidipine Hydrochloride as directed in the potassium chlo-
                                                               ride disc method under Infrared Spectrophotometry <2.25>,
Magnesium Sulfate Injection                                    and compare the spectrum with the Reference Spectrum or
                                                               the spectrum of Manidipine Hydrochloride Reference Stan-
硫酸マグネシウム注射液
                                                               dard: both spectra exhibit similar intensities of absorption at
                                                               the same wave numbers.
Add the following next to Extractable volume:                     (3) Add 10 mL of water to 0.1 g of Manidipine
Foreign insoluble matter <6.06>   Perform the test according   Hydrochloride, shake vigorously, and filter. Add 1 drop of
Supplement I, JP XV                                                                         Official Monographs             1901

ammonia TS to 3 mL of the filtrate, allow to stand 5               Hydrochloride, previously dried, and dissolve in a mixture
minutes, and filter. The filtrate responds to the Qualitative      of water and acetonitrile (1:1) to make exactly 50 mL. Pipet
Tests <1.09> (2) for chlorides.                                    5 mL of this solution, add exactly 5 mL of the internal stan-
                                                                   dard solution, add the mixture of water and acetonitrile (1:1)
Purity (1) Heavy metals <1.07>— Proceed with 1.0 g of
                                                                   to make 100 mL, and use this solution as the sample solu-
Manidipine Hydrochloride according to Method 2, and per-
                                                                   tion. Separately, weigh accurately about 25 mg of Manidi-
form the test. Prepare the control solution with 1.0 mL of
                                                                   pine Hydrochloride Reference Standard, previously dried,
Standard Lead Solution (not more than 10 ppm).
                                                                   and dissolve in the mixture of water and acetonitrile (1:1) to
   (2) Arsenic <1.11>—Prepare the test solution with 2.0 g
                                                                   make exactly 50 mL. Pipet 20 mL of this solution, add ex-
of Manidipine Hydrochloride according to Method 4, and
                                                                   actly 5 mL of the internal standard solution, add the mixture
perform the test (not more than 1 ppm).
                                                                   of water and acetonitrile (1:1) to make 100 mL, and use this
   (3) Related substances—Dissolve 20 mg of Manidipine
                                                                   solution as the standard solution. Perform the test with 20
Hydrochloride in 200 mL of a mixture of water and acetoni-
                                                                   mL each of the sample solution and standard solution as
trile (1:1), and use this solution as the sample solution. Pipet
                                                                   directed under Liquid Chromatography <2.01> according to
1 mL of the sample solution, add the mixture of water and
                                                                   the following conditions, and calculate the ratios, QT and
acetonitrile (1:1) to make exactly 100 mL, and use this solu-
                                                                   QS, of the peak area of manidipine to that of the internal
tion as the standard solution. Perform the test with exactly
                                                                   standard.
20 mL each of the sample solution and standard solution as
directed under Liquid Chromatography <2.01> according to                       Amount (mg) of C35H38N4O6.2HCl
the following conditions. Determine each peak area from                         = W S × ( Q T / Q S ) ×4
both solutions by the automatic integration method: the area
                                                                     WS: Amount (mg) of Manidipine Hydrochloride Refer-
of the peaks other than manidipine obtained from the sam-
                                                                         ence Standard
ple solution is not larger than 1/5 times the manidipine peak
area from the standard solution. Furthermore, the total of         Internal standard solution—A solution of butyl benzoate in
the areas of all peaks other than the manidipine peak from         acetonitrile (7 in 5000).
the sample solution is not larger than 7/10 times the peak         Operating conditions—
area of manidipine from the standard solution.                        Detector: An ultraviolet absorption photometer (wave-
Operating conditions—                                              length: 228 nm).
   Detector, column, column temperature, mobile phase,                Column: A stainless steel column 4.0 mm in inside di-
and flow rate: Proceed as directed in the operating condi-         ameter and 15 cm in length, packed with octadecylsilanized
tions in the Assay.                                                silica gel for liquid chromatography (5 mm in particle di-
   Time span of measurement: About 3.5 times as long as the        ameter).
retention time of manidipine, beginning after the solvent             Column temperature: A constant temperature of about
peak.                                                              259  C.
System suitability—                                                   Mobile phase: Dissolve 13.6 g of potassium dihydrogen
   Test for required detectability: Pipet 10 mL of the stan-       phosphate in water to make 1000 mL, and adjust to pH 4.6
dard solution, add a mixture of water and acetonitrile (1:1)       with diluted potassium hydroxide TS (1 in 10). To 490 mL of
to make exactly 100 mL. Confirm that the peak area of              this solution add 510 mL of acetonitrile.
manidipine obtained from 20 mL of this solution is equiva-            Flow rate: Adjust the flow rate so that the retention time
lent to 8 to 12z of that from 20 mL of the standard solution.      of manidipine is about 10 minutes.
   System performance: Dissolve 50 mg of Manidipine                System suitability—
Hydrochloride in a mixture of water and acetonitrile (1:1) to         System performance: When the procedure is run with 20
make 50 mL. To 10 mL of this solution add 5 mL of a solu-          mL of the standard solution under the above operating con-
tion of butyl benzoate in acetonitrile (7 in 5000) and the mix-    ditions, manidipine and the internal standard are eluted in
ture of water and acetonitrile (1:1) to make 100 mL. When          this order with the resolution between these peaks being not
the procedure is run with 20 mL of this solution under the         less than 5.
above operating conditions, manidipine and butyl benzoate             System repeatability: When the test is repeated 6 times
are eluted in this order with the resolution between these         with 20 mL of the standard solution under the above operat-
peaks being not less than 5.                                       ing conditions, the relative standard deviation of the ratio of
   System repeatability: When the test is repeated 6 times         the peak area of manidipine to that of the internal standard
with 20 mL of the standard solution under the above operat-        is not more than 1.0z.
ing conditions, the relative standard deviation of the peak
                                                                   Containers and storage Containers—Tight containers.
area of manidipine is not more than 2.0z.
                                                                     Storage—Light-resistant.
Loss on drying <2.41>    Not more than 1.5z (1 g, 1059 4
                                                      C,
hours).

Residue on ignition <2.44>    Not more than 0.2z (1 g).

Assay   Weigh accurately about 0.1 g of Manidipine
1902       Official Monographs                                                                      Supplement I, JP XV

Add the following:                                               in 45 minutes of Manidipine Hydrochloride Tablets is not
                                                                 less than 75z.
Manidipine Hydrochloride Tablets                                    Conduct this procedure using light-resistant vessels. Start
                                                                 the test with 1 tablet of Manidipine Hydrochloride Tablets,
マニジピン塩酸塩錠                                                        withdraw not less than 20 mL of the medium at the specified
                                                                 minute after starting the test, and filter through a membrane
  Manidipine Hydrochloride Tablets contain not less              filter with a pore size not exceeding 0.45 mm. Discard the
than 92.0z and not more than 108.0z of the labeled               first 10 mL of the filtrate, pipet V mL of the subsequent
amount of manidipine hydrochloride (C35H38N4O6.                  filtrate, and add the dissolution medium to make exactly V?
2HCl: 683.62).                                                   mL so that each mL contains about 5.6 mg of manidipine
                                                                 hydrochloride (C35H38N4O6.2HCl) according to the labeled
Method of preparation Prepare as directed under Tablets,
                                                                 amount. Pipet 2 mL of this solution, add exactly 2 mL of
with Manidipine Hydrochloride.
                                                                 methanol, and use this solution as the sample solution.
Identification To a quantity of powdered Manidipine              Separately, weigh accurately about 25 mg of Manidipine
Hydrochloride Tablets, equivalent to 10 mg of Manidipine         Hydrochloride Reference Standard, previously dried, dis-
Hydrochloride according to the labeled amount, add 5 mL          solve in a mixture of water and acetonitrile (1:1) to make ex-
of methanol, shake vigorously, centrifuge, and use the su-       actly 50 mL. Pipet 1 mL of this solution, and add the disso-
pernatant liquid as the sample solution. Separately, dissolve    lution medium to make exactly 100 mL. Pipet 2 mL of this
10 mg of Manidipine Hydrochloride Reference Standard in          solution, add exactly 2 mL of methanol, and use this solu-
5 mL of methanol, and use this solution as the standard so-      tion as the standard solution. Perform the test with exactly
lution. Perform the test with these solutions as directed un-    20 mL each of the sample solution and standard solution as
der Thin-layer Chromatography <2.03>. Spot 5 mL each of          directed under Liquid Chromatography <2.01> according to
the sample solution and standard solution on a plate of silica   the following conditions, and determine the manidipine
gel with fluorescent indicator for thin-layer chro-              peak areas, AT and AS, of both solutions.
matography. Develop the plate with a mixture of ethyl
                                                                 Dissolution rate (z) with respect to the labeled amount of
acetate and diethylamine (200:1) to a distance of about 10
                                                                 manidipine hydrochloride (C35H38N4O6.2HCl)
cm, and air-dry the plate. Examine under ultraviolet light
                                                                   =WS×(AT/AS)×(V?/V )×(1/C)×18
(main wavelength: 254 nm): the principal spot obtained
from the sample solution and the spot obtained from the            WS: Amount (mg) of Manidipine Hydrochloride Refer-
standard solution show the same Rf value.                              ence Standard
                                                                   C: Labeled amount (mg) of manidipine hydrochloride
Uniformity of dosage units <6.02> Perform the test accord-
                                                                      (C35H38N4O6.2HCl) in 1 tablet
ing to the following method: it meets the requirement of the
Content uniformity test.                                         Operating conditions—
   Conduct this procedure using light-resistant vessels. To 1       Detector: An ultraviolet absorption photometer (wave-
tablet of Manidipine Hydrochloride Tablets, add exactly 1        length: 228 nm).
mL of the internal standard solution per 1 mg of manidipine         Column: A stainless steel column 4.0 mm in inside di-
hydrochloride (C35H38N4O6.2HCl), disintegrate by adding a        ameter and 15 cm in length, packed with octadecylsilanized
mixture of water and acetonitrile (1:1) to make V mL so that     silica gel for liquid chromatography (5 mm in particle di-
each mL contains about 0.1 mg of manidipine hydrochloride        ameter).
(C35H38N4O6.2HCl), shake vigorously for 10 minutes, and             Column temperature: A constant temperature of about
filter through a membrane filter with a pore size not exceed-    259  C.
ing 0.45 mm. Discard the first 1 mL of the filtrate, and use        Mobile phase: A mixture of acetonitrile and a solution of
the subsequent filtrate as the sample solution. Then, proceed    potassium dihydrogen phosphate (681 in 100,000) (3:2).
as directed in the Assay.                                           Flow rate: Adjust the flow rate so that the retention time
                                                                 of manidipine is about 6 minutes.
        Amount (mg) of manidipine hydrochloride
                                                                 System suitability—
        (C35H38N4O6.2HCl)
                                                                    System performance: When the procedure is run with 20
          =WS×(QT/QS)×(V/250)
                                                                 mL of the standard solution under the above operating con-
  WS: Amount (mg) of Manidipine Hydrochloride Refer-             ditions, the number of theoretical plates and the symmetry
      ence Standard                                              factor of the peak of manidipine are not less than 1500 and
                                                                 not more than 1.5, respectively.
Internal standard solution—A solution of butyl benzoate in
                                                                    System repeatability: When the test is repeated 6 times
acetonitrile (7 in 10,000).
                                                                 with 20 mL of the standard solution under the above operat-
Dissolution <6.10> When the test is performed at 50 revolu-      ing conditions, the relative standard deviation of the peak
tions per minute according to the Paddle method, using 900       area of manidipine is not more than 2.0z.
mL of 0.05 mol/L acetic acid-sodium acetate buffer solu-
                                                                 Assay Conduct this procedure using light-resistant vessels.
tion, pH 4.0, as the dissolution medium, the dissolution rate
                                                                 Weigh accurately not less than 20 Manidipine Hydrochloride
Supplement I, JP XV                                                                         Official Monographs            1903

Tablets, and powder. Weigh accurately a portion of the             water.
powder, equivalent to about 10 mg of manidipine                      It gradually turns yellow on exposure to light.
hydrochloride (C35H38N4O6.2HCl), add exactly 10 mL of the
internal standard solution, add a mixture of water and             Change the Identification to read:
acetonitrile (1:1) to make 100 mL, shake vigorously for 10
                                                                   Identification (1) Dissolve 10 mg of Medazepam in 3 mL
minutes, and filter through a membrane filter with a pore
                                                                   of citric acid-acetic acid TS: a deep orange color develops.
size not exceeding 0.45 mm. Discard the first 1 mL of the
                                                                   Heat in a water bath for 3 minutes: the color changes to dark
filtrate, and use the subsequent filtrate as the sample solu-
                                                                   red.
tion. Separately, weigh accurately about 25 mg of Manidi-
                                                                      (2) Determine the absorption spectrum of a solution of
pine Hydrochloride Reference Standard, previously dried,
                                                                   Medazepam in methanol (1 in 100,000) as directed under
and dissolve in the mixture of water and acetonitrile (1:1) to
                                                                   Ultraviolet-visible Spectrophotometry <2.24>, and compare
make 50 mL. Pipet 20 mL of this solution, add exactly 10
                                                                   the spectrum with the Reference Spectrum: both spectra ex-
mL of the internal standard solution, add the mixture of
                                                                   hibit similar intensities of absorption at the same wave-
water and acetonitrile (1:1) to make 100 mL, and use this so-
                                                                   lengths.
lution as the standard solution. Then, proceed as directed in
                                                                      (3) Determine the infrared absorption spectrum of
the Assay under Manidipine Hydrochloride.
                                                                   Medazepam as directed in the potassium bromide disk
        Amount (mg) of manidipine hydrochloride                    method under Infrared Spectrophotometry <2.25>, and com-
        (C35H38N4O6.2HCl)                                          pare the spectrum with the Reference Spectrum: both spec-
          =WS×(QT/QS)×(2/5)                                        tra exhibit similar intensities of absorption at the same wave
                                                                   numbers.
  WS: Amount (mg) of Manidipine Hydrochloride Refer-
                                                                      (4) Perform the test with Medazepam as directed under
      ence Standard
                                                                   Flame Coloration Test <1.04> (2): a green color appears.
Internal standard solution—A solution of butyl benzoate in
acetonitrile (7 in 10,000).

Containers and storage Containers—Tight containers.                Mefruside Tablets
 Storage—Light-resistant.
                                                                   メフルシド錠

                                                                   Add the following next to Identification:
D-Mannitol           Injection
                                                                   Uniformity of dosage units <6.02> Perform the test accord-
D-マンニトール注射液                                                        ing to the following method: it meets the requirement of the
                                                                   Content uniformity test.
Add the following next to Extractable volume:                         To 1 tablet of Mefruside Tablets add 40 mL of methanol,
                                                                   disintegrate the tablet using ultrasonic waves with occasional
Foreign insoluble matter <6.06> Perform the test according
                                                                   stirring, then further treat with ultrasonic waves for 10
to Method 1: it meets the requirement.
                                                                   minutes, and add methanol to make exactly V mL of a solu-
Insoluble particulate matter <6.07>     It meets the require-      tion containing about 0.5 mg of mefruside (C13H19ClN2O5S2)
ment.                                                              per mL. Centrifuge the solution, pipet 5 mL of the super-
                                                                   natant liquid, add methanol to make exactly 20 mL, and use
Sterility <4.06> Perform the test according to the Mem-
                                                                   this solution as the sample solution. Then, proceed as direct-
brane filtration method: it meets the requirement.
                                                                   ed in the Assay.

                                                                          Amount (mg) of mefruside (C13H19ClN2O5S2)
Medazepam                                                                  =WS×(AT/AS)×(V/125)

                                                                     WS: Amount (mg) of mefruside for assay
メダゼパム

Change the origin/limits of content to read:
                                                                   Methyldopa Tablets
 Medazepam, when dried, contains not less than 98.5
                                                                   メチルドパ錠
z and not more than 101.0z of C16H15ClN2.
                                                                   Add the following next to Identification:
Change the Description to read:
                                                                   Uniformity of dosage units <6.02> Perform the test accord-
Description Medazepam occurs as white to light yellow
                                                                   ing to the following method: it meets the requirement of the
crystals or crystalline powder.
                                                                   Content uniformity test.
  It is freely soluble in methanol, in ethanol (99.5), in acetic
                                                                     To 1 tablet of Methyldopa Tablets add 50 mL of 0.05
acid (100) and in diethyl ether, and practically insoluble in
1904       Official Monographs                                                                      Supplement I, JP XV

mol/L sulfuric acid TS, shake for 15 minutes, then add 0.05     equivalent to 0.1 g (potency) of Minocycline Hydrochloride
mol/L sulfuric acid TS to make exactly 100 mL, and filter.      according to the labeled amount, dissolve in the mobile
Discard the first 20 mL of the filtrate, pipet V mL of the      phase to make 100 mL. Pipet 25 mL of this solution, add the
subsequent filtrate equivalent to about 5 mg of methyldopa      mobile phase to make exactly 50 mL, and use this solution as
(C10H13NO4), add exactly 5 mL of iron (II) tartrate TS, then    the sample solution. Perform the test with 20 mL of the sam-
add ammonia-ammonium acetate buffer solution, pH 8.5,           ple solution as directed under Liquid Chromatography
to make exactly 100 mL, and use this solution as the sample     <2.01> according to the following conditions, and determine
solution. Separately, weigh accurately about 0.11 g of          each peak area by the automatic integration method. Calcu-
Methyldopa Reference Standard (separately determine the         late the amounts of each peak by the area percentage
loss on drying <2.41> at 1259 for 2 hours), and dissolve in
                              C                                 method: the amount of epiminomycine, having the relative
0.05 mol/L sulfuric acid TS to make exactly 100 mL. Pipet 5     retention time of about 0.83 with respect to minocycline, is
mL of this solution, add exactly 5 mL of iron (II) tartrate     not more than 6.0z.
TS, then add ammonia-ammonium acetate buffer solution,          Operating conditions—
pH 8.5, to make exactly 100 mL, and use this solution as the       Detector, column, column temperature, mobile phase and
standard solution. Determine the absorbances at 520 nm, AT      flow rate: Proceed as directed in the operating conditions in
and AS, of the sample solution and standard solution as         the Assay.
directed under Ultraviolet-visible Spectrophotometry <2.24>.       Time span of measurement: About 2.5 times as long as the
                                                                retention time of minocycline, beginning after the solvent
        Amount (mg) of methyldopa (C10H13NO4)
                                                                peak.
         =WS×(AT/AS)×(5/V)
                                                                System suitability—
  WS: Amount (mg) of Methyldopa Reference Standard,                Test for required detectability: Pipet 2 mL of the standard
      calculated on the dried basis                             solution obtained in the Assay, add the mobile phase to
                                                                make exactly 100 mL, and use this solution as the solution
                                                                for system suitability test. Pipet 5 mL of the solution for sys-
Add the following:                                              tem suitability test, add the mobile phase to make exactly
                                                                100 mL. Confirm that the peak area of minocycline ob-
Minocycline Hydrochloride for                                   tained from 20 mL of this solution is equivalent to 3.5 to 6.5
                                                                z of that from 20 mL of the solution for system suitability
Injection                                                       test.
注射用ミノサイクリン塩酸塩                                                      System performance: Proceed as directed in the system
                                                                suitability in the Assay.
  Minocycline Hydrochloride for Injection is a prepa-              System repeatability: When the test is repeated 6 times
                                                                with 20 mL of the solution for system suitability test under
ration for injection which is dissolved before use.
  It contains not less than 90.0z and not more than             the above operating conditions, the relative standard devia-
110.0z of the labeled amount of minocycline                     tion of the peak area of minocycline is not more than 2.0z.
(C23H27N3O7: 457.48).                                           Water <2.48> Weigh accurately the mass of the content of
Method of preparation Prepare as directed under Injec-          one container of Minocycline Hydrochloride for Injection,
tions, with Minocycline Hydrochloride.                          dissolve in exactly 2 mL of methanol for water determina-
                                                                tion, and preform the test with exactly 1 mL of this solution
Description Minocycline Hydrochloride for Injection oc-         as directed in the Volumetric tiration (back tiration): not
curs as a yellow to yellow-brown powder or flakes.              more than 3.0z.
Identification Dissolve 4 mg of Minocycline Hydrochlo-          Bacterial endotoxins <4.01>    Less than 1.25 EU/mg (poten-
ride for Injection in 250 mL of a solution of hydrochloric      cy).
acid in methanol (19 in 20,000). Determine the absorption
spectrum of this solution as directed under Ultraviolet-visi-   Uniformity of dosage units <6.02>     It meets the requirement
ble Spectrophotometry <2.24>: it exhibits maxima between        of the Mass variation test.
221 nm and 225 nm, between 261 nm and 265 nm, and be-           Foreign insoluble matter <6.06> Perform the test according
tween 354 nm and 358 nm.                                        to Method 2: it meets the requirement.
pH <2.54> The pH of a solution, prepared by dissolving an       Insoluble particulate matter <6.07>     It meets the require-
amount of Minocycline Hydrochloride for Injection,              ment.
equivalent to 0.1 g (potency) of Minocycline Hydrochloride
according to the labeled amount, in 10 mL of water is 2.0 to    Sterility <4.06> Perform the test according to the Mem-
3.5.                                                            brane filtration method: it meets the requirement.

Purity Related substances—Conduct this procedure rapid-         Assay Weigh accurately an amount of Minocycline
ly after the preparation of the sample solution. Take an        Hydrochloride for Injection, equivalent to about 0.1 g
amount of Minocycline Hydrochloride for Injection,              (potency) of Minocycline Hydrochloride, dissolve in the mo-
Supplement I, JP XV                                                                       Official Monographs            1905

bile phase to make exactly 100 mL. Pipet 25 mL of this solu-      Identification Dissolve an amount of Mitomycin C for In-
tion, add the mobile phase to make exactly 50 mL, and use         jection, equivalent to 2 mg (potency) of Mitomycin C ac-
this solution as the sample solution. Separately, weigh ac-       cording to the labeled amount, in 200 mL of water, and de-
curately an amount of Minocycline Hydrochloride Refer-            termine the absorption spectrum of this solution as directed
ence Standard, equivalent to about 25 mg (potency), dis-          under Ultraviolet-visible Spectrophotometry <2.24>: it ex-
solve in the mobile phase to make exactly 50 mL, and use          hibits maxima between 216 nm and 220 nm, and between
this solution as the standard solution. Perform the test with     362 nm and 366 nm.
exactly 20 mL each of the sample solution and standard solu-
                                                                  pH <2.54> The pH of a solution, prepared by dissolving
tion as directed under Liquid Chromatography <2.01> ac-
                                                                  0.25 g of Mitomycin C for Injection in 20 mL of water, is 5.5
cording to the following conditions, and determine the peak
                                                                  to 8.5.
area of minocycline, AT and AS, of each solution.
                                                                  Loss on drying <2.41> Not more than 1.0z (0.4 g, in vacu-
   Amount [mg (potency)] of minocycline (C23H27N3O7)
                                                                  um not exceeding 0.67 kPa, phosphorus (V) oxide, 609 3
                                                                                                                       C,
    =WS×(AT/AS)×4
                                                                  hours).
  WS: Amount [mg (potency)] of Minocycline Hydrochlo-
                                                                  Bacterial endotoxins <4.01>   Less than 10 EU/mg (poten-
      ride Reference Standard
                                                                  cy).
Operating conditions—
                                                                  Uniformity of dosage units <6.02> Perform the test accord-
   Detector, column, column temperature, and flow rate:
                                                                  ing to the following method: it meets the requirement of the
Proceed as directed in the operating conditions in the Assay
                                                                  Content uniformity test.
under Minocycline Hydrochloride.
                                                                     To 1 container of Mitomycin C for Injection add exactly
   Mobile phase: Adjust the pH to 6.5 of a mixture of am-
                                                                  V mL of N,N-dimethylacetamide so that each mL contains
monium oxalate monohydrate solution (7 in 250), N,N-
                                                                  about 0.5 mg (potency) of Mitomycin C, shake, centrifuge,
dimethylformamide and 0.1 mol/L disodium dihydrogen
                                                                  and use the supernatant liquid as the sample solution.
ethylenediamine tetraacetate TS (11:5:4) with tetrabutylam-
                                                                  Separately, weigh accurately about 25 mg (potency) of
monium hydroxide TS.
                                                                  Mitomycin C Reference Standard, add N,N-dimethylaceta-
System suitability—
                                                                  mide to make exactly 50 mL, and use this solution as the
   System performance: Dissolve 50 mg of minocycline
                                                                  standard solution. Then, proceed as directed in the Assay
hydrochloride in water to make 25 mL. Heat 5 mL of this
                                                                  under Mitomycin C.
solution on a water bath for 60 minutes, then add water to
make 25 mL. When the procedure is run with 20 mL of this            Amount [mg (potency)] of mitomycin C (C15H18N4O5)
solution under the above operating conditions, epiminocy-            =WS×(AT/AS)×(V/50)
cline and minocycline are eluted in this order with the resolu-
                                                                    WS: Amount [mg (potency)] of Mitomycin C Reference
tion between these peaks being not less than 2.5.
                                                                        Standard
   System repeatability: When the test is repeated 6 times
with 20 mL of the standard solution under the above operat-       Foreign insoluble matter <6.06> Perform the test according
ing conditions, the relative standard deviation of the peak       to Method 2: it meets the requirement.
area of minocycline is not more than 2.0z.
                                                                  Insoluble particulate matter <6.07>   It meets the require-
Containers and storage    Containers—Hermetic containers.         ment.

                                                                  Sterility <4.06> Perform the test according to the Mem-
                                                                  brane filtration method: it meets the requirement.
Add the following:
                                                                  Assay Weigh accurately the mass of the contents of not
Mitomycin C for Injection                                         less than 10 containers of Mitomycin C for Injection. Weigh
                                                                  accurately an amount of the contents, equivalent to about 10
注射用マイトマイシン C                                                      mg (potency) of Mitomycin C, add exactly 20 mL of N,N-
                                                                  dimethylacetamide, shake, centrifuge, and use the super-
   Mitomycin C for Injection is a preparation for injec-          natant liquid as the sample solution. Separately, weigh ac-
tion, which is dissolved before use.                              curately an amount of Mitomycin C Reference Standard,
   It contains not less than 90.0z and not more than              equivalent to about 25 mg (potency), dissolve in N,N-
110.0z of the labeled amount of mitomycin C                       dimethylacetamide to make exactly 50 mL, and use this solu-
(C15H18N4O5: 334.33).                                             tion as the standard solution. Then, proceed as directed in
                                                                  the Assay under Mitomycin C.
Method of preparation Prepare as directed under Injec-
tions, with Mitomycin C.                                            Amount [mg (potency)] of mitomycin C (C15H18N4O5)
                                                                     =WS×(AT/AS)×(2/5)
Description Mitomycin C for Injection occurs as a blue-
purple powder.                                                      WS: Amount [mg (potency)] of Mitomycin C Reference
                                                                        Standard
1906      Official Monographs                                                                     Supplement I, JP XV

Containers and storage   Containers—Hermetic containers.       the sample solution are not larger than the mizoribine peak
                                                               area from the standard solution.
Add the following:                                             Operating conditions—
                                                                  Column, column temperature, mobile phase, and flow
Mizoribine                                                     rate: Proceed as directed in the operating conditions in the
                                                               Assay.
ミゾリビン                                                             Detector: An ultraviolet absorption photometer (wave-
                                                               length: 220 nm).
                                                                  Time span of measurement: About 3 times as long as the
                                                               retention time of mizoribine, beginning after the solvent
                                                               peak.
                                                               System suitability—
                                                                  Test for required detectability: Pipet 1 mL of the standard
                                                               solution, and add the mobile phase to make exactly 5 mL.
C9H13N3O6: 259.22                                              Confirm that the peak area of mizoribine obtained from 5
5-Hydroxy-1-b-D-ribofuranosyl-1H-imidazole-4-                  mL of this solution is equivalent to 14 to 26z of that from 5
carboxamide [50924-49-7]                                       mL of the standard solution.
                                                                  System performance: When the procedure is run with 5 mL
  Mizoribine contains not less than 98.0z and not              of the standard solution under the above operating condi-
more than 102.0z of C9H13N3O6, calculated on the               tions, the number of theoretical plates and the symmetry
anhydrous basis.                                               factor of the peak of mizoribine are not less than 10,000 and
                                                               not more than 1.4, respectively.
Description Mizoribine occurs as a white to yellowish             System repeatability: When the test is repeated 6 times
white crystalline powder.                                      with 5 mL of the standard solution under the above operat-
 It is freely soluble in water, and practically insoluble in   ing conditions, the relative standard deviation of the peak
methanol and in ethanol (99.5).                                area of mizoribine is not more than 2.0z.
Identification (1) Determine the absorption spectrum of        Water <2.48> Not more than 0.5z (0.5 g, volumetric titra-
a solution of Mizoribine (1 in 100,000) as directed under      tion, direct titration).
Ultraviolet-visible Spectrophotometry <2.24>, and compare
the spectrum with the Reference Spectrum or the spectrum       Residue on ignition <2.44>   Not more than 0.1z (1 g).
of a solution of Mizoribine Reference Standard prepared in     Assay Weigh accurately about 0.1 g of Mizoribine, and
the same manner as the sample solution: both spectra exhibit   dissolve in the mobile phase to make exactly 50 mL. Pipet 5
similar intensities of absorption at the same wavelengths.     mL of this solution, add the mobile phase to make exactly 50
  (2) Determine the infrared absorption spectrum of            mL, and use this solution as the sample solution. Separately,
Mizoribine as directed in the potassium bromide disk           weigh accurately about 10 mg of Mizoribine Reference Stan-
method under Infrared Spectrophotometry <2.25>, and com-       dard (separately determine the water <2.48> using the same
pare the spectrum with the Reference Spectrum or the spec-     manner as Mizoribine), dissolve in the mobile phase to make
trum of Mizoribine Reference Standard: both spectra exhibit    exactly 50 mL, and use this solution as the standard solu-
similar intensities of absorption at the same wave numbers.    tion. Perform the test with exactly 5 mL each of the sample
Optical rotation <2.49> [a]20: -25 – -279
                           D              (0.5 g calculated    solution and standard solution as directed under Liquid
on the anhydrous basis, water, 25 mL, 100 mm).                 Chromatography <2.01> according to the following condi-
                                                               tions, and determine the peak areas of mizoribine, AT and
Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of           AS, of both solutions.
Mizoribine according to Method 1, and perform the test.
Prepare the control solution with 2.0 mL of Standard Lead                      Amount (mg) of C9H13N3O6
Solution (not more than 20 ppm).                                                =WS×(AT/AS)×10
  (2) Related substances—Dissolve 0.10 g of Mizoribine in        WS: Amount (mg) of Mizoribine Reference Standard, cal-
the mobile phase to make 50 mL, and use this solution as the         culated on the anhydrous basis
sample solution. Pipet 5 mL of the sample solution, and add
the mobile phase to make exactly 50 mL. Pipet 1 mL of this     Operating conditions—
solution, add the mobile phase to make exactly 100 mL, and        Detector: An ultraviolet absorption photometer (wave-
use this solution as the standard solution. Perform the test   length: 279 nm).
with exactly 5 mL each of the sample solution and standard        Column: A stainless steel column 4.6 mm in inside di-
solution as directed under Liquid Chromatography <2.01>        ameter and 25 cm in length, packed with octadecylsilanized
according to the following conditions. Determine each peak     silica gel for liquid chromatography (5 mm in particle di-
area of both solutions by the automatic integration method:    ameter).
the areas of the peaks other than mizoribine obtained from        Column temperature: A constant temperature of about
                                                               259  C.
Supplement I, JP XV                                                                       Official Monographs            1907

   Mobile phase: Diluted phosphoric acid (1 in 1500).            solution by the automatic integration method: the area of
   Flow rate: Adjust the flow rate so that the retention time    the peak, having the relative retention time of about 0.3 with
of mizoribine is about 9 minutes.                                respect to mizoribine, obtained from the sample solution is
System suitability—                                              not larger than the peak area of mizoribine from the stan-
   System performance: When the procedure is run with 5 mL       dard solution, and the area of each peak other than mizori-
of the standard solution under the above operating condi-        bine from the sample solution is not larger than 2/5 times
tions, the number of theoretical plates and the symmetry         the peak area of mizoribine from the standard solution.
factor of the peak of mizoribine are not less than 10,000 and    Operating conditions—
not more than 1.4, respectively.                                    Column, column temperature, mobile phase, and flow
   System repeatability: When the test is repeated 6 times       rate: Proceed as directed in the operating conditions in the
with 5 mL of the standard solution under the above operat-       Assay under Mizoribine.
ing conditions, the relative standard deviation of the peak         Detector: An ultraviolet absorption photometer (wave-
area of mizoribine is not more than 1.0z.                        length: 220 nm).
                                                                    Time span of measurement: About 3 times as long as the
Containers and storage    Containers—Tight containers.
                                                                 retention time of mizoribine, beginning after the solvent
                                                                 peak.
                                                                 System suitability—
Add the following:
                                                                    Test for required detectability: To exactly 1 mL of the
                                                                 standard solution add the mobile phase to make exactly 5
Mizoribine Tablets                                               mL. Confirm that the peak area of mizoribine obtained
                                                                 from 5 mL of this solution is equivalent to 14 to 26z of that
ミゾリビン錠
                                                                 from 5 mL of the standard solution.
                                                                    System performance: When the procedure is run with 5 mL
  Mizoribine Tablets contain not less than 93.0z and
                                                                 of the standard solution under the above operating condi-
not more than 107.0z of the labeled amount of
                                                                 tions, the number of theoretical plates and the symmetry
mizoribine (C9H13N3O6: 259.22).
                                                                 factor of the peak of mizoribine are not less than 10,000 and
Method of preparation     Prepare as directed under Tablets,     not more than 1.4, respectively.
with Mizoribine.                                                    System repeatability: When the test is repeated 6 times
                                                                 with 5 mL of the standard solution under the above operat-
Identification To a quantity of powdered Mizoribine
                                                                 ing conditions, the relative standard deviation of the peak
Tablets, equivalent to 0.1 g of Mizoribine according to the
                                                                 area of mizoribine is not more than 2.0z.
labeled amount, add 5 mL of water, shake, filter, and use
the filtrate as the sample solution. Separately, dissolve 20     Uniformity of dosage units <6.02> Perform the test accord-
mg of Mizoribine Reference Standard in 1 mL of water, and        ing to the following method: it meets the requirement of the
use this solution as the standard solution. Perform the test     Content uniformity test.
with the sample solution and standard solution as directed          To 1 tablet of Mizoribine Tablets add 50 mL of water,
under Thin-Layer Chromatography <2.03>. Spot 1 mL each           shake until the tablet is disintegrated, and add water to make
of the sample solution and standard solution on a plate of       exactly 100 mL. Filter the solution, discard not less than 10
silica gel for thin-layer chromatography. Then develop the       mL of the first filtrate, pipet V mL of the subsequent
plate with a mixture of methanol, ammonia solution (28)          filtrate, add water to make exactly V? mL so that each mL
and 1-propanol (2:1:1) to a distance of about 10 cm, and air-    contains about 5 mg of mizoribine (C9H13N3O6), and use this
dry the plate. Allow the plate to stand in iodine vapor: the     solution as the sample solution. Separately, weigh accurately
principal spot from the sample solution and the spot from        about 25 mg of Mizoribine Reference Standard (separately
the standard solution show a red-brown color and the same        determine the water <2.48> in the same manner as Mizori-
Rf value.                                                        bine), and dissolve in water to make exactly 100 mL. Pipet 2
                                                                 mL of the solution, add water to make exactly 100 mL, and
Purity Related substances—To a quantity of powdered
                                                                 use this solution as the standard solution. Determine the ab-
Mizoribine Tablets, equivalent to 0.10 g of Mizoribine ac-
                                                                 sorbances, AT and AS, at 279 nm of the sample solution and
cording to the labeled amount, add 30 mL of the mobile
                                                                 standad solution as directed under Ultraviolet-visible Spec-
phase, shake, then add the mobile phase to make 50 mL.
                                                                 trophotometry <2.24>.
Filter the solution through a membrane filter with a pore size
not exceeding 0.5 mm and use the filtrate as the sample solu-                Amount of mizoribine (C9H13N3O6)
tion. Pipet 2 mL of the sample solution, add the mobile                       =WS×(AT/AS)×(V?/V)×(1/50)
phase to make exactly 20 mL. Pipet 1 mL of the solution,
                                                                   WS: Amount (mg) of Mizoribine Reference Standard, cal-
add the mobile phase to make exactly 20 mL, and use this so-
                                                                       culated on the anhydrous basis
lution as the standard solution. Perform the test with exactly
5 mL each of the sample solution and standard solution as        Dissolution <6.10> When the test is performed at 50 revolu-
directed under Liquid Chromatography <2.01> according to         tions per minute according to the Paddle method, using 900
the following conditions. Determine each peak area of each       mL of water as the dissolution medium, the dissolution rate
1908       Official Monographs                                                                       Supplement I, JP XV

in 45 minutes of Mizoribine Tablets is not less than 80z.        Content uniformity test.
   Start the test with 1 tablet of Mizoribine Tablets,              To 1 tablet of Morphine Hydrochloride Tablets add exact-
withdraw not less than 20 mL of the medium at the specified      ly 1 mL of the internal standard solution per 2 mg of mor-
minute after starting the test, and filter through a membrane    phine hydrochloride hydrate (C17H19NO3.HCl.3H2O), dis-
filter with a pore size not exceeding 0.5 mm. Discard not less   perse the tablet into a small particles using ultrasonic waves,
than 10 mL of the first filtrate, pipet V mL of the subse-       then treat with ultrasonic waves for 15 minutes with oc-
quent filtrate, add water to make exactly V? mL so that each     casional stirring, and add water to make V mL so that each
mL contains about 14 mg of mizoribine (C9H13N3O6) accord-        mL contains about 0.4 mg of morphine hydrochloride hy-
ing to the labeled amount, and use this solution as the sam-     drate (C17H19NO3.HCl.3H2O). Filter the solution, and use
ple solution. Separately, weigh accurately about 28 mg of        the filtrate as the sample solution. Then, proceed as directed
Mizoribine Reference Standard (separately determine the          in the Assay.
water <2.48> in the same manner as Mizoribine), and dissolve
                                                                      Amount (mg) of morphine hydrochloride hydrate
in water to make exactly 100 mL. Pipet 1 mL of this solu-
                                                                      (C17H19NO3.HCl.3H2O)
tion, add water to make exactly 20 mL, and use this solution
                                                                        =WS×(QT/QS)×(V/50)×1.1679
as the standard solution. Determine the absorbances, AT and
AS, at 279 nm of the sample solution and standard solution         WS: Amount (mg) of morphine hydrochloride for assay,
as directed under Ultraviolet-visible Spectrophotometry                calculated on the anhydrous basis
<2.24>.
                                                                 Internal standard solution—A         solution     of   etilefrine
Dissolution rate (z) with respect to the labeled amount of       hydrochloride (1 in 500).
mizoribine (C9H13N3O6)
  =WS×(AT/AS)×(V?/V)×(1/C)×45
                                                                 Add the following:
  WS: Amount (mg) of Mizoribine Reference Standard, cal-
       culated on the anhydrous basis
  C: Labeled amount (mg) of mizoribine (C9H13N3O6) in 1          Nabumetone
     tablet
                                                                 ナブメトン
Assay Weigh accurately not less than 20 Mizoribine
Tablets, and powder. Weigh accurately a portion of the
powder, equivalent to about 25 mg of mizoribine
(C9H13N3O6), add 50 mL of water and shake, then add water
to make exactly 100 mL. Filter the solution, discard not less
                                                                 C15H16O2: 228.29
than 10 mL of the first filtrate, pipet 2 mL of the subsequent
                                                                 4-(6-Methoxynaphthalen-2-yl)butan-2-one         [42924-53-8]
filtrate, add water to make exactly 100 mL, and use this so-
lution as the sample solution. Separately, weigh accurately
                                                                   Nabumetone contains not less than 98.0z and not
about 25 mg of Mizoribine Reference Standard (separately
                                                                 more than 101.0z of C15H16O2, calculated on the an-
determine the water <2.48> in the same manner as Mizori-
                                                                 hydrous basis.
bine), and dissolve in water to make exactly 100 mL. Pipet 2
mL of the solution, add water to make exactly 100 mL, and        Description Nabumetone occurs as white to yellowish
use this solution as the standard solution. Determine the ab-    white crystals or a crystalline powder.
sorbances, AT and AS, at 279 nm of the sample solution and         It is soluble in acetonitrile, sparingly soluble in methanol
standard solution as directed under Ultraviolet-visible Spec-    and in ethanol (99.5), and practically insoluble in water.
trophotometry <2.24>.
                                                                 Identification (1) Determine the absorption spectrum of
 Amount (mg) of mizoribine (C9H13N3O6)=WS×(AT/AS)                a solution of Nabumetone in methanol (1 in 30,000) as
                                                                 directed under Ultraviolet-visible Spectrophotometry <2.24>,
  WS: Amount (mg) of Mizoribine Reference Standard, cal-
                                                                 and compare the spectrum with the Reference Spectrum or
      culated on the anhydrous basis
                                                                 the spectrum of a solution of Nabumetone Reference Stan-
Containers and storage    Containers—Tight containers.           dard prepared in the same manner as the sample solution:
                                                                 both spectra exhibit similar intensities of absorption at the
                                                                 same wavelengths.
Morphine Hydrochloride Tablets                                     (2) Determine the infrared absorption spectrum of
                                                                 Nabumetone as directed in the potassium bromide disk
モルヒネ塩酸塩錠                                                         method under Infrared Spectrophotometry <2.25>, and com-
                                                                 pare the spectrum with the Reference Spectrum or the spec-
Add the following next to Identification:                        trum of Nabumetone Reference Standard: both spectra ex-
                                                                 hibit similar intensities of absorption at the same wave num-
Uniformity of dosage units <6.02> Perform the test accord-
                                                                 bers.
ing to the following method: it meets the requirement of the
Supplement I, JP XV                                                                         Official Monographs           1909

Melting point <2.60>     79 – 849C                                 with 10 mL of the standard solution under the above operat-
                                                                   ing conditions, the relative standard deviation of the peak
Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of
                                                                   area of nabumetone is not more than 5.0z.
Nabumetone according to Method 2, and perform the test.
Prepare the control solution with 1.0 mL of Standard Lead          Water <2.48> Not more than 0.2z (1 g, volumetric titra-
Solution (not more than 10 ppm).                                   tion, direct titration).
   (2) Related substances—Dissolve 20 mg of Nabumetone
                                                                   Residue on ignition <2.44>   Not more than 0.1z (1 g).
in 20 mL of acetonitrile, and use this solution as the sample
solution. Pipet 5 mL of the sample solution, add acetonitrile      Assay Weigh accurately about 20 mg each of Nabumetone
to make exactly 50 mL. Pipet 1 mL of this solution, add            and Nabumetone Reference Standard (separately determine
acetonitrile to make exactly 20 mL, and use this solution as       the water <2.48> in the same manner as Nabumetone), dis-
the standard solution. Perform the test with exactly 10 mL         solve them in acetonitrile to make exactly 20 mL, and use
each of the sample solution and standard solution as direct-       these solutions as the sample solution and the standard solu-
ed under Liquid Chromatography <2.01> according to the             tion, respectively. Perform the test with exactly 10 mL each
following conditions. Determine each peak area of both so-         of the sample solution and standard solution as directed un-
lutions by the automatic integration method: the peak area         der Liquid Chromatography <2.01> according to the follow-
of the related substance G obtained from the sample solu-          ing conditions, and determine the peak area of nabumetone,
tion is not larger than 3/5 times the peak area of nabume-         AT and AS, from each solution.
tone from the standard solution, and each peak area other
                                                                           Amount (mg) of C15H16O2=WS×(AT/AS)
than nabumetone and the related substance G from the sam-
ple solution is not larger than 1/5 times the peak area of           WS: Amount (mg) of Nabumetone Reference Standard,
nabumetone from the standard solution. Furthermore, the                  calculated on the anhydrous basis
total area of the peaks other than nabumetone from the sam-
                                                                   Operating conditions—
ple solution is not larger than 1.6 times the peak area of
                                                                      Detector: An ultraviolet absorption photometer (wave-
nabumetone from the standard solution. For these calcula-
                                                                   length: 254 nm).
tions, use each peak area of the related substances A, B, C,
                                                                      Column: A stainless steel column 4.6 mm in inside di-
D, E, F and G, which are having the relative retention time
                                                                   ameter and 15 cm in length, packed with octadecylsilanized
of about 0.73, 0.85, 0.93, 1.2, 1.9, 2.6 and 2.7 with respect
                                                                   silica gel for liquid chromatography (4 mm in particle di-
to nabumetone, after multiplying by their relative response
                                                                   ameter).
factors, 0.12, 0.94, 0.25, 0.42, 1.02, 0.91 and 0.1, respective-
                                                                      Column temperature: A constant temperature of about
ly.
                                                                   409  C.
Operating conditions—
                                                                      Mobile phase: To 600 mL of a mixture of water and acetic
   Detector, column, and column temperature: Proceed as
                                                                   acid (100) (999:1) add 400 mL of a mixture of acetonitrile
directed in the operating conditions in the Assay.
                                                                   and tetrahydrofuran (7:3).
   Mobile phase A: A mixture of water and acetic acid (100)
                                                                      Flow rate: Adjust the flow rate so that the retention time
(999:1).
                                                                   of nabumetone is about 10 minutes.
   Mobile phase B: A mixture of acetonitrile and tetra-
                                                                   System suitability—
hydrofuran (7:3).
                                                                      System performance: When the procedure is run with 10
   Flowing of mobile phase: Control the gradient by mixing
                                                                   mL of the standard solution under the above operating con-
the mobile phases A and B as directed in the following table.
                                                                   ditions, the number of theoretical plates and the symmetry
  Time after injection      Mobile phase       Mobile phase        factor of the peak of nabumetone are not less than 6000 and
    of sample (min)          A (volz)           B (volz)           not more than 1.5, respectively.
                                                                      System repeatability: When the test is repeated 6 times
          0 – 12                 60                 40
                                                                   with 10 mL of the standard solution under the above operat-
         12 – 28               60ª 20             40ª 80
                                                                   ing conditions, the relative standard deviation of the peak
   Flow rate: 1.3 mL per minute.                                   area of nabumetone is not more than 1.0z.
   Time span of measurement: About 3 times as long as the
                                                                   Containers and storage    Containers—Tight containers.
retention time of nabumetone, beginning after the solvent
peak.
System suitability—
   Test for required detectability: Pipet 2 mL of the standard
solution, and add acetonitrile to make exactly 10 mL. Con-
firm that the peak area of nabumetone obtained from 10 mL
of this solution is equivalent to 14 to 26z of that from 10 mL
of the standard solution.
   System performance: Proceed as directed in the system
suitability in the Assay.
   System repeatability: When the test is repeated 6 times
1910       Official Monographs                                                                       Supplement I, JP XV

Add the following:                                                  C: Labeled amount (mg) of nabumetone (C15H16O2) in 1
                                                                       tablet
Nabumetone Tablets                                                Assay Weigh accurately not less than 20 tablets of
                                                                  Nabumetone Tablets, and powder. Weigh accurately a por-
ナブメトン錠
                                                                  tion of the powder, equivalent to about 0.2 g of nabumetone
                                                                  (C15H16O2), add 10 mL of water and shake, add 40 mL of
  Nabumetone Tablets contain not less than 95.0z
                                                                  methanol, shake for 30 minutes, and then add methanol to
and not more than 105.0z of the labeled amount of
                                                                  make exactly 100 mL. Centrifuge this solution, pipet 5 mL
nabumetone (C15H16O2: 228.29).
                                                                  of the supernatant liquid, add exactly 5 mL of the internal
Method of preparation     Prepare as directed under Tablets,      standard solution, then add methanol to make 50 mL, and
with Nabumetone.                                                  use this solution as the sample solution. Separately, weigh
                                                                  accurately about 40 mg of Nabumetone Reference Standard
Identification To a quantity of powdered Nabumetone
                                                                  (separately determine the water <2.48> in the same manner as
Tablets, equivalent to 80 mg of Nabumetone according to
                                                                  Nabumetone), dissolve by adding 50 mL of methanol and
the labeled amount, add 50 mL of methanol, shake for 10
                                                                  exactly 20 mL of the internal standard solution, then add
minutes and centrifuge the solution. To 1 mL of the super-
                                                                  methanol to make 200 mL, and use this solution as the stan-
natant liquid, add methanol to make 50 mL, and determine
                                                                  dard solution. Perform t