Geneva by HC120916193544


									PHARM 462
2009    1/31
Good Manufacturing Practices (GMP)
       VALIDATION of

    2009                     2/31
       GMP - 4.11 Analytical methods,
     computers and cleaning procedures
 „It is of critical importance that particular attention is
  paid to the validation of analytical test methods,
  automated systems and cleaning procedures.”
 Validation of analytical procedures used in the
  examination of pharmaceutical materials (WHO Expert
  Committee on Specifications for Pharmaceutical Preparations.
  32nd Report. Geneva, WHO, 1992 (WHO Technical Report
  Series, No. 823).
 Text on Validation of Analytical Procedures Q2A
  (1994) Validation of Analytical Procedures:
  Methodology Q2B (1996) ICH Harmonized Tripartite

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         Characteristics of methods
 Accuracy (also termed trueness)
 Precision
      repeatability
      intermediate precision (intra-laboratory variation)
      reproducibility (inter-laboratory variation)
 Robustness, ruggedness
 Stability

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          Accuracy and precision

             Inaccurate &
 Inaccurate and imprecise

 Inaccurate but       Accurate but
    precise          Accurate
                       imprecise     Accurate and precise
         2009                                        5/31
       Accuracy (of any process)
The accuracy of an analytical  application of an analytical
procedure expresses the         procedure to an analyte of known
closeness of agreement          purity (e.g. reference material);
between the value, which is    comparison of the results of the
accepted either as a            proposed analytical procedure
                                with those of a second well-
conventional true value or an   characterized procedure, the
accepted reference value and    accuracy of which is stated
the value found (individual     and/or defined
observation or mean of         accuracy may be inferred once
measurements).                  precision, linearity and
                                specificity have been established.

        2009                                            6/31
        Precision (of any process)
Measured mean   Real mean
                              The precision (VARIABILITY)
                              of an analytical procedure is
                              usually expressed as the standard
                              deviation (S), variance (S2), or
                              coefficient of variation (= relative
                              standard deviation, R.S.D.) of a
                              series of measurements.

                              The confidence interval should be
                              reported for each type of
                              precision investigated.
         2009                                           7/31
Repeatability (of any process)
Measured mean
                           Repeatability expresses the
                           precision (spread of the data,
                           variability) under the same
                           operating conditions over a
                           short interval of time.
                           Repeatability is also termed
                           intra-assay precision.


    2009                                          8/31
           Intermediate Precision and
Measured means              Intermediate precision expresses
                            within-laboratories variations. #1,
                            #2 and #3: different days, different
                            analysts, different (manufacturing)
                            equipment, etc.

                            Reproducibility expresses the
                            precision between laboratories #1,
                            #2 and #3 (collaborative studies,
                            usually applied to standardization
                            of methodology). (Transfer of
   Intermediate precision
   or Reproducibility
    2009                                             9/31
       Sensitivity and robustness

                       Input-output relationship

2009                                      10/31
                           Stability (of the analytical
Stability Measured means
                           solution) expresses variation
                           of the measured mean as a
                           function of time.
                           (Manufacturing machines)

                           #1 … First mesurements
                           #2 … Second mesurements
                           of the same sample within a
                           relatively short period of

   2009                                          11/31
      Characteristics of analytical
 Linearity
 Range
 Specificity (selectivity)
 Sensitivity (versus robustness)
 Limit of detection
 Limit of quantitation

     2009                           12/31
Measured     Real               Linearity expresses
 mean        mean   Precision   differences in precision at
                                different points of a given
                                “The linearity of an
                                analytical procedure is its
                                ability (within a given
                                range) to obtain test
                                results, which are directly
                                proportional to the
                                concentration (amount) of
                                analyte in the sample.”

      2009                                          13/31
   Range (minimum requirements)
 Assay of an API or a FPP: ± 20% of the test concentration.
 Content uniformity: ± 30% of the test concentration (unless a
   wider more appropriate range, based on the nature of the dosage form
   (e.g., metered dose inhalers), is justified).
 Dissolution testing: ± 20 % over the specified range.
 Impurity: from the reporting level of an impurity to 120% of
  the specification. (Unusually potent or toxic impurities, LOD and LOQ
   should be commensurate with ICH requirement.)
 If assay and purity are performed together as one test and only
  a 100% standard is used, linearity should cover the range from
  the reporting level of the impurities to 120% of the assay

         2009                                                    14/31
       Specificity (selectivity)
 Specificity is the ability to assess unequivocally
  the analyte in the presence of components, which
  may be expected to be present. Typically these
  might include impurities, degradants, excipients,
 Stability indicating analytical methods should
  always be specific.

     2009                                    15/31
             LOD, LOQ and SNR

 Limit of Quantitation (LOQ)
 Limit of Detection (LOD)      Peak B
 Signal to Noise Ratio (SNR)

                       Peak A

 Baseline     noise

      2009                               16/31
    Classes of analytical tests
   „The objective of validation of an analytical procedure is to demonstrate that
    it is suitable for its intented purpose.”

 Class A: To establish identity
 Class B: To detect (Bd) and quantitate (Bq) impurities
 Class C: To determine quantitatively the concentration,
    or assay
 Class D: To assess characteristics
 Other classes not covered in the guides

        2009                                                             17/31
Criteria for analytical classes
Criteria                A    Bq   Bd   C           D
Accuracy                     X         X           X
Precision                    X         X           X
Robustness              X    X    X    X           X
Linearity and range          X         X           X
Specificity             X?   X    X    X           X
Limit of detection                X
Limit of quantitation        X
     2009                                  18/31
        General requirements
 Qualified and calibrated instruments
 Documented methods
 Reliable reference standards
 Qualified analysts
 Sample integrity
 Change control (e.g., synthesis, FPP composition)

     2009                                    19/31
HPLC Method Development and
 Validation for Pharmaceutical

         Pharmaceutical Technology
           Europe, 1 March 2004

  2009                               20/34
Prequalification requirements
 Analytical method validation is required by WHO
  for the prequalification of product dossiers. Non-
  compendial ARV APIs and FPPs were/are tested
  with methods developed by the manufacturer.
 Analytical methods should be used within GMP
  and GLP environments, and must be developed
  using the protocols and acceptance criteria set out
  in the ICH guidelines Q2A and Q2B.

     2009                                     21/31
       HPLC system

2009                 22/31
       Linearity and range

2009                         23/31
            ICH requirements
 Concentration range 0.025–0.15 mg/mL (25–150% of the
  theoretical concentration in the test preparation, n=3)
 Regression equation was found by plotting the peak area
  (y) versus the analyte concentration (x) expressed in
  mg/mL: y = 3007.2x + 4250.1 (r2 = 1.000).
 The regression coefficient demonstrates the excellent
  relationship between peak area and concentration of
 The analyte response is linear across 80-120% of the
  target progesterone concentration.

     2009                                          24/31

The data show that the recovery of analyte in spiked samples met the
evaluation criterion for accuracy (100 ± 2.0% across 80–120% of target

         2009                                                    25/31

2009                 26/31
 An example of specificity criterion for an assay
  method is that the analyte peak will have baseline
  chromatographic resolution of at least 2.0 minutes
  from all other sample components.
 In this study, a weight of sample placebo
  equivalent to the amount present in a sample
  solution preparation was injected to demonstrate
  the absence of interference with progesterone
  elution. Former slide demonstrates specificity.

     2009                                    27/31
              The repeatability
              precision obtained by
              one analyst in one
              laboratory was 1.25%
              RSD for the analyte and,
              therefore, meets the
              evaluation criterion of
              RSD ≤2%.

2009                            28/31
  Intermediate precision

2009                       29/31
              Limit of detection
 The limit of detection (LOD) is defined as the
  lowest concentration of an analyte in a sample that
  can be detected, not quantified. It is expressed as a
  concentration at a specified signal:noise ratio
  (SNR), usually between 3 and 2:1.
 In this study, the LOD was determined to be
  10ng/mL with a signal:noise ratio of 2.9:1.

      2009                                       30/31
             Limit of quantitation
 The limit of quantitation (LOQ) is defined as the
  lowest concentration of an analyte in a sample that
  can be determined with acceptable precision and
  accuracy under the stated operational conditions of
  the method. The ICH has recommended a signal :
  noise ratio (SNR) of 10:1.
 The LOQ was 20 ng/mL with a signal:noise ratio of
  10.2. The RSD for six injections of the LOQ
  solution was ≤2%.

      2009                                    31/31
 Analytical solution stability

Standard and sample solutions stored in a capped volumetric flask on
a lab bench under normal lighting conditions for 24 h were shown to
be stable with no significant change in progesterone concentration
during this period.
    2009                                                     32/31
               Main Points Again
 Validation of analytical procedures is a critical
  requirement in risk assessment and management:
      establishment of product-specific acceptance criteria, and
      stability of APIs and FPPs.
 Validation should demonstrate that the analytical
  procedure is suitable for its intented purpose.
 HPLC systems and method validation deserves
  special attention during the inspection of QC
        2009                                             33/31

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