Carbon Dioxide ( Carbon Dioxide (CO2) Reagent by steepslope9876

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									                 (CO2) Reagent
  Carbon Dioxide (
                                                                                   INTERFERENCES                                                                        When run as recommended the assay is linear from 1.0 to 50 mEq/L
                                                                                   Studies to determine the level of interference for hemoglobin, bilirubin, and
                                                                                   lipernia were carried out, the following results were obtained:                      Method Comparison:
                                                                                                                                                                        Studies performed between this procedure and a similar methodology yielded
                           Catalog #: 80130                                        Hemoglobin:
                                                                                   Do not use Hemolyzed samples.
                                                                                                                                                                        the following results:
                                                                                                                                                                        Number of samples pairs:            43
                                                                                                                                                                        Range of samples:                   15.0 – 39.0 (mEq/L)
                                                                                   Bilirubin:                                                                           Correlation Coefficient:            0.9786
                             for use with the                                      No significant interference (±10%) from bilirubin up to 8.4 mg/dL                    Slope:                              0.920
                                                                                                                                                                        Intercept:                          2.0 (mEq/L)
                                                                                   Lipemia:
   LIASYS-330 CLINICAL CHEMISTRY SYSTEM
   LIASYS-                                                                         No significant interference (±10%) from lipemia up to 185 mg/dL measured as          Precision:
                                                                                   triglycerides                                                                        Within Run               Level 1      Level 2      Level 3
                                                                                                                                                                        n=40
INTENDED USE                                                                       A number of drugs and substances may affect the accuracy of this test. See           Mean (mEq/L)             7.0          13.3         21.3
For the in vitro quantitative determination of Carbon Dioxide in serum.            Young, et al 5                                                                       S.D. (mEq/L)             0.00         0.32         0.5
                                                                                                                                                                        C.V. (%)                 0.0          2.4          2.6
SUMMARY AND EXPLANATION 1                                                          ADDITIONAL EQUIPMENT REQUIRED BUT NOT PROVIDED
Approximately ninety percent of Carbon Dioxide present in serum is in the form     1.   LIASYS 330 Clinical Chemistry System                                            Total
of bicarbonate. The measurement of bicarbonate, usually in conjuntion with tests   2.   Deionized water and related equipment, e.g.: pipettes                           n=40 (10 days / 2 runs per day / 2 replicates per run)
such as glucose, urea, sodium, potassium, and chloride is useful in the assess-    3.   Analyzer specific consumables, e.g.: sample cups
ment of acid-base balance resulting from metabolic or respiratory causes.          4.   Control, and Calibrator materials such as those provided by AMS Diag-           Mean (mEq/L)             7.0          13.3         21.3
                                                                                        nostics.                                                                        S.D. (mEq/L)             0.00         0.60         0.8
METHODOLOGY 2,3                                                                    5.                                                                                   C.V. (%)                 0.0          4.5          3.9
The AMS Diagnostics Carbon Dioxide procedure is based upon phosphoenolpy-          ASSAY PROCEDURE
ruvate caboxylase (PEPC) utilizing bicarbonate present in the sample to produce                                 System Parameters                                       Sensitivity / Limit of Detection:
oxaloacetate and phosphate. Malate dehydrogenase (MDH) then catalyses the                                                                                               A calibration factor of approximately 231.300 was obtained, which is equivalent
reduction of oxaloacetate to malate and the oxidation of NADH to NAD. The          Carbon Dioxide (CO2) Liquid                                                          to a sensitivity of 4.323 ∆Abs per mEq/L.
resulting decrease in absorbance can be measured at 380nm and is proportional      TEMPERATURE:                          37°C
to the amount of bicarbonate in the sample.                                        WAVELENGTH:                           380 nm                                         The lower Limit of Detection was found to be 1.0 mEq/L.
                                                                                   DIRECTION:                            Decrease
Principle                                                                          SAMPLE / RGT RATIO:                   1 : 100                                        REFERENCES
                                      PEPC                                         e.g. Sample Vol.                      0.01 mL (10mL)                                 1
                                                                                                                                                                            Zilv JF, Pannll PR: Hydrogen Ion Homeostasis: Blood Gas Levels, IN
            -
PEP + HCO 3                                        Oxaloacetate +   H2PO-4              Reagent Vol.                     1.00 mL                                            Clinical Chemistry in Diagnosis and Treatment. Lloyd-Luke, London, 1979,
                                                                                   REACTION TIME:                        5 min                                              pp 78-113
                                       MDH                                                                                                                              2
                                                                                                                                                                            Norris, K.A., Atkinson AR, Smith WG: Clin. Chem. 21:1093 (1975)
Oxaloacetate + NADH                                Malate + NAD                    Procedure Notes:                                                                     3
                                                                                                                                                                            Forrester RL, Wataji JJ, Silverman DA, Pierre JK Clin Chem 22:243-5,
                                                                                   1. The reagent and sample volumes may be altered proportionally to accom-                1978
                                                                                                                                                                        4
                                                                                      modate various instrument requirements.                                               Kaplan, L.A., Clinical Chemistry: Theory, Analysis and correlation,
                                                                                                                                                                            C.V.Mosby, St Louis, p872 (1989)
                                                                                                                                                                        5
REAGENT COMPOSITION                                                                Calculations:                                                                            Young, D.S., Effects of Drugs on Clinical Laboratory Tests, 3rd Ed. 1990:
Active Ingredients                    Concentration                                (A = Absorbance)                                                                         pp 57-59
Phosphoenolpyruvate                   8.0 mM                                                                                                                            6
                                                                                                                                                                            Tietz, N.W., Fundamentals of Clinical Chemistry, W.B. Saunders, Philadel-
NADH                                  1.4 mM                                       A (Reagent Blank) - A (Sample)         x Concentration of Standard     =     CO2         phia, PA., 1976 pp 885
Phosphoenolpyruvate carboxylase       >400 U/L                                     A (Reagent Blank) – A (Standard)             (mEq/L)                       (mEq/L)
(microbial)
MDH (microbial)                       >200 U/L                                     Example:                                                                                                                                            PI Rev 02/26/09
PH 7.9                                                                             A (Reagent Blank)                     = 1.500
                                                                                   A (Standard)                          = 1.200
                                                                                   A (Sample)                            = 1.240
Precautions                                                                        Concentration of Standard             = 30 mEq/L
1.   Reagent contains Sodium Azide. This may react with copper or lead
     plumbing to form explosive metal azides. Upon disposal, flush with a          1.500 – 1.240   x 30 mEq/L =      .260 x 30 mEq/L     =     26 mEq/L CO2
     large volume of water to prevent azide build up.                              1.500 – 1.200                     .300
2.   Reagents are for in vitro diagnostic use only.
3.   Do not ingest. Toxicity has not been established.                             Limitations
4.   Do not pipet by mouth to avoid CO2 contamination from the expired air.        1.    Samples exceeding linearity must be diluted 1:1 with saline, re-assayed,
                                                                                         and the result multiplied by two.
REAGENT PREPARATION                                                                2.    Carbon Dioxide contamination must be avoided.
Reagent is supplied ready to use.
                                                                                   CALIBRATION
REAGENT STORAGE                                                                    Use an aqueous CO2 standard or an appropriate serum calibrator. Refer to
1.  Store the reagent at 2-8°C (refrigerated)                                      appropriate instrument operator manual for recommended calibrator interval.
2.  The reagent is stable until the expiration date when stored at 2-8°C
                                                                                   QUALITY CONTROL                                                                                                      Manufactured for:
REAGENT DETERIORATION                                                              The integrity of the reagent should be monitored by use of a two level control
Do not use the reagent if:                                                         with known CO2 values.
1.   The absorbance of the reagent is less than 0.85 at 380nm
2.   Working reagent does not meet stated performance parameters                   EXPECTED VALUES 6
                                                                                   23 – 29 mEq/L
SPECIMEN COLLECTION AND STORAGE
1.  Fresh, unhemolyzed serum, collected under anaerobic conditions is the          It is strongly recommended that each laboratory determine its own reference                                   117 Old State Road
    recommended specimen                                                           range                                                                                            Brookfield, CT 06804 — Tel 1 866 419 7839
2.  The specimen stored tightly stoppered is stable for several days at 4°C.4
                                                                                                                                                                                                 Fax 1 843 277 0903
                                                                                   PERFORMANCE
                                                                                   Linearity:                                                                                                 www.amsdiagnostics.com

								
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