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    Bilirubin T+D

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    Munawwar Awa
    This document provides instructions for using a colorimetric method to measure total and direct bilirubin levels in serum or plasma samples. The method uses a reaction between bilirubin and diazonium salt of sulphanilic acid to form azobilirubin, which is measured spectrophotometrically. In the presence of DMSO, total bilirubin reacts, while only conjugated bilirubin reacts in its absence. Normal bilirubin levels are provided. The reagents, procedure, calculations, reference, and quality control are described to perform the assay.

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    Bilirubin T+D

    Uploaded by

    Munawwar Awa
    127 views2 pages
    This document provides instructions for using a colorimetric method to measure total and direct bilirubin levels in serum or plasma samples. The method uses a reaction between bilirubin and diazonium salt of sulphanilic acid to form azobilirubin, which is measured spectrophotometrically. In the presence of DMSO, total bilirubin reacts, while only conjugated bilirubin reacts in its absence. Normal bilirubin levels are provided. The reagents, procedure, calculations, reference, and quality control are described to perform the assay.

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    This document provides instructions for using a colorimetric method to measure total and direct bilirubin levels in serum or plasma samples. The method uses a reaction between bilirubin and diazonium salt of sulphanilic acid to form azobilirubin, which is measured spectrophotometrically. In the presence of DMSO, total bilirubin reacts, while only conjugated bilirubin reacts in its absence. Normal bilirubin levels are provided. The reagents, procedure, calculations, reference, and quality control are described to perform the assay.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    127 views2 pages

    Bilirubin T+D

    Uploaded by

    Munawwar Awa
    This document provides instructions for using a colorimetric method to measure total and direct bilirubin levels in serum or plasma samples. The method uses a reaction between bilirubin and diazonium salt of sulphanilic acid to form azobilirubin, which is measured spectrophotometrically. In the presence of DMSO, total bilirubin reacts, while only conjugated bilirubin reacts in its absence. Normal bilirubin levels are provided. The reagents, procedure, calculations, reference, and quality control are described to perform the assay.

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    BILIRUBIN (Total + Direct) STABILITY :

    Total 100 Tests The reagents are stable up to the expiry date
    Colorimetric Method specified when stored at +4 to +8 °C .
    Direct 100 Tests
    PROCEDURE :
    Direct Bilirubin Total Bilirubin
    PRINCIPLE :
    The reaction between bilirubin and the Blank Sample Blank Sample
    ( ml ) ( ml ) ( ml ) ( ml )
    diazonium salt of sulphanilic acid produced
    azobilirubin which shows a maximum Reagent 1 1.0 1.0 - -
    absorption at 535 nm in an acid medium. In Reagent 2 - - 1.0 1.0
    the presence of ( DMSO ) the total bilirubin
    Reagent 3 - 0.05 - 0.05
    participate in the reaction and in the absence
    of (DMSO ) only conjugated bilirubin react . Sample 0.1 0.1 0.1 0.1
    .
    SAMPLE : Mix. Incubate at room temperature for 10 min.
    Serum, plasma. Hemolysis will interfere with away from light. Read the absorbances of the
    the test .Samples should be kept away from sample (ASample ) against its blank at 535 nm.
    light. ( 530 – 540 nm ), using cuvettes 1 cm light path.
    Linearity up to 25 mg / dL (425 μmol/L). Color
    NORMAL VALUES : stable for one hour.
    Total bilirubin up to 1.0 mg / dL (17 μmol/L)
    Direct bilirubin up to 0.25 mg / dL (4.3 μmol/L)
    CALCULATION :
    REAGENTS : Total bilirubin (mg /dl) = ASample x 14
    1. Direct Bilirubin :
    Direct bilirubin (mg /dl) =
    ASample x 14
    Sulphanilic acid 60 mmol /L
    Hydrochloric acid 200 mmol /L Indirect bilirubin (mg / dl) = Total - Direct
    2. Total bilirubin :
    REFERENCE :
    Sulphanilic acid 60 mmol /L
    Hydrochloric acid 200 mmol /L Walter M. and Gerade H., (1970) Microchem. J.
    15. 231
    Dimethylsulfoxide ( DMSO ) 10 mol/L
    3. Sodium nitrite 90 mmol /L QUALITY CONTROL :
    For accuracy and reproducibility control:-
    Assayed Multi – Sera, Normal and Elevated
    BILIRUBIN
    ( Total + Direct )
    Colorimetric Method
    Direct 100 Tests
    +4 to +8˚C
    Total 100 Tests
    In vitro diagnostic use

    CAT. NO. BR 11 10

    REAGENTS
    R1 Sulphanilic acid 100 ml
    R2 Sulphanilic acid –DMSO 100 ml
    R3 Sodium nitrite 10 ml

    CONTACTS
    Tele: 02-33385184
    Mobil: 0109 – 349 20 77
    Fax : 02-33385184 (102)
    e.maile : info@bio-diagnostic.com
    Website: www.bio-diagnostic.com
    Adress: 29 Tahreer St., Dokki, Giza, Egypt

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