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    (Mod. IFCC Method) For The Determination of SGPT (ALT) Activity in Serum. (For in Vitro Diagnostic Use Only)

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    Dinesh Sreedharan
    SGPT (ALT) is an enzyme found mainly in the liver that is involved in the transfer of amino groups. Increased levels of SGPT (ALT) are found in conditions affecting the liver such as hepatitis, cirrhosis, and obstructive jaundice. The document provides the procedure, calculations, reference values, and specifications for determining SGPT (ALT) activity levels in serum samples.

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    (Mod. IFCC Method) For The Determination of SGPT (ALT) Activity in Serum. (For in Vitro Diagnostic Use Only)

    Uploaded by

    Dinesh Sreedharan
    458 views1 page
    SGPT (ALT) is an enzyme found mainly in the liver that is involved in the transfer of amino groups. Increased levels of SGPT (ALT) are found in conditions affecting the liver such as hepatitis, cirrhosis, and obstructive jaundice. The document provides the procedure, calculations, reference values, and specifications for determining SGPT (ALT) activity levels in serum samples.

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    SGPT

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    SGPT (ALT) is an enzyme found mainly in the liver that is involved in the transfer of amino groups. Increased levels of SGPT (ALT) are found in conditions affecting the liver such as hepatitis, cirrhosis, and obstructive jaundice. The document provides the procedure, calculations, reference values, and specifications for determining SGPT (ALT) activity levels in serum samples.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    458 views1 page

    (Mod. IFCC Method) For The Determination of SGPT (ALT) Activity in Serum. (For in Vitro Diagnostic Use Only)

    Uploaded by

    Dinesh Sreedharan
    SGPT (ALT) is an enzyme found mainly in the liver that is involved in the transfer of amino groups. Increased levels of SGPT (ALT) are found in conditions affecting the liver such as hepatitis, cirrhosis, and obstructive jaundice. The document provides the procedure, calculations, reference values, and specifications for determining SGPT (ALT) activity levels in serum samples.

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    SGPT (ALT) ASSAY PROCEDURE:

    (Mod. IFCC method) Wavelength/ Filter : 340 nm


    For the determination of SGPT (ALT) activity in serum. Temperature : 37°C
    (For In vitro Diagnostic Use Only) Light Path : 1 cm

    SUMMARY: Pipette into clean dry test tube labeled as Test(T)


    SGPT is found in a variety of tissues but is mainly found in the liver. Increased levels are found in Addition Sequence Test (T)
    hepatitis, cirrhosis, obstructive jaundice and other hepatic diseases. Slight elevation of the Working Reagent 1000µl
    enzymes is also seen in myocardial infarction.
    Sample 100µl
    PRINCIPLE: Mix well and read the initial absorbance A after 1 min. and repeat the absorbance reading
    SGPT (ALT) catalyzes the transfer of amino group between L-Alanine and a-Ketoglutarate to after 1,2 & 3 minutes. Calculate the mean absorbance change per min.( A/min).
    form Pyruvate and Glutamate. The Pyruvate formed reacts with NADH in the presence of Lactate
    Dehydrogenase to form NAD. The rate of oxidation of NADH to NAD is measured as a decrease CALCULATIONS:
    in absorbance which is proportional to the SGPT (ALT) activity in the sample. SGPT (ALT) Activity in IU/L 37°C = A/ min. x 1746 x tf
    SGPT
    L-Alanine + a-Ketoglutarate Pyruvate+L-Glutamate TEMPERATURE CONVERSION FACTORS:
    LDH Assay Desired Reporting Temp 37°C
    Pyruvate + NADH + H Lactate + NAD 25°C 1.00
    30°C 0.73
    CONTENTS:
    37°C 0.48
    PACK SIZE ENZYME REAGENT (A1) DILUENT (A2)
    NORMAL REFERENCE VALUES:
    6x10ml 6x10ml 60ml Serum (males) : Up to 40 IU/L at 37°C
    5x20ml 5x20ml 100ml (females) : Up to 31 IU/L at 37°C

    REAGENT PREPARATION: LINEARITY:


    WORKING REAGENT : For sample start assays a single reagent is required. Reconstitute one The procedure is linear up to 450 IU/L at 37°C, If the absorbance change (? A/ min.) exceeds
    vial of enzyme reagent (A1) with equivalent Volume of diluent (A2) (mentioned on the Enzyme 0.250, use only the value of the first two minutes to calculate the result, or dilute the sample
    Reagent (A1) labels 1+9 with normal saline (NaCl 0.9%) and repeat the assay (Results x 10).

    STORAGE AND STABILITY: NOTE:


    Working reagent: This working reagent is stable for at least 4 weeks when stored at 2-8°C. Samples having a very high activity show a very low initial absorbance as most of the NADH
    is consumed prior to the start of measurement. If this is suspected then dilute the sample and
    SAMPLE MATERIAL: repeat the assay. The working reagent or the combined reagent should have an absorbance
    Serum. Free from hemolysis. SGPT (ALT) is report to be stable in serum for 3 days at 2-8°C. above 1.0 against distilled water at 340 nm. Discard the reagent if the absorbance is below
    1.0.
    GENERAL SYSTEM PARAMETER:
    It is recommended that each laboratory establish its own normal range representing its patient
    Reaction Mode U.V Kinetic Sample Volume 100 µl population.
    Wavelength 340 nm Reagent volume 1000 µl
    Blank Distilled Water Factor 1746 REFERENCES:
    IFCC methods for the measurements of catalytic concentrations of enzymes, J. Clin. Chem.
    Incubation 37°C Reaction Slope Decreasing
    Clin. Biochem. (1986) 24: 481
    Delay Time 60 sec Linearity 450 IU/L
    Read Time 180 sec Units IU/L

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