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

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    Dinesh Sreedharan
    SGOT is an enzyme found mainly in heart, liver, skeletal muscle and kidneys. Elevated levels are found with injuries to these tissues like myocardial infarction, hepatitis, and muscle diseases. The assay measures SGOT activity by its ability to catalyze the transfer of an amino group from L-aspartate to α-ketoglutarate, forming oxaloacetate and glutamate. Oxaloacetate reacts with NADH and malate dehydrogenase to form NAD, whose oxidation rate is measured as a decrease in absorbance proportional to SGOT activity. Normal reference ranges are up to 37 IU/L at 37°C for males and 31 IU/L for females.

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

    Uploaded by

    Dinesh Sreedharan
    778 views1 page
    SGOT is an enzyme found mainly in heart, liver, skeletal muscle and kidneys. Elevated levels are found with injuries to these tissues like myocardial infarction, hepatitis, and muscle diseases. The assay measures SGOT activity by its ability to catalyze the transfer of an amino group from L-aspartate to α-ketoglutarate, forming oxaloacetate and glutamate. Oxaloacetate reacts with NADH and malate dehydrogenase to form NAD, whose oxidation rate is measured as a decrease in absorbance proportional to SGOT activity. Normal reference ranges are up to 37 IU/L at 37°C for males and 31 IU/L for females.

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    SGOT

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    SGOT is an enzyme found mainly in heart, liver, skeletal muscle and kidneys. Elevated levels are found with injuries to these tissues like myocardial infarction, hepatitis, and muscle diseases. The assay measures SGOT activity by its ability to catalyze the transfer of an amino group from L-aspartate to α-ketoglutarate, forming oxaloacetate and glutamate. Oxaloacetate reacts with NADH and malate dehydrogenase to form NAD, whose oxidation rate is measured as a decrease in absorbance proportional to SGOT activity. Normal reference ranges are up to 37 IU/L at 37°C for males and 31 IU/L for females.

    Copyright:

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

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

    Uploaded by

    Dinesh Sreedharan
    SGOT is an enzyme found mainly in heart, liver, skeletal muscle and kidneys. Elevated levels are found with injuries to these tissues like myocardial infarction, hepatitis, and muscle diseases. The assay measures SGOT activity by its ability to catalyze the transfer of an amino group from L-aspartate to α-ketoglutarate, forming oxaloacetate and glutamate. Oxaloacetate reacts with NADH and malate dehydrogenase to form NAD, whose oxidation rate is measured as a decrease in absorbance proportional to SGOT activity. Normal reference ranges are up to 37 IU/L at 37°C for males and 31 IU/L for females.

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    SGOT (AST) ASSAY PROCEDURE:

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


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

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


    SUMMARY:
    SGOT is an enzyme found mainly in heart muscle, liver cells, skeletal muscle and kidneys. Injury to Addition Sequence Test (T)
    these tissues results in the release of the enzyme in blood. Elevated levels are found in myocardial Working Reagent 1000µl
    infarction, Cardiac operations, Hepatitis, Cirrhosis, acute pancreatitis, acute renal diseases, Sample 100µl
    primary muscle diseases. Decreased levels may be found in Pregnancy, Beri Beri and Diabetic
    ketoacidosis. Mix well and read the initial absorbance A after 1 min. and repeat the absorbance reading after
    1,2 & 3 minutes. Calculate the mean absorbance change per min.( A/min).
    PRINCIPLE:
    SGOT (AST) catalyzes the transfer of amino group between L-Aspartate and a-Ketoglutarate to CALCULATIONS:
    form Oxaloacetate and Glutamate. The Oxaloacetate formed reacts with NADH in the presence of SGOT (AST) Activity in IU/L 37°C = A /min. x 1746 x tf
    Malate Dehydrogenase to form NAD. The rate of oxidation of NADH to NAD is measured as a
    decrease in absorbance which is proportional to the SGOT (AST) activity in the sample. TEMPERATURE CONVERSION FACTORS:
    SGOT Desired Reporting Temperature(tf)
    L-Aspartate acid + a-Ketoglutarate Oxaloacetate+L-Glutamate Assay
    25°C 30°C 37°C
    MDH
    25°C 1.00 1.37 2.08
    Oxaloacetate + NADH + HMalate + NAD
    30°C 0.73 1.00 1.54
    CONTENTS: 37°C 0.48 0.65
    PACK SIZE ENZYME REAGENT(A1) DILUENT (A2)
    NORMAL REFERENCE VALUES:
    6x10ml 6x10ml 60ml Serum (males) : Up to 37 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 1+9
    Reagent (A1) labels 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. SGOT (AST) 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
    population.
    Reaction Mode U.V Kinetic Sample Volume 100 µl
    Wavelength 340 nm Reagent volume 1000 µl REFERENCES:
    Blank Distilled Water Factor 1746 IFCC methods for the measurements of catalytic concentrations of enzymes, J. Clin. Chem.
    Clin. Biochem. (1986) 24: 497
    Incubation 37°C Reaction Slope Decreasing
    Delay Time 60 sec Linearity 450 IU/L
    Read Time 180 sec Units IU/L

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