Estimation of Albumin in serum - liver function tests


    Liver Function Tests

    Estimation of Albumin in serum

    Estimation of Albumin in serum
    Introduction
    ·       Albumin accounts 60% of human plasma protein
    ·       It binds to water-insoluble substances (as bilirubin, fatty acids , or hormones)
    ·       Regulate the osmotic pressure of blood
    ·       Albumin is synthesized by liver using dietary proteins
    Principle
    ·       Under acidic conditions , Albumin present in serum sample binds to bromo cresol green to give a blue - green colored complex which read at 630 nm
    ·       The intensity of color is directly proportional to Albumin concentration
    Albumin + bromo cresol green → Albumin – bromo cresol green complex (blue green color)
    Normal range
    ·       Albumin : 3.5 – 5.5 g %


    Reagents
    ·       Citrate buffer :  give acidic medium
    ·       Bromo cresol green : color reagent
    ·       Bovine serum albumin (BSA) : standard protein
    ·       Detergent : it prevent the conversion of bromo cresol green from green to dark blue , Buffer isn’t completely soluble ,but with detergent it becomes completely soluble
    Procedure
    Depend on colorimetric technique
    Reagent
    Test
    Standard
    Blank
    Serum
    20µ
    -
    -
    St-protein
    -
    20µ
    -
    Color reagent
    3ml
    3ml
    3ml

    Mix well ,stand for 5 min at RT , then read at 630 nm

    Calculations
    Calculations
    Serum Albumin level increased in:
    •        Dehydration
    •        Poor protein utilization
    Serum Albumin level decreased in:
    ·       Malnutrition
    ·       Liver dysfunction (Ascites)
    Procedure with kits in lab :
    •        Depend on colorimetric technique
    •        1000µ Albumin reagent + 10µ serum , Mix well, stand for 5 min at RT then read 

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