liver function testsMedical analysis

Estimation of Albumin in serum – 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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