1. Determination Of Albumin In Serum By Bromocresol Green Method
By: Dr. Mohammed Golam Rasul

2. السلام عليكم ورحمة الله وبركاته

3. Objectives
To understand the importance of measuring blood albumin level.
To determine the concentration of albumin in serum sample by using known standard.

4. Introduction
Albumin is made in the liver and is responsible for maintaining proper fluid balances. Decreased amounts of albumin can occur when the liver is not making enough or if albumin is being lost through the kidneys. Increases in albumin do not occur naturally but can be seen in patients who had received albumin suspensions.

5. Functions of Albumin • Principal functions – Maintaining fluid balance
– Carrier
– Anti-oxidant activity
– Buffer

6. Methods of Albumin estimation
1-Precipitation method
2-Electrophoresis
3-Globulin Tryptophan content method
4-Immunochemical methods.
5-Dye binding methods

7. Types of Samples Plasma Serum
Water + solids (e.g. glucose, urea, albumin, fibrinogen)
No cells
Serum
Serum = plasma – clotting factors

8. Preparation of sample: Serum
Venous
Blood
Transfer the
clear
supernatant
to specimen
tube
Allow blood to clot
(20min.)
Remove the clot
and centrifuge
cells
Cent. tube

9. Determination of Serum albumin
Requirements:
Automatic pipettes
Tips
Cuvettes
Spectrophotometer
Reagent
Specimen: 
Serum, heparinised or EDTA-plasma

10. Determination of Serum albumin
Principle:
Bromocresol green with albumin in citrate buffer forms a colored complex.
The intensity of the color is directly proportional to the albumin concentration in the sample.

11. Determination of Serum albumin
Procedure
Blank
Standard
Sample

12. Determination of Serum albumin
Procedure (kit)
Reagents
Blank
Standard
Sample
Standard (ml) - 20
Sample (ml)
Reagent (mll)
2000

13. Procedure Add the reagent, sample & standard according to instruction.
Mix and let stand at room temperature for 5 min.
Read the absorbance at 630 nm.
Zero the spectrophotometer with the blank reagent.
Read the absorbances of the standard and samples within 30 min.
Enter the absorbances readings and calculated values in the table above.

14. Calculation
Concentration of albumin (g/dl) = Sample Absorbance x 4 g/dl
Stand. Absorbance
Normal level
g/dl

15. Clinical Significance
One of the most important serum proteins produced in the liver is albumin. Variation in albumin from normal level results malnutrition, Liver diseases ( for example hepatitis or cirrhosis) skin lesions such as dermatitis and burns or dehydration. Clinical diagnosis should not be made on a single test result; it should integrate clinical and other laboratory data.