VLD 421: Veterinary Clinical Biochemistry and Laboratory Diagnosis-II (0+1)

Estimation of Albumin

METHODS OF ESTIMATION OF ALBUMIN: BCG DYE METHOD

Sample : Serum or Plasma 

Principle:

• Albumin in serum binds with the dye Bromocresol Green (BCG) at pH 3.8 to form a green colored complex, the absorbance of which is measured at 600 nm.

Reagents:

• Albumin Reagent (BCG Reagent – Succinate buffer, Bromocresol green)
• Albumin standard (3g%)

Procedure:

 Mark three test tubes as Blank, Standard and Test and proceed as follows:

S.No.	Reagents	Blank (B)	Standard (S)	Test (T)
1.	BCG Reagent	1.0 ml	1.0 ml	1.0 ml
2.	Albumin Standard	-	10 m l	-
3.	Serum	-	-	10 m l
4.	Distilled Water	2.0 ml	2.0 ml	2.0 ml

• Mix well and allow the tubes to stand at R.T for 1 min.
• Measure the O.D of all the Standard and Test against Blank at 600nm or using a red filter.

Calculations :

Albumin (g/dL)  =                 OD of (T)

                                               ------------ x 3

                                                OD of (S)

Globulins (g/dL) = (Total protein ) – (Albumin)

 

A/G Ratio =                      (Albumin)

                                  ---------- ------------

                               (Total protein) – (Albumin)