Veterinary Clinical Biochemistry and Laboratory Diagnosis

PROCEDURE

• Specimen: Serum/Plasma
• Reagents: (Biosystem diagnostics Kit)
• (1) Diazo-A (2) Diazo-B (3) Diazo Blank (4) Methanol (5) Artificial standard (=10mg% Bilirubin)
• Preparation of Diazo Reagent: Just before use, mix 3.3ml of Diazo-A with 0.1 ml of Diazo-B.

Procedure :[reagents in the kit vary from  manufacturer to

                    manufacturer so the procedure in  the kit used may be followed.]

• Mark four test tubes as Total (T1), Total Blank(T2), Direct (D1) and Direct Blank (D)

S. No.	Reagents (ml)	T1	T2	D1	D2
1	Serum/Plasma	0.2	0.2	0.2	0.2
2	Distilled Water	1.8	1.8	-	-
3	Diazo Blank	-	0.5	-	0.5
4	Diazo Reagent	0.5	-	0.5	-
5	Distilled Water	-	-	2.5	2.5
6	Methonol	2.5	2.5	-	-

• Mix well and keep the tubes T1 and T2 in dark at room temperature for 30 minutes.
• Read the OD of all the tubes at 540 nm or using an yellow green filter.

Calculation