1-Total Bilirubin:

Procedure:
sample serum or plasma

DON'T HAEMOLYSIS

1. Prepare two test tubes mark one as Blank tube and the other as Test tube .
2. put in the blank tube : 100 µ reagent 1 + 500 µ reagent 3 + 100 µ specimen mix and leave for 10 min. at room temp.
3. at same time put in test tube : 100 µ reagent 1 + 25 µ reagent 2 + 500 µ reagent 3 + 100 µ specimen mix and leave for 10 min. in room temp.
4. then put 500 µ reagent 4 in each tube mix and leave for 5 min. in room temp.
5. MEASURING AGAINST BLANK at wave length 580 nm.

Calculation:
reading of test x 10.8 = ......mg / dl .

Normal values up to 1.0 mg / dl .

2-Direct Bilirubin:

1. Prepare two test tubes mark one as Blank tube and the other as Test tube .
2. put in the blank tube : 100 µ reagent 1 + 1 ml saline + 100 µ specimen mix and leave for 5 min.         
3. put in test tube at same time : 100 µ reagent 1 + 25 µ reagent 2 + 1 ml saline + 100 µ specimen mix and leave for 5 min.
4. MEASURE AGAINST BLANK at wave length 580 nm .

Calculation:
Reading of test x 14.4 = ....... mg / dl .

Normal values up to 0.25 mg / dl