DETERMINATION OF TOTAL AND DIRECT BILURUBIN

1. Specimen Collection
(Syringe Venipuncture
Procedure)
*Note: Bilirubin is light
sensitive so we must cover the
syringe barrel and EDTA Tube
to avoid false results.

2. Inversion EDTA Tube (8x)

3. Total And Direct Bilirubin

3.1 Direct Bilirubin
3.1.a.

Aspirate 100ul of
unhemolyzed Test
Sera/Controls in a test tube.

3.1.b.

Add 1400ul of Bilirubin

Reagent to 100ul of
unhemolyzed Test
Sera/Controls in a test tube

3.1.c.

Add ONE DROP of Bilirubin

Accelerator
3.1.d.

Mix well and read

%T/Absorbance of each tube at
550 nm against Water Blank
after exactly ONE MINUTE.

3.2. Total Bilirubin

3.2.a

Add 1500ul of Absolute

Methyl Alcohol to each of the
mixture of Direct Bilirubin

3.2.b

Shake VIGOROUSLY and

read %T/Absorbance of each
tube at 550 nm against Water
Blank after exactly FIVE
MINUTES. CALIBRATOR:
Place the Bilirubin Calibrator
into the cuvette and read
/absorbance at 550 nm against
Water Blank.
3.3. Absorbance Mode Manual
Computation
3.3.a

The normal value of direct bilirubin (0.10-0.60 mgs/dl; SI unit 1.71-

10.26 mmol/l), total bilirubin (0.30-1.30 mgs/dl;SI unit 5.13-22.23
mmol/l), and indirect bilirubin ( 0.2-0.7 mgs/dl; SI unit 3.42-11.97
mmol/l. The result for Direst Bilirubin of the patient adult female,
Ms. Keyt Ambros is 0.388 mgs/dl or 6.63 mmol/ l in SI unit and for
total bilirubin is 0.832 mgs/dl or 14.23 mmol/l in SI unit. Therefore
the patient has a normal value of direct bilirubin and total bilirubin.
And also has a normal value of indirect bilirubin, 0.444 mgs/dl or
7.6 mmol/l in SI unit.