BILIRUBIN DETERMINATION

This module aims to focus on determining the qualitative examination of bilirubin

in a urine specimen. The students are expected to know the principle involved in each
procedure as well as to identify some clinical indications if the sample is positive for
bilirubin.

Learning Objectives:

At the end of the module, the students will be able to:

1. Know the principle involved on the following procedures

2. Know how the procedure is done
3. Properly identify the result obtained
4. Indicate possible causes of the presence of bilirubin in urine

The principal pigment of bile is bilirubin which is the degradation product of

haemoglobin. Tests for bile depend upon the oxidation of bile pigments by acids with
formation of a series of colored derivatives of bilirubin such as biliverdin (green),
bilicyanide (blue), and choletellin (yellow) with bilirubin as the most important of all.

A. Smith’s test
Procedure:
1. Take 5mL of urine in a test tube.
2. Overlay the urine with the tincture of alcoholic iodine
3. Observe the result and report as positive or negative.

Result:

An emerald green ring at the point of contact shows the presence of bile
pigments.

B. Harrison’s Spot Test

Procedure:
1. Take 5mL of urine and 5mL of barium chloride in a test tube.
 2. Mix and stand for a few minutes.
3. Filter and spread the filter paper bearing the precipitate on another piece of
dry filter paper.
4. Place 3 drops of Fouchet’s reagent on the precipitate.
5. Observe the result and report as positive or negative.

Result:

A positive result is indicated by a blue to green color.

Clinical Conditions:

1. Hepatitis A, Hepatitis B, and Hepatitis C

2. Fatty liver disease and cirrhosis
3. Liver cancer
4. Hemochromatosis
5. Wilson disease