BILE PIGMENT , BILE

SALTS
Sanjana
BMLT21108
BILE PIGMENT (BILIRUBIN)
 Bilirubin is a breakdown product of hemoglobin.
Undetectable in urine of normal person
Conjugated bilirubin appears in the urine.
Bilirubinuria : presence of bilirubin in urine.
INDICATIONS
1. Acute viral hepatitis
2. In fever unknown origin
3. Differential diagnosis of jaundice

Hemolytic Hepatocellular Obstructive

Urine test
jaundice jaundice jaundice
• Bilirubin • absent • present • present
TESTS FOR DETECTION OF
BILIRUBIN
Sample : fresh , kept protected from light

Methods for detection :

• Foam test
• Gmelin`s test
• Lugol iodine test
• Fouchet`s test
• Reagent strip
FOAM TEST

About 5ml of urine in a test tube is

shaken and observed for development
of yellowish foam
 Normal urine : white foam
GMELIN`S TEST

Take 3ml of concentrated

nitric acid in a test tube and
slowly place equal quantity
of urine over it .
The tube is shaken gently
Play of colors indicates
positive test
LUGOL IODINE TEST

Take 4ml of Lugol iodine solution in a test tube

 add 4 drops of urine.
Mix by shaking.
Positive test : Development of green color
FOUCHET`S TEST

Take 5ml of fresh urine in a test tube

Add 2.5ml of 10% barium chloride, mix well
A precipitate of sulfates appears
Filter to obtain the precipitate on a filter paper
To the filtered precipitate , add 1 drop of Fouchet`s reagent
Immediate development of blue-green color around the drop
indicates presence of bilirubin.
REAGENT STRIP TEST

These tests are based on the reaction of bilirubin with diazo

reagent.
Color change is proportional to the concentration of bilirubin.
BILE SALTS
Bile salts are the biological detergents synthesized from
cholesterol in the liver.
The conjugated bile salts synthesized in the liver accumulate in
gall bladder.
From there they are secreted into the small intestine where they
serve as emulsifying agents for the digestion and absorption of fats
and fat soluble vitamin
Bile salts along with bilirubin can be detected in obstructive
jaundice
 HAY`S SURFACE TENSION
TEST
Take some fresh urine in a conical glass
Sprinkle on the surface particles of sulfur
If bile salts are present, sulfur particles
sink to the bottom.
REFERENCE
•Essentials of Clinical Pathology – Shirish M Kawthalkar
•BIOCHEMISTRY - Satyanarayan –
Dr. U. Satyanarayana
Dr. U. Chakrapan
PRINCIPLE
 Oxidation test-where bilirubin is oxidised to green biliverdin
 Barium chloride precipitates the sulphate radicals present in urine
to form precipitate of barium sulphate. If bile pigments are present
in urine, they adhere to these molecules. Ferric chloride present in
Fouchet’s reagent then oxidises yellow bilirubin, in the presence of
trichloroacetic acid to green biliverdin.