1.

Detection of Albumin in Urine

Intro: Albumin is a protein produced by the liver and is mainly found in the blood. A trace of protein
is found in normal urine daily. Under pathological conditions like Albuminuria, the level of albumin
found in urine will be way above the normal level.

In the kidney disturbance and in high blood pressure, albumin level in urine is significantly high.

To detect albumin in the urine, Sulphosalicylic Acid test and Heller’s test are done.

Sulphosalicylic Acid Test

Materials Required: Urine sample in a test tube

30% sulphosalicylic acid

Dropper

Procedure: Using a dropper, take a small quantity of sulphosalicylic acid.

Now, add it to the test tube containing the urine sample.

NOTE: A cloudy turbid solution or precipitate appearing in the solution indicates the presence of
albumin urine sample.

Conclusion: When the sulphosalicylic acid reacts with albumin, it causes denaturation of protein in
the sample that appears in the forms of precipitate.

HELLER’s Test

Materials required:
 Concentrated Nitric Acid (HNO3) in a test tube
 Urine Sample
 Dropper

Procedure:

 Using a dropper, take a small quantity of urine from the urine sample bottle.
 Take the test tube containing conc. HNO3
 Incline the test tube and add the sample of urine using a dropper along the inner side of the
test tube.
 NOTE: As the Urine comes in contact with the nitric acid, a white ring is formed at the point
of contact. The presence of a white ring at the junction of two layers indicates the presence
of albumin in the sample.

Conclusion: Nitric Acid causes denaturation of proteins with the formation of a white precipitate. As
urine that contains albumin comes in contact with the nitric acid, it forms white ring at the point of
contact.
2. Detection of sugar in Urine

Intro: Sugar is ordinarily not present in normal urine. When the sugar level in blood rises above normal
level, the kidney eliminates the extra sugar through the urine. Sugar then starts to appear in urine.
Generally Benedict’s and Fehling’s test are used to detect the presence of sugar in urine. These test give
coloured precipitates such as: light green, yellow, and brick red, depending on the concentration of
sugar present in urine.

Benedict’s test

 Materials required:
 Urine sample in a test tube
 Benedict’s reagent
 Burner
 Test tube holder
 Dropper
Procedure:

 Using a dropper take a small quantity of Benedict’s reagent.

 Now, add the reagent to the test tube containing urine sample.
 Boil the sample over a burner for 2minutes holding the test tube firmly with a test tube holder.
 Keep shaking the test tube as it is being heated
 Note: A bricked red precipitate appears, indicating the presence of sugar in the urine
Conclusion: On boiling the Urine sample with the Benedict’s reagent, the cupric ion present in the
Benedict’s reagent is reducing agent sugar, to form a brick red coloured precipitate of cuprous oxide.
Therefore, sugar is present in the urine sample.

Fehling’s test

Materials required:

 Urine sample in a test tube

 Test tube holder
 Fehling’s solution A
 Fehling’s Solution B
 Dropper
 Burner
Procedure:

 Using a dropper take a small quantity of Fehling’s solution A.

 Now, add the Fehling’s Solution A to the test tube containing urine sample.
 Using a dropper, take a small quantity of Fehling’s Solution B.
 Add Fehling’s solution B to the test tube containing urine sample.
 Boil the sample over a burner for two minutes, holding the test tube firmly with a test tube
holder
 Keep shaking the test tube while heating.
 Note: A brick red precipitate appears indicates the presence of sugar in the urine.
Conclusion: The cupric ion present in the Fehling’s solution is reduced on boiling by the reducing
substance sugar, to form the brick red coloured precipitate of cuprous oxide. Therefore, sugar is present
in the urine sample.

3. Detection of Bile Salt in Urine

Intro: Bile is a yellow-green fluid that contains water and organic molecules such as cholesterol, bile
acids, and bilirubin. In humans and most vertebrates, bile is produced by the liver. The gall bladder holds
the bile produced in the liver and when the organisms eats, bile is discharged into the duodenum. To
detect bile salt in urine, Smith’s test and Pettenkofer’s test are done.

Smith’s Test

Materials required: Urine Sample

Smith’s reagent in a test tube

Dropper

Procedure: Using a dropper, take a small quantity of urine from the urine sample bottle.

Take the test tube containing Smith’s reagent.

Now, Incline the test tube and add the sample of urine by means of a dropper along the
inner side of the test tube.

As urine comes in contact with the smith’s reagent, a green ring is formed at the point
of contact

NOTE: The presence of a green ring at the junction of two layers indicates the presence of bile salt in the
sample.

Pettenkofer’s Test:

Materials required: Urine Sample in a test tube

Conc. H2SO4

Dropper

Sucrose

Spatula

Procedure: Using a spatula, take some sucrose powder from the watch glass.

Add sucrose powder to the test tube contain urine and shake it well.

Using a dropper, take a small quantity of H2SO4.

Now, add the H2SO4 to the test tube containing urine sample.

NOTE: Red color appearing in the solution indicates the presence of bile salts in urine.