Detection of Urea adulteration in liquid Milk

Principle: Urea is hydrolyzed by urease enzyme. During this, ammonia liberated will form
ammonium hydroxide (base) with water. Phenol red indicator changes its color in basic medium
(pH reaches 8.4 by liberation of ammonium hydroxide. the tube will turn from an orange color to
a pink color.

Reagents Required:

1. Urease solution (0.2 %, freshly prepared)

2. Phenol red indicator.

1. 10 ml Pipette or any available.

2. Incubator
3. Test tube
Procedure :

1. Take 5 ml of sample in two test tubes namely A and B each.

2. Add 5 mL urease solution in each of the test tubes by using pipette.
3. Add 1 - 2 drops of phenol red indicator in both tubes.
4. Place test tube A in incubator at 40ºC for 15 min. while other will remain in shelf
5. Observe the color of solution in test tube A and compare its color with that of B to evaluate
status of milk.

Results

Color changes from B urea +ve

Color remains same as that of B urea - ve.