1.2.1.

3 Urea spot test

Procedure
A) Weigh 2.5 g urease powder with small amount of distilled water, stir into paste and
dilute to 50 ml with distilled water.
B) Rub 0.15 g bromothymol blue indicator powder in mortar with 2.4 ml 0.1 N NaOH
(Sodium Hydroxide) solution. After indicator dissolves, wash mortar and pestle with
distilled water and dilute to 50 ml with distilled water. Solution should be green; pH
approx. 7.0.
C) Mix solution (A) and (B) in equal proportion.
• Take 90 ml mixture solution of (C) and mix with 10 ml glycerol.
• Pour mixture into watch glass. Dip pieces of heavy filter paper in solution.
• Allow to dry papers in a place free from NH3 fumes, strong air currents or heat.
• Paper should be orange when dried.
• Make strips of these papers and store in amber coloured glass bottle in a cool
place.

Testing
• Dissolve small quantity of feed/ feed ingredients with distilled water in a beaker.
• Deep the strip in a beaker and allow to dry it.
• Appearance of blue colour indicates the presence of urea.

1.2.1.4 Identification of plant and animal protein in feed

1. Mix 1-2 g test sample with 100 ml boiling water or boil the mixture for 2-3 min. Place
a few ml of the cooled mixture in test tube and add 5-6 drops of iodine solution. If
starch is present, the mixture turns blue.
2. Spread 1-2 g test sample into a petri dish. Add 5-6 drops of iodine solution and
let stands for 10 min. A purple brown colour indicated the presence of plant fibre,
whereas yellow indicated animal fibre (protein) using a microscopic examination.

1.2.1.5 Spot tests for minerals in finished feed

Preparation of sample
Usually the minerals in mixed feed are powder or fine particles. Sieve the sample through
fine sieve and put the fine portion in chloroform or carbon tetrachloride in a 100 ml beaker.
Shake thoroughly and keep it for 5 minutes. Pour off the floating feed material. Sprinkle the
residue mineral portion over the paper with a small spatula for spot test.

Test for cobalt (Co), copper (Cu) and iron (Fe)

Reagents
Solution A: Sodium potassium tartrate (Rochelle salt, KNaC4.H4O6.H2O). Dissolve 100 g of
Rochelle salt and make volume 500 ml with distilled water.
Solution B: Dissolve 1 g of 1-nitroso-2-hydroxynaphthalene-3, 6 g disulfonic acid and make
volume 500 ml with distilled water.

Procedure
Moisten the filter paper with 3 to 4 drops of solution A and B then sprinkle the feed sample
on filter paper. Allow the filter paper to dry and check closely under low power stereoscopic
microscope.