Protein determination in milk sample (Kjeldhal method)

Principle
The sample was digested with sulphuric acid and nitrogenous compound of the sample was
converted into ammonium sulphate. Ammonium sulphate was further decomposed with the 40%
sodium hydroxide and ammonia collected in the boric acid solution and ammonium borate was
formed. Ammonium borate was titrated against an acid and the nitrogen contents of the sample
were calculated.
Digestion:
Take 10ml of milk sample in digestion flask carefully. 5 gram of digestion mixture (100g K 2SO4,
10g CuSO4 and 5g FeSO4) was also added into the digestion flask. At the end 35ml sulphuric
acid was added and placed the digestion flask into the fume hood for digestion until the mixture
becomes light green color. The mixture was cooled for some time then by adding distilled water
150ml volume was made. Digestion flask was rinsed 2 or 3 times for complete removal of
digested sample from the digestion flask.
Distillation:
After digestion, distillation was done by taking 10ml digested sample along with 10ml NaOH
(40%) into the conical flask. The distillate was collected with 10ml boric acid (4%) solution by
using methyl red as an indicator. When the color of boric acid was changed from red to yellow
distillation was continued for 2-3 minutes until bubbling stopped and maximum ammonia was
trapped.

Titration:
The material retained in the flask after distillation was titrated against 0.1N H 2SO4 until pink
colour appeared.

% Nitrogen = Vol. of H2SO4 used (ml) ×250 ×0.0014 × 100

Vol. of sample ×Sample used for distillation

Total Protein (%) = % Nitrogen × 6.38