Estimation of Nitrogen BY Kjeldahl Method

The Kjeldahl method in analytical chemistry is a method for the quantitative determination of nitrogen contained in organic substances plus the nitrogen in inorganic ammonia and ammonium (NH₃/NH₄⁺). Other forms of inorganic nitrogen, for instance nitrate, are not included in this measurement. This method was developed by Johan Kjeldahl in 1883

The method consists of heating a substance with sulphuric acid, which decomposes the organic substance by oxidation to liberate the reduced nitrogen as ammonium sulphate. In this step potassium sulphate is added to increase the boiling point of the medium (from 337 °C to 373 °C) . Chemical decomposition of the sample is complete when the initially very dark-coloured medium has become clear and colourless. The solution is then distilled with a small quantity of sodium hydroxide, which converts the ammonium salt to ammonia. The amount of ammonia present, and thus the amount of nitrogen present in the sample, is determined by back titration. The end of the condenser is dipped into a solution of boric acid. The ammonia reacts with the acid and the remainder of the acid is then titrated with a sodium carbonate solution by way of a methyl orange pH indicator.

Degradation: Sample + H₂SO₄ → (NH₄)₂SO₄(aq) + CO₂(g) + SO₂(g) + H₂O(g)

Liberation of ammonia: (NH₄)₂SO₄(aq) + 2NaOH → Na₂SO₄(aq) + 2H₂O(l) + 2NH₃(g)

Capture of ammonia: B(OH)₃ + H₂O + NH₃ → NH₄⁺ + B(OH)₄⁻

Back-titration: B(OH)₃ + H₂O + Na₂CO₃ → NaHCO₃(aq) + NaB(OH)₄(aq) + CO₂(g) + H₂O

In practice, this analysis is largely automated; specific catalysts accelerate the decomposition. Originally, the catalyst of choice was mercuric oxide. However, while it was very effective, health concerns resulted in it being replaced by cupric sulfate. Cupric sulfate was not as efficient as mercuric oxide, and yielded lower protein results. It was soon supplemented with titanium dioxide, which is currently the approved catalyst in all of the methods of analysis for protein in the Official Methods and Recommended Practices of AOAC International

By Roshan220195 - Own work, CC BY-SA 3.0, Link

Reference: https://en.wikipedia.org/wiki/Kjeldahl_method