Question
Describe principle and procedure of the Kjeldahal method used for protein determination.
Answer :
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Skip in 10
The protein content of foods can be determined by numerous methods. The Kjeldahl method and the nitrogen combustion (Dumas) method for protein analysis are based on nitrogen determination. Both methods are official for the purposes of nutrition labeling of foods. While the Kjeldahl method has been used widely for over a hundred years, the recent availability of automated instrumentation for the Dumas method in many cases is replacing use of the Kjeldahl method.
Principle Of Kjeldahl Method
The Kjeldahl procedure measures the nitrogen content of a sample. The protein content then, can be calculated assuming a ratio of protein to nitrogen for the specific food being analyzed. The Kjeldahl procedure can be basically divided into three parts:
(1) Digestion
(2) Distillation
(3) Titration
In the digestion step, organic nitrogen is converted to an ammonium in the presence of a catalyst at approximately 370°C. In the distillation step, the digested sample is made alkaline with NaOH and the nitrogen is distilled off as NH3 . This NH3 is “trapped” in a boric acid solution. The amount of ammonia nitrogen in this solution is quantified by titration with a standard HCl solution. A reagent blank is carried through the analysis and the volume of HCl titrant required for this blank is subtracted from each determination.
Reagents
Sulfuric Acid (concentrated, N-Free)
Catalyst/Salt
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