Kjeldahl method

Kjeldahl method
The Kjeldahl method or Kjeldahl digestion (Danish pronunciation:[kldl]) in analytical chemistry is a method for the quantitative determination of nitrogen in chemical substances developed by Johan Kjeldahl in 1883.[1]

Method
The method consists of heating a substance with sulfuric acid, which decomposes the organic substance by oxidation to liberate the reduced nitrogen as ammonium sulfate. In this step potassium sulfate is added to increase the boiling point of the medium (from 337F to 373F / 169C to 189C). Chemical decomposition of the sample is complete when the initially very dark-colored medium has become clear and colorless. 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 (mercury oxide or copper sulfate) accelerate the decomposition.

Applications
The Kjeldahl method's universality, precision and reproducibility have made it the internationally-recognized method for estimating the protein content in foods and it is the standard method against which all other methods are judged. It does not, however, give a measure of true protein content, as it measures nonprotein nitrogen in addition to the nitrogen in proteins. This is evidenced by the 2007 pet food incident and the 2008 Chinese milk powder scandal, when melamine, a nitrogen-rich chemical, was added to raw materials to fake high protein contents. Also, different correction factors are needed for different proteins to account for different amino acid sequences. Additional disadvantages, such as the need to use concentrated sulfuric acid at high temperature and the relatively long testing time (an hour or more), compare unfavorably with the Dumas method for measuring crude protein content.[2]

Kjeldahl method

Total Kjeldahl nitrogen

Total Kjeldahl nitrogen or TKN is the sum of organic nitrogen, ammonia (NH3), and ammonium (NH4+) in the chemical analysis of soil, water, or wastewater (e.g. sewage treatment plant effluent). To calculate Total Nitrogen (TN), the concentrations of nitrate-N and nitrite-N are determined and added to TKN. Today, TKN is a required parameter for regulatory reporting at many treatment plants, and as a means of monitoring plant operations.

Conversion factors
TKN is often used as a surrogate for protein in food samples. The conversion from TKN to protein depends on the type of protein present in the sample and what fraction of the protein is composed of nitrogenous amino acids, like arginine and histidine. However, the range of conversion factors is relatively narrow. Example conversion factors, known as N factors, for foods range from 6.38 for dairy and 6.25 for meat, eggs, maize (corn) and sorghum to 5.83 for most grains; 5.70 for wheat flour, and 5.46 for peanuts.[3]

References
[1] Julius B. Cohen Practical Organic Chemistry 1910 Link to online text (http:/ / www. archive. org/ details/ PracticalOrganicChemistry) [2] Dr. D. Julian McClements. "Analysis of Proteins" (http:/ / www-unix. oit. umass. edu/ ~mcclemen/ 581Proteins. html). University of Massachusetts Amherst. . Retrieved 2007-04-27. [3] http:/ / www. fao. org/ docrep/ 006/ y5022e/ y5022e03. htm

Wastewater Engineering: Treatment and Reuse, Metcalf & Eddy

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