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Determination of Crude Protein

Determination of Crude Protein

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Published by kolita kamal
determination of protein in foods products by using kjeldhal method
determination of protein in foods products by using kjeldhal method

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Categories:Types, Research, Science
Published by: kolita kamal on Jan 22, 2010
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03/03/2013

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Determination of CrudeProtein
 
B.K.K.K.Jinadasa
 
12/13/2009
 
(GS/M.Sc./FOOD/3608/08)
 
 
200
2
In Kjeldahl method a food is digested with a strong acid so that it releases nitrogen which can bedetermined by a suitable titration technique. The amount of protein present is then calculatedfrom the nitrogen concentration of the food. It is usually considered to be
the
standard method of determining protein concentration. Because the Kjeldahl method does not measure the proteincontent directly a conversion factor
(F)
is needed to convert the measured nitrogen concentrationto a protein concentration. A conversion factor of 6.25 (equivalent to 0.16 g nitrogen per gram of protein) is used for many applications, however, this is only an average value, and each proteinhas a different conversion factor depending on its amino-acid composition. The Kjeldahl methodcan conveniently be divided into three steps: digestion, neutralization and titration. In thedigestion step nitrogen in protein is oxidized to ammonium sulphate by using sulphuric acid andcatalyst.
 
200
3
Kjeldhal digestion kit (Kjeldhal digestion flasks, Kjeldhal heater, fume trap, distillation unit(Pranas Wagner still)Titration flasksWeighing balanceSodium sulphate solution(Dissolve 50 g of NaOH pellets and 8g of sodiumthiosulphate in 100 mL distilled water)100 mL distilled water4% Boric acid solution0.02 M HCl standard hydrochloric acid solutionsKjeldhal catalyst tabletsIndicators Methyl redMethylene blue(Prepare a solution by mixing two parts of 2% methyl red (alcoholic) solution with one part of 0.2% (alcoholic) methylene blue solution.)conc. H
2
SO
4
 
3.2. Methods
Weigh 0.05 g of prepared sample was transferred on a tissue paper to the kjeldhal digestion flask.Catalyst powder and 2 mL of conc. H
2
SO
4
were added to the flask. Then the flask was connectedto fume trap and attached to the pump. Sample was allowed to digest for four hours, until a clearsolution without black particles was obtained. Then the sample was cooled for one hour. Blank digestion was also carried out.Sample was dissolved in a minimum amount of ammonia free distilled water and transferred to asemi micro kjeldhal distillation apparatus which has been previously conditioned by passingsteam for several minutes.Add 8 mL of NaOH/Na
2
SO
3
solution to the kjeldhal apparatus.5mL of 4% boric acid solution and 3 drops of indicator were added into a titration flask and keptat the end of the apparatus to trap the ammonia liberated.Steam was passed through the flask until about 15 mL of distillate was received.This solution was collected at the titration flask and titrated with standard HCl solution. The endpoint is pink colour.Same procedure was applied to a blank sample as well.

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