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Sunday, December 13, 2009

Estimation of α - amino nitrogen by Pope Stevensons’ method

How to estimate alfa - amino nitrogen of given sample of protein hydrolyses.

This method is based on the formation of soluble complex between amino acid and cu++ ions present in the form of cupric phosphate Cu3 (PO4 )2 . The sample is treated with excess of cupric phosphate and the excess of cupric phosphate is filtered off. The amount of cu++ ions taken up for the formation of complex is determined idiomatically.

Requirement:- N / 100 Na2S2O3, starch indicator, Glacial acetic acid, 1N NaOH, Thymolpthalein indicator, copper suspension reagent, KI.

Procedure: - Take 10 ml of protein hydrolyses in a 100 ml vol. flask. And add 4 drops of thymolpthalein indicator. Add 1N NaOH till a taint blue co lour is obtained [ PH745 – 10.2]. Add 30 ml of copper suspension reagent and make up the volume to 100 ml with H2O. Mix well and filter Acidify 10 ml of filtrate with 0.5 ml of glacial acetic acid and approximately 1 gm of kI crystals (or add 10 ml of 10% Ki ). Titrate the liberated I2 immediately against N/100 Na2S2O3 solution using starch as indicator; End point is blue to colourless
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