Lab 2 Color Tests for Proteins and Amino Acids

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Lab 2 Color Tests for Proteins and Amino Acids

Color Tests for Proteins and Amino Acids B)Color Test for Specific Amino Acids 1/Millon's Test: *Used for detecting the presence of monohydroxybenzene derivatives (e.g: Tyrosine, Tyrosine derivatives, Phenol). *Millon's reagent is a solution of mercuric and mercurous ions in nitric and nitrous acids. *Phenolic group of tyrosine ( OH ) reacts with Millon's reagent and give a red color, which is due to a mercury salt of nitrated tyrosine.

N.AL-Mebairik

2/Hopkins-Cole Test:. principle of Tryptophan 2/Hopkins-Cole Test: *principle of Tryptophan. Tryptophan + Hopkins-Cole reagent purple ring *Hopkins-Cole reagent contain magnesium glyoxylate (Magnesium powder and Oxalic acid). *Purple ring is formed by the reaction of glyoxylic acid ( CHO-COOH ) with the indole ring of tryptophan in presence of H2SO4.

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3/Sakaguchi Test: *This test is specific for the guanidine group of arginine (H2N-C-NH), but a non-amino acid like creatine which contain this group will also answer this test. *This test positive for all proteins containing arginine and for arginine itself. *When arginine reacts with α-Naphthol and Sodium hypobromite (NaOBr) a red color result. *Red color is due to a reaction between the hypobromite and the –NH2 group of the guanidino part of arginine. N.AL-Mebairik

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4/Lead Acetate Test: *Used for detecting the presence of Sulfur group in Cystine and methionine . *Proteins or peptides containing these amino acid heated with NaOH to split the sulfide group from amino acids. *Then, lead acetate react with free sulfide ions resulting in the formation of lead sulfide ( pbs ) a brown to black color. N.AL-Mebairik

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5/Folin's Test: *This test for free cystine, therefore protein or peptide must be first hydrolyzed by sodium carbonate and sodium sulfite. *Then, Folin's uric acid reagent (phosphotungstic acid) is added which is reduced to tungsten blue. *This can be used for the quantitative determination of cystine.

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