Color Reactions of Amino Acids PDF

Color reactions of amino acids Expt 6: Color Reactions Amino Acids and Proteins Slideshow Test Biuret Composition of Reagent + Result 10% NaOH and 0.01M CuSO Grp Responsible Positive Violet solution Peptide Bond Albumin or any peptide Ninhydrin Ninhydrin solution Dark blue/violet soluti...

Color reactions of amino acids Expt 6: Color Reactions Amino Acids and Proteins Slideshow Test Biuret Composition of Reagent + Result 10% NaOH and 0.01M CuSO Grp Responsible Positive Violet solution Peptide Bond Albumin or any peptide Ninhydrin Ninhydrin solution Dark blue/violet solution Amines (amino part in the AA) Albumin, any protein or free amino acids Xanthoprot Conc HNO, eic NaOH Yellow Solution Aromatic ringWY, For protein w/ WYF amino acids Millon Nasse Millon Nasse Reagent Pink to Red Phenolic complex group +Y or protein with Y Test Composition of Reagent Grp Responsible Positive Unoxidized 10% NaOH, Sulfur Black Thiol group +Cysteine Hopkins Gole + Result saturated lead precipitate- acetate PbS lead Pb(CH,COO), (II) sulfide Hppkins Cole Violet Reagent, conc cyclic/ring Indole group + Tryptophan H₂SO₄ solution Proteins - Biuret Test 5 drops of the supernate + 5 drops 10% NaOH + 0.5% CuSO4 Control: 1% Albumin Benedict's Reagent: Sodium Hydroxide and Copper Sulfate Alkaline solution of proteins treated with copper sulfate results in the production of a rose-pink to violet, then purple Test used to detect the presence of peptide bonds by hydrolysis (+) a copper(II) ion is reduced to copper(1), which forms a complex with the nitrogens and carbons of the peptide bonds in an alkaline solution Discussion Biuret Test Test to determine the presence of peptide bonds in protein. In a positive test, a copper(II) ion forms a violet-colored coordination complex with peptide bonds in alkaline conditions. The intensity of the violet color is proportional to the number of peptide bonds participating in the reaction. Ninhydrin Test Chemical that used to detect protein or specific to detect ammonia and amine. These reaction will form a dark blue or purple colour QUALITATIVE TESTS for PROTEINS Millon's test (a) Milon's reagent consists of mercury dissolved in mitic acid (forming a misture of mercuric & mercurous nibrates) (b) When mbank reagent is added to a proteer, a white ppt is formed, which han brick red on heating. (c) This test is given by protein which yield tyrosine on hydrolysis (das to the presence of phenolic group), Nihydrin test This test is given by all pratens () when protein is boiled with a dute solution of nrhydien, a viclet, colour is produced Millon-Nasse Test Specific to phenolic group of tyrosine. The hydroxy benzene group of tyrosine reacts with Millon's reagent to form a red-colored complex. Xanthoproteic Test This test is positive for proteins and amino acids containing an aromatic side chain(phenylalanine, tyrosine, and tryptophan). The benzene ring undergoes nitration with concentrated HNO3 giving nitro derivatives which are yellow in colour. Phenylalanine does not response readily to this test and requires H₂SO₄ as catalyst Xanthoproteic Test The amino acids that contain benzene ring like tyrosine and tryptophan undergo nitration in this test and gives yellow colour Hopkins_cole Test It is specific for tryptophan, the only amino acid containing an indole group. The indole ring reacts with glyoxylic acid in the presence of a strong acid to form a voilet cyclic product. SAKAGUCHI TEST The Sakaguchi Test is used to test for a certain amino acid and proteins. The amino acid that is detected in this test is arginine. The sample to be tested is treated with alpha-naphthol and sodium hypochlorite. A positive result yields a reddish wine color when arginine is present. BROMINE WATER TEST Pyrrole Ring of TRYPTOPHAN (W) undergoes halogenation with Bromine in water in the presence of amyl alcohol A positive result yields a pink/ lavender color UNOXIDIZED SULFUR TEST For Sulfur Containing Amino Acids like CYSTEINE. Liberated sulfur reacts with Lead Acetate to form Black Precipitate of Lead Sulfide.