Test Ninhydrin

Color Reactions of Proteins Composition of + Result Grp. Responsible Reagent Triketohydrin Hydrate Lavander Free amino and free COOH NaOH + CuSO4 Hg in HNO3 Conc. HNO3 Glyoxylic acid and conc. H2SO4 Conc HCl , sucrose Pb(OAc)2 Sullfanilic acid in HCl NH4OH 10% NaOH, naphtol, alkaline hypobromite HCHO conc. H2SO4 KOH, Pb(OAc)2 Br.H2O, amyl alcohol naphtol in alcoholic H2SO4 Glacial HOAc and conc. H2SO4 Violet Red Lemon yellow Violet ring Violet Red orange lighter orange Peptide linkages Hydroxyphenyl group Benzene ring Indole group Indole group

Importance Test for a.a. peptides in determining amino acids + to tripeptides up to protein + to tryptophan + Tyr, Phe, Trp + trp + trp + his and tyr + arginine +trp + cystine, Cystein and methionine + trp Glycoprotein + trp

Biuret Millons Xanthroproteic Hopkins-Cole Liebermann Erlichs Diazo Sakaguchi AcreeRosenheim Reduced Sulfur Br water Molisch Adamskiewez

Intense red color Guanidine Violet ring Black ppt Pink Violet ring Reddish violet ring at the junction Indole group Sulfur Indole group Carbohydrates Indole group

Test for Carbohydrates Test Composition of Reagent Molisch naphtol in alcoholic H2SO4 Seliwanoff Bials Mucic Iodine test Resorcinol in HCl Orcinol in HCl Conc. HNO3 Iodine in KI

+ Result Purple ring Red color + brown ppt Blue green White ppt / colorless / red like crystals Deep blue starch Purple amylodextrin Red- erythrodextrin Colorless achroodextrin Mahogany red inulin Yellow ppt Brick red ppt Brick red ppt of Cu2O Black ppt of Bi Silver mirror Caramel odor, brown color

Importance Gen. Test for carbohydrates Test for ketoses Test for pentoses Test for galactose and lactose Test for polysaccharides; differentiate from helical polyssacharide

Osazone Barfoeds Benedicts Nylanders Tollens Moores

Phenylhydrazine + NaOAc Cu(OAc)2, HOAc CuSO4, Na2CO3, Sodium citrate (not strongly alkaline) Bismuth subnitrate + KOH + Rochelle salt AgNO3 + xss NH3 NaOH

Test to identify sugars in microscopic appearance Distinguish mono from disaccharides Test for reducing sugars Test for reducing sugars Test for reducing sugars Test for reducing sugar