Practical # 1 Scheme for detection of Proteins y General reactions of proteins 1. Lead acetate test 2. Sulpho salicyclic test 3.

Esback s test

Color Reaction of Protein 1. 2. 3. 4. 5. Biuret test Xanthoprotiein test Millon s test Formaline test Sulphur test Salicyclic acid test EXPERIMENT 2ml (0.5) + 2 drops of sulphosalicyclic acid OBSERVATION If ppt dissolve, clear solution If ppt persists INTEREFERENCE BJP +ve Other Proteins present (albumin etc)

Color Reaction of Proteins Biuret test:EXPERIMENT 2ml (5%) NaOH + 2 drops of 1% CuSO4. Divide the solution into 2 parts, T.T.A (test tube A) and T.T.B(test tube B). Add 1ml of distilled water(d/w) in TTA Add 1 ml of original solution in TTB OBSERVATION Solution containing protein turns purple violet INTEREFERENCE Peptide linkage protein are present

Coagulation Test:EXPERIMENT 3ml (OS) and heat on low flame OBSERVATION If coagulation appears INTEREFERENCE Albumin and globulin present

Half Saturation Test EXPERIMENT Take 3ml of given solution in test tube and add equal amount (3ml) of saturated ammonium(SO4). Now the OBSERVATION If white ppt. appears INTEREFERENCE Globulin is present

solution becomes half saturated

Full Saturation Test EXPERIMENT Take 5ml of given solution in a test tube and saturate it with solid ammounium sulphate OBSERVATION If white ppt. appears INTEREFERENCE Albumin is present

Precipitation of Protein Acetic Acid test for Casein Reagent :- 1% Acetic Acid solution EXPERIMENT 5ml (OS) + 1% Acetic acid drop by drop till solution changes to white ppt. OBSERVATION White ppt +ve INTEREFERENCE Casein +ve

Principle:Casein is precipitated out at its iso electric PH(4.6) when 1% Acetic acid is added dropwise. CT(confirmation test) for Casein(Neuman s test):Reagent:conc. H2SO4 : Con. HNO3 : Ammonium molybedate solution 5% : Methyl Red Indicator Principle:- On heating e conc. H2SO4 e HNO3. Casein is digested and phosphorous is released, NH3 is added to provide alkaline media. Ammonium molybedate react e phosphorous in alkaline media form yellow ppt of phospho-ammonium molybedate EXPERIMENT Ppt. of acetic acid test + 1 drop of HNO3 + 12 drops of H2SO4, then heat till white fumes appear. Add 1 drop of HNO3, heat till clear solution appear, then cool & dilute e 5ml D/w Add 1 drop of methyl red of strong NH3 till color becomes light yellow + 2ml of ammonium molybedate OBSERVATION Yellow ppt. of phosphoammonium molybedate appear INTEREFERENCE Casein Confirmed

Proteinuria:Pre Renal, Renal (Glomerulus) and Post Renal (bladder trouble, could be stones / trauma)