Clinical Pharmacy Biochemistry-1 PB302 (Practical) PDF

Scheme for unknown identification Biuret test Violet Color No Violet Color Protein Carbohydrate...

Scheme for unknown identification Biuret test Violet Color No Violet Color Protein Carbohydrate or NPN Heat coagulation test Molisch’s test opaque mass insoluble in dil. No opaque mass Violet Ring No Violet Ring acetic acid Red litmus Albumin Carbohydrate NPN paper Remains Red Turns Blue Neutral Proteins Alkaline Proteins Peptone Gelatin m-protein Casein Biuret test using Rose or Violet pale yellow color yellowish Sulfur test very dilute CuSo4 pink color color brown color Half saturation No ppt White ppt Coagulation true coagulum false coagulum test at IEP 69  Clinical Pharmacy 2024/2025 Level two PharmD Biochemistry-1 PB302 (Practical course) No violet color with Biuret test & violet ring with Molisch’s test Carbohydrate Iodine test Violet color Blue color No characteristic color Not Dextrin Starch polysaccharide Fehling’s test Red ppt No Red ppt Reducing sugar Sucrose Monosaccharides Disaccharides Apply Fehling’s after hydrolysis and ketose test Ketose test Glucose Fructose No reddish brown color reddish brown color 70 Clinical Pharmacy 2024/2025 Level two PharmD Biochemistry-1 PB302 (Practical course) No violet color with Biuret test & No violet ring with Molisch’s test NPN Red litmus paper Remains Red Turns Blue Urea Na Urate Apply biuret after Apply Follin’s & fusion and sodium Fehling’s tests hypobromide tests Positive Biuret after fusion Positive Follin’s test 71  Chemical reactions procedures 1 -Molisch test: Procedure:- 1 ml sample + 8 drops of alcoholic alpha naphthol, shake well, then add 1 ml conc. sulphuric on the wall of the test tube carefully (no Shaking) 2 -Biuret Test: Procedure:- 1 ml Sample + 1 ml 10% NaOH, shake well, then add few drops of dil. CuSO4 3- Iodine test Procedure Fill half the test tube with sample then add one drop of dil. iodine 4- Fehling test Procedure: In a test tube: 1 ml Fehling A+ 1 ml Fehling B, Shake well till dark clear blue solution = Fehling reagent In another test tube: add 1ml sample + 1 ml Fehling reagent, Heat for 2-5 mins on direct flame 6- Ketose test: Procedure:- 1ml Sample + 3ml conc HCl, Heat on water bath for 3 mins 7- Fehling after hydrolysis: Procedure:- 3 ml sample + 2 ml dil. H2SO4, boil 2-3 min, cooling and neutralize xss. acid with 2ml 30 % NaOH Add 2ml of previously mixed Fehling reagents (1 ml Fehling A+ 1 ml Fehling B) to 2 ml of the hydrolyzed sugar, Heat for 2-5 mins on direct flame 8- Fehling after prolonged heating: Procedure:- In a test tube: 1 ml Fehling A+ 1 ml Fehling B, Shake well till dark clear blue solution = Fehling reagent In another test tube: add 1ml sample + 1 ml Fehling reagent, Heat for 5 mins on direct flame 9- Heat coagulation test: Procedure:- Fill 3/4 the test tube with sample then heat the top of the solution on direct flame. 10-Sulfur test: Procedure:- 1 ml sample + 1 ml 30% NaOH, boil on direct flame for 5 minutes then add few drops  of lead acetate. 11- Coagulation at isoelectric point (IEP): Procedure:- 2 ml sample then Add dil. acetic acid stepwise till the formation of precipitate (IEP), Heat the precipitate for 5 min on direct flame ✓ If the opaque mass does not dissolve, it is a true coagulum ✓ If soluble, it is a false coagulum 12-Biuret using very dil.CuSO4: Procedure:- Preparation of very diluted CuSo4: 1ml of dil. CuSO4 then fill the test tube with water 1 ml Sample + 1 ml 10% NaOH, shake well, then add drop by drop of the very dil. CuSO4 till the first color appeared 13-Follin's test: Procedure:- 1 ml sample + 1 ml Follin's reagent 14-Biuret after fusion: Procedure:- Evaporate 2 ml of sample till dryness and formation of white residue (biuret residue) dissolve this residue in the least volume of 10% NaOH add 2 drops of dil. CuSO4