Determination of Blood Glucose By Alkaline Copper Reduction Method LAB # 2

Determination of Blood Glucose (BG) 2- Alkaline copper reduction method (Asatoor & King method): Principle Glucose + cupric cuprous + gluconic acid Cuprous + phosphomolybdic acid cupric + molybdenum blue

Determination of Blood Glucose (BG) Procedure 1- Deproteinization 1-Add 0.1 ml Of Na tungstate 3- add 0.1 ml serum 2- add 3.8 ml of Isotonic Na2SO4/CuSO4 Soln. -Mix well -Label centrifuge Clean & Dry Cent. tube

Procedure: cont. Transfer clear supernatant Label as to clean dry t.t Using Pasteur pipette Label as supernatant Test tube

Procedure: cont. Test Standard Blank T B S Add 1 ml of supernatant +1 ml of alkaline tararate Put in boiling water bath for 10 mins Mix well and close them with cotton wool Add 1ml Standard Glucose +1 ml alkaline tararate Cool under tap water Add 3 ml of phosphomolybdic reagent to each tube Shake well 1 ml isotonic Na2SO4/CuSO4 Soln. + 1 ml alkaline tararate Mix well to remove all air bubbles Let the tubes stand for 5 mins. Measure the absorbance of test and stand. against blank at 680 nm Add 3 ml of distilled water to each tube

Calculation Glucose conc. (mg/dl) = Test abs. X Stand. conc. Stand. abs.