Estimation of Blood Glucose by Folin – wu method Glucose quantitatively reduces ul baric ions to cuprous ions in a boiling alkaline solution. It gets oxidized to glycolic acid, consequently reduce phosphomolybdic acid [Mo +6 Mo +4] are themselves oxidized back to cubic ions. The reduced form phosphonolybdic acid imparts a blue co lour which is read calorimetrically.
Requirements: – Blood, 10% Sodium turnstile, 0.6 NH2SO4, std glucose solution [ 5mg / ml], alkaline cuso4 soln, phosphomolybdic acid.
Procedure: – Protein free filtrate of blood; to 1 ml of distilled water in a c.f. add 0.2 ml blood, wash ppt with 1 ml water and add to centrifuge. Add 0.2ml 10% Na- tang state and 0.2 ml 0.6N H2804 centrifuge or filter.
Standard curve: – Prepare a series of test tube with graded amount of glucose [0.05 to 0.2 mgs]. Add 2 ml of alkaline cu reagent. Heat tubes in a boiling water bath for 10 mins. Cool tubes to R.T.treat with 2 ml phosphomolybdic acid [from burette]. Make up volume to 10 ml with distilled water. Mix solution in each test tube and measure extinction at 420 nm against blank. Treat 1 ml sample in same manner and determine extinction determine glucose cone by extrapolation.
Express cone. of sugar in mg / 100 ml of glucose.
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