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.
Boiling sugar is mainly glucose normal blood sugar levels in adult human subject in the post absorptive absorptive state (overnight fast) varies from 60 – 110 mg / 100 ml. No difference in blood sugar is found with r.t. sex. A blood sugar level is found to be raised by 10 – 20 mg / 100 mg in state of anger fear anxiety and pain. After carbohydrate digestion, blood sugar level may rise to 130 – 140 mg / 100 ml and after 1 – 3 hrs. Come back to normal. This fall is due to consumption thro glycolysis. TCA cycle, HMP pathway, glycogen sis and biogenen’s of lipids and proteins. As blood proteins interfere c estimation they must be ppted before proceeding for determination. Blood sugar must be level are normally under normal control.
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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