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Download Test for Ketone Bodies
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.Faculty of pharmacy/Third class/Second course/Biochemistry Lab Test for Ketone Bodies ketone bodies include acetoacetic acid B-hydroxybutyric and acetone which are formed during the lipid metabolism. the presence of urinary keton bodies is detected by colour tests , such as Rothera s and Gerhardt test. Ketone bodies do not appear in urine because acetoacetic acid which is produced normally in liver is completely oxidised in tissues .but if fats are metabolised excessively in liver as in diabetes and starvation there will be over production of acetoacetic acid. the tissues are unable to oxidise the excessive amount of acetoacetic acid with the limited supply of oxygen. Apart of excess acetoacetic acid is decarboxylated to acetone and remaining circulates in blood as acetoacetic acid and B-hydroxyl .butyric acid , when are reversible in equilibrium. Rothera s test Principel: Ketone bodies react with sodium nitroprusside under alkaline condition to yield a purple colour. Reagents: 1-sodume nitroprusside (W% w/v) solution 2-Chilled liquor ammonia 3-Ammomium sulphate(solid) Procedure: 1-Saturate 5 ml of the urine sample with solid ammonium sulpate in a test tube 2-Add 2-3 drops of sodium nitroprusside reagent 3-folloewd by addition of few drops of chilled liquor ammonia and mix Result : Observation Purple ring Inference Acetone or acetoacetic acid or may present Q/Acetone or acetoacetic acid or may present? why Msc pure Biochemistry –Ahmed kareem thamer
