CHOLESTROL ESTIMATION DR. BEENISH ZAKI INSTRUCTOR DEPARTMENT OF BIOCHEMISTRY

OBJECTIVES SKILL: At the end of the practical session, the student shall be able to demonstrate the ability to estimate serum Cholesterol through quantitative method. KNOWLEDGE At the end of the session the student shall be able to explain and interpret the principle and its clinical application.

Prerequisite For Test: 12 hours fasting from 8pm to 8 am. Non-hemolyzed blood sample.

Method This is an enzymatic colorimetric test using CHOD- PAP method(Cholesterol Oxidase- Phenol 4-Aminoantipyrine Peroxidase)

Principle Cholesterol esters+ H 2 0 Cholesterol esterase Cholesterol + fatty acid Cholesterol + H Cholesterol oxidase Cholestenone + H H Aminoantipyrine + 3, 5- Dichlorophenol Peroxidase Coloured quinonic derivative + 4 H 2 0

Procedure: Take 1 cuvette. Label as test (T). Mix well and let stand for 2 minutes at room temperature. Read optical density using green filter. Working Reagent Test Sample20 ul Reagent2000ul

CALCULATION O.D of Test x 200 = Concentration of test O.D of standard The unit is mg/dl

Normal Values of Serum Cholesterol National Cholesterol Education Programme guidelines: The Normal recommended level <170 mg/dl Borderline mg/dl High> 200 mg/dl

Cause of Hypercholesterolemia: Increase in the amount of serum Cholesterol caused by because of: Nephrotic Syndrome Myxodema Coronary Artery Thrombosis

Causes of Hypocholesterolemia: Decreased in amount of cholesterol can be due to: Hyperthyroidism Malabsorption syndrome Hemolytic Jaundice

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