This is hetrogeneous group of compounds which are important constituents
of all cells the nerve cell are highly enriched with lipids The lipids are insoluble water and therefore present in serum either in combination with certain proteins (Lipoproteins) or in an emulsified form as chylomicrons. Plasma lipoprotein are classified into 4 major groups according to density 1. hylomicrons composed of trigycerides! Transport trigycerides from the gut ". #L$L (very low density lipoproteins) composed of trigycerides! 1%& Transport trigycerides from liver to peripheral tissues. '. L$L (low density lipoproteins) ()& composed of cholesterol esters. Transport cholesterol to tissues 4. *$L (*igh density lipoproteins) "%& composed of cholesterol esters. Transport cholesterol from tissues to liver. CLINICAL PRESENTATION +stimation of lipoprotein *$L and L$L is playing greater role in diagnosis, management and prognosis of patients with coronary artery disease. -ormal value HDL 35- 75 mg/dl -ormal value LDL 130-160 mg/dl Patients suffering from coronary artery disease have low level of *$L below '%mg.dl and high level of L$L above 1()mg.dl
HDL OBJECT: determination of high density lipoprotein cholesterol Principles: Chylomicrons, VLDL and LDL are precipitated by adding phosphotngstic acid and magnesim ions to the sample! Centrifgation lea"es only the HDL in the spernatant! Reagents: Concentrations of the reagents #hospotngstic acid $! mmol%L &agnesim chloride '!( mmol%L Procedure: sample / standard 200 ul Regent 500 ul Mix and incubate for 5 minute t!en centrifuge for 20 minutes at 2500g" #it!in 2 !ours after centrifugation transfer 0"ml of clear supernatant to t!e reaction solution for determination of c!olesterol" $a%elengt! 500nm"
)tandard )ample )pernatant * $++ ,L )tandard $+++ , L * Cholesterol reagent $+++ , L $+++ , L &i- and incbate for $+ min at room temperatre or . min and at /0 o C! Then measre the absorbance of the sample or the standard against the reagent blan1 "ale 2ithin 3. min! &alculation: 4ith standard HDL 5 cholesterol 6mg % dL7 8 9 : sample ; conc! )tandard 6mg % dL7 9 : standard LDL OBJECT: determination of lo2 density lipoprotein cholesterol Principle: Lo2 density lipoproteins <LDL= are precipitated by addition of heparin! High density lipoproteins <HDL= and "ery lo2 density lipoproteins <VLDL= remain in the spernatant after centrifgation and are measred en>ymatically by the CHOD* #:# method! The concentration of LDL cholesterol is calclated as the difference of total cholesterol and cholesterol in the spernatant! Reagents: Concentration of the reagents Heparin $++ +++ ?%L )odim citrate (3 mmol%L Procedure: sample / standard 00 ul Regent 000 ul Mix and incubate for 5 minute t!en centrifuge for 20 minutes at 2500g" #it!in !ours after centrifugation transfer 00 ul of clear supernatant to t!e reaction solution for determination of c!olesterol" $a%elengt! 500nm"
)tandard )ample 'upernatant * $++ ,L 'tandard $++ , L * &!olesterol reagent $+++ , L $+++ , L &i- and incbate for $+ min at room temperatre or . min and at /0 o C!, read absorbance of the sample for the standard 2ithin 3. min! against reagent blan1 &alculation: Cholesterol in spernatant Cholesterol spernatant 6mg % dL7 8 9 : sample ; conc! )tandard 6mg % dL7 9E standard