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The Serum Triglyceride Determination Kit can be used for the measurement of glycerol, true triglycerides, or total triglycerides in serum or plasma. The procedure involves enzymatic hydrolysis by lipase of the triglycerides to glycerol and free fatty acids. The glycerol produced is then measured by coupled enzyme reactions. Many of the triglyceride reagents which are commercially available, do not differentiate between endogenous glycerol and glycerol derived by hydrolytic action of lipase on glycerides.
Triglycerides are first hydrolyzed by lipoprotein lipase to glycerol and free fatty acids. Glycerol is then phosphorylated by adenosine-5′-triphosphate (ATP) forming glycerol-1-phosphate (G-1-P) and adenosine-5′-diphosphate (ADP) in the reaction catalyzed by glycerol kinase (GK). G-1-P is then oxidized by glycerol phosphate oxidase (GPO) to dihydroxy-acetone phosphate (DAP) and hydrogen peroxide (H
2O
2). Peroxidase (POD) catalyzes the coupling of H
2O
2 with 4-aminoantipyrine (4-AAP) and sodium
N-ethyl-N-(3-sulfopropyl)
m-anisidine (ESPA) to produce a quinoneimine dye that shows an absorbance maximum at 540 nm. The increase in absorbance at 540 nm is directly proportional to triglyceride concentration of the sample.
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