Preksha Meditation & Human Health: 5.3.1 Estimation of Triglycerides in Serum (Henry J.B., 1991)

(a) Principle

Glycerol released from hydrolysis of triglycerides by lipoprotein lipase is converted by glycerol kinase into glycerol-3-phosphate which is oxidized by glycerol phosphate oxidase to dihydroxyacetone phosphate and hydrogen peroxide. In presence of peroxidase, hydrogen peroxide oxidizes phenolic chromogen to a red colored compound. The intensity of this colored compound was to be measured which is quantitative index of triglyceride.

* GPO = Glycerol phosphate oxidase

(b) Specimen collection

Arterial blood of subject was collected by sterilized syringe, in a clean and dry glass container. Serum was separated from the cells with the help of centrifuge machine at the earliest possible (within 30 minutes).

(c) Procedure

Pipette into clean, dry tubes labeled Blank (B), Standard (s) and Test (T) and add the reagents in following order.

Incubate the assay mixture for 10 minutes at 37° C. After incubation, absorbance value of the Colour against blank was measured at 510 nm with the help of autoanalyser.

(d) Calculation

$\mathrm{Concentration\; (mg\; \%)}=\frac{\mathrm{Absorbance\; of\; sample}}{\mathrm{Absorbance\; of\; standard}} \mathrm{X\; 200}$