(a) Principle
Glucose is oxidized by the enzyme Glucose Oxidase (GOD) to give D-gluconic acid and hydrogen peroxide. Hydrogen peroxide in presence of enzyme Peroxidase (POD) oxidizes phenol, which combines with 4-Aminoantipyrine to produce a red coloured quinoneimine dye. The intensity of the colour developed is proportional to glucose concentration in the sample.
(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.
B | S | T | |
Reagent | 1.0 ml | 1.0 ml | 1.0 ml |
Distilled Water | 0.01ml | ||
Standard | 0.01ml | ||
Serum | 0.01ml |
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