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 coloured compound.
Procedure
Reaction type ………… ………. End-Point
Reaction time …. 10 mins. at 37°C/20 mins. at R.T. (25-30°C)
Wavelength ……….. ………… 510 nm. (500 – 530 nm.)
Zero setting with ……….. ………… Reagent Blank
Blank absorbance limit … ……… < 0.300 Abs.
Sample volume …………………………… 0.01 ml (10 jul)
Reagent volume ……….. …………. 1.0 ml
Standard concentration .. …………… 200 mg%
Linearity ……………. ….. 800 mg/dl
Quality Control
To ensure adequate quality control, it is recommended that
each laboratory should use a normal and an abnormal
commercial reference control material. It should be realized
that the use of quality control material checks both reagent
and instrument functions together. Factors which might affect
the performance of this test include instrument function,
temperature control, cleanliness of glasswares and accuracy
of pipetting.
Other Details
The kit should be stored at 2-8°C and is stable till the expiry date indicated on the label.
The reagent and standard are ready-to-use and are stable til expiry, when stored at 2- 8°C. DO NOT FREEZE THE REAGENT.
The reagent should be stored only in the amber bottle provided to protect it from direct light. Before using, swirl in the reagent gently. DO NOT SHAKE VIGOROUSLY.
Over time, the reagent may develop a light pink colour. This is expected and does not affect the reagent performance. Discard the reagent if the absorbance of the same exceeds 0.300 O.D. against distilled water at 510 nm.
Contamination of the reagent should be strictly avoided. Should the
reagents develop turbidity and discard the reagent.
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