Glycerol released from hydrolysis of triglycerides by lipoprotein lipase is converted by glycerol kinase into glycerol - 3 - phosphate which is oxidised by glycerol phosphate oxidase to dihydroxyacetone phosphate and hydrogen peroxide. In presence of peroxidase, hydrogen peroxide oxidizes phenolic chromogen to a red coloured compound.
REAGENT STORAGE, STABILITY & HANDLING
COMPONENTS & CONCENTRATION OF WORKING SOLUTION
Component |
Concentration |
Buffer, pH 7.2 |
50 mmol/I |
Lipase |
2000 IU/I |
Glycerol kinase |
300 IU/I |
Glycerol phosphate oxidase |
1000 IU/I |
Peroxidase |
500 IU/I |
ATP |
1 mmol/I |
Chromogen |
2 mmol/I |
NOTE :
1.0 ml procedure
|
Serum / Plasma |
Standard |
Blank |
Reagent |
0.01 ml |
0.01 ml |
|
1.0 ml |
1.0 ml |
1.0 ml |
Incubation
Incubate the assay mixture for 10 minutes at 37 degree C or 20 minutes at
R.T. (25 -30 degree C). After incubation, measure the absorbance against blank at 510 nm. (500-530 nm.). Final colour is stable for 30 minutes if not exposed to direct light.
EXPECTED VALUES
Upto 170 mg%
NOTE :
Expected range varies from population to population therefore each laboratory should establish its own normal range.
PROCEDURE LIMITATIONS
QUALITY CONTROL
To ensure adequate quality control, it is recommended that each batch should include a normal and an abnormal commercial reference control serum. It should be realised that the use of quality control material checks both instrument and reagent functions together. Factors which might affect the performance of this test include proper instrument function, temperature control, cleanliness of glassware and accuracy of pipetting
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