Principle
The chromogenic lipase substrate 1, 2-0-dilauryl-rac-glycero-3-glutaric acid-(6-methyl resorufin) ester is cleaved by the catalytic action of lipase to form 1,2-0-dilauryl-rac-glycerol and an unstable intermediate glutaric acid-(6-methyl resorufin) ester. This decomposes spontaneously in alkaline medium to form glutaric acid and methyl resorufin. The lipase activity in the specimen is proportional to the rate of formation of methyl resorufin in the reaction and can be determined photometrically at 578 nm.
Procedure
Reaction type ………………… Kinetic
Reaction direction ……………. Increasing
Wavelength ………………….. 578 nm.
A Flow Cell temperature …. 37°C
Zero setting with ………….. Reagent Blank
Delay time. ………… 120 seconds
No. of readings …… 3
Interval ……….. 60 seconds
Sample volume ………… 0.01 ml (10 pl)
R1 volume …. 0.8 ml (800 pl)
R2 volume .. 0.2 ml (200 ul)
Linearity …… 300 10/1
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 reagents R1 & R2 are ready-to-use.The reagent kit should be stored at 2- 8°C and is stable till the expiry date indicated on the label.
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