LIPASE
Methyl Resorufin Method:-
INTENDED USE:
The reagent kit is intended for the in vitro quantitative determination of lipase in serum/plasma.
PRINCIPLE:
In the presence of colipase and bile acids, lipase splits the synthetic substrate (1,2-O-dilauryl-rac-glycero-3-glutaric acid 6-methylresorufin ester) to form lauric acid and methylresorufin. The rate of hydrolysis is measured photometrically and is proportional to the catalytic activity of lipase present in the sample.
CONTENTS:
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Reagent 1: Lipase Reagent R1
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Reagent 2: Lipase Reagent R2
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Reagent 3: Lipase Calibrator
SAMPLE:
Serum or plasma with sodium citrate, EDTA, or heparin.
PRECAUTION:
To avoid contamination, use clean laboratory materials, use clear reagents and glassware. Avoid direct exposure of reagents to light.
PROCEDURE:
Pipette into clean dry test tubes labeled as Blank (B), Calibrator (C), and Test (T):
| Component | Blank (B) | Calibrator (C) | Test (T) |
|---|---|---|---|
| Reagent 1 | 1.0 ml | 1.0 ml | 1.0 ml |
| Calibrator | — | 20 µl | — |
| Sample | — | — | 20 µl |
Mix carefully (do not vortex). Incubate for 1–5 min at 37°C and read absorbance at 580 nm.
Add Reagent 2: 250 µl to each tube, mix gently and read absorbance after exactly 120 sec (A2).
CALCULATION:
For kinetic:
Lipase U/L = (ΔOD/min Sample / ΔOD/min Blank) × Calibrator Concentration
For fixed time:
Lipase U/L = (Abs (A2–A1) Sample / Abs (A2–A1) Calibrator) × Calibrator Concentration
NORMAL RANGE:
Up to 60 U/L
(Each laboratory should establish its own normal range.)
CLINICAL SIGNIFICANCE:
Lipase is a pancreatic enzyme necessary for the absorption and digestion of nutrients. The deficiency of lipase or disorders of fat digestion may result in the abnormal digestion of fats and malabsorption. Clinical diagnosis necessitates the presence of pancreatic duct obstruction, acute pancreatitis, and other conditions.
GENERAL SYSTEM PARAMETERS:
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Reaction type: Kinetic
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Reaction slope: Increasing
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Wavelength: 580 nm
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Cuvette: 1 cm
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Reaction temperature: 37°C
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Dead time: 60 sec
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No. of readings: 3
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Sample volume: 20 µl
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Calibrator: 2 points
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Working reagent volume: 1.25 ml (1000 + 250)
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Blank: Reagent
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Assay procedure: 1 cm light path