Lipase 1 x 24 mL, 1x48 mL

Kinetic Colorimetric

Clinical Significance
Lipase (LPS) is a pancreatic enzyme necessary for the Materials required
absorption and digestion of nutrients that catalyzes the - Spectrophotometer or colorimeter measuring at 580 nm.
hydrolysis of glycerol esters of fatty acids. Determination - Thermostatic bath at 37º C (± 0.1ºC).
of LPS is used for diagnosis of diseases of pancreas such - Matched cuvettes 1.0 cm light path.
as acute and chronic pancreatitis and obstruction of the - General laboratory equipment.
pancreatic duct1,7,8. Clinical diagnosis should not be
made on a single test result; it should integrate clinical Specimen and Stability
and other laboratory data. Principle Serum or plasma with sodium citrate, EDTA or heparin.
The pancreatic lipase in presence of colipase, Avoid repeated frozen and unfrozen.
desoxycholate and calcium ions, hydrolyses the substrate Stability: 2 days at 2-8ºC.
1-2-O-dilauryl-rac-glycero-3-glutaric acid-(6' - Assay Procedure
methylresorufin)-ester. The sequence of reactions
involved in the enzymatic direct lipase determination is 1. Assay conditions:
the following: Wavelength: . . . . . . . . . . . . . . . . . . . . . . . 580 nm
Cuvette: . . . . . . . . . . . . . . . . . . . . . 1 cm light path
1-2-O-dilauryl-rac-glycero-3-glutaric-(6' -methylresorufin)-ester Constant temperature . . . . . . . . . . . . . . . . . . .37ºC
Lipase 2. Adjust the instrument to zero with distilled water.
1-2-O-dilauryl-rac-glycerol+ Glutaric-6'-methylresorufin 3. Pipette into a cuvette:
- ester Blank Sample or Calibrator
OH-- R1 1000 µl 1000 µl
(no stable) Glutaric acid + Methylresorufin
R2 200 µl 200 µl
The rate of methylresorufin formation, measured Distilled Water 10 µl --
photometrically, is proportional to the catalytic Sample / Calibrator -- 10 µl
concentration of lipase present in the sample.
4. Mix, incubate at 37ºC for 1 minute and read initial
Reagents absorbance (A1) of the sample.
R1 TRIS pH 8.3 40 mmol/L 5. Thereafter read the absorbance once again after 2
minutes (A2).
Buffer Colipase > 1 mg/L 6. Calculate the difference between absorbances A2-A1 =ΔA.
Desoxycholate 1.8 mmol/L
Application sheets for automated systems are
Taurodesoxycholate 7.2 mmol/L
available on request.
R2 Tartrate pH 4.0 15 mmol/L
Calculation
Substrate Lipase > 0.7
(microemulsion) mmol/L ∆A of Sample x Calibrator Concentration= U/L Lipase in Sample
Calcium Chloride 0.1 mmol/L ∆A of Calibrator
(CaCl2)
Lipase Cal. Calibrator. Lyophilized human serum Units: One international unit (IU) is the amount of
The LPS activity (U/L methylresorufin enzyme that transforms 1 μmol of substrate per minute,
at 370 C) is indicated on the vial label in standard conditions. The concentration is expressed in
units per litre of sample (U/L).
Warnings and Precautions
LIPASE CAL Components from human origin have been Quality Control
tested and found to be negative for the presence of Commercially available normal and pathological control
HBsAg, HCV, and antibody to HIV (1/2). However handle sera are recommended to monitor the performance of the
cautiously as potentially infectious. procedure.
If the controls are found outside the defined range, check
the instrument, reagents and calibrator for problems.
Storage Instructions and Reagent Stability Serum controls are recommended for internal Quality
All the components of the kit are stable until the control. Each laboratory should establish its own Quality
expiration date on the label when stored tightly closed at Control scheme and corrective actions
2-8ºC, protected from light and contaminations prevented
during their use. Do not use reagents over the expiration Reference Range
date. < 38 U/L (U/L methylresorufin at 37ºC).
Signs of reagent deterioration:
- Presence of particles and turbidity. These values are for orientation purpose; each laboratory
- Blank absorbance (A) at 580 nm ≥ 1.00. should establish its own reference range.
- R 2 is a turbid orange-colored micro-emulsion, discard if
turning to red. Performance Characteristics
Reagent Preparation Measuring range
Measuring range: From detection limit of 5 U/L to linearity
- R1 – R2 Ready to use. Stability after opening 90 days
limit of 250 U/L.
at 2-8ºC.
If the results obtained were greater than linearity limit,
- R2 Mix gently before use (Note 1).
dilute the sample 1/10 with NaCl 9 g/L and multiply the
- LIPASE CAL: Dissolve with 1 mL of distilled water.
result by 10.
Cap and mix gently to dissolve contents. Stability: 7
Accuracy: Results obtained using this reagent (y) did not
days at 2-8ºC or 3 months at –20ºC; aliquote into
show systematic differences when compared with other
small volumes and freeze.
commercial reagents (x).
The results obtained using 100 samples were the Literature
following: 1. McNeely M. Lipase. Kaplan A et al. Clin Chem The
Correlation coefficient (r):0.997. C.V. Mosby Co. St Louis. Toronto. Princeton 1984;
Regression equation: y= 0.50054x + 3.9443. 1130-1134, 892.
The results of the performance characteristics depend on
the analyzer used. 2. Neumann U et al. Comptes Rend. 4 colloque de Pont-
a-Musson, Masson 627-634 (1979) .
Specificity / Interferences 3. Junge W et al. J.Clin.Chem.Clin.Biochem., 21 445-451
Triglycerides at 300 mg/dL interfere on determination (1983).
reducing the activity of enzyme of 6%. Hemoglobin 4. Neumann U et al. Methods of Enzymatics Analysis,
concentration lower than 150 mg/dL and Bilirubin lower 3rd ed. Vol.4, 26-34 (1984) .
than 20 mg/dL do not interfere. A list of drugs and other 5. Young DS. Effects of drugs on Clinical Lab. Tests, 4th
interfering substances with lipase determination has been ed AACC Press, 1995.
reported by Young et. al2,3. 6. Young DS. Effects of disease on Clinical Lab. Tests,
4th ed AACC 2001.
SYSTEM PARAMETERS
7. Burtis A et al. Tietz Textbook of Clinical Chemistry,
3rd ed AACC 1999.
Mode : Two Point/Fixed time
Reaction : Ascending 8. Tietz N W et al. Clinical Guide to Laboratory Tests,
Wavelength : 580 nm 3rd ed AACC 1995.
Blank with : Distilled water
Sample Volume : 10 µL Manufactured by: Spinreact, S.A.
Reagent Volume : 1200 µL Marketed By: Euro Diagnostic Systems Pvt. Ltd.,
Delay Time : 60 (Sec) Millennium House, M.K.Srinivasan Nagar Main Road,
Read Time : 120 (Sec) No. 144, Old Mahabalipuram Road, Perungudi,
Interval time : 60 (Sec) Chennai – 600 096; Email: [email protected],
Calibrator : Stated on the vial www.eurods.in
Normal range : < 38
Sensitivity : 5 U/L
Linearity limit : 250
Unit : U/L

NOTES
1. In some storage conditions (i.e. storage at a
temperature lower that the one indicate) a precipitate
may appear in the vial that will not influence that the
reagent performance; however, it is recommended to
resuspend the product with a slight rotation.
2. In order to avoid contamination it is recommended to
use disposable material.