03029590322V1

LIPC
Lipase colorimetric assay

• Indicates cobas c systems on which reagents can be used

Order information Roche/Hitachi cobas c systems
Lipase colorimetric assay cobas c 501
200 tests Cat. No. 03029590 322 System-ID 07 5900 7 •
Calibrator f.a.s. (12 x 3 mL) Cat. No. 10759350 190 Code 401
Calibrator f.a.s. (12 x 3 mL for USA) Cat. No. 10759350 360 Code 401
Precinorm U plus (10 x 3 mL) Cat. No. 12149435 122 Code 300
Precipath U plus (10 x 3 mL) Cat. No. 12149443 122 Code 301
Precinorm U (20 x 5 mL) Cat. No. 10171743 122 Code 300
Precipath U (20 x 5 mL) Cat. No. 10171778 122 Code 301

English Reagents - working solutions

System information R1 BICIN* buffer: 50 mmol/L, pH 8.0; colipase (porcine pancreas): ≥0.9 mg/L;
LIPC: ACN 731 Na-deoxycholate: 1.6 mmol/L; calcium chloride: 10 mmol/L; detergent;
preservative
SLIPC: ACN 733 (STAT, reaction time: 5)
R2 Tartrate buffer: 10 mmol/L, pH 4.0; 1,2-O-dilauryl-rac-glycero-3-glutaric
Intended use
Enzymatic in vitro test for the quantitative determination of lipase in human acid-(6-methylresorufin) ester: 0.27 mmol/L; taurodeoxycholate:
serum and plasma on Roche/Hitachi cobas c systems. 8.8 mmol/L; detergent; preservative
* BICIN = N,N-bis(2-hydroxyethyl)glycine
Summary1,2,3,4,5,6,7
Lipases are glycoproteins with a molecular weight of 47000 daltons. They
Precautions and warnings
are defined as triglyceride hydrolases which catalyze the cleavage of
For in vitro diagnostic use.
triglycerides to diglycerides with subsequent formation of monoglycerides
Exercise the normal precautions required for handling all laboratory reagents.
and fatty acids. In addition to α-amylase, pancreatic lipases have for many
Safety data sheet available for professional user on request.
years been undeniably the most important clinical chemistry parameters for
Disposal of all waste material should be in accordance with local guidelines.
the differential diagnosis of diseases of the pancreas. The lipase activity
determination has gained increasing international recognition because of its Reagent handling
high specificity and rapid response. After acute pancreatitis the lipase activity Ready for use.
increases within 4-8 hours, reaches a peak after 24 hours and decreases
after 8 to 14 days. However, there is no correlation between the lipase activity Storage and stability
determined in serum and the extent of damage to the pancreas. LIPC
Numerous methods have been described for the determination of Shelf life at 2-8°C: See expiration date on
lipase which determine the decrease in substrate turbidimetrically or cobas c pack label.
nephelometrically or determine degradation products.
On-board in use and refrigerated on the analyzer: 4 weeks
This method is based on the cleavage of a specific chromogenic lipase
substrate 1,2-O-dilauryl-rac-glycero-3-glutaric acid-(6-methylresorufin) ester
emulsified with bile acids. The pancreatic enzyme activity is determined Specimen collection and preparation
specifically by the combination of bile acid and colipase used in this assay. For specimen collection and preparation, only use suitable
Virtually no lipase activity is detected in the absence of colipase. Colipase only tubes or collection containers.
activates pancreatic lipase, but not other lipolytic enzymes found in serum. The Only the specimens listed below were tested and found acceptable.
high amount of cholates ensure that the esterases present in the serum do not Serum.
react with the chromogenic substrate due to the highly negative surface charge. Plasma: Li-heparin plasma.
Test principle8,9,10,11 The sample types listed were tested with a selection of sample collection tubes
Enzymatic colorimetric assay 1,2-O-dilauryl-rac-glycero-3-glutaric that were commercially available at the time of testing, i.e. not all available
acid-(6-methyl-resorufin) ester as substrate. tubes of all manufacturers were tested. Sample collection systems from
The chromogenic lipase substrate 1,2-O-dilauryl-rac-glycero-3-glutaric various manufacturers may contain differing materials which could affect
acid-(6-methylresorufin) ester is cleaved by the catalytic action of alkaline the test results in some cases. When processing samples in primary tubes
lipase solution to form 1,2-O-dilauryl-rac-glycerol and an unstable intermediate, (sample collection systems), follow the instructions of the tube manufacturer.
glutaric acid-(6-methylresorufin) ester. This decomposes spontaneously in Centrifuge samples containing precipitates before performing the assay.
alkaline solution to form glutaric acid and methylresorufin. Addition of detergent
Stability:12 1 week at 15-25°C
and colipase increases the specificity of the assay for pancreatic lipase.
1 week at 2-8°C
lipase 1 year at (-15)-(-25)°C
1,2-O-dilauryl-rac-glycero-3-glutaric acid-(6-methylresorufin) ester
1,2-O-dilauryl-rac-glycerol + glutaric acid-(6-methylresorufin) ester Materials provided
spontaneous
See “Reagents - working solutions” section for reagents.
glutaric acid-(6-methylresorufin) ester glutaric acid +
methylresorufin Materials required (but not provided)
decomposition
See “Order information” section.
The color intensity of the red dye formed is directly proportional to the Distilled water
lipase activity and can be determined photometrically. General laboratory equipment
Assay
For optimal performance of the assay, follow the directions given in this
document for the analyzer concerned. Refer to the appropriate operator
manual for analyzer-specific assay instructions.

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LIPC
Lipase colorimetric assay
The performance of applications not validated by Roche is not Measuring range
warranted and must be defined by the user. 3-300 U/L (0.05-5.01 µkat/L)
Application for serum and plasma Extended measuring range (calculated)
3-300 U/L (0.05-5.01 µkat/L)
cobas c 501 test definition
Lower detection limit
Assay type Rate A
3 U/L (0.05 µkat/L)
Reaction time / Assay points 10 / 22-31 (STAT 5 / 22-31) The lower detection limit represents the lowest measurable analyte
Wavelength (sub/main) 700/570 nm level that can be distinguished from zero. It is calculated as the value
Reaction direction Increase lying three standard deviations above that of the lowest standard
Units U/L (µkat/L) (standard 1 + 3 SD, within-run precision, n = 21).
Reagent pipetting Diluent (H2O) Expected values15
R1 80 µL 20 µL Adults: 13-60 U/L (0.22-1.00 µkat/L)
R2 48 µL – Each laboratory should investigate the transferability of the expected values to
Sample volumes Sample Sample dilution its own patient population and if necessary determine its own reference ranges.
Sample Diluent (H2O) Specific performance data
Normal 2 µL – – Representative performance data on the analyzers are given below.
Decreased 2 µL – – Results obtained in individual laboratories may differ.
Increased 4 µL – –
Precision
Calibration Reproducibility was determined using human samples and controls in
Calibrators S1: H2O an internal protocol (within-run n = 21, total n = 63).
S2: C.f.a.s. The following results were obtained:
Calibration mode Linear Within-run Mean SD CV
Calibration frequency 2-point calibration U/L (µkat/L) U/L (µkat/L) %
- after reagent lot change Precinorm U 62.2 (1.04) 0.4 (0.01) 0.7
- as required following quality control Precipath U 102 (1.70) 1 (0.01) 0.7
procedures Human serum 1 30.1 (0.50) 0.3 (0.01) 1.0
Traceability: This method has been standardized manually against Roche Human serum 2 231 (3.86) 2 (0.03) 0.9
reagent using calibrated pipettes together with a manual photometer providing
Total Mean SD CV
absolute values and the substrate-specific absorptivity, ε.
U/L (µkat/L) U/L (µkat/L) %
Quality control Precinorm U 61.0 (1.02) 0.9 (0.02) 1.5
For quality control, use control materials as listed in the Precipath U 99.3 (1.66) 1.9 (0.03) 1.9
“Order information” section.
Human serum 3 28.8 (0.48) 0.6 (0.01) 2.1
Other suitable control material can be used in addition.
Human serum 4 320 (5.34) 6 (0.09) 1.7
The control intervals and limits should be adapted to each laboratory’s
individual requirements. Values obtained should fall within the defined Method comparison
limits. Each laboratory should establish corrective measures to be Lipase values for human serum and plasma samples obtained on a
taken if values fall outside the limits. Roche/Hitachi cobas c 501 analyzer (y) were compared with those determined
Calculation using the same reagent on a Roche/Hitachi 917 analyzer (x).
Roche/Hitachi cobas c systems automatically calculate the Sample size (n) = 185
analyte activity of each sample. Passing/Bablok16 Linear regression
Conversion factor: U/L x 0.0167 = µkat/L y = 0.982x - 0.25 U/L y = 0.962x + 1.32 U/L
τ = 0.935 r = 0.998
Limitations - interference13 The sample activities were between 9.4 and 299 U/L (0.16 and 4.99 µkat/L).
Criterion: Recovery within ± 10% of initial values at a lipase
activity of 60 U/L (1.00 µkat/L). References
Icterus: No significant interference up to an I index of 60 (approximate 1. Greiling H, Gressner AM, eds. Lehrbuch der Klinischen Chemie und
conjugated and unconjugated bilirubin concentration: 1026 µmol/L (60 mg/dL)). Pathobiochemie, 3rd ed. Stuttgart/New York: Schattauer Verlag, 1995.
Hemolysis: No significant interference up to an H index of 1000 (approximate 2. Keller H, ed. Klinisch-chemische Labordiagnostik für die Praxis, 2nd ed.
hemoglobin concentration: 620 µmol/L (1000 mg/dL)). Stuttgart/New York: Georg Thieme Verlag, 1991:354-361.
Lipemia (Intralipid): No significant interference up to an L index 3. Kazmierczak S, Catrou P, Van Lente F. Diagnostic accuracy
of 2000. There is poor correlation between the L index (corresponds of pancreatic enzymes evaluated by use of multivariate data
to turbidity) and triglycerides concentration. analysis. Clin Chem 1993;39:1960-1965.
Drugs: No interference was found using common drug panels.14 4. Steinberg WM, Goldstein SS, Davies ND et al. Diagnostic assays in
Exception: Calcium dobesilate causes artificially low lipase acute pancreatitis [Review]. Ann Intern Med 1985; 102:576-580.
results at high drug concentrations. 5. Panteghini M et al. Diagnostic value of four assays for lipase determination
in serum: A comparative reevalution. Clin Biochem 1991;24:497-503.
In very rare cases gammopathy, in particular type IgM (Waldenström’s
6. Tietz NW et al. Lipase in serum - the elusive enzyme: An overview.
macroglobulinemia), may cause unreliable results.
Clin Chem 1993;39:746-756.
For diagnostic purposes, the results should always be assessed in conjunction 7. Tietz NW. Clinical Guide to Laboratory Tests, 3rd ed. Philadelphia, PA:
with the patient’s medical history, clinical examination and other findings. WB Saunders, 1995:865.
Special wash requirements 8. Neumann U et al. New substrates for the optical determination
The determination of certain analytes interferes with this assay requiring of lipase. EP 207252 (1987).
a special wash step. Refer to the NaOHD/SMS/Multiclean method sheet 9. Borgström B. The action of bile salts and other detergents on
and the operator manual for further instructions. pancreatic lipase and the interaction with colipase. Biochimica
et Biophysica Acta 1977;488:381-391.

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LIPC
Lipase colorimetric assay
10. Gargouri Y, Julien R, Bois A et al. Studies on the detergent inhibition of
pancreatic lipase activity. J of Lipid Research 1983;24:1336-1342.
11. Leybold A, Junge W. Importance of colipase for the measurement of
serum lipase activity. Adv clin Enzymol 1986;4:60-67.
12. Guder W, Fonseca-Wollheim W, Heil O et al. Maximum
permissible transport and storage times for analysis of blood
(serum, plasma), urine and cerebrospinal fluid. DG Klinische
Chemische Mitteilungen 1995;26:207-224.
13. Glick MR, Ryder KW, Jackson SA. Graphical Comparisons of Interferences
in Clinical Chemistry Instrumentation. Clin Chem 1986;32:470-474.
14. Report on the Symposium “Drug effects in clinical chemistry methods”,
Breuer J, Eur J Clin Chem Clin Biochem 1996;34:385-386.
15. Junge W, Abicht K, Goldman J et al. Evaluation of the Colorimetric Liquid
Assay for Pancreatic Lipase on Hitachi Analyzers in 7 Clinical Centers in
Europe, Japan and USA. Clin Chem Lab Med 1999;37, Special Suppl:469.
16. Bablok W et al. A General Regression Procedure for Method
Transformation. J Clin Chem Clin Biochem 1988;26:783-790.
FOR US CUSTOMERS ONLY: LIMITED WARRANTY
Roche Diagnostics warrants that this product will meet the specifications
stated in the labeling when used in accordance with such labeling and
will be free from defects in material and workmanship until the expiration
date printed on the label. THIS LIMITED WARRANTY IS IN LIEU OF ANY
OTHER WARRANTY, EXPRESS OR IMPLIED, INCLUDING ANY IMPLIED
WARRANTY OF MERCHANTABILITY OR FITNESS FOR PARTICULAR
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INCIDENTAL, INDIRECT, SPECIAL OR CONSEQUENTIAL DAMAGES.

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©2006 Roche Diagnostics

Roche Diagnostics GmbH, D-68298 Mannheim

for USA: US Distributor:
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US Customer Technical Support 1-800-428-2336

2006-06, V 1 English 3/3 cobas c systems