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    Lipase (S.L) : ADL/V.02/060114

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    Aniket dubey
    This document provides instructions for using a lipase reagent kit to quantify lipase levels in human serum or plasma samples. The kit uses a methyl resorufin method to measure the rate at which lipase hydrolyzes a synthetic substrate. The rate of methylresorufin formation is proportional to the lipase concentration in the sample. The reagent consists of two parts that are mixed with the sample and calibrator. The absorbance is then read to determine the lipase level using a calculation comparing the sample and calibrator readings. Normal lipase levels are up to 60 U/L but each laboratory should establish its own reference range.

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    Lipase (S.L) : ADL/V.02/060114

    Uploaded by

    Aniket dubey
    73 views1 page
    This document provides instructions for using a lipase reagent kit to quantify lipase levels in human serum or plasma samples. The kit uses a methyl resorufin method to measure the rate at which lipase hydrolyzes a synthetic substrate. The rate of methylresorufin formation is proportional to the lipase concentration in the sample. The reagent consists of two parts that are mixed with the sample and calibrator. The absorbance is then read to determine the lipase level using a calculation comparing the sample and calibrator readings. Normal lipase levels are up to 60 U/L but each laboratory should establish its own reference range.

    Original Title

    11417001

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    This document provides instructions for using a lipase reagent kit to quantify lipase levels in human serum or plasma samples. The kit uses a methyl resorufin method to measure the rate at which lipase hydrolyzes a synthetic substrate. The rate of methylresorufin formation is proportional to the lipase concentration in the sample. The reagent consists of two parts that are mixed with the sample and calibrator. The absorbance is then read to determine the lipase level using a calculation comparing the sample and calibrator readings. Normal lipase levels are up to 60 U/L but each laboratory should establish its own reference range.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    73 views1 page

    Lipase (S.L) : ADL/V.02/060114

    Uploaded by

    Aniket dubey
    This document provides instructions for using a lipase reagent kit to quantify lipase levels in human serum or plasma samples. The kit uses a methyl resorufin method to measure the rate at which lipase hydrolyzes a synthetic substrate. The rate of methylresorufin formation is proportional to the lipase concentration in the sample. The reagent consists of two parts that are mixed with the sample and calibrator. The absorbance is then read to determine the lipase level using a calculation comparing the sample and calibrator readings. Normal lipase levels are up to 60 U/L but each laboratory should establish its own reference range.

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    © All Rights Reserved
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    of 1

    1 x 25 mL

    LIPASE (S.L) 11417001


    INTENDED USE GENERAL SYSTEM PARAMETER
    This reagent is intended for in vitro quantitative determination of lipase in human Mode of Reaction Kinetic Fixed Time
    serum or plasma. Slope of reaction Increasing Increasing
    - Methyl resorufin method Wavelength 580 nm 580 nm
    - Linear up to 300 U/L Temperature 37 0C 37 0C
    - Reagent is ready for use Calibrator concentration As on the vial As on the vial
    CLINICAL SIGNIFICANCE Linearity 300 U/L 300 U/L
    Lipase is a pancreatic enzyme necessary for the absorption and digestion of Blank Reagent Reagent
    nutrients that catalyses the hydrolysis of glycerol esters of fatty acids. Determination Delay time 120 sec. 120 sec
    of lipase is used for diagnosis of diseases such as acute and chronic pancreatitis No of reading 2 -
    and obstruction of the pancreatic duct. Clinical diagnosis should not be made on a Interval 60 sec 120 sec
    single test result; it should integrate clinical and other laboratory data.
    Sample volume 20 μL 20 μL
    PRINCIPLE Reagent volume 1250 μL (1000+250) 1250 μL (1000 + 250)
    In the presence of colipase and bile acids lipase splits the synthetic substrate (1,2- Cuvette 1cm light path 1 cm light path
    O-Dilauryl-rac-glycero-3-glutaricacid (6-methyl-resorufin-ester) to glycerol and LABORATORY PROCEDURE
    methylresorufin-ester, which is spontaneously degraded to glutaric acid and
    methylresorufin. The rate of methylresorufin formation, measured photometrically Blank Calibrator Sample
    is proportional to the catalytic concentration of lipase present in the sample. Reagent 1 1000 μL 1000 μL 1000 μL
    Calibrator - 20 μL -
    REAGENT COMPOSITION
    Sample - - 20 μL
    LIPASE (S.L) R1 2 x 10 mL Dist. water 20 μL - -
    Goods Buffer (pH 8.0) 40 mmol/L Mix carefully (do not vortex); incubate for 1-5 minutes at 370C. Then add
    Taurodeoxycholate 3.4 mmol/L Reagent 2 250 μL 250 μL 250 μL
    Deoxycholate 6.4 mmol/L Mix and incubate for 2 min at 37oC, read absorbance against reagent blank. Measure
    Calcium chloride 12 mmol/L the change in absorbance per minute ( OD/min) during 2 min.
    Colipase 1.7 mg/dL or
    Mix and read the optical density (T1) 120 seconds after the Reagent 2 addition. Take
    LIPASE(S.L) R2 1 x 5 mL second reading (T 2) exactly after 120 seconds.
    Tartrate Buffer (pH 4.0) 1.5 mmol/L
    Taurodeoxycholate 3.4 mmol/L CALCULATION
    Color substrate 0.13 mmol/L Lipase U/L =
    LIPASE CALIBRATOR 1 x 3 mL ( OD/min) Sample - ( OD/min) Blank
    Lipase calibrator concentration is stated on the vial label. ---------------------------------------------------- x Calibrator concentration
    ( OD/min) calibrator - ( OD/min) Blank
    STORAGE & STABILITY or
    The sealed reagents are stable up to the expiry date stated on the label, when stored
    at 2 - 80C, protected from light.
    (T2-T1 ) of sample
    LINEARITY ------------------------- x Calibrator concentration
    This reagent is linear up to 300 U/L. (T2-T1 ) of standard
    If the concentration is greater than linearity (300 U/L), dilute the sample with
    normal saline and repeat the assay. Multiply the result with dilution factor. BIBLIOGRAPHY
    1. Mc Neely , M. ; Lipase. Kaplan, A. et al.; Clin. Chem. The C.V.Mosby Co. St Louis,
    NORMAL RANGE Toronto. Princeton 1984, 1130-1135
    It is recommended that each laboratory establish its own reference values. 2. Burtis, A., et al. ; Tietz Textbook of Clinical chemistry, 3rd ed AACC
    The following value may be used as guide line. 3. Neumann, U., et al.; Methods of Enzymatic Analysis, Vol 4, 3rd Ed.
    Serum / Plasma : Up to 60 U/L
    PREPARATION AND STABILITY OF REAGENT
    Lipase R1 and Lipase R2 are ready to use.
    Calibrator : Reconstitute with 3 mL of distilled water. Dissolve the content of the
    vial by swirling gently to avoid the formation of foam.
    Stability : Reconstituted calibrator is stable only for 7 days at 2-80C and 3 months
    at -200C.
    PRECAUTION
    To avoid contamination, use clean laboratory wares. Use clean dry disposable
    pipette tips for dispensing. Close reagent and calibrator bottles immediately after
    Use. Avoid direct exposure of reagent to light.
    SAMPLE
    Serum or plasma with sodium citrate, EDTA or heparin.

    ADL/V.02/060114

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