Kit CompositionR1 1 x 50 mL + R2 1 x 50 mL + STD 1 x 5 mLR1 1 x 125 mL + R2 1 x 125 mL + STD 1 x 5 mL

POCT Services Private Limited, Plot No. – 3, Pocket – H, Sector – 5, DSIIDC, Bawana Industrial Area, New Delhi – 110 039, India

INTENDED USEQ-Line™ CREATININE JAFFE is intended for the quantitative in vitro diagnostic determination of creatinine in human serum, plasma and urine.

METHOD Colorimetric, Jaffe - Kinetic.

(3-4)PRINCIPLE The rate of formation of a coloured complex between creatinine and alkaline picrate is measured. The effect of interfering substances are reduced using the kinetic procedure.

REAGENTS COMPOSITIONReagent 1: R1Picric acid 8.73 mmol/LReagent 2: R2Sodium hydroxide 312.5 mmol/LDisodium phosphate 12.5 mmol/LStandard: StdCreatinine 2 mg/dL 177 µmol/L

MATERIALS REQUIRED BUT NOT PROVIDED- CALI0550ET ELICAL 2 4 x 3 mL- CONT0060ET ELITROL I 10 × 5 mL- CONT0160ET ELITROL II 10 × 5 mL- General Laboratory equipment - Do not use materials that are not required as indicated above.

WARNINGS AND PRECAUTIONS- This reagent is for professional in vitro diagnostic use only.- The reagent R2 is classified as hazardous:Warning : May be corrosive to metals. Causes skin irritation. Causes serious eye

irritation. Wear protective gloves/protective clothing/eye protection face protection. Do not breathe mist/vapours/spray. IF IN EYES : Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do. Continue rinsing. If eye irritation persists: Get medical advice/attention. Absorb spillage to prevent material damage.

- For more information, refer to the Safety Data Sheet (SDS).- Take normal precautions and adhere to good laboratory practice.- The standard solution be immediately & tightly capped to prevent contamination &

evaporation.- Use clean or single use laboratory equipment only, to avoid contamination.- Do not interchange reagent vials from different kits.

STABILITY OF REAGENTSStore at 2-25 °C and protect from light. Do not freeze.The reagents are stable until the expiry date stated on the labels.

On board stability: The on-board stability is specific for each analyser (Refer to § PERFORMANCE DATA)

PREPARATION AND STABILITY OF WORKING REAGENTThe reagents & standard are ready to use.

PRODUCTS DETERIORATIONThe reagent and standard solution should be clear. Cloudiness would indicate deterioration. Do not use the product if there is visible evidence of biological, chemical or physical deterioration.

DAMAGED PACKAGING Do not use the reagent if the damages of packing might have an effect on the product performance (leakage, pierced vial)

(5)SAMPLES SpecimenSerum.24h-Urine to be diluted 1/25 with distilled water before analysis (when there is no pre-dilution by the analyser).Do not use other specimens.

WARNINGS AND PRECAUTIONSAccording to Good Laboratory Practice, venipuncture should be performed prior to the administration of drugs.

CREATININE JAFFE

(1-2)CLINICAL SIGNIFICANCE Creatinine is the waste spontaneous product of creatine metabolism. It is an xcellent

marker of the renal function. The serum creatinine rate tends to remain constant. A high

serum creatinine rate (associated to a high urea rate) corresponds to a decrease in renal

glomerular filtration (FGR). The serum creatinine test is more reliable than the urea test.

Indeed, the urea serum rate is affected by factors such as diet, dehydratation degree and

protein metabolism (the serum creatinine rate is not influenced by these factors). The test

of creatinine clearance can also be used to measure the FGR. In the case of renal

transplantation, any increase in serum creatinine, as little as it may be, can reflect the

rejection of the transplant. An increase of creatinine serum and urine can be the sign of

muscular necrosis.

STORAGE AND STABILITYSerum is stable for 24 hours at 15 - 25 °C.7 days at 2 - 8 °C and 3 months at -20 °CUrines are stable for 4 days at 2-8 °C.

(6)REFERENCE VALUES

Men Women

Serum 0.8 - 1.3 0.6 - 1.2 mg/dL 71 - 115 53 - 106 µmol/L

Urine 0.8 - 2 0.6 - 1.8 g/24h 7.1 - 17.7 53 - 15.9 mmol/24h 53 - 133 40 - 120 mg/dL* 4.7 - 11.8 3.5 - 10.6 mmol/L* *For an urinary volumes of 1.5 L per 24 hours.Note: The quoted range should serve as a guide only. It is recommended that each laboratory verifies this range or establishes a reference interval for the intended population.

PROCEDUREThese reagents can be used on most Fully Auto-analysers, semi-automated analysers and manual method.

MANUAL METHOD ( For Semi-automatic Analyzers)

CALCULATION

∆A (OD2 - Od1)

xn where n = Standard conc. (2 mg/dL) ∆A (OD2 – Od1)

Conversion factor : mg/dL × 88.40 = µmol/L mg/dL × 10 = mg/L

Take dilution factor in account for calculation of creatinine concentration in urine as underUrine Creatinine (mg/dL) = Obtained Value x 25 mg/dL

CALIBRATIONFor calibration, Creatinine Standard 2 mg/dL must be used. Its concentration value is traceable to the reference method ID-MS (Isotope Dilution – Mass Spectrometry)

Calibration Frequency : The calibration is specific for each analyser (Refer to § PERFORMANCE DATA)

PROCEDURE for ELItech Clinical Systems Selectra AnalyzersThe applications are available on request.

CALCULATION

Sample

Calibrator

Creatinine (mg/dL) =

Wavelength : 505 nmTemperature : 37 °CRead against distilled water.

Reagent R1

Reagent R2

Test

500 µL

500 µL

100 µL

-

Standard

Sample

Standard

500 µL

500 µL

-

100 µL

Mix and read the optical density (OD1) 10 seconds after the sample or standard addition. Exactly 2 minutes after first reading, take second reading (Od2).

Read against distilled water.

Test

125 µL

25 µL

-

125 µL

-

25 µL

Calibration

Reagent R1

Calibrator

Sample

125 µLReagent R2

Mix and wait 4 minutes and 43 seconds, then add

125 µL

Mix and after 24 seconds of incubation, read the variation of the absorbance (∆A) during106 seconds.

Sample

Calibrator

CRTG0100PS 2 x 50 mLCRTG0250PS 2 x 125 mL

Reference / Pack Sizes

∆A

x n where n = Calibrator concentration

∆A

POCT Services Private Limited, Plot No. – 3, Pocket – H, Sector – 5, DSIIDC, Bawana Industrial Area, New Delhi – 110 039, India

Take dilution factor in account for calculation of creatinine concentration in urine.

Conversion factor:� mg/dL × 88.4 = µmol/L� � � mg/dL × 10 = mg/L

CALIBRATIONFor calibration, multiparametric calibrator ELICAL 2 must be used. Its value is traceable to ID-MS (Isotope Dilution - Mass Spectrometry) reference method.

Calibration frequency: The calibration is specific for each analyzer. (Refer to § PERFORMANCE DATA).

QUALITY CONTROLTo ensure adequate quality, control sera such as ELITROL I (normal control) and ELITROL II (abnormal control) should be used. These controls must be performed & validated before the patient samples are assayed. The control frequency must be at least once a day, after each calibration and should be adapted to Quality Control procedures of each laboratory and the regulatory requirements. Results should be within the defined ranges. If values fall outside of the defined ranges, each laboratory should take corrective measures. Quality control material should be used in accordance with local guidelines.

WASTE MANAGEMENTDisposal of all waste material should be in accordance with local and legal requirements.

PERFORMANCE DATA at 37 °C on ELITech Clinical Systems Selectra E AnalyzerMeasuring rangeThe reagent is linear from 0.5 to 15 mg/dL (44 to 1326 μmol/L).

(7)Detection limit Determined according to SFBC protocol, the detection limit is equal to 0.1 mg/dL (9 μmol/L).

CorrelationA comparative study has been performed on this reagent between Elitech Clinical Systems Selectra E and Cobas Mira analysers (Jaffe Method) on 30 human serum samples.The sample concentrations were between 0.47 and 18.26 mg/dl (42 and 1614 μmol/L)The parameters of linear regression are as follows:Correlation coefficient : (r) = 0.9998Linear regression : (y) = 0.9786 x + 0.06 mg/dL (5 μmol/L)

(7)Limitations/InferencesDo not report results outside of the usable range.According to SFBC recommendations, some studies have been performed to determine the level of interference from different components:Conjugated Bilirubin : Negative bias from 51.3 μmol/L (2.3 mg/dL).Unconjugated Bilirubin : Negative bias from 50 μmol/L (2.9 mg/dL).Haemoglobin : Negative bias from 150 mg/dL (1.5 g/L).Turbidity : No significant interference up to 580 mg/dL (6.55 mmol/L) Triglycerides equivalent.

In very rare cases, monoclonal gammopathies (multiple myeloma), in particular IgM type (8)(Waldenstrom'smacroglobulinemia) can cause unreliable results.

Many other substances and drugs may interfere.(9-10)Some of them are listed in reviews published by Young.

The results of this assay should only be interpreted in conjunction with other diagnostic test results, clinical findings and the patient's medical history.

On board stability / calibration frequency On-board stability : 3 daysCalibration frequency : 3 days

Recalibrate when reagent lots change, when quality control results fall outside the established range, and after a maintenance operation.

Precision

Within-run reproducibility

Between-run reproducibility

n

Mean

CV (%)

n

Mean CV (%)

mg/dL µmol/L mg/dL µmol/L

Level 1 20 0.57 50 2.7 20 0.59 52 4.1

Level 2 20

1.60

141

0.4

20 1.60

141

2.8

Level 3 20 6.57 581 1.0 20 6.72 594 2.3

BIBLIOGRAPHY

1. Allston, C.A., Non protein nitrogenous compounds and renal function.

Clinical Chemistry: Concepts and Application,Anderson, S.C., Cockayne,

S. (W.B. Saunders eds. Philadel-phiaUSA), (1993), 369.

2. Newman, D.J., Price C.P., Non protein nitrogen metabolite. thTietzFundamentals of Clinical Chemistry, 5 Ed., Burtis, C.A. &Ashwood,

E.R. (W.B. Saunders eds. Philadelphia USA), (2001), 414.

3. Butler, A.R., The Jaffe reaction. Identification of the coloured species. Clin.

Chim. Acta., (1975), 59, 227.

4. Vasiliades, J., Reaction of alkaline picrate with creatinine. 1. Kinetics and

mechanism of formation of the mono-creatinine picric acid complex. Clin.

Chem., (1976), 22, 1664.

5. Guder,W.G.,et al., Use of anticoagulant in diagnostic laboratory

investigations and stability of blood, plasma and serum samples. (2002).

WHO/DIL/LAB/99.1 Rev 2.rd6. Tietz, N.W. Clinical guide to laboratory tests, 3 Ed, (W.B. Saunders eds.

Philadelphia USA), (1995), 186.

7. Vassault A., et al., Ann. Biol. Clin., (1986), 44, 686.

8. Berth, M. &Delanghe, J., Protein precipitation as a possible important pitfall

in the clinical chemistry analysis of blood samples containing monoclonal

immunoglobulins: 2 case reports and a review of literature, ActaClin Belg.,

(2004), 59, 263.

9. Young, D.S., Effects of preanalytical variables on clinical laboratory tests, nd2 edition, AACC Press (1997).

th10. Young, D.S., Effects of drugs on clinical laboratory tests, 4 edition, AACC

Press (1995).

Temperature Limitation

General Technical ParametersTwo Point / Fixed Time

505 nm

None / No

Increasing / Positive

No

No

100 µL

1000 µL

10 Seconds

120 Seconds

NA

1- Point

NA (Calculate by System)

2.0 mg/dL

15 mg/dL

2

37 °C

mg/dL

0.8 / 0.6 mg/dL

1.3 / 1.2 mg/dL

Mode

Wavelength (Filter)

Secondary Wavelength / Bichromatic

Reaction Direction / Type

Reagent Blank

Sample Blank

Sample Vol.

Reagent Vol.

Delay Time / Lag Time

Measuring Time / Interval

Reagent Blank Abs.

Calibration Method

Factor

Standard (Conc.)

Linearity

Decimal Places

Temp.

Unit

Ref. Low (Male/Female)

Ref. High (Male/Female)