CREATININE REAGENT COMPOSITION

Wavelength: 492 (490-510 nm)

Contents Initial Concentrations of Solutions Cuvette: 1 cm light path
INTENDED USE Temperature: 25/30/37oC
For the quantitative determination of creatinine in serum, Measurement: against air
plasma or urine. 1. Standard 177 µmol/l (2 mg/dl)
2. Picric acid 35 mmol/l Pipette into cuvette:
Cat. No. Surfactant
3. Sodium hydroxide 0.32 mol/l Standard Sample
CR 510 200 ml 1. Standard 1x 5 ml Macro Semi Macro Semi
2. Picric acid 1 x 100 ml Micro Micro
3. Sodium hydroxide 1 x 100 ml PRECAUTIONS AND WARNINGS
For in vitro diagnostic use only. Do not pipette by mouth. Working reagent 2.0 ml 1.0 ml 2.0 ml 1.0 ml
CR 524 6 x 500 ml 1. Standard 1 x 30 ml Exercise the normal precautions required for handling Standard solution 1 0.2 ml 0.1 ml - -
2. Picric Acid 3 x 500 ml laboratory reagents. Sample - - 0.2 ml 0.1 ml
3. Sodium hydroxide 3 x 500 ml
Solution 2 contains picric acid which is poisonous. Mix and after 30 seconds read the absorbance A1 of the
CLINICAL SIGNIFICANCE standard and sample. Exactly 2 minutes later, read
Solution 3 contains sodium hydroxide which is caustic.
Creatinine is derived from creatine and creatine phosphate absorbance A2 of standard and sample.
in muscle tissue and may be defined as a nitrogenous Health and Safety data sheets available on request. CALCULATION
waste product. Creatinine is not reutilised but is excreted
from the body in the urine via the kidney. It is produced STABILITY AND PREPARATION OF REAGENTS A2 - A1 = ∆A sample or ∆A standard
and excreted at a constant rate which is proportional to the All reagents are supplied ready to use. Stable to expiry
body muscle mass. As a consequence of the way in which date when stored at +15 to +25°C. Concentration of creatinine in serum or plasma.
creatinine is excreted by the kidney, creatinine
measurement is used almost exclusively in the assessment
PREPARATION OF WORKING REAGENT ∆Asample
of kidney function. Creatinine is regarded as the most
Mix equal volumes of Solutions 2 + 3. Stable for 3 days at x 177 = µmol/l
useful endogenous marker in the diagnosis and treatment
+15 to +25°C. ∆Astandard
of kidney disease.
Creatinine is measured primarily to assess kidney function ∆Asample
PROCEDURE
and has certain advantages over the measurement of urea. x 2 = mg/dl
Materials Provided
The plasma level of creatinine is relatively independent of Creatinine Standard ∆Astandard
protein ingestion, water intake, rate of urine production and Creatinine Picric Acid
exercise. Since its rate of production is constant, elevation Creatinine Sodium Hydroxide Concentration of creatinine in urine.
of plasma creatinine is indicative of under-excretion,
suggesting kidney impairment. Depressed levels of Materials Required But Not Provided ∆Asample
x 8.85 = mmol/l
plasma creatinine are rare and not clinically significant. Pipetting devices for the delivery of 100µl, 200µl, 1ml and
2ml. ∆Astandard
COLORIMETRIC METHOD(1) Timing device and water bath or heating block to maintain
temperature at 25, 30 or 37°C. ∆Asample
Spectrophotometer with wavelength capability of 490 to x 100 = mg/dl
PRINCIPLE
510nm. ∆Astandard
Creatinine in alkaline solution reacts with picric acid Randox Assayed Human Multisera, HN1530, HE1532. mg creatinine/dl urine
to form a coloured complex. The amount of the complex Creatinine x ml urine 24 hrs
formed is directly proportional to the creatinine Clearance = [ml/min]
concentration. mg creatinine/dl serum x 1440

SAMPLE COLLECTION AND PREPARATION

Serum or plasma
Urine: diluted 1 + 49 with double distilled water.

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MANUAL - CREATININE - CR510 Page 2 of 2

QUALITY CONTROL SPECIAL PERFORMANCE CHARACTERISTICS METHOD COMPARISON (Plasma)

Randox Assayed Human Multisera, Level I and Level II are The following performance data were obtained using a The Randox method (Y) was compared to another
recommended for daily quality control. One control (Level I COBAS MIRA automated analyser @ 37°C. For commercially available method (X). Thirty patient samples,
or Level II) should be assayed after every 10 samples. The information on other automated analysers, please contact with values spanning the range 46.7 to 259.4 µmol/l were
values obtained for these controls should fall within a Randox Laboratories Ltd. tested. Linear regression analysis of the data resulted in the
specified range. If control values fall outside the ranges following equation,
and repetition excludes technical error the following steps SENSITIVITY
should be taken: The minimum detectable level has been determined as Y = 0.93 X - 1.05 with a correlation coefficient r = 0.999
14µmol/l.
1. Check wavelength setting and light source. METHOD COMPARISON (Serum)
2. Check cleanliness of glassware and pipettes. PRECISION (Plasma) The Randox method (Y) was compared to another
3. Check water, contaminants i.e. bacterial growth may commercially available method (X). Thirty nine patient
contribute to inaccurate results. Intra - Assay Level l Level ll samples, with values spanning the range 68.2 to 216.7µmol/l
4. Check reaction temperature. Mean (µmol/l) 76.7 448.6 were tested. Linear regression analysis of the data resulted
5. Check expiration date of kit and contents. SD 2.08 3.93 in the following equation,
6. Contact Randox Laboratories Customer Technical CV (%) 2.7 0.88
Support, Northern Ireland (028) 94422413. n 20 20 Y = 0.949 X - 1.02 with a correlation coefficient r = 0.99

NORMAL VALUES(2) Inter - Assay Level l Level ll REFERENCE

Serum: Men 53 - 97 µmol/l (0.6 - 1.1 mg/dl) Mean (µmol/l) 75.9 454.7 1. Bartels, H., Bohmer, M., (1972) Clin. Chem. Acta 37:
Women 44 - 80 µmol/l (0.5 - 0.9 mg/dl) SD 2.2 7.28 193.
Urine: 8.84 - 13.3 mmol/24 hrs CV (%) 2.9 1.6 2. Schirmeister, J., H. Willmann, and H. Kiefer. (1964).
1 - 1.5 g/24 hrs n 40 40 Dtsch. Med. Wschr. 89: 1018.
It is recommended that each laboratory establish its own PRECISION (Serum)
reference range to reflect the age, sex, diet and geographic
location of the population. Intra - Assay Level l Level ll
Mean (µmol/l) 75.9 149.5
LINEARITY SD 1.45 2.91
If the concentration exceeds 884 µmol/l (10 mg/dl) in CV (%) 1.91 1.95
serum or plasma or 44.2 mmol/l (500 mg/dl) in urine, dilute n 20 20
serum, plasma or diluted urine 1+4 with 0.9% (w/v) NaCl
and repeat the assay. Multiply the result by 5. Inter - Assay Level l Level ll
Mean (µmol/l) 75.6 148.0
PROCEDURE NOTES SD 1.8 3.05
Reaction rate and absorbance of the reaction product are CV (%) 2.39 2.06
very sensitive to temperature. The specified temperature n 20 20
must therefore be maintained.
Haemolysis interferes with the test. Do not use lipaemic
sera. The method is subject to interferences from high
levels of reducing substances. Boiling urine before testing
may help reduce this interference.

Revised 16/10/97

RANDOX Laboratories Ltd., Ardmore, Diamond Road, Crumlin, Co. Antrim, United Kingdom, BT29 4QY

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Tel:CRUMLIN (028)94422413 Fax.No.INT.44 (028)94452912 UK (028)94452912
Email: applications@randox.com Website: www.randox.com