BIOLABO

www.biolabo.fr CREATININE
MANUFACTURER:
BIOLABO SAS, K inetic method
Les Hautes Rives
02160, Maizy, France Reagent for quantitative determination of creatinine
in human serum, plasma, or urines

REF 80107 R1 1 x 125 mL R2 1 x 125 mL R3 1 x 10 mL

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TECHNICAL SUPPORT AND ORDERS
Tel : (33) 03 23 25 15 50
IVD IN VITRO DIAGNOSTIC USE
Fax: (33) 03 23 256 256

CLINICAL SIGNIFICANCE (1) STABILITY AND STORAGE

Interconversion of phosphocreatine and creatine is a particular feature Store at 18-25°C, well cap in the original vial and away from light.
of the metabolism processes of muscle contraction. Creatine and • Standard (vial R3): Transfer the requested quantity, recap and store
phosphocreatine partially convert to a waste product, creatinine. Thus, at 18-25°C
the amount of creatinine produced each day is related to the muscle • Reagents are stable until expiry date stated on the label of the kit
mass (and body weight), age, sex, diet or exercise and does not when free from contamination, stored and used as described in the
greatly vary from day to day. Because creatinine is endogenously insert.
produced and released into body fluids at a constant rate and its • Once reconstituted, working reagent is stable for 30 days at 2-8°C
plasma levels are maintained within narrow limits, its clearance can be when free from contamination.
measured as an indicator of glomerular filtration rate (GFR). • Discard any reagent if cloudy or if the absorbance of working reagent
is > 0.300 at 490 nm.
PRINCIPLE (4)(5) • Don’t use working reagent after expiry date stated on the label of the
Colorimetric reaction (Jaffe reaction) of creatinine with alkaline picrate Kit.
measured kinetically at 490 nm (490-510), without any pre-treatment
step. This reaction has been improved (specificity, speed and SPECIMEN COLLECTION AND HANDLING (2)
adaptability) by the development of an initial-rate method. Serum or heparinised plasma.

REAGENTS COMPOSITION Urines: Collect during precisely timed interval’s (4, 12 or 24 h).
Dilute 1+19 in demineralised water before determination.
Vial R1 BASE • Creatinine is stable in specimen:
for 24 h at 2-8°C (freeze for longer storage).
Xi: IRRITANT ,
R36/38: Irritating to eyes and skin.
S36/37/39: Wear suitable protective clothing, gloves and eyes/face INTERFERENCES (1) (2) (3) (5)
protection Procedure n°1:
Disodium Phosphate 6.4 mmol/L Creatinine (µmol/L) Interferent Results
Sodium hydroxide 150 mmol/L In specimen
Vial R2 DYE 249 µmol/L Glucose No interference up to 1200 mg/dL
Sodium dodecyl sulfate 0.75 mmol/L 115 µmol/L Proteins Positive interference above 4000
Picric acid 4.0 mmol/L mg/dL
pH 4.0 99 µmol/L Ascorbic No interference up to 25 mg/dL
acid
Vial R3 STANDARD
106 µmol/L Bilirubin Negative interference above
177 µmol/L (2 mg/dL) 20µmol/L
96 µmol/L Haemoglobin No interference up to 250µmol/L
SAFETY CAUTIONS 105 µmol/L Lipemia No interference up to 0.320 abs
BIOLABO reagents are designated for professional, in vitro diagnostic (measured at 600nm)
use.
Procedure n°2: No interference of Bilirubin
• Verify the integrity of the contents before use.
Some antibiotics interfere also with the determination of creatinine
• Use adequate protections (overall, gloves, glasses).
according to Jaffe method.
• Do not pipette by mouth.
• In case of contact with skin or eyes, thoroughly wash affected areas For a more comprehensive review of factors affecting this assay refer
with plenty of water and seek medical advice. to the publication of Young D.S.
• Material Safety Data Sheet is available upon request.
• Waste disposal: Respect legislation in force in the country. CALIBRATION
All specimens should be handled as potentially infectious, in • Kit Standard (vial R3) or BIOLABO Multicalibrator, REF 95015
accordance with good laboratory practices using appropriate traceable to SRM 909b (ID-MS) or SRM914a/SRM967a validated
precautions. Respect legislation in force in the country. according to the recommendations of AFSSAPS (1 zero point, 1
point within normal level, 1 point within high level).
MATERIAL REQUIRED BUT NOT PROVIDED • Or any calibrator traceable to a reference method or material.
1. Basic medical analysis laboratory equipment. The calibration frequency depends on proper instrument functions and
2. Normal and pathological control sera. on the preservation of the reagent.
It is recommended to calibrate in the following cases :
REAGENTS PREPARATION 1. When changing batch of reagent.
Mix vial R1 and vial R2 contents (1 volume/1 volume). A graduated 2. After maintenance operations on the instrument .
test-tube may be used. • When control values obtained are out of range, even after using a
Automated instrument: reagent R1 and R2 may be added separetely new vial of fresh serum
(see § MANUAL PROCEDURE).

IVD REF LOT →

Manufacturer Use by In vitro diagnostic Temperature limitation Catalogue number See insert Batch number Store away from light sufficient for dilute with

Made in France Latest revision : www.biolabo.fr Revision : 26/07/2011

QUALITY CONTROL MANUAL PROCEDURE
• BIOLABO EXATROL-N Level I REF 95010. Let stand reagents and specimens at temperature of measurement.
• BIOLABO EXATROL-P Level II REF 95011. Perform all the assays at constant temperature (see note 4).
• Assayed control sera referring to the same method. Procedure n°1: For non icteric specimen using “Working reagent”
• AFSSAPS recommends to use low control, subnormal control and
pathological control Pipette in a 1 cm path Blank Standard Assay
length cuvette: (optional)
• External quality control program.
It is recommended to control in the following cases: Working reagent (R1 + R2) 1 mL 1 mL 1 mL
• At least once a run.
Demineralised water 100 µL
• At least once within 24 hours.
• When changing vial of reagent. Standard 100 µL
• After maintenance operations on the instrument.
If control is out of range, apply following actions: Specimen (Note 1) 100 µL
1. Repeat the test with the same control.
2. If control is still out of range, prepare a fresh control serum and Mix well. After 30 seconds, record absorbance A1 at 490 nm (490-510) against
repeat the test. reagent blank or distilled water. Exactly 2 minutes after the first reading, record
3. If control is still out of range, use a new vial of calibrator or a fresh absorbance A2.
calibrator and repeat the test.
4. If control is still out of range, calibrate with a new vial of reagent. Procedure n°2: For icteric specimen using “Bi-reagent”
5. If control is still out of range, please contact BIOLABO technical Pipette in a 1 cm path Blank Standard Assay
support or your local Agent. length cuvette: (optional)
Reagent R1 0.5 mL 0.5 mL 0.5 mL
EXPECTED VALUES (2)
Demineralised water 100 µL
Serum or plasma
Creatinine [ µmol / L ] mg/dL Standard 100 µL
Male [ 80-115 ] 0.9 to 1.3
Specimen (Note 1) 100 µL
Female [ 53-97]] 0.6 to 1.1
Urines Incubate for 5 minutes at constant temperature, then add:

Creatinine [ µmol / kg / 24 h]] mg / kg / 24 h Reagent R2 0.5 mL 0.5 mL 0.5 mL

Male [ 124-230 ] 14 to 26
Female [ 97-177 ] 11 to 20 Mix well. After 30 seconds, record absorbance A1 at 490 nm (490-510) against
reagent blank or distilled water. Exactly 2 minutes after the first reading, record
absorbance A2.
GFR (Glomerular filtration rate) mL per minute
Adult < 40 years 120 (100 – 140) Notes:
Adult > 40 years Physiologically decreased approx. 1% every year.
1. Specimen: serum, plasma, or diluted urines 1+19 in distilled water.
2. Reading interval is the main determinant for the specificity of the
Each laboratory should establish its own normal ranges for the Jaffe reaction; some interferents act quickly (acetoacetate) and
population that it serves. others slowly (proteins). The majority of kinetic methods recommend
a reading interval between 30 and 150 seconds.
PERFORMANCES (PROCEDURE N°1) 3. Specific procedures are available upon request for automated
instruments. Please contact BIOLABO technical support.
Low Medium High Between run Low Medium High
Within run 4. Perform this test at 37°C to optimise the sensitivity.
n = 20 level level level n = 20 level level level
Mean 54,4 117 323 Mean 69,7 96,4 409 CALCULATION (6)
µmol/L µmol/L
Calculate the result as follows:
S.D. 2,12 1,41 2,65 S.D. 2,04 5,75 11,1
(A2 - A1) Assay
µmol/L µmol/L
Serum or plasma: Result = x Standard Concentration
(A2 - A1) Standard
C.V. % 3.9 1,2 0,8 C.V. % 2,9 5,9 2,7
Urines diluted with 1+19: Multiply the above result by dilution factor 20.
Detection limit: approximately 18 µmol/L at 37°C.
GFR (by creatinine clearance determination):
Sensitivity for 1 mg/dL: approximately 18 mAbs/min at 37°C.
Comparison study with commercially available reagent Using 24 h urine and serum creatinine

Corrected Creatinine Clearance (mL/min) = UCr x V x 1.73

(Jaffé Kinetic Method):
SCr x BSA
60 sera within 44.2 and 884 µmol/L have been evaluated with both
reagents: y = 1.06 x – 5.4 r = 0.9981 UCr = Urine Creatinine in mg/dL or µmol/L
SCr = Serum Creatinine in mg/dL or µmol/L
Units (µmol/L) Y calculated Observed Acceptable V= Urine volume excreted in mL/min (24 h urine volume/1440)
value Inaccuracy Inaccuracy BSA = Body Surface Area in m2
50,4 48,7 -1,7 7,97 OR
139,8 143,4 3,8 14,2
Using only serum creatinine (by Cockcroft and Gault formula)
593 624,8 31,8 47,8 140 – age in years x 2.12 x weight in Kg x K
Creatinine Clearance =
Serum Creatinine (µmol/L) x BSA (m2)
Units (µmol/L) Reference BIOLABO Difference
Mean n=60 104,8 106,3 +1,5 K = 1.00 for men or K = 0.85 for women
Standard Deviation 79,1 83,9 +4,8
REFERENCES
(1) TIETZ N.W. Text book of clinical chemistry, 3rd Ed. C.A. Burtis, E.R.
LINEARITY Ashwood, W.B. Saunders (1999) p. 1241-1245.
th
The assay is linear up to 1327 µmol/L (15 mg/dL). Above, dilute the (2) Clinical Guide to Laboratory Test, 4 Ed., N.W. TIETZ (2006) p. 316-321
th
specimen (1/4) with saline solution and reassay taking into account the (3) YOUNG D.S., Effect of Drugs on Clinical laboratory Tests, 4 Ed. (1995)
dilution factor. Linearity limit depends on specimen/reagent ratio. p.3-190 to 3-211
(4) Fabiny D. L., et Ertingshausen G., Clin. Chem. ( 1971), 17, p.696-700.
(5) D. Labbé et al., Ann. Biol. Clin. (1996), 54, p. 285 – 298
(6) SRM: Standard Reference Material ®

Made in France Latest revision : www.biolabo.fr Revision : 26/07/2011