REAGENT COMPOSITION no interference up to:

Ascorbic Acid 10 mM
COMPONENTS CONCENTRATION Bilirubin 40 mg/dL
Liquid Reagents – ready to use Reagent1: (R1) Bilirubin, conjugated 30 mg/dL
Good’s Buffer, pH 7-8 Hemoglobin 500 mg/dL
Creatinase 12 – 60 KU/L Triglycerides 1000 mg/dL
CREATININE Sarcosine oxidase (SOD) 4 – 17 KU/L
Enzymatic, PAP TOOS 0.07 – 0.21 g/L
2 Reagents Ascorbate oxidase
Reagent 2: (R2) MANUAL TEST PROCEDURE
Good’s Buffer, pH 7-8
Bring reagents and samples to room temperature.
Creatininase 135 – 670 KU/L
Diagnostic reagent for quantitative in vitro Peroxidase Substrate Start
determination of creatinine in human serum or urine 4-Aminoantipyrine (4-AA) 0.3 – 0.9 g/L
on photometric systems Sodium azide < 0.1 % Pipette into test tubes Blank Std/Cal. Sample
Reagent 1 900 µL 900 µL 900 µL
Sample - - 20 µL
REAGENT PREPARATION
Std./Cal. - 20 µL -
Substrate Start Dist. water 20 µL - -
D06430 4 x 50 ml 4 x 37.5 ml Reagent 1 The reagents are ready to use.
1 x 50 ml Reagent 2 Mix. Incubate 5 min. at 37°C and read absorbance A1
Sample Start: at 550 nm against the blank. Then add:
D06450 4 x 10 ml 4 x 7.5 ml Reagent 1 Not possible (elimination of endogenous creatine).
Reagent 2 300 µL 300 µL 300 µL
1 x 10 ml Reagent 2
Mix. Incubate 5 min. at 37°C and read absorbance A2
Additionally offered: REAGENT STABILITY AND STORAGE at 550 nm against the blank.
D94592 1 x 3 ml Creatinine Standard Conditions: Reagents are light-sensitive.
Calc.: ΔA = [(A2 – 0.754 A1) sample or standard]
D98485 5 x 3 ml Calibrator Diacal Auto → Protect from light!
D98481 12 x 5 ml Control normal Diacon N Close immediately after use.
D98482 12 x 5 ml Control abnormal Diacon P Do not freeze the reagents!
D08581 12 x 5 ml Diacon Urine Level 1 Storage: at 2 – 8°C CALCULATION (light path 1 cm)
D08582 12 x 5 ml Diacon Urine Level 2 Stability (unopened): up to the expiration date
Serum:
On board stability: ΔA Sample
Creatinine (mg/dL) = x Conc. Std/Cal (mg/dL)
30 days ΔA Std/Cal
TEST PARAMETERS Urine:
ΔA Sample
Method: Colorimetric, enzymatic, endpoint, SAMPLE PREPARATION Creatinine (mg/dL) = x Conc. Std/Cal (mg/dL) x 10
ΔA Std/Cal
increasing reaction
Urine: Dilute urine 1 + 9 with dist. water.
Wavelength: 550 nm
Temperature: 37°C UNIT CONVERSION
STANDARD
Sample: Serum, urine (has to be ordered separately) mg/dL x 88.4 = µmol/L
Linearity: up to 30 mg/dL Concentration 2 mg/dL (177 μmol/L)
Storage: 2 – 25°C
Sensitivity: The lower limit of detection is 0.14 mg/dL Stability: up to the expiration date
CLOSE IMMEDIATELY AFTER USE!
INTERFERING SUBSTANCES

S:\pm\allg\inserts\inserts_word\clinical chemistry\creatinine_enz-PAP_rev04.doc Page 1 of 2 DI M. Wagner Rev. 04, 2009.04.15.

REFERENCE RANGE [6] * Within run precision Mean SD CV serum samples with a serum based calibrator.
n = 21 [mg/dL] [mg/dL] [%]
Serum:
Sample 1 29.09 0.10 0.36 AUTOMATION
Females 0.51 – 0.95 mg/dl
Sample 2 87.1 0.27 0.31
Males 0.67 – 1.17 mg/dl Sample 3 196.7 0.90 0.46 Special adaptations for automated analyzers can be made on
First morning urine: request.
Females 29 – 226 mg/dL Total precision Mean SD CV
n = 20 [mg/dL] [mg/dL] [%]
Males 40 – 278 mg/dL
Sample 1 29.9 0.79 2.64 WARNINGS AND PRECAUTIONS
* These values are for orientation purpose. Each laboratory Sample 2 87.7 0.67 0.76
Sample 3 195 1.19 0.60 1. The reagent contains sodium azide (< 0.95 g/l) as
should establish its own reference range.
preservative which may react with lead and copper plumbing
LINEARITY to form highly explosive metal azide. On disposal, flush with a
TEST PRINCIPLE large volume of water to prevent azide buildup. Do not
The test has been developed to determine creatinine
swallow! Avoid contact with skin and mucous membranes.
concentrations within a measuring range from 0.14 – 30 mg/dL.
The enzymatic assay for creatinine involves a series of coupled 2. Take the necessary precautions for the use of laboratory
If values exceed this range samples should be diluted with NaCl
enzymatic reactions including creatininase enzymatic conversion of reagents.
solution (9 g/L sodium chloride in dest. water) and the result
creatinine into the product creatine which itself is converted to
multiplied by the dilution factor.
sarcosine by creatinase, followed by oxidation of sarcosine by
sarcosine oxidase (SOD) producing hydrogen peroxide. In the
WASTE MANAGEMENT
METHOD COMPARISON
presence of peroxidase (POD) the hydrogen peroxide is quantified Please refer to local legal requirements.
at 550 nm by the formation of a colored dye.
[4] This assay (y) was compared with a legally marketed creatinine
Creatininase assay (x) using serum samples (range 0.2 to 13.51 mg/dL) and
creatinine + H2O < > creatine urine samples (range 0.14 to 141 mg/dL): REFERENCES
Creatinase
creatine + H2O < > sarcosine + urea Serum samples: y = 0.946 x + 0.0643; r= 0.9981 1. Tietz, N.W. (Ed.): Fundamentals of Clinical Chemistry, W.B. Saunders
Sarcosine oxidase Urine samples: y = 1.005 x – 0.2979 ; r = 0.9969 Co., philadelphia, 865 (1982)
sarcosine + O2 + H2O < > glycine + HCHO + H2O2
Peroxidase 2. National Kidney Foundation K/DOQI. Clinical Practice Guidelines for
2 H2O2 + 4-AA + TOOS < > quinone dye chronic kidney disease: evaluation, classification, and stratification.
The absorbance of the produced red dye (λmax 556 nm) is QUALITY CONTROL Am J Kidney Dis 2002; 39:S1-S200.
proportional to the creatinine concentration in the sample. 3. Badiou S, Dupuy AM, Descomps B, Cristolead, JP. Comparison
All control sera and urine controls with Creatinine values between the enzymatic vitros assay for creatinine determination and
Any endogenous creatine present in the sample is removed by determined by this method can be used. We recommend: three other methods adapted on the Olympus analyzer, Journal of
creatinase and sarcosine oxidase during preincubation. Clinical Laboratory Analysis 2003:17,235-240
4. Hayes AW. Principles and Methods of Toxicology, Taylor & Francis,
1028 (2001).
D98481 12 x 5 ml DIACON N Control Serum Normal
5. Cristenson RH, Johnson LJ, Gregory, LC. Appleton and Lange’s
PERFORMANCE CHARACTERISTICS D98482 12 x 5 ml DIACON P Control Serum Abnormal Outline Review Clinical Chemistry, McGraw-Hill Professional, 118
D08581 12 x 5 ml Diacon Urine Level 1 (2001)
PRECISION (at 37°C) 12 x 5 ml
D08582 Diacon Urine Level 2 6. Mazzachi BC, Peake MJ, Ehrhardt V. Reference Range and Method
Serum Testing Comparison Studies for Enzymatic and Jaffé Creatinine Assays in
Within run precision Mean SD CV plasma and Serum and Early Morning Urine. Clin Lab 2000; 46;53-55.
n = 80 [mg/dL] [mg/dL] [%] CALIBRATION
Sample 1 0.74 0.015 2.1 The assay requires the use of a Creatinine Standard or Calibrator.
Sample 2 1.38 0.015 1.1 We recommend:
Sample 3 4.04 0.029 0.7
DIALAB Produktion und Vertrieb von chemisch – technischen
Total precision Mean SD CV Produkten und Laborinstrumenten Gesellschaft m.b.H.
n = 80 [mg/dL] [mg/dL] [%] A – 2351 Wiener Neudorf, Austria
D94592 1 x 3 ml CREATININE STANDARD IZ-NÖ Süd, Hondastrasse, Objekt M55
Sample 1 0.74 0.022 3.0 D98485 5 x 3 ml DIACAL AUTO Assayed Multi Phone: ++43 (0) 2236 660910-0
Sample 2 1.38 0.026 1.9 Calibration Serum Fax: ++43 (0) 2236 660910-30 e-mail: [email protected]
Sample 3 4.04 0.058 1.4 NOTE: calibration of serum samples with an aqueous standard
Urine Testing may cause matrix related bias. It is recommended to calibrate

S:\pm\allg\inserts\inserts_word\clinical chemistry\creatinine_enz-PAP_rev04.doc Page 2 of 2 DI M. Wagner Rev. 04, 2009.04.15.