AU/DxC AU CREA

Instructions For Use Creatinine (Enzymatic)

© 2022 Beckman Coulter, Inc. All rights reserved.
OSR61204 4 x 45 mL R1, 4 x 15 mL R2

For in vitro diagnostic use only.

PRINCIPLE
INTENDED USE

Enzymatic assay for the quantitative determination of creatinine in human serum, plasma and urine on Beckman Coulter
AU analysers.

SUMMARY AND EXPLANATION

Reference1,2,3
Creatinine is a metabolic product of creatine and phosphocreatine, which are both found almost exclusively in muscle.
Thus, creatinine production is proportional to muscle mass and varies little from day to day.
Measurements of creatinine are used in the diagnosis and treatment of renal disease and prove useful in the evaluation
of kidney glomerular function and in monitoring renal dialysis. However, the serum level is not sensitive to early renal
damage and responds more slowly than blood urea nitrogen (BUN) to haemodialysis during treatment of renal failure.
Both serum creatinine and BUN are used to differentiate prerenal and postrenal (obstructive) azotemia. An increase
in serum BUN without concomitant increase of serum creatinine is key to identifying prerenal azotemia. In post renal
conditions where obstruction to the flow of urine is present e.g. malignancy, nephrolithiasis and prostatism, both the
plasma creatinine and urea levels will be increased; in these situations the rise is disproportionately greater for BUN due
to the increased back diffusion of urea.
Serum creatinine varies with the subject’s age, body weight, race and sex. It is sometimes low in subjects with relatively
small muscle mass, cachectic patients, amputees, and in older persons. A serum creatinine level that would usually be
considered normal does not rule out the presence of impaired renal function.

METHODOLOGY

Creatinine is hydrolysed by creatininase to creatine. The creatine formed is hydrolysed by creatinase to sarcosine
and urea. Sarcosine oxidase catalyzes the oxidative demethylation of the sarcosine to yield glycine, formaldehyde and
hydrogen peroxide. In the presence of peroxidase (POD), the hydrogen peroxide formed reacts by quantitative oxidation
condensation with N-(3-sulfopropyl)-3-methoxy-5-methylaniline (HMMPS) and 4-aminoantipyrine to yield a blue pigment.
The creatinine concentration is proportional to the change in absorbance at 600/700 nm.

CHEMICAL REACTION SCHEME

Creatininase
Creatinine + H2O Creatine
Creatinase
Creatine + H2O Sarcosine + Urea
Sarcosine Oxidase

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 Sarcosine + H2O + O2 Glycine + Formaldehyde + H2O2
POD
H2O2 + 4-aminoantipyrine + HMMPS Blue Pigment

SPECIMEN
TYPE OF SPECIMEN

Serum and heparinised plasma.

Stable in serum and plasma for 7 days when stored at 2…25°C.4
Urine: Collect urine without using preservatives. Store at 2…8°C. 5

REAGENTS
WARNING AND PRECAUTIONS

Exercise the normal precautions required for handling all laboratory reagents.
Dispose of all waste material in accordance with local guidelines.

REACTIVE INGREDIENTS

Contents, Reagent Composition in the Test

Good's Buffer 50 mmol/L

Creatinase 56.3 IU/mL
Sarcosine oxidase 15 IU/mL
HMMPS 0.68 mmol/L
Creatininase 100 IU/mL
Peroxidase 12.5 U/mL
4-Aminoantipyrine 1.53 mmol/L
Preservative
The concentrations of the reactive components of the reagents shown on the kit label are the actual concentrations in
the individual R1/R2 vials. The reagent composition which is shown in the Instructions For Use is the final concentration
of these components in the reaction cuvette after addition of R1, Sample, and R2.

CAUTION
Sodium azide preservative may form explosive compounds in metal drain lines.
See NIOSH Bulletin: Explosive Azide Hazard (8/16/76).
To avoid the possible build-up of azide compounds, flush wastepipes with
water after the disposal of undiluted reagent. Sodium azide disposal must be in
accordance with appropriate local regulations.

GHS HAZARD CLASSIFICATION

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 Creatinine (Enzymatic) R1 DANGER

H360 May damage fertility or the unborn child.

P201 Obtain special instructions before use.
P280 Wear protective gloves, protective clothing and eye/face
protection.
P308+P313 IF exposed or concerned: Get medical advice/attention.
Boric Acid 0.1 - 1%

Safety Data Sheet is available at beckmancoulter.com/techdocs

REAGENT PREPARATION

The reagents are ready for use and can be placed directly on board the instrument.

STORAGE AND STABILITY

The reagents are stable, unopened, up to the stated expiry date when stored at 2…8°C. Once open, reagents stored on
board the instrument are stable for 60 days.

CALIBRATION
CALIBRATOR REQUIRED

Use System Calibrator Cat. No. 66300 for serum and plasma application and Urine Calibrator Cat. No. B64606 for
urine application.
The serum calibrator creatinine value is traceable to the Isotope Dilution Mass Spectroscopy (IDMS) method via National
Institute of Standards and Technology (NIST) Standard Reference Material (SRM) 967.
The urine calibrator B64606 creatinine value is traceable to NIST SRM 3667.
Recalibrate the serum and plasma application every 14 days and the urine application every 30 days, or when the
following occur:
Change in reagent lot or significant shift in control values;
Major preventative maintenance was performed on the analyser or a critical part was replaced.

QUALITY CONTROL
Controls Cat. No. ODC0003 and ODC0004 or other control materials with values determined by this Beckman Coulter
system may be used for the serum/plasma application.
Biorad Liquichek Urine Chemistry Controls Cat. No. 397 and 398 or other control materials with values determined by
this Beckman Coulter system may be used for the urine application.
Each laboratory should establish its own control frequency however good laboratory practice suggests that controls be
tested each day patient samples are tested and each time calibration/blanking is performed.

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The results obtained by any individual laboratory may vary from the given mean value. It is therefore recommended that
each laboratory generates analyte specific control target values and intervals based on multiple runs according to their
requirements. These target values should fall within the corresponding acceptable ranges given in the relevant product
literature.
If any trends or sudden shifts in values are detected, review all operating parameters.
Each laboratory should establish guidelines for corrective action to be taken if controls do not recover within the specified
limits.

TESTING PROCEDURE(S)
Refer to the appropriate Beckman Coulter AU analyser User Guide/Instructions For Use (IFU) for analyser-specific assay
instructions for the sample type as listed in the Intended Use statement.

CALCULATIONS
The Beckman Coulter analysers automatically compute the creatinine concentration of each sample.

REPORTING RESULTS
REFERENCE INTERVALS

Serum/Plasma 6
Male 64 – 104 µmol/L (0.72 – 1.18 mg/dL)
Female 49 –90 µmol/L (0.55 – 1.02 mg/dL)
Neonate 22 – 90 µmol/L (0.26 – 1.01 mg/dL)
Infant (2 months – < 3 years) 11 – 34 µmol/L (0.15 – 0.37 mg/dL)
Child (3 – < 15 years) 21 – 65 µmol/L (0.24 – 0.73 mg/dL)
Urine 7
Male 124 – 230 µmol/kg/day (14 – 26 mg/kg/day)
Female 97 – 177 µmol/kg/day (11 – 20 mg/kg/day)
Expected values may vary with age, sex, sample type, diet and geographical location. Each laboratory should verify
the transferability of the expected values to its own population, and if necessary determine its own reference interval
according to good laboratory practice. For diagnostic purposes, results should always be assessed in conjunction with
the patient's medical history, clinical examinations and other findings.
Data contained within this section is representative of performance on Beckman Coulter systems. Data obtained in your
laboratory may differ from these values.

PROCEDURAL NOTES
INTERFERENCES

Results of serum studies conducted to evaluate the susceptibility of the method to interference were as follows:

Lipemia: Interference less than 10% or 14 µmol/L up to 1,000 mg/dL Intralipid.

Icterus: Interference less than 7% or 9.9 µmol/L up to 40 mg/dL or 684 µmol/L bilirubin
Haemolysis: Interference less than 5% or 7 µmol/L up to 5 g/L haemoglobin

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 Ascorbate : Interference less than 5% or 7 µmol/L up to 20 mg/dL ascorbate
Creatine: Interference less than 5% or 7 µmol/L up to 30 mg/dL creatine
Results of urine studies conducted to evaluate the susceptibility of the method to interference were as follows:

Icterus: Interference less than 5% or 354 µmol/L up to 50 mg/dL conjugated bilirubin

Ascorbate : Interference less than 5% or 354 µmol/L up to 20 mg/dL ascorbate
Glucose: Interference less than 5% or 354 µmol/L up to 3,000 mg/dL glucose
Patients treated with N-Acetyl Cysteine (NAC) for a Paracetamol overdose may generate a false low result for creatinine.
Venipuncture immediately after or during the administration of Metamizole (Dipyrone) may lead to falsely low results for
Enzymatic Creatinine. Venipuncture should be performed prior to the administration of Metamizole.
N-acetyl-p-benzoquinone imine (metabolite of Paracetamol) will generate erroneously low results in samples for patients
that have taken toxic doses of paracetamol.
In very rare cases gammopathy, especially monoclonal IgM (Waldenström’s macroglobulinemia), may cause unreliable
results.
Refer to Young 8 for further information on interfering substances.

PERFORMANCE CHARACTERISTICS
LINEARITY

The test is linear within a concentration range of 4.4 – 4,420 µmol/L (0.05 – 50.0 mg/dL) for serum and plasma.
The test is linear within a concentration range of 88 – 44,200 µmol/L (1 – 500 mg/dL) for urine.

SENSITIVITY

The limit of detection for creatinine using serum settings on an AU640 analyser was established at 0.88 µmol/L; limit of
blank = 0.11 µmol/L.
The limit of detection for creatinine using urine settings on an AU640 analyser was established at 13.9 µmol/L; limit of
blank = 3.1 µmol/L.
The limit of detection was determined consistent with the guidelines in the NCCLS protocol EP17-A 9 with proportions
of false positives less than 5% and false negatives less than 5%; based on 150 determinations, with 60 blank and 90
low-level samples.

METHODS COMPARISON

Patient serum samples were used to compare this Creatinine (Enzymatic) OSR61204 assay on the AU2700 against
another commercially available enzymatic creatinine assay. Results of linear regression analysis were as follows:

y = 1.014x + 1.768 r = 1.000 n = 237 Sample range = 12.4 – 1,966 µmol/L

Patient urine samples were used to compare this Creatinine (Enzymatic) OSR61204 assay on the AU2700 against
another commercially available IDMS traceable creatinine assay. Results of linear regression analysis were as follows:

y = 0.986x - 104.577 r = 0.997 n = 151 Sample range = 707 – 19,793 µmol/L

PRECISION

The following data was obtained on an AU2700 using 3 serum pools analysed over 20 days.

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 n = 80 Within-run Total
Mean µmol/L SD CV% SD CV%
62.1 0.7 1.2 1.4 2.3
180.2 1.2 0.6 2.9 1.6
908.3 6.7 0.7 14.0 1.5
The following data was obtained on an AU2700 using 3 urine pools analysed over 20 days.

n = 80 Within-run Total
Mean µmol/L SD CV% SD CV%
2335 37 1.6 51 2.2
8601 111 1.3 189 2.2
15146 159 1.1 331 2.2

ADDITIONAL INFORMATION
DxC 700 AU requires that each reagent application has a standard format of abbreviated Closed Test Name. This Closed
Test Name is required to allow automated loading of the calibrator information for each application as part of the DxC
700 AU Closed System. Refer to the table below for the Closed Test Name assigned to each application for this assay.

Test Name Description

CRE3N Creatinine (Serum)
CRE3N Creatinine (Urine)

Setting Sheet Footnotes

# User defined
† System Calibrator Cat. No.: 66300
† Urine Calibrator Cat. No: B64606. Ensure relevant value sheet is used.
*Values set for working in SI units (µmol/L). To work in mg/dL divide by 88.4.

REVISION HISTORY

Remove CE mark.

Preceding version revision history

Revised Interferences section.

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REFERENCES
1. Newman DJ, Price CP. Renal function and nitrogen metabolites. In: Burtis CA, Ashwood ER, eds. Tietz textbook
of clinical chemistry. Philadelphia: WB Saunders Company, 1999;1241-1246.

2. Mayne PD, ed. Clinical chemistry in diagnosis and treatment, 6th ed. London: Arnold,1994:18pp.

3. Thomas L. Creatinine. In: Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical laboratory
results. Frankfurt/Main: TH-Books Verlagsgesellschaft, 1998:366-371.

4. Ehret W, Heil W, Schmitt Y, Töpfer G, Wisser H, Zawta B, et al. Use of Anticoagulants in Diagnostic Laboratory
Investigations and Stability of Blood, Plasma and Serum Samples. WHO/DIL/LAB/99.1 Rev.2:28pp.

5. NCCLS. Urinalysis and collection, transportation, and preservation of urine specimens; approved guideline.
NCCLS Document GP16-A2, 2nd ed. Pennsylvania: NCCLS, 2001.

6. Ceriotti F, Boyd JC, Klein G, Henny J, Queraltó J, Kairisto V, Panteghini M. Reference intervals for serum creatinine
concentrations: Assessment of available data for global application. Clin Chem. 2008; 54(3): 559-566.

7. Painter PC, Cope JY, Smith JL. Reference information for the clinical laboratory. In: Burtis CA, Ashwood ER, eds.
Tietz textbook of clinical chemistry. Philadelphia:WB Saunders Company, 1999;1809pp.

8. Young DS. Effects of drugs on clinical laboratory tests, 5thed. AACC Press, 2000.

9. NCCLS. Protocols for determination of limits of detection and limits of quantitation; approved guideline. NCCLS
Document EP17-A. Pennsylvania: NCCLS, 2004.

Beckman Coulter Ireland Inc., Lismeehan, O’Callaghan’s Mills, Co. Clare, Ireland +(353) (0) 65 683 1100
www.beckmancoulter.com

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