2015/09/01

Clinical Chemistry A93A01276FEN

A11A01933

27.5 mL
ABX Pentra 8 mL

Creatinine 120 CP

ABX Pentra 400

Diagnostic reagent for quantitative in vitro determination of creatinine in

serum, plasma and urine by colorimetry.

Application Release Jaffe chemistry is a kinetic procedure which does not

require deproteinization of the sample and is formulated
to reduce the interference in serum proteins.
Serum, plasma: CREA3 (not for use in the USA)
Creatinine + alkaline picrate creatininepicrate complex
3.xx
At an alkaline pH, creatinine reacts with picrate to form
Urine: CREA_U3 Janovsky complex.
The rate of increase in absorbance at 510 nm due to the
3.xx
formation of creatinine-picrate complex is directly
proportional to the creatinine concentration present in the
sample.
Intended Use
ABX Pentra Creatinine 120 CP reagent is a diagnostic
reagent for quantitative in vitro determination of
Reagents
Creatinine in human serum, plasma and urine based on a
kinetic method using alkaline picrate (Jaff method). ABX Pentra Creatinine 120 CP is ready-to-use.
Creatinine measurements are used in the diagnosis and
treatment of renal diseases, in monitoring renal dialysis, Reagent 1:
and as a calculation basis for measuring other urine Sodium hydroxide 0.25 mol/L
analytes. Surfactants

Reagent 2:
Clinical Interest
Picric acid 20.5 mmol/L
Creatinine measurements are used in the diagnosis and
treatment of renal diseases, in monitoring renal dialysis ABX Pentra Creatinine 120 CP should be used
and as a calculation basis for measuring other urine according to this reagent notice. The manufacturer
analytes. cannot guarantee its performance if used otherwise.

Method Handling

In 1886, Jaffe devoloped an assay for creatinine based 1. Remove both caps of the cassette.
upon the reaction between creatinine and sodium picrate
2. If present, remove foam by using a plastic pipette.
(1). In 1904, Folin (2) used this reaction for the quantitative
QUAL-QA-TEMP-0846 Rev.9

determination of creatinine in urine. Kinetic procedures 3. Position a protective cap ref. GBM0969 on Reagent 1
based on the observed reaction rates various substances, and on Reagent 2.
including creatinine, with alkaline picrate have been
proposed by Fabing (3) and Soldin (4). This improved

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 Clinical Chemistry

ABX Pentra
Creatinine 120 CP

4. Place the cassette into an ambient ABX Pentra 400 24h urine has to be collected without additive.
reagent compartment.
Stability:

Serum, plasma (5):

Calibrator
At 20-25C: 7 days
For calibration, use: At 4-8C: 7 days
ABX Pentra Multical, Ref. A11A01652 (not included) At -20C: 3 months
10 x 3 mL (lyophilisate)
Urine (6):

Control At 20-25C: 2 days

At 4-8C: 6 days
For internal quality control, use: At -20C: 6 months
ABX Pentra N Control, Ref. A11A01653 (not included)
10 x 5 mL (lyophilisate)
ABX Pentra P Control, Ref. A11A01654 (not included) Reference Range
10 x 5 mL (lyophilisate)
ABX Pentra Urine Control L/H, Ref. A11A01674 (not Each laboratory should establish its own reference
included) ranges. The values given here are used as guidelines only.
1 x 10 mL + 1 x 10 mL
Each control should be assayed daily and/or after a Serum/Plasma (7):
calibration.
The frequency of controls and the confidence intervals Men Women
should correspond to laboratory guidelines and country-
8 - 13 mg/L 6 - 12 mg/L
specific directives. You should follow federal, state and
local guidelines for testing quality control materials. The 0.8 - 1.3 mg/dL 0.6 - 1.2 mg/dL
results must be within the range of the defined confidence 71 - 115 mol/L 53 - 106 mol/L
limits. Each laboratory should establish a procedure to
follow if the results exceed these confidence limits. Urine (24 hours) (8):

Men Women
Materials Required but not Provided 14 - 26 mg/kg/day 11 - 20 mg/kg/day
124 - 230 mol/kg/day 97 - 177 mol/kg/day
Automated clinical chemistry analyzer: ABX Pentra 400
Calibrator: ABX Pentra Multical, Ref. A11A01652
Controls:
ABX Pentra N Control, Ref. A11A01653, and Storage and Stability
ABX Pentra P Control, Ref. A11A01654
ABX Pentra Urine Control L/H, Ref. A11A01674 Reagents, in unopened cassettes, are stable up to the
Standard laboratory equipment. expiry date on the label if stored between 18-26C.

Stability after opening: refer to the paragraph

Specimen "Performance on ABX Pentra 400".

Fresh, clear serum.

Plasma in lithium heparin or EDTA. Waste Management
Fresh centrifuged urine.
Please refer to local legal requirements.
Anticoagulants other than those listed have not been
tested by HORIBA Medical and are therefore not
recommended for use with this assay.

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 Clinical Chemistry

ABX Pentra
Creatinine 120 CP

General Precautions a Sample Volume: 10 L/test

Limit of Quantitation:
This reagent is for professional in vitro diagnostic use
only. The limit of quantitation is determined according to CLSI
For prescription use only. (NCCLS), EP17-A protocol (9) and equals 12.2 mol/L
This reagent is classified as hazardous in compliance (0.14 mg/dL).
with regulation (EC) N.1272/2008.
Reagent 1 (R1): Accuracy and Precision:
Warning
H290: May be corrosive to metals. Repeatability (within-run precision)
H315: Causes skin irritation.
H319: Cause serious eye irritation. 4 specimens of low, medium and high concentration and
P234: Keep only in original container. 2 controls are tested 20 times according to the
P264: Wash hands thoroughly after handling. recommendations found in the Valtec protocol (10).
P280: Wear protective gloves/protective clothing/eye
protection/face protection. Mean value CV %
P302 + P352: IF ON SKIN: Wash with plenty of soap
mol/L mg/dL
and water.
P305 + P351 + P338: IF IN EYES: Rinse cautiously Control 94.52 1.07 1.58
specimen 1
with water for several minutes. Remove contact lenses,
if present and easy to do. Continue rinsing. Control 333.01 3.76 0.83
P321: Specific treatment (see [***] on this label). specimen 2
P332 + P313: If skin irritation occurs: Get medical Specimen 1 49.37 0.56 2.59
advice/attention. Specimen 2 142.00 1.60 1.46
P337 + P313: If eye irritation persists: Get medical
Specimen 3 597.28 6.75 0.72
advice/attention.
P362 + P364: Take off contaminated clothing and Specimen 4 1314.55 14.85 0.59
wash it before reuse.
P390: Absorb spillage to prevent material damage. Reproducibility (total precision)
P406: Store in corrosive resistant container with a
resistant inner liner. 4 specimens of low, medium and high levels and 2
Observe the standard laboratory precautions for use. controls are tested in duplicate for 20 days (2 series per
The reagent cassettes are disposable and should be day) according to the recommendations found in the CLSI
disposed of in accordance with the local legal (NCCLS), EP5-A2 protocol (11).
requirements.
Please refer to the MSDS associated with the reagent. Mean value CV %
Do not use the product if there is visible evidence of
mol/L mg/dL
biological, chemical or physical deterioration.
It is the user's responsibility to verify that this Control 90.84 1.03 2.88
document is applicable to the reagent used. specimen 1
Control 333.02 3.76 1.84
specimen 2
Performance on ABX Pentra 400 Specimen 1 48.81 0.55 5.03

The performance data listed below have been obtained Specimen 2 125.08 1.41 2.21
on the ABX Pentra 400 analyzer. Specimen 3 575.30 6.50 2.07
Specimen 4 1299.53 14.68 1.99
Serum, plasma (not for use in the USA)
Number of Tests: 120 tests
Measuring Range:
On Board Reagent Stability: The assay confirmed a measuring range from 12.2 to
Once opened, the reagent cassette placed in the ambient 1600 mol/L (0.14 to 18.08 mg/dL), with an automatic
ABX Pentra 400 compartment is stable for 19 days. post-dilution up to 4800 mol/L (54.24 mg/dL).

aModification: modification of classification.

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 Clinical Chemistry

ABX Pentra
Creatinine 120 CP

The reagent linearity has been assessed up to On Board Reagent Stability:

1600 mol/L (18.08 mg/dL) according to the
recommendations found in the CLSI (NCCLS), EP6-A Once opened, the reagent cassette placed in the ambient
protocol (12). ABX Pentra 400 compartment is stable for 19 days.

Sample Volume: 15 L/test

Correlation:
168 patient samples (serum) are correlated with a Limit of Quantitation:
commercial reagent taken as reference according to the
The limit of quantitation is determined according to CLSI
recommendations found in the CLSI (NCCLS), EP9-A2
(NCCLS), EP17-A protocol (9) and equals 261 mol/L
protocol (13). Values ranged from 45.39 to
(2.9 mg/dL).
1542.18 mol/L (0.51 to 17.43 mg/dL).
The equation for the allometric line obtained using
Passing-Bablock regression procedure (14) is: Accuracy and Precision:
Y = 1.02 X - 6.73 (mol/L) Repeatability (within-run precision)
Y = 1.02 X - 0.08 (mg/dL)
with a correlation coefficient r2 = 0.999. 3 specimens of low, medium and high concentration and
2 controls are tested 20 times according to the
Interferences: recommendations found in the Valtec protocol (10).
Haemoglobin: No significant influence is observed up
to 261 mol/L (450 mg/dL). Mean value CV %
Triglycerides: No significant influence is observed up mol/L mg/dL
to an Intralipid concentration
Control 5305 60.0 0.53
(representative of lipemia) of specimen 1
7.0 mmol/L (612.5 mg/dL).
Control 11829 133.7 0.61
Total Bilirubin: No significant influence is observed up
specimen 2
to 57 mol/L (3.3 mg/dL).
Direct Bilirubin: No significant influence is observed up Specimen 1 993 11.2 1.28
to 50 mol/L (2.9 mg/dL). Specimen 2 8585 97.0 0.56
Glucose: No significant influence is observed up Specimen 3 21899 247.5 0.52
to 18.4 mol/L (332 mg/dL).
Ascorbic Acid: No significant influence is observed up
Reproducibility (total precision)
to 340 mol/L (5.98 mg/dL).
Total Proteins: No significant influence is observed up
to 34.2 to 149 g/L. 3 specimens of low, medium and high levels and 2
Other limitations are given by Young as a list of drugs and controls are tested in duplicate for 20 days (2 series per
preanalytical variables known to affect this methodology day) according to the recommendations found in the CLSI
(15, 16). (NCCLS), EP5-A2 protocol (11).

Calibration Stability: Mean value CV %

mol/L mg/dL
The reagent is calibrated on Day 0. The calibration
stability is checked by testing 2 control specimens. Control 5307 60.0 1.32
specimen 1
The calibration stability is 3 days.
Note: A recalibration is recommended when reagent lots Control 11784 133.2 1.50
change, and when quality control results fall outside the specimen 2
range established. Specimen 1 925 10.5 2.30
Specimen 2 8681 98.1 2.21
Application release: (not for use in the USA) Specimen 3 20380 230.3 1.59
3.xx
Measuring Range:

Urine The assay confirmed a measuring range from 261 to

25000 mol/L (2.9 to 282.5 mg/dL), with an automatic
Number of Tests: 120 tests post-dilution up to 75000 mol/L (847.5 mg/dL).

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 Clinical Chemistry

ABX Pentra
Creatinine 120 CP

The reagent linearity has been assessed up to 2. Folin O. Beitrag zur Chemie des Kreatinins und
25000 mol/L (282.5 mg/dL) according to the Kreatins im Harne. Z. Physiol. Chem. (1904) 41:
recommendations found in the CLSI (NCCLS), EP6-A 223-242.
protocol (12). 3. Fabing DL, Ertingshausen G. Automated Reaction
Rate Method for the Determination of Serum
Correlation: Creatinine with the Centrifichem. Clin. Chem. (1971)
17: 391.
109 patient samples (urine) are correlated with a 4. Soldin S, Henderson L, Hill G. The Effect of Bilirubin
commercial reagent taken as reference according to the and Ketones on Reaction Rate Methods for the
recommendations found in the CLSI (NCCLS), EP9-A2 Measurement of Creatinine. Clin. BioChem. (1978):
protocol (13). Values ranged from 284.86 to 82-86.
23919.5 mol/L (3.22 to 270.29 mg/dL). 5. Use of anticoagulants in diagnostic laboratory
The equation for the allometric line obtained using investigations. WHO publication WHO/DIL/LAB/99.1
Passing-Bablock regression procedure (14) is: Rev. 2 (2002): 28.
Y = 1.04 X - 38.87 (mol/L) 6. Use of anticoagulants in diagnostic laboratory
Y = 1.04 X - 0.44 (mg/dL) investigations. WHO publication WHO/DIL/LAB/99.1
with a correlation coefficient r2 = 0.9987. Rev. 2 (2002): 46.
7. Tietz NW. Clinical guide to laboratory tests, 3rd Ed,
Interferences: (WB. Saunders eds. Philadelphia USA), (1995): 186.
8. Roberts WL, McMillin GA, Burtis CA, Bruns DE.
Haemoglobin: No significant influence is observed up Reference Information for the Clinical Laboratory,
to 210 mol/L (362 mg/dL). TIETZ Textbook of Clinical Chemistry and Molecular
Triglycerides: No significant influence is observed up Diagnostics. 4th Ed; Burtis CA, Ashwood ER, Bruns
to an Intralipid concentration DE, (Elsevier Saunders eds. St Louis, USA), (2006):
(representative of lipemia) of 2264.
612.5 mg/dL. 9. Protocols for determination of limits of detection and
Direct Bilirubin: No significant influence is observed up limits of quantitation. Approved Guideline, CLSI
to 500 mol/L (29.3 mg/dL). (NCCLS) document EP17-A (2004) 24 (34).
Ascorbic Acid: No significant influence is observed up 10. Vassault A, Grafmeyer D, Naudin C et al. Protocole
to 340 mol/L (5.99 mg/dL). de validation de techniques (document B). Ann. Biol.
Other limitations are given by Young as a list of drugs and Clin. (1986) 44: 686-745.
preanalytical variables known to affect this methodology 11. Evaluation of Precision Performance of Quantitative
(15, 16). Measurement Method. Approved Guideline, CLSI
(NCCLS) document EP5-A2 (2004) 24 (25).
Calibration Stability: 12. Evaluation of the Linearity of Quantitative Analytical
Methods. Approved Guideline, CLSI (NCCLS)
The reagent is calibrated on Day 0. The calibration document EP6-A (2003) 23 (16).
stability is checked by testing 2 control specimens. 13. Method Comparison and Bias Estimation Using
The calibration stability is 3 days. Patient Samples. Approved Guideline, 2nd ed., CLSI
Note: A recalibration is recommended when reagent lots (NCCLS) document EP9-A2 (2002) 22 (19).
change, and when quality control results fall outside the 14. Passing H, Bablock W. A new biometrical procedure
range established. for testing the equality of measurements from two
different analytical methods. J. Clin. Chem. Clin.
Application release: Biochem. (1983) 21: 709-20.
15. Young DS. Effects of Drugs on Clinical Laboratory
3.xx
Tests. 4th Edition, Washington, DC, AACC Press
(1997) 3: 143-163.
Conversion factor: 16. Young DS. Effects of Preanalytical Variables on
mol/L x 0.0113 = mg/dL Clinical Laboratory Tests. 2nd Edition, Washington,
DC, AACC Press (1997) 3: 120-132.

Reference
1. Jaffe M. Hoppe Selyer's. Z. Physiol. Chem. (1886) 10:
391-400.

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