2014/08/20

Clinical Chemistry A93A01295BEN

A11A01642

29 mL
ABX Pentra
Urinary Proteins CP

■ Pentra C400

Diagnostic reagent for quantitative in vitro determination of Total Proteins in

urine by colorimetry.

Application Release nm. When this complex is combined with protein in acidic
conditions, a blue-purple color develops with an increase
in absorption at 598 nm (3).
Urine: TPU The color is directly proportional to the protein
1.xx concentration.

Intended Use Reagents

ABX Pentra Urinary Proteins CP reagent is intended for
the quantitative in vitro diagnostic determination of urinary ABX Pentra Urinary Proteins CP is ready-to-use.
proteins in urine.
Reagent:
Identification of urinary protein is used in the diagnosis
and treatment of disease conditions such as renal or Pyrogallol red 60 µmol/L
heart diseases or thyroid disorders, which are Sodium molibdate 40 µmol/L
characterized by proteinuria or albuminuria. Detergents

ABX Pentra Urinary Proteins CP should be used

Clinical Interest (1, 2)
according to this reagent notice. The manufacturer
Elevated concentration of total protein in urine cannot guarantee its performance if used otherwise.
(proteinuria) can be detected in the majority of kidney
diseases. Primary and secondary nephropathies may
cause increased glomerular permeability or decreased Handling
tubular reabsorption. Post-renal causes of proteinuria are
infections, bleedings or malignant diseases of the urinary 1. Remove the cap of the cassette.
tract. Elevated urine protein levels can also be related to
2. If present, remove foam by using a plastic pipette.
other acute disorders like fever.
3. Position the protective cap ref. GBM0969 on the
cassette.
Method 4. Place the cassette into the refrigerated Pentra C400
reagent compartment.
The total protein test for urine is based on the procedure
developed by Watanabe et al. (3) which is a dye-binding
colorimetric method utilizing pyrogallol red-molybdate
complex. This photometric test which provides good Calibrator
QUAL-QA-TEMP-0846 Rev.9

precision and linearity, has been modified to equalize the

reactivity of albumin and gamma-globulin (4). For calibration, use:
The pyrogallol red is combined with molybdenum acid, ABX Pentra TPU Cal, Ref. A11A01898 (not included)
forming a red complex with maximum absorbance at 467 3 x 3 mL

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 Clinical Chemistry

ABX Pentra
Urinary Proteins CP

Control Do not freeze the reagents.

For internal quality control, use:

ABX Pentra Urine Control L/H, Ref. A11A01674 (not Waste Management
included)
1 x 10 mL + 1 x 10 mL Please refer to local legal requirements.
Each control should be assayed daily and/or after a
calibration.
The frequency of controls and the confidence intervals General Precautions a
should correspond to laboratory guidelines and country-
specific directives. You should follow federal, state and ■ This reagent is for professional in vitro diagnostic use
local guidelines for testing quality control materials. The only.
results must be within the range of the defined confidence ■ For prescription use only.
limits. Each laboratory should establish a procedure to ■ Do not swallow. Avoid contact with skin and mucous
follow if the results exceed these confidence limits. membranes.
■ Observe the standard laboratory precautions for use.
■ The reagent cassettes are disposable and should be
Materials Required but not Provided disposed of in accordance with the local legal
requirements.
■ Automated clinical chemistry analyzer: Pentra C400 ■ Please refer to the MSDS associated with the reagent.
■ Calibrator: ABX Pentra TPU Cal, Ref. A11A01898 ■ Do not use the product if there is visible evidence of
■ Control: ABX Pentra Urine Control L/H, Ref. biological, chemical or physical deterioration.
A11A01674 ■ It is the user's responsibility to verify that this
■ Standard laboratory equipment. document is applicable to the reagent used.

Specimen Performance on Pentra C400

■ Urine. The performance data listed below are representative of

performance on HORIBA Medical Systems.
Stability (5): 1 day at 20-25°C
7 days at 4-8°C Number of Tests: 100 tests b
1 month at -20°C
On Board Reagent Stability:
Once opened, the reagent cassette placed in the
Reference Range (6) refrigerated Pentra C400 compartment is stable for 30
days.
Each laboratory should establish its own reference
ranges. The values given here are used as guidelines only. Sample Volume: 5.0 µL/test

Urine (Excretion): Detection Limit:

Adult: < 100 mg/day (< 0.10 g/day)
Pregnancy: < 150 mg/day (< 0.15 g/day) The detection limit is determined according to the Valtec
protocol (7) and equals 0.027 g/L (2.7 mg/dL).

Storage and Stability

Reagents, in unopened cassettes, are stable up to the

expiry date on the label if stored between 2-8°C.

Stability after opening: refer to the paragraph

"Performance on Pentra C400".

aModification: general precautions modification.

bModification: recommendation removed.

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 Clinical Chemistry

ABX Pentra
Urinary Proteins CP

Accuracy and Precision: Interferences:

■ Repeatability (within-run precision) Haemoglobin: Do not use hemolysed samples.

Direct Bilirubin: No significant influence is observed up
3 specimens of low, medium and high concentration and to 80 µmol/L (4.8 mg/dL).
2 controls are tested 20 times according to the Other limitations are given by Young as a list of drugs and
recommendations found in the Valtec protocol (7). preanalytical variables known to affect this methodology
(12, 13).
Mean value CV %
Calibration Stability:
g/L mg/dL
Control specimen 1 0.19 19 2.50 The reagent is calibrated on Day 0. The calibration
stability is checked by testing 2 control specimens.
Control specimen 2 0.37 37 1.24
The calibration stability is 21 days.
Specimen 1 0.19 19 2.67 Note: A recalibration is recommended when reagent lots
Specimen 2 0.59 59 0.87 change, and when quality control results fall outside the
Specimen 3 1.16 116 0.44
range established.

Application release:
■ Reproducibility (total precision)
1.xx
2 specimens (low and high levels) and 2 controls are
tested in duplicate for 20 days (2 series per day) Conversion factor:
according to the recommendations found in the CLSI
(NCCLS), EP5-A protocol (8). g/L x 100.0 = mg/dL

Mean value CV %
Reference
g/L mg/dL
1. Johnson AM, Rohlfs EM, Silverman LM. Proteins. In:
Control specimen 1 0.18 18 8.77
Burtis CA, Ashwood ER, editors. Tietz Textbook of
Control specimen 2 0.36 36 2.86 Clinical Chemistry. 3rd ed. Philadelphia: WB
Specimen 1 0.24 24 4.25 Saunders Company (1999): 477-540.
Specimen 2 1.28 128 2.61 2. Felgenhauer K. Laboratory diagnosis of neurological
diseases. In: Thomas L. Clinical Laboratory
Measuring Range: Diagnostics. 1st ed. Frankfurt: THBooks
Verlagsgesellschaft (1998): 1308-26.
The assay confirmed a measuring range from 0.03 to 3. Watanabe N, Kamei S, Ohkubo A, Yamanaka M,
2.70 g/L (3 to 270 mg/dL), with an automatic post-dilution Ohsawa S, Makino K, Tokuda K. Urinary protein as
up to 5.4 g/L (540 mg/dL). measured with a pyrogallol redmolybdate complex.
The reagent linearity has been assessed up to 2.70 g/L Manually and in a Hitachi 726 automated analyzer.
(270 mg/dL) according to the recommendations found in Clin. Chem. (1986) 32 (8): 1551-4.
the CLSI (NCCLS), EP6-A protocol (9). 4. Orsonneau JL, Douet P, Massoubre C, Lustenberger
P, Bernard S. An improved pyrogallol red-molybdate
Correlation: method for determining total urinary protein. Clin.
Chem. (1989) 35: 2233-6.
115 patient samples (urine) are correlated with a
5. Guder WG, Zawta B. The Quality of Diagnostics
commercial reagent taken as reference according to the
Samples. Samples: From the Patient to the
recommendations found in the CLSI (NCCLS), EP9-A2
protocol (10). Values ranged from 0.04 to 2.64 g/L (4 to Laboratory. 1st Ed. Guder WG, Narayanan S, Zawta
264 mg/dL). B. (WHILEY-VCH, Darmstadt, Germany), (2001):
The equation for the allometric line obtained using 52-53.
Passing-Bablock regression procedure (11) is: 6. Roberts WL, McMillin GA, Burtis CA, Bruns DE.
Y = 1.05 X - 0.03 (g/L) Reference Information for the Clinical Laboratory,
Y = 1.05 X - 2.73 (mg/dL) TIETZ Textbook of Clinical Chemistry and Molecular
with a correlation coefficient r2 = 0.9886. Diagnostics. 4th Ed., Burtis CA, Ashwood ER, Bruns
DE, (Elsevier Saunders eds., St Louis, USA), (2006):
2293.

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 Clinical Chemistry

ABX Pentra
Urinary Proteins CP

7. Vassault A, Grafmeyer D, Naudin C et al. Protocole

de validation de techniques (document B). Ann. Biol.
Clin. (1986) 44: 686-745.
8. Evaluation of Precision Performance of Clinical
Chemistry Devices. Approved Guideline, CLSI
(NCCLS) document EP5-A (1999) 19 (2).
9. Evaluation of the Linearity of Quantitative Analytical
Methods. Approved Guideline, CLSI (NCCLS)
document EP6-A (2003) 23 (16).
10. Method Comparison and Bias Estimation Using
Patient Samples. Approved Guideline, 2nd ed., CLSI
(NCCLS) document EP9-A2 (2002) 22 (19).
11. Passing H, Bablock W. A new biometrical procedure
for testing the equality of measurements from two
different analytical methods. J. Clin. Chem. Clin.
Biochem. (1983) 21: 709-20.
12. Young DS. Effects of Drugs on Clinical Laboratory
Tests. 4th Edition, Washington, DC, AACC Press
(1997) 3: 143-163.
13. Young DS. Effects of Preanalytical Variables on
Clinical Laboratory Tests. 2nd Edition, Washington,
DC, AACC Press (1997) 3: 120-132.

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