BIOLABO
www.biolabo.fr Inorganic PHOSPHORUS U . V . M e t h o d
MANUFACTURER: Reagent for quantitative determination of inorganic phosphate
BIOLABO S.A.S in human plasma and serum or urines
Les Hautes Rives REF 80015 R1 2 x 125 mL R2 1 x 5 mL
02160, Maizy, France
TECHNICAL SUPPORT AND ORDERS
Tel: (33) 03 23 25 15 50
| IVD
Made In France
support@biolabo.fr
Latest revision : www.biolabo.fr I: corresponds to significant modifications
INTENDED USE I SAFETY CAUTIONS
This reagent is designated for professional use in laboratory (manual or · Refer to current Material Safety Data Sheet available on request or on
automated method). www.biolabo.fr
It allows the quantification of phosphorus in human serum and plasma, · Verify the integrity of the contents before use.
or urines to assess phosphorous homeostasis. · Waste disposal: Respect legislation in force in the country.
· All specimens or reagents of biological origin should be handled as
GENERALITIES (1) (2) potentially infectious. Respect legislation in force in the country.
An adult human body contains approximately 600 g of phosphates Any serious incident that has occurred in connection with the device is
expressed as phosphorus, of which about 85% is bound to calcium in notified to the manufacturer and the competent authority of the Member
bones and the rest principally in other tissue cells. Most phosphate State in which the user and/or patient is based.
present in cells is organic and incorporated into phospholipids, nucleic
acids, and high energy compounds. Serum/plasma contains
approximately 1% of total phosphate as inorganic phosphate, the REAGENT PREPARATION
fraction measured in routine biochemical analysis. Ready for use
Elevation of phosphorus in serum/plasma is often associated with bone
diseases, renal failures, hypoparathyroidism, hypervitaminosis D. STABILITY AND STORAGE
Decreased serum/plasma phosphorus concentrations are found in Stored away from light, well caped in the original vial at 18-25°C,
osteomalacia, vitamin D deficiency, primary hyperparathyroidism. when used and stored as described, reagents are stable:
Unopened:
PRINCIPLE (4) (5) · Until expiry date stated on the label
Once opened
Method without deproteinisation described by Daly and al. modified by
Gamst O.K. and Try K. · Standard (vial R2): transfer requested quantity, well recap the vial and
In an acid medium, phosphate ions form a phosphomolybdic complex store at 18-25°C.
with the ammonium molybdate.The absorbance measured at 340 nm is · Reagent is stable at least 6 months.
proportional to the concentration of phosphate ions in the specimen. · Discard any reagent if cloudy or if absorbance at 340 nm > 0.800.
· Do not use after expiry date
I REAGENTS
R1 Phosphorus Molybdate Reagent SPECIMEN COLLECTION AND HANDLING (2)
Ammonium Molybdate 0.63 mmol/L Unhemolysed serum or plasma:
Sulfuric acid 210 mmol/L Separate from erythrocytes within 1 hour after collection.
Surfactant · Phosphorus is stable in serum for:
ü several days at 2-8°C.
EUH210: Safety Data Sheet (MSDS) available on request ü several months at –15°C.
R1 Phosphorus Standard 24h Urines:
Collect in acid washed, detergent free container. Acidified urines (pH <
Phosphorus 5 mg/dL (1.61 mmol/L)
3 with concentrated hydrochloric acid) should be diluted (1+9) in
According to 1272/2008/EC regulation, these reagents are not demineralised water free from phosphorus before performing the assay.
classified as dangerous · Phosphorus is stable in acidified urines for:
ü 6 months.
LIMITS (3)
To avoid contamination with environmental phosphorus, it is
recommended to use disposal glass or plastic ware, carefully cleaned
material with hydrochloric acid 1 N and well rinsed with distilled water.
For a more comprehensive review of factors affecting this assay refer to
the publication of Young D.S.
I MATERIALS REQUIRED BUT NOT PROVIDED
1. Basic medical analysis laboratory equipment.
2. Spectrophotometer or Biochemistry Clinical Analyzer
3. Saline (Specimen Blank)
IVD H2O
Manufacturer Expiry date In vitro diagnostic Storage temperature Dematerialized water Biological risk
REF LOT ®
Product Reference See Insert Batch number Store away from light Sufficient for Dilute with
PH_220E_IFU_80015_V03_20241025
CALIBRATION (6) QUALITY CONTROL
· REF 95015 BIOLABO Multicalibrator traceable to SRM 3186 · REF 95010 EXATROL-N level I
· REF 80015 (vial R2) for urines and manual method only · REF 95011 EXATROL-P level II
The calibration frequency depends on proper instrument functions and · REF 95012 Urinary controls
on the preservation of the reagent. · External quality control program.
It is recommended to control in the following cases:
· At least once a run.
REFERENCE INTERVALS (2)
· At least once within 24 hours.
· When changing vial of reagent.
In serum or plasma (mg/dL) (mmol/L)
· After maintenance operations on the instrument.
In cord 3.7-8.1 [1.20-2.62] If control is out of range, apply following actions:
Premature 5.4-10.9 [1.74-3.52] 1. Prepare a fresh control serum and repeat the test
2. If control is still out of range, use a new vial of fresh calibrator
0-10 days 4.5-9.0 [1.45-2.91] 3. If control is still out of range, use a new vial of reagent and reassay
10 days-24 months 4.5-6.7 [1.45-2.16] If control is still out of range, please contact BIOLABO technical support
or your local Agent.
24 months-12 years 4.5-5.5 [1.45-1.78]
12-60 years 2.7-4.5 [0.87-1.45]
I PROCEDURE
> 60 years, male 2.3-3.7 [0.74-1.20] Manual method
> 60 years, female 2.8-4.1 [0.90-1.32] Pipette into well identified test tubes:
Reagent Specimen Standard Assay
In 24 h Urines (mg/24 h) (mmol/24 h) Blank blank
Constant daily diet (*) < 1000 [< 32.3]
Reagent 1 mL 1 mL 1 mL
Non-restricted diet 400-1300 [12.9-42.0]
Saline Solution 1 mL
Each laboratory should establish its own normal ranges for the
population that it serves. Demineralised water 20 µL
(*)Constant daily diet: 0.9 at 1.5 g P (29-48 mmol P) and 10 mg Ca/Kg
Standard 20 µL
(0.25 mmol Ca/Kg).
Specimen 20 µL 20 µL
I PERFORMANCES
Mix well. Incubate for 2 minutes at room temperature.
Serum: KENZA 240TX with specimen blank, at 37°C, 340 nm Use a 1 cm path length cuvette and read Standard and assays
Linearity Range: between 0.92 (LOQ) and 10 mg/dL absorbance at 340 nm (334-366) against Reagent blank.
Above, dilute specimen with demineralised water free from phosphorus Read Specimen blank against saline solution.
and re-assay taking into account dilution factor to calculate the result. 1. Urines:
Precision: - Dilute specimen 1+9 in demineralized water
Within-run Low Normal High Between run Normal High - Use standard of the kit to calibrate (do not dilute)
N = 20 level level level N = 12 level level - Control with REF 95012 (to dilute as patient’s urines)
Mean (mg/dL) 1.86 3.89 7.15 Mean (mg/dL) 3.88 8.05 2. Performances with manual procedure should be validated by user
S.D. mg/dL 0.02 0.06 0.08 S.D. mg/dL 0.05 0.14 3. KENZA applications and other applications proposal are available
C.V. % 0.9% 1.5% 1.2% C.V. % 1.3% 1.7% on request
4. Specimen blanking is recommended
Analytical Sensitivity: approx. 0.1079 abs for 1 mg/dL 5. Values of control sera and calibrator are indicated with or without
Comparison study with commercially available method on human specimen blank.
specimens between 1,5 and 10,8 mg/dL (n=74):
y = 0.990 x + 0.0504 r = 0.9948 CALCULATION
Interferences: Serum or plasma:
No interference up to 0,240OD Result = Abs (Assay)- Abs (Specimen blank) x Standard concentration
Turbidity
(7 mmol/L triglycerides) Abs (Standard) – Abs (Standard blank)
Ascorbic acid No interference up to 2500 mg/dL
Total bilirubin No interference up to 450 µmol/L Urines diluted 1+9: Multiply the result by 10 (dilution factor).
Direct bilirubin No interference up to 90 µmol/L
Haemoglobin Positive interference from 170 µmol/L REFERENCES
Glucose No interference up to 1000 mg/dL
(1) TIETZ N.W. Text book of clinical chemistry, 3rd Ed. C.A. Burtis, E.R.
Other substances may interfere (see § Limits) Ashwood, W.B. Saunders (1999) p. 1406-1457.
(2) Clinical Guide to Laboratory Test, 4th Ed., N.W. TIETZ (2006) p.852-855
On the board stability: 1 month (3) YOUNG D.S., Effect of Drugs on Clinical laboratory Tests, 4th Ed. (1995)
Calibration Stability: 1 month p.3-456 to 3-462
(4) DALY J. A. and ERTINGSHAUSEN G.,Clin. Chem., Direct method for
Make a new calibration when changing reagent batch, if quality control inorganic phosphate determination, (1972), 18, p.263-265
results are found out of the established range and after maintenance (5) GAMST O.K., TRY K., Scand. J. Clin. Lab. Invest. (1980), 40, p.483-486
operations (6) SRM: Standard Reference Material
Performances and stability data on KENZA 450TX/ISE and KENZA
ONE are available on request.
PH_220E_IFU_80015_V03_20241025
