BIOLABO

www.biolabo.fr ALKALINE PHOSPHATASE

MANUFACTURER:
BIOLABO SAS,
C o l o r i m e t r i c M et h o d
Les Hautes Rives Reagent for quantitative determination of alkaline phosphatase activity
02160, Maizy, France [EC 3.1.3.1] in human serum and plasma.

REF 80014 R1 2 x 56 mL R2 1 x 2,5 mL

R3 1 x 27 mL R4 1 x 27 mL

TECHNICAL SUPPORT AND ORDERS IVD

Tel : (33) 03 23 25 15 50
Made In France
[email protected]
Latest revision : www.biolabo.fr I: corresponds to significant modifications

I 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 quantitative determination of alkaline phosphatase activity · Verify the integrity of the contents before use.
[EC 3.1.3.1]in human serum and plasma to screen its level. · Waste disposal: Respect legislation in force in the country.
· All specimens or reagents of biological origin should be handled as
I GENERALITIES (1) potentially infectious. Respect legislation in force in the country.
Alkaline phosphatase (ALP) is found in many tissues, including bone, Any serious incident that has occurred in connection with the device is
liver, intestine, kidney, and placenta. Serum ALP measurements are of notified to the manufacturer and the competent authority of the Member
particular interest in the investigation of two groups of conditions : State in which the user and/or patient is based.
hepatobiliary diseases and bone diseases associated with increased
osteoblastic activity.
ALP determined reflects total serum levels and does not distinguish the REAGENTS PREPARATION
source of the isoenzyme. Clinicians must therefore rely on other Reagents are ready to use.
parameters of liver or other organ function or a more specific
determination of ALP to assess its source.
STABILITY AND STORAGE
PRINCIPLE (4) (5) Store at 2-8°C and away from light.
Colorimetric determination of the ALP activity as follows : · Unopened :
Reagents are stable upon expiry date stated on the label.
Alkaline phosphatase
· Once reconstituted :
Phenylphosphate Phenol + Phosphate
Reagents are stable at least for 3 months when free from
Free phenol liberated by hydrolysis of the substrate reacts then with contamination.
4-amino-antipyrine in the presence of alkaline potassium ferricyanide to
form a red-coloured complex which absorbance measured at 510 nm is SPECIMEN COLLECTION AND HANDLING (2)
directly proportional to the ALP activity in the specimen. Unhemolysed serum or heparinised plasma, immediately refrigerated.
ALP activity is stable in the specimen for :
REAGENTS · 2-3 days at 2-8°C.
R1 ALKALINE PHOSPHATASE Substrate-Buffer · 1 month at –25°C.
Disodium Phenyl phosphate 5 mmol/L
Carbonate-bicarbonate buffer pH 10 50 mmol/L LIMITS (3)
Stabilizer Avoid haemolysed serum.
For a more comprehensive review of factors affecting this assay refer to the
R2 ALKALINE PHOSPHATASE Standard
publication of Young D.S.
Phenol corresponding to 20 U King and Kind
R3 ALKALINE PHOSPHATASE Blocking Reagent MATERIALS REQUIRED BUT NOT PROVIDED
4-Amino-antipyrine 60 mmol/L 1. Basic medical analysis laboratory equipment.
Sodium arsenate 240 mmol/L 3.Spectrophotometer and water bath thermostated at 37°C
DANGER: Acute Tox. 4: H302 - Harmful if swallowed.
Aquatic Chronic 2: H411 - Toxic to aquatic life with long lasting effects.
Carc. 1A: H350 - May cause cancer.
P264: Wash thoroughly after handling, P273: Avoid release to the
environment, P280: Wear protective gloves/face protection/protective
clothing/respiratory protection/protective footwear, P308+P313: IF
exposed or concerned: Get medical advice/attention, P501: Dispose of
contents/container in accordance with regulations on hazardous waste or
packaging and packaging waste respectively. Classification is due to
sodium arsenate 2,5 - < 10%. For more details, see Safety Data Sheet
(SDS)
R4 ALKALINE PHOSPHATASE Dye Reagent
Potassium ferricyanide 150 mmol/L
R1, R2, R4: These reagents are not classified as hazardous according to
CLP Regulation (EC) No 1272/2008.

ALP_DT_220_IFU_80014_EN_V02_20220707
QUALITY CONTROL CALIBRATION
· External quality control program. · Standard enclosed in the Kit (vial R2)
It is recommended to control in the following cases :
· At least once a run. PROCEDURE
· At least once within 24 hours.
Manual method
· When changing vial of reagent.
· After maintenance operations on the instrument. Let stand reagent and specimens at room temperature.
If control is out of range, apply following actions : Reagent Specimen
1. Check the temperature and repeat the test with the same control. Prepare tubes as follows : Standard Assay
blank blank
2. If control is still out of range, prepare a fresh control serum and repeat
the test. Reagent R1 2 mL 2 mL 2 mL 2 mL
3. If control is still out of range, use a new vial of Standard and repeat Incubate 5 minutes at 37°C.
the test.
4. If control is still out of range, calibrate with a new vial of reagent. Specimen 50 µL
5. If control is still out of range, please contact BIOLABO technical Reagent R2 (Standard) 50 µL
support or your local Agent.
Let stand exactly 15 minutes at 37°C.

REFERENCE INTERVALS (2) Reagent R3 0,5 mL 0,5 mL 0,5 mL 0,5 mL

Mix well.
Kind and King Units /dL IU/L (37°C)
Reagent R4 0,5 mL 0,5 mL 0,5 mL 0,5 mL
Birth 5-15 [36-107]
Specimen 50 µL
Premature (1.5 to 2 x adult value)
Demineralised water 50 µL
1 month 10-30 [71-213]
Mix. Incubate 10 minutes at room temperature and away from light. Read
3 years 10-20 [71-142]
absorbances of the blank specimen, standard and assay at 510 nm against
10 years 15-30 [107-213] reagent blank.
Adults 4.5-13 [32-92] Coloration is stable for 45 minutes away from light.

Children : Values may be increased (up to threefold during puberty) Notes:

Each laboratory should establish its own normal ranges for the 1. Performances with manual procedure should be validated by user.
population that it serves.

CALCULATION
1) Results (Kind and King unit):
Quantity of enzyme which, on reaction’s conditions, liberates 1 mg of
phenol in 15 minutes at 37°C.
ALP activity
(Kind and King units/ 100 mL) = Abs Assay - Abs Specimen blank x 20
Abs Standard

2) Result (IU/L) = 7,09 x Result (Kind and King Unit/100 mL)

REFERENCES
(1) TIETZ N.W. Text book of clinical chemistry, 3rd Ed. C.A. Burtis, E.R.
Ashwood, W.B. Saunders (1999) p. 676-684 et p.1429-1431.
(2) Clinical Guide to Laboratory Test, 4th Ed., N.W. TIETZ (2006) p. 78-83
(3) YOUNG D.S., Effect of Drugs on Clinical laboratory Tests, 4th Ed. (1995)
P.3-26 à 3-35
(4) Kind P. R. N., King E. J, Estimation of plasma Phosphatase by
determination of hydrolysed phenol with amino-anti-antipyrine, J. Clin. Path.
(1954), 7, p.322-326
(5) Belfield A, Goldberg D. M., Revised assay for serum phenylphosphatase
activity using 4-amino-antipyrine, Enzyme (1971), 12, p.561-573

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

ALP_DT_220_IFU_80014_EN_V02_20220707