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IVD
ALKALIN PHOSPHATASE
ALP (DGKG)
Kinetic determination of
alkalin phosphatase activity (EC 3.1.3.1)
FT En 14 Date of updating: 10/ 2020 Version B
IN VITRO DIAGNOSTIC USE
Manufacturer Use by In Vitro Diagnostic Temperature
Limitation
Catalogue number See insert Store away from light Sufcient
for < n > essais
Batch number
CLINICAL SIGNIFICANCE
Alkalin phosphatases (ALPs) are enzymes found in most body tissues, particularly in bone, liver,
intestine, kidney, and placenta. Approximately 80% of circulating PAL activity comes from liver and
bone isoforms. The PAL activity assay is often prescribed in cases of suspected liver or bone disease.
An increase in PAL activity is seen in cholestasis or biliary obstruction, or in bone diseases such as
rickets, Paget’s disease, osteomalacia, and bone metastases.
PRINCIPLE
This is the kinetic determination of alkalin phosphatase activity (PAL) according to the method re-
commended by the German Society for Clinical Chemistry (DGKG).
In an alkalin medium, alkalin phosphatases catalyze the hydrolysis of Nitrophenylphosphate to Ni-
trophenol and Phosphate.
The kinetic of nitrophenol’s formation is proportional to the PAL activity in the sample.
REAGENT COMPOSITION
Reagent 1
Buffer
Buffer diethanolamine pH 9,8
Magnésium chloride
1 mmol/l
0,5 mmol/l
Reagent 2
Substrate
Nitrophenylphosphate 10 mmol/l
SAFETY CAUTIONS
Biomaghreb reagents are intended for use by qualified personnel for in vitro use (do not pipette by
mouth).
• Refer to the current SDS available on request or at www.biomaghreb.com;
• Verify the integrity of the reagents before use; and
• Disposal of waste: comply with the legislation in force.
For safety reasons, treat any specimen or reagent of biological origin as potentially infectious.
Respect the legislation in force.
REAGENT PREPARATION
Working solution:
Take the bottle R1 with the necessary quantity of R2, listed below:
REF
13019 and
REF
13040 : R1...................................... 3 ml
R2....................................0,3 ml
REF
13026 : R1.....................................10 ml
R2.......................................1 ml
REF
13033 : R1.....................................50 ml
R2.......................................5 ml
SAMPLE COLLECTION AND HANDLING
Heparinized serum or plasma without hemolysis.
PRESERVATION AND STABILITY
Stored in the original, tightly stoppered bottle at 2-8°C, the reagents are stable if used and stored
under the recommended conditions:
• Before opening: Until the expiry date indicated on the label of the box at 2-8°C;
• After opening : (Working Solution):
5 days at 15 -25°C;
15 days at 2-8°C.
ADDITIONAL EQUIPMENT
• Basic equipment of the medical analysis laboratory ;
• Spectrophotometer or Clinical Biochemistry Analyzer.
QUALITY CONTROL
External quality control program.
It is recommended to control in the following cases:
• At least one test per series.
• Change of reagent bottle.
• After maintenance work on the analyzer.
If a control value is outside the confidence limits, repeat the procedure using the same control.
Use normal and pathological control sera.
LINEARITY
If the ΔO.D./min > 0.250 repeat the test by diluting the sample 1/5 in a 9g/l NaCl solution and multiply
the result by 5.
PROCEDURE
Wavelength: 405 nm;
Temperature: 25- 30 or- 37°C;
Tank: 1 cm thick;
Adjust the spectrophotometer zero to air or distilled water.
Working solution
1 ml
Incubate at 25 - 30 or 37°C
Sample
20 µl
Mix and introduce into a thermostatically controlled tank.
Wait 1 minute and then measure the average increase in optical density per minute for 1
to 3 minutes.
Note: Measurement of enzyme activity is better within four hours of sampling.
CALCULATION
At 405 nm ...............PAL (UI/L) = ΔD.O/min x 2750;
At 410 nm ...............PAL (UI/L) = ΔD.O/min x 2910.
REFERENCE VALUES
25°C 30°C 37°C
Children
400 UI/l 500 UI/l 650 UI/l
Adults
40 -190 UI/l 50 - 230 UI/l 70 - 300 UI/l
REFERENCES
Haussamen T.U. et al. Clin. Chim. Acta. 35, 271-273 (1977).
