Quantitative Determination of Alkaline Phosphatase (ALP)
Quantitative Determination of Alkaline Phosphatase (ALP)
Quantitative Determination of Alkaline Phosphatase (ALP)
Alkaline phosphatase
p-Nitrophenylphosphate. Kinetic. DGKC
Quantitative determination of alkaline phosphatase (ALP) 5. Read initial absorbance (A) of the sample, start the stopwatch and
IVD read absorbances at 1 minute intervals thereafter for 3 minutes.
6. Calculate the difference between absorbances and the average
Store at 2-8ºC absorbance differences per minute ( A/min).
SAMPLES NOTES
1
Serum or heparinzed plasma . Use unhemolyzed serum, separated SPINREACT has instruction sheets for several automatic analyzers.
from the clot as soon as possible. Stability: 3 days at 2-8ºC. Instructions for many of them are available on request.
BIBLIOGRAPHY
PROCEDURE 1. Wenger C. et al. Alkaline phosphatase. Kaplan A et al. Clin Chem The C.V.
1. Assay conditions: Mosby Co. St Louis. Toronto. Princeton 1984; 1094-1098.
Wavelength: . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .405 nm 2. Rosalki S et al. Clin Chem 1993; 39/4: 648-652.
Cuvette: . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . 1 cm light path 3. Young DS. Effects of drugs on Clinical Lab. Tests, 4th ed AACC Press, 1995.
Constant temperature . . . . . . . . . . . . . . . . . . 25ºC / 30ºC / 37ºC 4. Young DS. Effects of disease on Clinical Lab. Tests, 4th ed AACC 2001.
2. Adjust the instrument to zero with distilled water or air. 5. Burtis A et al. Tietz Textbook of Clinical Chemistry, 3rd ed AACC 1999.
3. Pipette into a cuvette: 6. Tietz N W et al. Clinical Guide to Laboratory Tests, 3rd ed AACC 1995.
BEIS07-I 26/02/13 SPINREACT,S.A./S.A.U. Ctra.Santa Coloma, 7 E-17176 SANT ESTEVE DE BAS (GI) SPAIN
Tel. +34 972 69 08 00 Fax +34 972 69 00 99. e-mail: [email protected]
ALP
Fosfatasa alcalina
p-Nitrofenilfosfato. Cinético. DGKC
Determinación cuantitativa de fosfatasa alcalina (FAL) 5. Leer la absorbancia (A) inicial de la muestra, poner en marcha el
IVD cronometro y leer la absorbancia cada minuto durante 3 minutos.
6. Calcular el promedio del incremento de absorbancia por minuto
Conservar a 2-8ºC ( A/min).
PRINCIPIO DEL MÉTODO
CÁLCULOS
La fosfatasa alcalina (FAL) cataliza la hidrólisis del p-nitrofenilfosfato
(pNPP) a pH 10,4 liberando p-nitrofenol y fosfato, según la siguiente A/min x 3300 = U/L de FAL
reacción: Unidades: La unidad internacional (UI) es la cantidad de enzima que convierte 1
FAL mol de substrato por minuto, en condiciones estándar. La concentración se
p-Nitrofenilfosfato + H2O p-Nitrofenol + Fosfato expresa en unidades por litro (U/L).
BEIS07-E 26/02/13 SPINREACT,S.A./S.A.U. Ctra.Santa Coloma, 7 E-17176 SANT ESTEVE DE BAS (GI) SPAIN
Tel. +34 972 69 08 00 Fax +34 972 69 00 99. e-mail: [email protected]