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DISCRETPAK™ REAGENTS - Alkaline Phosphatase (ALP)

Intended Use

For IN VITRO quantitative determination of Alkaline Phosphatase in serum using manual or automated applications.

Method Principle

Alkaline Phosphatase (ALP) catalyzes the conversion of p-Nitrophenyl Phosphate (pNPP) to p-Nitrophenol. P-Nitrophenol is a bright yellow-colored compound which has maximum absorbance at 405 nm. The rate of increase in absorbance from p-Nitrophenyl Phosphate (colorless) to p-Nitrophenol (color) is directly proportional to the ALP enzyme activity in the serum sample. The reaction scheme below illustrates the reaction that occurs in this method.

           ALP
 pNPP + H3PO4 + H2O ────────> p-Nitrophenol
             Mg++

 

Preparation Of Working Reagent

Catachem ALP Reagents are packaged ready for use.

Reagent Storage And Stability

ALP Reagents are stored at 2-8°C. When stored as directed, the reagents are stable until the expiration date stated on the label.

Method Performance Characteristics

Sensitivity: The sensitivity of this method is 0.00060-0.00080 absorbance units per u/L.
Linear Range: In this method there is no significant nonlinearity over the range of 0-1200 u/L.
Precision: Within-run and day-to-day precision is tabulated on the tale below:

Alkaline Phosphatase Within-Run Precision Total Precision

MEAN

SD

CV

SD

CV

U/L

U/L

%

U/L

%

49

1.48

3.00

3.8

7.8

261

0.71

0.27

2.2

0.80

500

2.1

0.40

6.5

1.30

 

Correlation

A comparison of this method using an automated analyzer and a reference method resulted in the following regression statistics.

Correlation Data

Parameter

Data Observed

N

153

Range

30-665 U/L

Regression

Y = 0.948x + 34

Correlation

0.999

Sy,x

4.7