Diagnostic Laboratory Section

METHOD OF ESTIMATION OF ALP ( DGKC - SCE PROCEDURE)

Principle

• Kinetic determination of Alkaline Phosphatase (ALP) based upon the following reactions.

Reagent composition

• Alkaline phosphatase (S.L) R1
• Diethanolamine Buffer, (pH 10.2) - 125 mmol/L
• Magnesium Chloride - 0.625 mmol/Lab
• Alkaline phosphatase (S.L) R2
• P-Nitrophenyle phosphate - 50mmol/L

Laboratory procedure

• Mix reagent R1 (4 volume) and R2 (1volume) to get  the working solution
• Working Reagent - 1000 µL
• Sample - 20 µL
• Mix and incubate for 1 minute at 37 ° C. Measure the change in absorbance per minute (OD /min) during 3 minutes.

Calculation

• ALP Activity (U/L) = ------------- (OD/ min ) X 2750