An assay for the determination of alkaline phosphatase activity using disodium p-acetylphenylphosphate as a substrate / 临床检验杂志

ABSTRACT

Objective To establish a new method to measure alkaline phosphatase (ALP) activity using disodium p-acetylphenylphosphate (PAP-PNa2) as substrate.Methods Experimental parameters were set up with continuous-monitoring procedures on a semiautomatic analyzer.Results The wavelength of maximum absorption of PAP was 325 nm,and the Km of ALP was 0.376 mmol/L.The molecular extinction coefficient of PAP at 340 nm was 23 390 L?mol-1?cm-1 and the concentration of citrate buffer was 0.438 mol/L.In the measurement,the optimum pH was 10.4,and the optimum concentration of substrate was 5.0 mmol/L.The delay time was 60 sec and the time keeping the linear reaction was 15 min,and the linear range was 0~1 110 U/L.The measurement was not interfered by the hemoglobin