Electroanalytical determination of acid phosphatase activity by monitoring p-nitrophenol

Abstract

The development of a modified electrode for the selective determination of p-nitrophenol (PNP) to be applied in the determination of the enzymatic activity (EA) of acid phosphatase using p-nitrophenylphosphate (PNPP) as substrate is described. Elimination of the interference of PNPP was made by covering the electrode surface using 25 μl of a 5% (m/v) Nafion ® solution. In comparison to the glassy carbon (GC) electrode without modification, the presence of the membrane promoted a reduction in the sensitivity and an increase in the detection limit for the determination of PNP, in addition to a decrease of the electron transfer constant. The GC electrode covered with a Nafion ® membrane showed a linear response range between 20 and 230 μmol l −1 for PNP, adjusted by the equation I P=0.157+0.0089 [PNP], for N=22 with a correlation coefficient of 0.9984. This electrode was used for the determination of the EA of acid phosphatase from castor bean seeds and an activity of 38.40 U mg −1 was found.