Catalytic spectrofluorimetric determination of superoxide anion radical and superoxide dismutase activity using N, N-dimethylaniline as the substrate for horseradish peroxidase (HRP)

Abstract

The coupled reaction of N, N-dimethylaniline (DMA) with 4-aminoantipyrine (4-AAP) using superoxide anion radical (O 2 −) as oxidizing agent under the catalysis of horseradish peroxidase (HRP) was studied. Based on the reaction, O 2 − produced by irradiating Vitamin B 2, (V B2) was spectrophotometricly determined at 554 nm. The linear range of this method was 1.8×10 −6–1.2×10 −4 mol l −1 with a detection limit of 5.3×10 −7 mol l −1. The effect of interferences on the determination of O 2 − was investigated. The proposed method was successfully applied to the determination of superoxide dismutase (SOD) activity in human blood and mouse blood.