Spectrophotometric determination of hydrogen peroxide by using the cleavage of Eriochrome black T in the presence of peroxidase

Abstract

A new hydrogen donor for peroxidase, Eriochrome black T, was reported for the first time. Steady-state catalytic velocity depends upon enzyme and substrate concentrations, and a Michaelis Menten K(m) value of 1.72 x 10-5 mol l-1 and a V(max) value of 4.43 x 10-3 s-1 were measured at pH 8.6. Trace amount of hydrogen peroxide (2 x 10-7 1.0 x 10-5 mol-1) was determined in aqueous solution by using the cleavage of Eriochrome black T catalyzed by peroxidase. The method is simple and practical, with high sensitivity and enzymatic activity.