Assay for Peroxidase Using 1, 2-Diarylethylenediamines and Catechol Compounds as Fluorogenic Substrates

Abstract

A novel method for the fluorometric assay of horseradish peroxidase and microperoxidase activities is described based on their catalytic reactions between 1, 2-diarylethylenediamines and catechol compounds as fluorogenic substrates in the presence of hydrogen peroxide. meso-1, 2-Diphenylethylenediamine and epinephrine as a couple of substrates are most recommendable in practical use. This method is highly sensitive: the detection limits (S/N=2) are 10μU tube-1 for horseradish peroxidase and 500fmol tube-1 for microperoxidase.