Open AccessA hydrogen peroxide assay based on the peroxidase‐oxidase reaction
Author(s) -
BOUTELET Isabelle,
ALEXANDRE Stéphane,
VINCENT JeanClaude
Publication year - 1994
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1994.tb19017.x
Subject(s) - hydrogen peroxide , horseradish peroxidase , superoxide dismutase , chemistry , peroxidase , superoxide , nuclear chemistry , enzyme , biochemistry
In order to explain an assay method for H 2 O 2 , based on the horseradish peroxidase/NADH/H 2 O 2 system, we performed a numerical simulation of this complex reaction mechanism. For this purpose, we applied a simplified version of a model designed by Yokota and Yamazaki to our experimental conditions [Yokota, K. & Yamazaki, I. (1977) Biochemistry 16 , 1913–1918]. This modification leads to a kinetic behaviour of the system very similar to our experimental results when the presence of superoxide dismutase is simulated. These simulations confirm that hydrogen peroxide concentrations are proportional to the total amount of NADH oxidized. We also showed that superoxide dismutase activity can be determined by this assay method. Moreover, in samples containing superoxide dismutase and hydrogen peroxide, the simulations showed that both superoxide dismutase and hydrogen peroxide can be determined using our assay method.