H. Yoshioka et al., A novel method of measuring hydroxyl radical-scavenging activity of antioxidants using gamma-irradiation, FREE RAD RE, 35(3), 2001, pp. 265-271
A new method using ESR spin trapping was proposed for measuring the scaveng
ing activity of antioxidants for the hydroxyl (OH) radical. (-)-Epigallocat
echin gallate (EGCg) and 5,5-dimethyl-1-pyrroline N-oxide (DMPO) were used
as the antioxidant and spin trapping agent, respectively. The conventional
method using a Fenton reaction had problems associated with the estimation
of activity, because the antioxidant disturbs the system for generating OH
radical by coordinating on Fe2+ and by consuming H2O2, besides scavenging t
he spin adduct (DMPO-OH). Intense, gamma -irradiation was therefore used to
generate OH radicals, and the intensity decrease in DMPO-OH after irradiat
ion was followed to obtain the rate constant for the scavenging of DMPO-OH
by EGCg. The intensities were extrapolated to zero time to estimate the qua
ntity of DMPO-OH formed during gamma -irradiation. By using these values, t
he reaction rate constant between OH radical and EGCg was calculated as a r
atio to that of DMPO. It was shown that this method is useful for comparing
the OH radical-scavenging activity of various antioxidants.