The ability of scavengers to distinguish OH. production in the iron-catalyzed Haber-Weiss reaction: comparison of four assays for OH.

Kinetic analysis has been used to access how well scavenger inhibition can characterize the reactivity of oxidants produced in the iron-catalyzed reaction of H2O2 with xanthine oxidase-derived O2-.. Formate oxidation to CO2, deoxyribose oxidation, benzoate hydroxylation, and ethylene production from alpha-keto-gamma-methiolbutyric acid (KMB) were measured. With Fe(EDTA) as catalyst, inhibition by most scavengers was quantitatively as expected for OH. involvement. Exceptions were urate and thiourea, which inhibited excessively and appeared to scavenge intermediates of the detection reactions. With nonchelated iron, there was minimal formate oxidation, but benzoate, KMB, and deoxyribose gave, respectively, 17%, 25%, and approximately the same product yield as with Fe(EDTA). Deoxyribose oxidation was not inhibited by some scavengers and excessively inhibited by others. However, scavengers that did not inhibit deoxyribose oxidation did inhibit with KMB and benzoate, and differences in scavenger effects in the presence and absence of EDTA in these assays were relatively minor. The results with formate and deoxyribose, but not KMB and benzoate, can therefore exclude free OH. as a significant oxidant product of the nonchelated iron-catalyzed Haber-Weiss reaction. It is proposed that the different patterns of scavenger inhibition arise in the different assays because scavengers can react with intermediates in the detection reactions, all of which are multistep chains. Thus, inhibition may not signify OH. involvement, and similarities with inhibition expected for OH. my be fortuitous.