Hydroxyl radicals (•OH) can be generated via Fenton chemistry catalyzed by transition metals. An in vitro Fenton system was developed to test both the inhibition and stimulation of •OH formation, by monitoring salicylate aromatic hydroxylation derivatives as markers of •OH production. The reaction was optimized with either iron or copper, and target analytes were determined by means of an original HPLC method coupled to coulometric detection. The method granted good sensitivity and precision, while method applicability was tested on antioxidant compounds with and without chelating properties in different substance to metal ratios. This analytical approach shows how Fenton's reaction can be monitored by HPLC coupled to coulometric detection, as a powerful tool for studying molecules' redox behavior.

Department for Life Quality Studies Alma Mater Studiorum University of Bologna Corso d'Augusto 237 47921 Rimini Italy

Department of Pharmacology and Toxicology Faculty of Pharmacy in Hradec Králové Charles University Heyrovského 1203 50005 Hradec Králové Czech Republic

Department of Pharmacy and Biotechnology Alma Mater Studiorum University of Bologna Via Belmeloro 6 40126 Bologna Italy

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