Photosensitization by Harmine: An ESR Spin Trapping Study on the Generation of the Superoxide Anion Radical

Abstract

Harmine was in situ UV-irradiated in the presence of 5,5-dimethyl-1-pyrroline-N-oxide (DMPO), α-phenyl-N-tert-butyl nitrone (PBN), or α-(4-pyridyl-1-oxide) N-tert-butyl nitrone (4-POBN) as spin traps and observed by electron spin resonance (ESR) spectroscopy. The superoxide radical (O₂·⁻) was detected as the corresponding DMPO, PBN, or 4-POBN spin adduct in dimethylsulfoxide (DMSO) or acetonitrile solution. The detection of these spin adducts was prevented by the addition of superoxide dismutase (SOD) to the solution. Also, the O₂·⁻ adduct formation was inhibited by the addition of antioxidants as cysteine methyl ester or ascorbic acid, in a dose-dependent manner. The studies carried out in aqueous buffered solution did not allow superoxide radical adduct detection. In the presence of DMPO as a spin trap, the DMPO–OH spin adduct was detected. On the other hand, classical scavengers of the hydroxyl radical such as mannitol or glycerol abolished DMPO–OH spin adduct detection. Other scavengers such as ethanol, DMSO, or sodium azide inhibited the trapping of hydroxyl radicals and resulted in the formation of new radical adducts. No PBN or 4-POBN spin adducts were detected in aqueous solution. The origin of the hydroxyl radical is discussed.