Structure Effect on Antioxidant Activity of Catecholamines toward Singlet Oxygen and Other Reactive Oxygen Species in vitro.

The reactivity of catecholamine neurotransmitters and the related metabolites were precisely investigated toward 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals and reactive oxygen species. Catecholamines reacted immediately with DPPH radicals, their reactivity being stronger than that of ascorbic acid as a reference. Superoxide scavenging activities of catecholamines determined by WST-1 and electron spin resonance (ESR) spin trapping methods were also high. Whereas tyrosine, the dopamine precursor showed no reactivity toward superoxide. The reactivity toward singlet oxygen was evaluated by observing specific photon emission from singlet oxygen. The results revealed that reactivity of catecholamines was markedly higher than that of sodium azide, and catechin as catechol reference. The reaction of catecholamines and singlet oxygen was further studied by ESR using 55-dimethyl-1-pyrroline N-oxide (DMPO) as a spin trapping reagent and rose bengal as photosensitizer. DMPO-OH signal of epinephrine was significantly small compared to other catecholamines, catechin, and 4-methylcatechol as a reference compound and was as small as that of tyrosine. The signal formation was totally dependent on singlet oxygen, and the presence of catechol compounds. These results indicated that epinephrine is the most potent singlet oxygen quencher than other catecholamines, and the secondary amino group in its alkyl side chain could play a role in unique singlet oxygen quenching property of epinephrine.

Free radical scavenging and antioxidant activity of ascorbigen versus ascorbic acid: studies in vitro and in cultured human keratinocytes.

Ascorbigen (ABG) is formed by indole-3-carbinole and ascorbic acid in brassica vegetables. In the present study, ABG has been systematically analyzed for its free radical scavenging and antioxidant capacity. To directly measure the free scavenging activity of ABG and ascorbic acid (used as a positive control), electron spin resonance and spin trapping experiments were performed. Ascorbic acid exhibited a potent free radical scavenging activity, whereas ABG did not scavenge superoxide anion free radicals and showed only little scavenging activity toward 2,2-diphenyl-1-picrylhydrazyl free radicals. Similar data were obtained for the ferric reducing ability of plasma and trolox equivalent antioxidant capacity assays. In cultured human keratinocytes, ABG counteracted tert-buthylhydroperoxide-induced cytotoxicity, whereas ascorbic acid did not exhibit any protective activity. Furthermore, in ABG-treated human keratinocytes, a decrease in tert-buthylhydroperoxide-induced lipid peroxidation was detected, whereas an ascorbic acid pretreatment did not result in the prevention of lipid peroxidation. These data indicate that ascorbic acid seems to be a more potent free radical scavenger than ABG in vitro, while ABG prevented tert-buthylhydroperoxide cytotoxicity more effectively as compared to ascorbic acid in cultured cells.

Electron spin resonance spectroscopy studies on the free radical scavenging activity of wine anthocyanins and pyranoanthocyanins.

Anthocyanins are a group of natural occurring pigments responsible for the red-blue color of grapes and many fruits and vegetables. Anthocyanins and derived pigments are of double interest, one technological, as they can be used as natural colorants, and another one due to their implication on human health through their antioxidant activity. Although there are numerous studies regarding the antioxidant activity of grape extracts as well as red wine, the free radical scavenging activity of purified anthocyanins and pyranoanthocyanins is largely unknown. In the present study, the hydroxyl and superoxide anion scavenging activities of anthocyanins and their pyruvic acid adducts were systematically investigated by electron spin resonance spectroscopy and spin trapping. The 3-glucosides of delphinidin, cyanidin, petunidin, pelargonidin and malvidin, and the pyruvic adduct of the 3-glucoside of delphinidin exhibited a potent superoxide anion radical scavenging and, to a lesser extent hydroxyl anion radical scavenging activity. The pyranoanthocyanins of cyanidin, petunidin, malvidin and pelargonidin showed a high capacity to scavenge superoxide anion radicals but did not scavenge hydroxyl radicals. Current data indicate that formation of anthocyanin adducts with pyruvic acid, which may occur during wine ageing or fruit juice processing, decreases the hydroxyl and superoxide anion scavenging and thus could decrease the antioxidant potential of these compounds.