Riboflavin-induced Type 1 photo-oxidation of tryptophan using a high intensity 365 nm light emitting diode

The mechanism of photo-oxidation of tryptophan (Trp) sensitized by riboflavin (RF) was examined employing high concentrations of Trp and RF, with a high intensity 365 nm light emitting diode (LED) source under N₂, 20% and 100% O₂ atmospheres. Dimerization of Trp was a major pathway under the N₂ atmosphere, though this occurred with a low yield (^Dφ_Trp = 5.9 × 10^-3), probably as a result of extensive back electron transfer reactions between RF^•- and Trp(H)^•+. The presence of O₂ decreased the extent of this back electron transfer reaction, and the extent of Trp dimerization. This difference is attributed to the formation of O₂^•- (generated via electron transfer from RF^•- to O₂) which reacts rapidly with Trp^• leading to extensive consumption of the parent amino acid and formation of peroxides and multiple other oxygenated products (N-formylkynurenine, alcohols, diols) of Trp, as detected by LC-MS. Thus, it appears that the first step of the Type 1 mechanism of Trp photo-oxidation, induced by this high intensity 365 nm light source, is an electron transfer reaction between the amino acid and ³RF, with the presence of O₂ modulating the subsequent reactions and the products formed, as a result of O₂^•- formation. These data have potential biological significance as LED systems and RF-based treatments have been proposed for the treatment of pathological myopia and keratitis.