Kinetics-Based Quantification of Peroxynitrite Using the Oxidative Decarbonylation of Isatin

Peroxynitrite and its radical decomposition products are highly reactive nitrogen and oxygen species that can influence the balance between health and disease in multiple organ systems. Despite vigorous research activity, real-time quantitative monitoring of peroxynitrite generated by donor compounds remains challenging. Here, we report a kinetics-based fluorescence method for quantitative tracking of peroxynitrite generation using the oxidative decarbonylation of isatin to form anthranilic acid as a fluorescent probe. This method relies on knowledge of the rate of the reaction of peroxynitrite with the probe, which we measure using stopped-flow fluorescence techniques. To the best of our knowledge, this is the first optical method capable of providing real-time quantitative measures of peroxynitrite concentrations generated from donor compounds, as demonstrated herein for SIN-1 and Angeli’s salt.