Detection and identification of oxidants formed during •NO/O2•⁻ reaction: a multi-well plate CW-EPR spectroscopy combined with HPLC analyses.

ABSTRACT: New techniques and probes are routinely emerging for detecting short-lived free radicals such as superoxide radical anion (O₂(•-)), nitric oxide ((•)NO), and transient oxidants derived from peroxynitrite (ONOO(-)/ONOOH). Recently, we reported the profiles of oxidation products (2-hydroxyethidium, ethidium, and various dimeric products) of the fluorogenic probe hydroethidine (HE) in the (•)NO/O₂(•-) system (Zielonka et al. 2012). In this study, we used HPLC analyses of HE oxidation products in combination with continuous wave electron paramagnetic resonance (CW-EPR) spin trapping with 5-tert-butoxycarbonyl-5-methyl-1-pyrroline N-oxide (BMPO) to define the identity of the oxidizing species formed in the (•)NO/O₂(•-) system. EPR spin-trapping technique is still considered as the gold standard

SUBMITTER: Koto T 

PROVIDER: S-EPMC4780754 | biostudies-literature | 2014 Apr

REPOSITORIES: biostudies-literature