Identification of Peroxynitrite by Profiling Oxidation and Nitration Products from Mitochondria-Targeted Arylboronic Acid

Methods Mol Biol. 2021:2275:315-327. doi: 10.1007/978-1-0716-1262-0_20.

Abstract

The development of boronic probes enabled reliable detection and quantitative analysis of hydrogen peroxide , other nucleophilic hydroperoxides, hypochlorite , and peroxynitrite . The major product, in which boronate moiety of the probe is replaced by the hydroxyl group, is, however, common for all those oxidants. Here, we describe how ortho-isomer of mitochondria-targeted phenylboronic acid can be used to detect and differentiate peroxynitrite-dependent and independent probe oxidation. This method highlights detection and quantification of both the major, phenolic product and the minor, peroxynitrite-specific cyclic and nitrated products of probe oxidation.

Keywords: Boronic probes; HPLC-MS; Hydrogen peroxide; Mitochondria-targeted probes; Peroxynitrite; o-MitoPhB(OH)2.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Boronic Acids / chemistry*
  • Chromatography, High Pressure Liquid
  • Hydrogen Peroxide
  • Isotope Labeling
  • Mice
  • Mitochondria / chemistry*
  • Oxidation-Reduction
  • Peroxynitrous Acid / analysis*
  • RAW 264.7 Cells
  • Tandem Mass Spectrometry

Substances

  • Boronic Acids
  • Peroxynitrous Acid
  • Hydrogen Peroxide