Issue 5, 2011

GC-MS and HPLC methods for peroxynitrite (ONOO and O15NOO) analysis: a study on stability, decomposition to nitrite and nitrate, laboratory synthesis, and formation of peroxynitrite from S-nitrosoglutathione (GSNO) and KO2

Abstract

Nitric oxide (˙NO) and superoxide (O2˙) are ubiquitous in nature. Their reaction product peroxynitrite (ONOO) and notably its conjugated peroxynitrous acid (ONOOH) are highly unstable in aqueous phase. ONOO/ONOOH (referred to as peroxynitrite) isomerize and decompose to NO3, NO2 and O2. Here, we report for the first time GC-MS and HPLC methods for the analysis of peroxynitrite in aqueous solution. For GC-MS analysis peroxynitrite in alkaline solution was derivatized to a pentafluorobenzyl derivative using pentafluorobenzyl bromide. O15NOO was synthesized from H2O2 and 15NO2 and used as internal standard. HPLC analysis was performed on stationary phases consisting of Nucleosil® 100–5C18AB or Nucleodur® C18 Gravity. The mobile phase consisted of a 10 mM aqueous solution of tetrabutylammonium hydrogen sulfate and had a pH value of 11.5. UV absorbance detection at 300 nm was used. HPLC allows simultaneous analysis of ONOO, NO2 and NO3. The GC-MS and HPLC methods were used to study stability, synthesis, formation from S-[15N]nitrosoglutathione (GS15NO) and KO2, and isomerization/decomposition of peroxynitrite to NO2 and NO3 in aqueous buffer.

Graphical abstract: GC-MS and HPLC methods for peroxynitrite (ONOO− and O15NOO−) analysis: a study on stability, decomposition to nitrite and nitrate, laboratory synthesis, and formation of peroxynitrite from S-nitrosoglutathione (GSNO) and KO2

Article information

Article type
Paper
Submitted
12 Aug 2010
Accepted
22 Nov 2010
First published
20 Dec 2010

Analyst, 2011,136, 979-987

GC-MS and HPLC methods for peroxynitrite (ONOO and O15NOO) analysis: a study on stability, decomposition to nitrite and nitrate, laboratory synthesis, and formation of peroxynitrite from S-nitrosoglutathione (GSNO) and KO2

D. Tsikas, Analyst, 2011, 136, 979 DOI: 10.1039/C0AN00625D

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