Direct and simultaneous determination of reduced and oxidized glutathione and homoglutathione by HPLC-ESI/MS (TOF) in plant tissue extracts

Abstract

Glutathione and homoglutathione as well as ascorbic acid, are low-moLecular mass antioxidants involved in celluLar redox homeostasis in pLants (Noctor and Foyer, 1998). Efforts are being made to deveLop methods to determine the concentrations of oxidized and reduced forms of these compounds and their ratio (Camera et al, 2002). Many of the methodologies developed so far, however, are time-consuming and complex, and therefore analytes can decompose and/or their redox status can change during the anaLysis process. We have developed a method that allows for the simultaneous determination of reduced (GSH and hGSH) and oxidized forms (GSSG) of these compounds, and is aLso suitable for the determination of ascorbic acid (ASA) and S- nitrosoglutathione (GSNO).