Simultaneous detection of reduced and oxidized glutathione in tissues and mitochondria by capillary electrophoresis

C. Muscari, M. Pappagallo, D. Ferrari, E. Giordano, C. Capanni, C. M. Caldarera, C. Guarnieri

Research output: Contribution to journalArticlepeer-review

Abstract

We have developed a rapid and precise method for glutathione quantitation by capillary electrophoresis, that allows a low amount of both redox forms to be measured. Small fragments of rat heart or liver tissues (20 mg wet weight) and the corresponding mitochondria (1 mg protein) were homogenized in 1% perchloric acid and the acid-soluble phase ultrafiltered by centrifugation with a microconcentrator (Mr cut-off 3000 Da). The analysis was performed at a constant temperature (28°C) using a Beckman P/ACE System 2100, equipped with a UV absorbance detector set to 200 nm. The limit of quantitation in heart tissue was 1.8 μM for GSH and 1.2 μM for GSSG. Myocardial concentrations of GSH and GSSG were 8.1 ± 2.6 and 0.45 ± 0.15 (nmol/mg protein ± S.D.), respectively. The ratio of GSH to GSSG was 17.8 ± 1.3 for heart tissue, whereas it was much higher (> 100) in the mitochondria. An oxidative stress decreased the myocardial tissue GSH/GSSG ratio, indicating that the CE analysis of both glutathione forms is also a useful method to study biological redox modification.

Original languageEnglish
Pages (from-to)301-307
Number of pages7
JournalJournal of Chromatography B: Biomedical Applications
Volume707
Issue number1-2
DOIs
Publication statusPublished - Apr 10 1998

Keywords

  • Glutathione
  • Heart
  • Liver
  • Mitochondria

ASJC Scopus subject areas

  • Chemistry(all)

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