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Simultaneous determination of reduced and oxidized glutathione in freshly isolated rat hepatocytes and cardiomyocytes by HPLC with electrochemical detection
Author(s) -
Remião Fernando,
Carmo Helena,
Carvalho Félix,
Bastos Maria L.
Publication year - 2000
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/1099-0801(200011)14:7<468::aid-bmc10>3.0.co;2-c
Subject(s) - glutathione , chemistry , chromatography , oxidative stress , high performance liquid chromatography , coulometry , redox , thiol , quantitative analysis (chemistry) , biochemistry , electrochemistry , enzyme , organic chemistry , electrode
Glutathione and glutathione disulphide constitute an essential thiol redox system present in the cell. The balance in favour of the latter is an indication of oxidative stress. Glutathione and glutathione disulphide quantification in isolated cells may therefore be essential for the evaluation of mechanistic and comparative studies of toxic xenobiotics. In this study, a rapid and sensitive isocratic reverse‐phase high‐performance liquid chromatographic method using coulometric detection was implemented for the simultaneous detection of glutathione and glutathione disulphide, in freshly isolated hepatocytes and cardiomyocytes of the rat. The method implemented proved to be effective for the measurement of glutathione and glutathione disulphide in control conditions and for the detection of variations in this redox system, induced by tert ‐butylhydroperoxide. tert ‐Butylhydroperoxide is an organic peroxide, which has been used as a model molecule for inducing oxidative stress in isolated cells. A comparative study with a previously published HPLC‐electrochemical detection method was performed. Copyright © 2000 John Wiley & Sons, Ltd. Abbreviations used: GSH GSSG t ‐BHP tert ‐butylhydroperoxide