The simultaneous determination of oxidized and reduced glutathiones in liver tissue by ion pairing reverse phase high performance liquid chromatography with a coulometric electrochemical detector

P. R.C. Harvey; R. G. Ilson; S. M. Strasberg

doi:10.1016/0009-8981(89)90001-6

The simultaneous determination of oxidized and reduced glutathiones in liver tissue by ion pairing reverse phase high performance liquid chromatography with a coulometric electrochemical detector

P. R.C. Harvey, R. G. Ilson, S. M. Strasberg

Department of Surgery

Research output: Contribution to journal › Article › peer-review

84 Scopus citations

Abstract

The oxidized (GSSG) and reduced (GSH) forms of glutathione were quantified simultaneously in rat liver tissues by ion-pairing reverse phase high pressure liquid chromatography coupled to a coulometric electrochemical detector (HPLC-EC). Other biological thiols namely cysteine, cystathionine, homocysteine and methionine were shown to be well resolved from the glutathiones. Standard curves for GSH and GSSG were linear over the range of glutathione concentrations found in biological tissues with a correlation coefficient (r) > 0.999. Rat liver tissue content of GSH (16.96 ± 4.29, n = 5 and GSSG (0.467 ± 0.188 n = 5) found in this study are similar to results reported by other investigators. This method is also applicable to determine glutathione levels in rat bile samples. The advantages and disadvantages of employing coulometric over amperometric, as well as precautions in establishing HPLC-EC for the detection of oxidized and reduced glutathiones is discussed.

Original language	English
Pages (from-to)	203-212
Number of pages	10
Journal	Clinica Chimica Acta
Volume	180
Issue number	3
DOIs	https://doi.org/10.1016/0009-8981(89)90001-6
State	Published - Apr 14 1989

Keywords

Coulometric electrochemical detector
Glutathione
HPLC

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10.1016/0009-8981(89)90001-6

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title = "The simultaneous determination of oxidized and reduced glutathiones in liver tissue by ion pairing reverse phase high performance liquid chromatography with a coulometric electrochemical detector",

abstract = "The oxidized (GSSG) and reduced (GSH) forms of glutathione were quantified simultaneously in rat liver tissues by ion-pairing reverse phase high pressure liquid chromatography coupled to a coulometric electrochemical detector (HPLC-EC). Other biological thiols namely cysteine, cystathionine, homocysteine and methionine were shown to be well resolved from the glutathiones. Standard curves for GSH and GSSG were linear over the range of glutathione concentrations found in biological tissues with a correlation coefficient (r) > 0.999. Rat liver tissue content of GSH (16.96 ± 4.29, n = 5 and GSSG (0.467 ± 0.188 n = 5) found in this study are similar to results reported by other investigators. This method is also applicable to determine glutathione levels in rat bile samples. The advantages and disadvantages of employing coulometric over amperometric, as well as precautions in establishing HPLC-EC for the detection of oxidized and reduced glutathiones is discussed.",

keywords = "Coulometric electrochemical detector, Glutathione, HPLC",

author = "Harvey, {P. R.C.} and Ilson, {R. G.} and Strasberg, {S. M.}",

note = "Funding Information: This study was supportedb y the Medical Research Council of Canada. Dr. P.R.C. Harvey is a Career Scientist for the Ontario Ministry of Health, Health PersonnelD evelopmentP rogram.",

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doi = "10.1016/0009-8981(89)90001-6",

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The simultaneous determination of oxidized and reduced glutathiones in liver tissue by ion pairing reverse phase high performance liquid chromatography with a coulometric electrochemical detector. / Harvey, P. R.C.; Ilson, R. G.; Strasberg, S. M.

In: Clinica Chimica Acta, Vol. 180, No. 3, 14.04.1989, p. 203-212.

Research output: Contribution to journal › Article › peer-review

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T1 - The simultaneous determination of oxidized and reduced glutathiones in liver tissue by ion pairing reverse phase high performance liquid chromatography with a coulometric electrochemical detector

AU - Harvey, P. R.C.

AU - Ilson, R. G.

AU - Strasberg, S. M.

N1 - Funding Information: This study was supportedb y the Medical Research Council of Canada. Dr. P.R.C. Harvey is a Career Scientist for the Ontario Ministry of Health, Health PersonnelD evelopmentP rogram.

PY - 1989/4/14

Y1 - 1989/4/14

N2 - The oxidized (GSSG) and reduced (GSH) forms of glutathione were quantified simultaneously in rat liver tissues by ion-pairing reverse phase high pressure liquid chromatography coupled to a coulometric electrochemical detector (HPLC-EC). Other biological thiols namely cysteine, cystathionine, homocysteine and methionine were shown to be well resolved from the glutathiones. Standard curves for GSH and GSSG were linear over the range of glutathione concentrations found in biological tissues with a correlation coefficient (r) > 0.999. Rat liver tissue content of GSH (16.96 ± 4.29, n = 5 and GSSG (0.467 ± 0.188 n = 5) found in this study are similar to results reported by other investigators. This method is also applicable to determine glutathione levels in rat bile samples. The advantages and disadvantages of employing coulometric over amperometric, as well as precautions in establishing HPLC-EC for the detection of oxidized and reduced glutathiones is discussed.

AB - The oxidized (GSSG) and reduced (GSH) forms of glutathione were quantified simultaneously in rat liver tissues by ion-pairing reverse phase high pressure liquid chromatography coupled to a coulometric electrochemical detector (HPLC-EC). Other biological thiols namely cysteine, cystathionine, homocysteine and methionine were shown to be well resolved from the glutathiones. Standard curves for GSH and GSSG were linear over the range of glutathione concentrations found in biological tissues with a correlation coefficient (r) > 0.999. Rat liver tissue content of GSH (16.96 ± 4.29, n = 5 and GSSG (0.467 ± 0.188 n = 5) found in this study are similar to results reported by other investigators. This method is also applicable to determine glutathione levels in rat bile samples. The advantages and disadvantages of employing coulometric over amperometric, as well as precautions in establishing HPLC-EC for the detection of oxidized and reduced glutathiones is discussed.

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KW - HPLC

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The simultaneous determination of oxidized and reduced glutathiones in liver tissue by ion pairing reverse phase high performance liquid chromatography with a coulometric electrochemical detector

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