Evaluation ofo-phthalaldehyde as bifunctional fluorogenic post-column reagent for glutathione in LC |
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Authors: | P. Leroy A. Nicolas M. Wellmann F. Michelet T. Oster G. Siest |
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Affiliation: | (1) Laboratoire de Chimie Analytique et Bromatologie, URA CNRS 597, Faculté de Sciences Pharmaceutiques et Biologiques, B.P. 403, 54001 Nancy Cedex, France;(2) Centre du Médicament, URA CNRS 597, Faculté de Sciences Pharmaceutiques et Biologiques, B.P. 403, 54001 Nancy Cedex, France |
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Abstract: | Summary A method for the assay of various forms of glutathione in human plasma has been developed. The reduced form was measured after direct injection of deproteinized plasma with perchloric acid. The unbound oxidized forms were assayed in deproteinized plasma after reduction with dithiothreitol. The total amount of glutathione was determined by reduction before protein precipitation. Separation of reduced glutathione from other endogenous thiols was by ion-pair, reversed-phase, high-performance liquid chromatography. Post-column derivatization withortho-phthalaldehyde made the assay more selective than when using pyrenemaleimide as thiol fluorogenic reagent. The linear range was 0.02 to 2.0 g mL–1 with good repeatability (relative standard deviation less than 5% for the lowest concentration measured). |
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Keywords: | Column liquid chromatography Glutathione o-Phthalaldehyde Post-column derivatization Plasma |
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