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Derivatization of GSSG by pHMB in alkaline media. Determination of oxidized glutathione in blood |
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| Authors: | Valeria Angeli Zoltan Mester Alessandro D’Ulivo |
| Institution: | a Italian National Research Council-Istituto per i Processi Chimico-Fisici, Laboratory of Instrumental Analytical Chemistry, Via G. Moruzzi 1, 56124 Pisa, Italy b Key Laboratory of Biogeology and Environmental Geology of Chinese Ministry of Education & School of Environmental Studies & Sino-Hungarian Joint Laboratory of Environmental Science and Health, China University of Geosciences, 430074 Wuhan, PR China c Institute for National Measurement Standards, National Research Council Canada, 1200 Montreal Rd., Ottawa, ON K1A 0R6, Canada d Center for Instrumental Analysis, China Pharmaceutical University, Nanjing 210009, PR China |
| Abstract: | Chromatographic determination of glutathione disulfide (GSSG) without any preliminary reduction has been presented using GSSG derivatization by p-hydroxymercuribenzoate (pHMB) in strong alkaline medium followed by the determination of GS-pHMB complex by reversed phase chromatography coupled to chemical vapour generation and atomic fluorescence detector (RPC-CVGAFS). A detection limit of 35 nM for GSSG (corresponding to 1.8 pmol) detected as GS-pHMB species was achieved based on a signal-to-noise ratio of 3 in buffer and in blood. The proposed method was applied to the determination of GSSG in whole blood and validated by the classical determination of GSSG by derivatization after reduction with dithiothreitol (DTT). |
| Keywords: | Oxidized glutathione GSSG Liquid chromatography Atomic fluorescence spectrometry |
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