Talanta 2010-07-15

Derivatization of GSSG by pHMB in alkaline media. Determination of oxidized glutathione in blood.

Valeria Angeli, Huilun Chen, Zoltan Mester, Yulan Rao, Alessandro D'Ulivo, Emilia Bramanti

Index: Talanta 82 , 815-20, (2010)

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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).Copyright 2010 Elsevier B.V. All rights reserved.