Derivatization of GSSG by pHMB in alkaline media. Determination of oxidized glutathione in blood.
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).
Angeli
Valeria
V
Italian National Research Council-Istituto per i Processi Chimico-Fisici, Laboratory of Instrumental Analytical Chemistry, Pisa, Italy.
Chen
Huilun
H
Mester
Zoltan
Z
Rao
Yulan
Y
D'Ulivo
Alessandro
A
Bramanti
Emilia
E
D000468:Alkalies / Q000737:chemistry; D019803:Glutathione Disulfide / Q000097:blood* / Q000737:chemistry*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D006902:Hydroxymercuribenzoates / Q000737:chemistry*; D057230:Limit of Detection; D010084:Oxidation-Reduction; D021241:Spectrometry, Mass, Electrospray Ionization
D000468:Alkalies; D006902:Hydroxymercuribenzoates; C030601:4-hydroxymercuribenzoate; D019803:Glutathione Disulfide