Catalytic cathodic stripping voltammetry of oxidized glutathione at a hanging mercury drop electrode in the presence of nickel ion

Abstract

Oxidized glutathione (GSSG) can be determined after previous accumulation on the HMDE at E > −0.2 V (vs. the reference electrode). GSH is formed during the accumulation, possibly by a mercury-ion-assisted hydrolytic disproportionation of GSSG. In the subsequent cathodic scan GSH is released and catalyses the reduction of nickel ion, giving a peak located at −0.6 V. This enables the determination of GSSG by differential-pulse cathodic stripping voltammetry at pH 7.0 in the phosphate acetate or MOPS buffer containing 0.5−1.0 mM Ni(II). The detection limit is 10 nM. The calibration graph is linear even in the presence of small amounts of human serum albumin, HSA. However, HSA increases the detection limit (20 nM for 3 × 10−4% HSA). Acetyl-cysteine in small excess or Cu(II) present as reagent impurity do not interfere. Glutathione, cysteine and similar compounds, which accumulate as mercury salts and form stable nickel complexes, will interfere. The method is put forward as a novel alternative stripping voltammetric method to those involving accumulation and determination as mercury or copper salts and complexes, in the knowledge that it may have advantages in particular analytical situations. In particular the method discriminates against compounds which accumulate as mercury salts but which do not form stable nickel complexes.