Determination of glutathione and glutathione disulfide in hepatocytes by liquid chromatography with an electrode modified with functionalized carbon nanotubes.
Glutathione (GSH) and glutathione disulfide (GSSG) are important thiols, which provide defence against oxidative stress by scavenging free radicals or causing the reduction of hydrogen peroxide. The ratio GSH/GSSG is often used as a sensitive index of oxidative stress in vivo. In this paper, a direct electrochemical method using an electrode modified with functionalized carbon nanotubes as electrochemical detector (ED) for liquid chromatography (LC) was described. The electrochemical behaviors of GSH and GSSG on this modified electrode were investigated by cyclic voltammetry and it was found that the functionalized carbon nanotubes exhibited efficiently electrocatalysis on the current responses of GSH and GSSG. In LC-ED, both of the analytes showed good and stable current responses. The detection limit of GSH was 0.2 pmol on column and that of GSSG was 1.2 pmol on column, which were low enough for the analysis of real small samples. The method was sensitive enough to detect difference in concentration of GSH and GSSG in hepatocytes from animals with and without introduction of oxidation stress by glucose or hydrogenperoxide.
Zhang
Wen
W
Department of Chemistry, East China Normal University, Shanghai 200062, PR China. wenzhang26@163.com
Wan
Fangli
F
Zhu
Wei
W
Xu
Haihong
H
Ye
Xiyun
X
Cheng
Rongyi
R
Jin
Li-Tong
LT
D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods*; D003921:Diabetes Mellitus, Experimental; D004563:Electrochemistry; D004566:Electrodes; D005947:Glucose / Q000494:pharmacology; D005978:Glutathione / Q000032:analysis*; D019803:Glutathione Disulfide / Q000032:analysis*; D022781:Hepatocytes / Q000737:chemistry* / Q000187:drug effects; D006861:Hydrogen Peroxide / Q000494:pharmacology; D051379:Mice; D037742:Nanotubes, Carbon; D018384:Oxidative Stress; D015203:Reproducibility of Results
D037742:Nanotubes, Carbon; D006861:Hydrogen Peroxide; D005978:Glutathione; D005947:Glucose; D019803:Glutathione Disulfide