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基于还原氧化石墨烯修饰微电极的毛细管电泳法测定单个HepG2细胞中的谷胱甘肽

Determination of glutathione in single HepG2 cells by capillary electrophoresis with reduced graphene oxide modified microelectrode.

作者信息

Wang Xiaolei, Wang Jun, Fu Hongyan, Liu Dongju, Chen Zhenzhen

机构信息

College of Chemistry, Chemical Engineering and Materials Science, Shandong Normal University, Jinan, P. R. China.

出版信息

Electrophoresis. 2014 Dec;35(23):3371-8. doi: 10.1002/elps.201400155. Epub 2014 Oct 27.

Abstract

Determination of intracellular bioactive species will afford beneficial information related to cell metabolism, signal transduction, cell function, and disease treatment. In this study, the electrochemically reduced graphene oxide modified carbon fiber microdisk electrode (ER-GOME) was used as a detector of CZE-electrochemical detection and developed to detect glutathione (GSH). The electrocatalytic activity of the modified microelectrode was characterized by cyclic voltammetry. Under optimized experimental conditions, the concentration linear range of GSH was from 1 to 60 μM. When the S/N ratio was 3, the concentration detection limit was 1 μM. Compared with the unmodified carbon fiber microdisk electrode, the sensitivity was enhanced more than five times. With the use of this method, the average contents of GSH in single HepG2 cells were found to be 7.13 ± 1.11 fmol (n = 10). Compared with gold/mercury amalgam microelectrode, which was usually used in determining GSH, the electrochemically reduced graphene oxide modified carbon fiber microdisk electrode was friendly to environment for free mercury. Furthermore, there were several merits of the novel electrochemical detector coupled with CE, such as comparative repeatability, easy fabrication, and high sensitivity, hold great potential for the single-cell assay.

摘要

细胞内生物活性物质的测定将提供与细胞代谢、信号转导、细胞功能及疾病治疗相关的有益信息。在本研究中,电化学还原氧化石墨烯修饰的碳纤维微盘电极(ER-GOME)被用作毛细管区带电泳-电化学检测的检测器,并用于检测谷胱甘肽(GSH)。通过循环伏安法对修饰微电极的电催化活性进行了表征。在优化的实验条件下,GSH的浓度线性范围为1至60μM。当信噪比为3时,浓度检测限为1μM。与未修饰的碳纤维微盘电极相比,灵敏度提高了五倍以上。使用该方法,发现单个HepG2细胞中GSH的平均含量为7.13±1.11 fmol(n = 10)。与通常用于测定GSH的金/汞齐微电极相比,电化学还原氧化石墨烯修饰的碳纤维微盘电极对环境友好,不含游离汞。此外,这种新型的电化学检测器与毛细管电泳联用具有相对重复性好、易于制备和灵敏度高等优点,在单细胞分析方面具有巨大潜力。

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