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中国科学院大学学报 ›› 2014, Vol. 31 ›› Issue (1): 38-45.DOI: 10.7523/j.issn.2095-6134.2014.01.007

• 化学与生命科学 • 上一篇    下一篇

基于微芯片电泳-电化学方法的细胞内谷胱甘肽分析

姚琳, 吴亮其, 丁永胜   

  1. 中国科学院大学生命科学学院, 北京 100049
  • 收稿日期:2013-04-07 修回日期:2013-05-22 发布日期:2014-01-15
  • 通讯作者: 丁永胜,E-mail:dingysh@ucas.ac.cn
  • 基金资助:

    国家自然科学基金(21075135)和中国科学院生物局“十二五”基础前沿专项(KSCX2-EW-J-29)资助

Analysis of glutathione in cells by microchip electrophoresis coupled with electrochemical detection

YAO Lin, WU Liangqi, DING Yongsheng   

  1. College of Life Sciences, University of Chinese Academy of Sciences, Beijing 100049, China
  • Received:2013-04-07 Revised:2013-05-22 Published:2014-01-15

摘要:

建立微芯片电泳-电化学检测技术分析细胞内还原型(GSH)和氧化型(GSSG)谷胱甘肽的新方法. 分别考察缓冲液pH值、缓冲液浓度、SDS浓度、分离电压、进样时间、检测电位等因素对GSH、GSSG分离检测的影响. 在最优实验条件下,在3 min内实现GSH和GSSG的有效分离和检测. GSH和GSSG的线性范围分别为5.0~200.0 μmol/L和2.0~100.0 μmol/L(R2>0.99),最低检测限(S/N≥3)分别为4.87 μmol/L和1.98 μmol/L. 最终将所建立的方法应用于细胞样品中2种物质含量的测定,结果令人满意.

关键词: 合金修饰电极, 谷胱甘肽, 微芯片电泳, 电化学检测

Abstract:

We established a new method for simultaneous determination of reduced and oxidized glutathione. The separation and detection conditions were optimized in terms of the buffer pH, buffer concentration, SDS concentration, separation potential, injection time, and detection potential. Under the optimized condition, GSH and GSSG were separated within 3 min, and the satisfactory linear ranges were 5~200 and 2~100 μmol/L for GSH and GSSG, respectively. The detection limits of GSH and GSSG were 4.87 and 1.98 μmol/L, respectively. Quantitative analyses of GSSG and GSH in cells were performed with satisfactory results.

Key words: alloy modified electrode, glutathione, microchip electrophoresis, electrochemical detection

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