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将碳纤维纳米电极应用于微芯片电泳安培法检测大鼠嗜铬细胞瘤(PC12)细胞中的神经递质多巴胺。

Carbon fiber nanoelectrodes applied to microchip electrophoresis amperometric detection of neurotransmitter dopamine in rat pheochromocytoma (PC12) cells.

作者信息

Cheng Han, Huang Wei-Hua, Chen Rong-Sheng, Wang Zong-Li, Cheng Jie-Ke

机构信息

Chemistry Department, Wuhan University, Wuhan, PR China.

出版信息

Electrophoresis. 2007 May;28(10):1579-86. doi: 10.1002/elps.200600603.

Abstract

Microelectrodes have been adopted in electrochemical detection for CE or microchip CE in recent years. In this paper, the use of nanoelectrodes (with tip diameter of 100-300 nm) as the electrochemical detector in microchip CE is firstly reported. The experimental results indicated that both the sensitivity and resolution of microchip CE with the carbon fiber nanoelectrode (CFNE) amperometric detection have been improved markedly comparing with the traditional microelectrodes. The detection limit of dopamine (S/N = 3) is 5.9x10(-8) M, which is one or two orders of magnitude lower than that reported so far, and the resolution of dopamine (DA) and isoprenaline (IP) has also improved from 0.6 (using 7 mum carbon fiber microelectrodes, CFME) to 1.0. We assembled a novel and easily operated microchip CE system with end-column amperometric detection, which allows the convenient and fast replacement of the passivated electrodes. Under the optimized condition, the RSDs of peak height and migration time are 1.47 and 0.31%, respectively (n = 40), indicating that the system displays excellent reproducibility. The nanoelectrode-based microchip CE system has been successfully applied to the determination of DA in cultured rat pheochromocytoma (PC12) cells, and the average content of DA in an individual PC12 cell is 0.54 +/- 0.07 fmol, which is in good agreement with that reported in the literature.

摘要

近年来,微电极已被应用于毛细管电泳(CE)或微芯片CE的电化学检测中。本文首次报道了将纳米电极(尖端直径为100 - 300 nm)用作微芯片CE中的电化学检测器。实验结果表明,与传统微电极相比,采用碳纤维纳米电极(CFNE)安培检测的微芯片CE的灵敏度和分辨率均有显著提高。多巴胺的检测限(信噪比S/N = 3)为5.9×10⁻⁸ M,比迄今报道的值低一到两个数量级,多巴胺(DA)和异丙肾上腺素(IP)的分离度也从0.6(使用7μm碳纤维微电极,CFME)提高到了1.0。我们组装了一种新型且易于操作的具有柱端安培检测功能的微芯片CE系统,该系统可方便快捷地更换钝化电极。在优化条件下,峰高和迁移时间的相对标准偏差(RSD)分别为1.47%和0.31%(n = 40),表明该系统具有出色的重现性。基于纳米电极的微芯片CE系统已成功应用于培养的大鼠嗜铬细胞瘤(PC12)细胞中多巴胺的测定,单个PC12细胞中多巴胺的平均含量为0.54±0.07 fmol,与文献报道结果吻合良好。

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