Electrocatalytic microelectrode detectors for choline and acetylcholine following separation by capillary electrophoresis.
Electrocatalytic microelectrode detectors for choline and acetylcholine following separation by capillary electrophoresis.
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DOI：
10.1021/ac9010843
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发表时间：
2009-08-15
影响因子：
7.4
通讯作者：
Kirchhoff, Jon R.
中科院分区：
文献类型：
--
作者：
Mukherjee, Jhindan;Kirchhoff, Jon R.
通过毛细管电泳 (CE) 分离后，采用两个电催化酶修饰的微电极系统作为胆碱 (Ch) 和乙酰胆碱 (ACh) 的端柱安培检测器。将基于 Os 的氧化还原聚合物水凝胶 (HRP-Os) 中交联的辣根过氧化物酶物理吸附在 Au 微电极上，然后对乙酰胆碱酯酶 (AChE) 和胆碱氧化酶 (ChO) 进行化学交联。另一种方法是利用过渡金属催化剂普鲁士蓝 (PB) 在 Pt 微电极上的沉积作为电催化剂。使用丁酰胆碱 (BuCh) 作为内标，HRP-Os/AChE-ChO 和 PB/AChE-ChO 电极对 Ch 和 ACh 分别在 2-2000 μM 和 10-2000 μM 范围内表现出优异的线性响应。 HRP-Os/AChE-ChO 电极的检测限为 0.1 μM 或 38 amol。 PB/AChE-ChO 电极上 ACh 和 Ch 的检测限为 5 μM 或 9.5 fmol。电极分别在 +0.10 V 和 -0.10 V vs. Ag/AgCl (3M NaCl) 的电位下工作，从而最大限度地减少了可氧化干扰的电位响应。此外，两种电催化电极在超过70小时内都表现出良好的运行稳定性。 HRP-Os/AChE-ChO 和 PB/AChE-ChO 电极增强的检测能力与 Ch 和 ACh 的高效 CE 分离相结合，为监测和定量生物体液中的这些胆碱能生物标志物提供了一种新的灵敏和选择性策略。
Two electrocatalytic enzyme modified microelectrode systems were employed as end-column amperometric detectors of choline (Ch) and acetylcholine (ACh) following separation by capillary electrophoresis (CE). Horseradish peroxidase crosslinked in an Os based redox polymer hydrogel (HRP-Os) was physically adsorbed on Au microelectrodes followed by chemical crosslinking of the enzymes acetylcholinesterase (AChE) and choline oxidase (ChO). An alternative approach utilized the deposition of the transition metal catalyst, Prussian Blue (PB), on Pt microelectrodes as the electrocatalyst. Utilizing butyrylcholine (BuCh) as an internal standard, the HRP-Os/AChE-ChO and PB/AChE-ChO electrodes exhibited excellent linear responses from 2–2000 μM and 10–2000 μM, respectively, for both Ch and ACh. Detection limits of 0.1 μM or 38 amol were determined for the HRP-Os/AChE-ChO electrode. The limit of detection for ACh and Ch at the PB/AChE-ChO electrode was 5 μM or 9.5 fmol. The electrodes were operated at potentials of +0.10 and −0.10 V vs. Ag/AgCl (3M NaCl), respectively, and thus minimized the potential response from oxidizable interferences. In addition, both electrocatalytic electrodes showed good operational stability for more than 70 hours. The enhanced detection capability of the HRP-Os/AChE-ChO and PB/AChE-ChO electrodes in combination with efficient CE separation of Ch and ACh provides a new sensitive and selective strategy for monitoring and quantifying these cholinergic biomarkers in biological fluids.
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影响因子：
1.7
作者：
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