Department of Applied Chemistry, Graduate School of Science and Technology, Tokushima University, Tokushima 770-8506, Japan.
Faculty of Pharmaceutical Sciences, Tokushima University, 1-78-1 Shomachi, Tokushima 770-8505, Japan.
J Chromatogr B Analyt Technol Biomed Life Sci. 2024 Oct 15;1247:124318. doi: 10.1016/j.jchromb.2024.124318. Epub 2024 Sep 14.
A capillary high-performance liquid chromatography (HPLC) system equipped with a dual-electrode detector utilizing track-etched membrane electrodes (TEMEs) was combined with a microdialysis sampling setup. The electrochemical detector benefits from the high electrolysis efficiency of TEMEs, allowing for calibration-free coulometric detection and simplifying data analysis to determine the dopamine recovery through a dialysis probe. Additionally, this system was used for in vivo monitoring of dopamine in the right striatum of a mouse brain. Temporal changes in dopamine levels, including an exponential decay immediately after the dialysis probe insertion and an excess release of dopamine induced by a high concentration of potassium ions, confirmed the system's proper operation. Furthermore, subsequent measurements following the intraperitoneal injection of mirtazapine showed no increase in dopamine levels in the right dorsal striatum. The dual-electrode system displayed characteristic dopamine detection behavior, with anodic and cathodic peak pairs indicative of reversible electrochemical reactions. This capability facilitated the identification of the dopamine peak within the complex chromatogram of the mouse brain dialysate. The consistency between dopamine collection efficiency from standard solutions and dialysate indicated the absence of interfering electroactive substances overlapping with the dopamine peak in the chromatogram. This integrated analysis system successfully tracked temporal fluctuations in dopamine concentration within the mouse brain.
一种配备双电极检测器的毛细管高效液相色谱(HPLC)系统,采用了带有刻蚀膜电极(TEME)的双电极检测器,与微透析采样装置相结合。电化学检测器受益于 TEME 的高电解效率,实现了无校准库仑检测,并简化了数据分析,通过透析探针来确定多巴胺的回收率。此外,该系统还用于在体监测小鼠大脑右侧纹状体中的多巴胺。多巴胺水平的时间变化,包括透析探针插入后立即呈指数衰减,以及高浓度钾离子引起的多巴胺过量释放,证实了系统的正常运行。此外,在腹腔注射米氮平后进行的后续测量显示,右侧背侧纹状体中的多巴胺水平没有增加。双电极系统表现出特征性的多巴胺检测行为,具有指示可逆电化学反应的阳极和阴极峰对。这一特性有助于在小鼠脑透析液的复杂色谱图中识别出多巴胺峰。从标准溶液和透析液中收集的多巴胺效率之间的一致性表明,在色谱图中没有与多巴胺峰重叠的干扰电化学物质。该集成分析系统成功地跟踪了小鼠大脑内多巴胺浓度的时间波动。