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一种高通量微透析-固相萃取联用-电感耦合等离子体质谱联用系统,用于连续监测活体大鼠脑外细胞金属离子。

A high-throughput microdialysis-parallel solid phase extraction-inductively coupled plasma mass spectrometry hyphenated system for continuous monitoring of extracellular metal ions in living rat brain.

机构信息

Department of Biomedical Engineering and Environmental Sciences, National Tsing-Hua University, 30013 Hsinchu, Taiwan.

Department of Biomedical Engineering and Environmental Sciences, National Tsing-Hua University, 30013 Hsinchu, Taiwan.

出版信息

J Chromatogr A. 2014 Jan 24;1326:73-9. doi: 10.1016/j.chroma.2013.12.047. Epub 2013 Dec 25.

Abstract

To significantly improve the temporal resolving power of in vivo trace brain metal monitoring system, in this paper we describe a novel analytical configuration combining the dual functions of online segmentation of the rat brain microdialysate and parallel solid phase extraction (SPE) of multiple segmented samples. In contrast to traditional SPE procedures, in this study the three pumped media-the buffered rat brain microdialysate, the eluent, and the air stream-were converted to a series of segmented streams through the manipulation of a flow-through stream selector. After optimizing this online automatic MD/parallel poly(vinyl chloride) SPE/inductively coupled plasma mass spectrometry hyphenated system for the analysis of ultra-trace metal ions, the sample volume of the microdialysate was set at 0.83μL, the analytical sequence was repeatable every 20s, and the detection limits were in the range 0.03-0.24μgL(-1), with spike analyses of Mn, Co, Ni, Cu, and Zn in a rat brain ECF sample agreeing well with expected values (88-107%). To further examine the system's practicability, we also performed (i) in vivo dynamic monitoring of these trace metal ions in living rat brain extracellular fluid post-probe implantation (the basal values for Mn, Co, Ni, Cu, and Zn were 1.17±0.18, 1.27±0.36, 2.46±0.62, 0.86±0.37, and 2.35±0.55μgL(-1), respectively) and (ii) real-time visualization of the physiological response to acute neural depolarization elicited upon perfusing a high-K(+) medium through the MD probe.

摘要

为了显著提高活体示踪脑内金属监测系统的时间分辨率,本文描述了一种将大鼠脑微透析液在线分段和多分段样品固相萃取(SPE)的双重功能相结合的新型分析配置。与传统的 SPE 程序不同,在本研究中,通过流通流选择器的操作,将三种泵送介质——缓冲的大鼠脑微透析液、洗脱液和气流——转化为一系列分段流。在线自动 MD/平行聚氯乙烯 SPE/电感耦合等离子体质谱联用系统优化后,用于分析超痕量金属离子,微透析液的样品体积设定为 0.83μL,分析序列可每 20s 重复一次,检测限在 0.03-0.24μgL(-1)范围内,对大鼠脑 ECF 样品中 Mn、Co、Ni、Cu 和 Zn 的加标分析与预期值吻合良好(88-107%)。为了进一步考察该系统的实用性,我们还进行了(i)探针植入后活体大鼠脑细胞外液中这些痕量金属离子的动态监测(Mn、Co、Ni、Cu 和 Zn 的基础值分别为 1.17±0.18、1.27±0.36、2.46±0.62、0.86±0.37 和 2.35±0.55μgL(-1)),以及(ii)通过微透析探针灌注高 K(+) 介质引起的急性神经去极化的实时可视化。

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