High temporal resolution monitoring of glutamate and aspartate in vivo using microdialysis on-line with capillary electrophoresis with laser-induced fluorescence detection
High temporal resolution monitoring of glutamate and aspartate in vivo using microdialysis on-line with capillary electrophoresis with laser-induced fluorescence detection
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DOI：
10.1021/ac970518u
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发表时间：
1997-11-15
影响因子：
7.4
通讯作者：
Kennedy, RT
中科院分区：
文献类型：
--
作者：
Lada, MW;Vickroy, TW;Kennedy, RT
建立了一种利用毛细管电泳-激光诱导荧光检测技术（CE-LIF）高时间分辨率自动化检测神经递质谷氨酸和天冬氨酸的方法。微透析探针放置在纹状体的麻醉大鼠耦合在线与CE系统的自动流量门控接口。分析物用邻苯二甲醛/β-巯基乙醇在线衍生化，并使用He-Cd激光器的354 nm线（7 mW）激发通过LIF检测。在透析速率为1.2 μ L/min的情况下，透析探针处的谷氨酸盐和天冬氨酸盐的检测限为200 nM。谷氨酸和天冬氨酸可在不到5 s的时间内分离，理论塔板数超过20万。采样时间受到分离时间的限制，而时间分辨率被限制为12 s，因为在探头及其相关管路内会发生谱带展宽。高时间分辨率允许在大鼠脑中的急性电刺激期间第一次同时监测谷氨酸和天冬氨酸溢出。
An automated method for high temporal resolution monitoring of the neurotransmitters glutamate and aspartate in vivo using capillary electrophoresis (CE) with laser-induced fluorescence (LIF) detection was developed. Microdialysis probes placed in the striatum of anesthetized rats were coupled on-line with the CE system by an automated flow-gated interface. Analytes were derivatized on-line with o-phthaldialdehyde/beta-mercaptoethanol and detected by LIF using the 354 nm line (7 mW) of a He-Cd laser for excitation. With dialysis now rates of 1.2 mu L/min, the detection limit at the dialysis probe was 200 nM for glutamate and aspartate. Glutamate and aspartate could be resolved in less than 5 s with over 200 000 theoretical plates. The sampling time was limited by the separation time while the temporal resolution was limited to similar to 12 s because of band broadening that occurs within the probe and its associated tubing. The high temporal resolution allowed the first simultaneous monitoring of glutamate and aspartate overflow during acute electrical stimulation in the rat brain.