Chen Hong-Li, Zhang Xiao-Jun, Qi Sheng-Da, Xu Hong-Xi, Sung Joseph J Y, Bian Zhao-Xiang
School of Chinese Medicine, Hong Kong Baptist University, Kowloon Tong, Hong Kong SAR, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2009 Oct 1;877(27):3248-52. doi: 10.1016/j.jchromb.2009.08.006. Epub 2009 Aug 12.
This study presented a capillary zone electrophoresis (CZE) method for the analysis of trace amount of neurotransmitters glutamate (Glu) and aspartate (Asp) in the microdialysates of rat periaqueductal gray matter (PAG). Glu and Asp were derivatized with the fluorescent agent 5-carboxyfluorescein N-succinimidyl ester (CFSE) for the first time and detected by laser-induced fluorescence (LIF) after separation by CZE. The concentration detection limits (S/N=2) were 6.9x10(-10)M and 8.1x10(-10)M for Glu and Asp, respectively. The repeatability (expressed as RSD) of the migration times of CFSE-amino acid were better than 0.5%, and not higher than 1.5% even over the period of 1 month. This method was applied to quantify Glu and Asp in rat PAG microdialysates with the treatment of formalin injection, and the measured basal concentrations of Glu and Asp were (22.4+/-1.6)x10(-6)M and (0.9+/-0.1)x10(-6)M, respectively.
本研究提出了一种毛细管区带电泳（CZE）方法，用于分析大鼠中脑导水管周围灰质（PAG）微透析液中痕量神经递质谷氨酸（Glu）和天冬氨酸（Asp）。首次用荧光剂5-羧基荧光素N-琥珀酰亚胺酯（CFSE）对Glu和Asp进行衍生化，并在CZE分离后通过激光诱导荧光（LIF）进行检测。Glu和Asp的浓度检测限（S/N = 2）分别为6.9×10⁻¹⁰M和8.1×10⁻¹⁰M。CFSE-氨基酸迁移时间的重复性（以相对标准偏差表示）优于0.5%，即使在1个月的时间内也不高于1.5%。该方法应用于福尔马林注射处理的大鼠PAG微透析液中Glu和Asp的定量分析，测得的Glu和Asp基础浓度分别为（22.4±1.6）×10⁻⁶M和（0.9±0.1）×10⁻⁶M。
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