Li Hua, Wang Hong, Chen Jin-he, Wang Li-hua, Zhang Hua-shan, Fan Yun
College of Chemistry, Wuhan University, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2003 May 5;788(1):93-101. doi: 10.1016/s1570-0232(02)01032-2.
An efficient, sensitive and rapid analysis of the amino acid neurotransmitters in the cerebral cortex of rats was developed by capillary electrophoresis with laser-induced fluorescence detection and fluorescein isothiocyanate (FITC) derivatization. This method was used to investigate the pharmacological effect of baicalin during cerebral ischemia. Different parameters which influenced derivatization and separation were optimized. The separation of amino acids was carried out in an uncoated fused-silica capillary (57 cm x 75 microm I.D.) with a buffer of 15 mM borate at pH 9.2 and an applied voltage of 17.5 kV. The detection limits for six amino acids were in the range of 2.1 x 10(-11)-6.3 x 10(-10) M. The changes in the level of amino acid neurotransmitters in brain cortex of three experimental rat groups were studied by this capillary electrophoresis-laser-induced fluorescence detection method. The results show that cerebral ischemia can cause a significant elevation in the concentrations of Glu, Asp, GABA, and Gly in cerebral cortex. Baicalin administration can attenuate the elevations of Glu and Asp induced by cerebral ischemia. This research demonstrates that baicalin may act as a neuroprotectant during cerebral ischemia.
采用毛细管电泳-激光诱导荧光检测法及异硫氰酸荧光素(FITC)衍生化技术,建立了一种高效、灵敏、快速分析大鼠大脑皮质中氨基酸神经递质的方法。该方法用于研究黄芩苷在脑缺血过程中的药理作用。对影响衍生化和分离的不同参数进行了优化。氨基酸的分离在未涂层熔融石英毛细管(57 cm×75μm内径)中进行,缓冲液为pH 9.2的15 mM硼酸盐,施加电压为17.5 kV。六种氨基酸的检测限在2.1×10⁻¹¹ - 6.3×10⁻¹⁰ M范围内。采用该毛细管电泳-激光诱导荧光检测法研究了三个实验大鼠组大脑皮质中氨基酸神经递质水平的变化。结果表明,脑缺血可导致大脑皮质中Glu、Asp、GABA和Gly浓度显著升高。给予黄芩苷可减轻脑缺血诱导的Glu和Asp升高。本研究表明,黄芩苷在脑缺血期间可能起到神经保护剂的作用。