高效液相色谱荧光法同时测定小鼠脑组织中4种.doc

高效液相色谱荧光法同时测定小鼠脑组织中4种氨基酸类神经递质( 桂 莉1(, 田 洪1,郑 健1,段 炜1,叶 枫2 （第三军医大学：1新桥医院神经内科，重庆 400037，2军事预防医学院防化教研室，重庆 400038） 摘 要:目的　采用高效液相-荧光检测技术建立一种稳定的用于测定脑组织中多种氨基酸类神经递质的方法。方法　用微透析技术收集小鼠脑组织中多种氨基酸类神经递质，用邻苯二甲醛(OPA)柱前衍生高效液相色谱（high performance liquid chromatography，HPLC）法测定微透析液中天门冬氨酸（Asp）、谷氨酸(Glu)、甘氨酸(Gly)、γ-氨基丁酸(GABA)的含量。结果　兴奋性Asp、Glu及抑制性Gly、GABA在18 min内完全分离，在0.625～5μmol/L浓度范围内与峰面积有良好的线性关系。结论　HPLC法快速、准确、灵敏度高，适用于脑组织中氨基酸类神经递质含量的测定。 关键词:高效液相色谱；柱前衍生；微透析；氨基酸；神经递质 中图法分类号： 文献标识码：A Simultaneously determining 4 amino acid neurotransmitters in mice brain by high-performance liquid chromatography with fluorescence detector GUI Li1, TIAN Hong1, ZHENG Jian1, DUAN Wei1, YE Feng2 (1Department of Neurology, Xinqiao Hospital, Third Military Medical University, Chongqing 400037; 2Department of Chemical Defense, College of Military Preventive Medicine, Third Military Medical University, Chongqing 400038, China) Abstract: Objective To establish a stable method of HPLC-FL to determine multi-amino acids in the mice microdialysate. Methods The sample was collected by microdialysis. Four amino acid neurotransmitters, Asp, Glu, Gly and GABA in the microdialysate were determined by, reverse phase HPLC with fluorescence detection after o-Phthaldehyde (OPA) pre-column derivatization. Results Excitatory amino acids, Asp and Glu, and inhibitory amino acids, Gly and gamma-aminobutyric acid (GABA) were totally separated and determined within 18 min. A good linear response was obtained for the 4 standards over the range of 0. 625 to 5 μmol/L with correlation coeffi- cients. Conclusion Our method is quick, reliable and sensitive to measure the amino acid neurotransmitters in mice brain. Key words: high performance liquid chromatography; precolumn derivatization; microdialysis; amino acid; neurotransmitters 中枢神经系统内大部分神经递质是氨基酸类神经递质,其中谷氨酸(Glu)和天门冬氨酸(Asp)是脑部的重要兴奋性神经递质,γ-氨基丁酸(GABA)、甘氨酸(Gly)是脑部重要的抑制性神经递质，这些氨基酸类神经递质存在于脑组织和脑脊液中，其含量及比例的变化伴随着多种中枢神经系统疾病[1]，还反映出许多疾病的病理、生理过程。因此，测定这些氨基酸类神经递质含量变化对于神