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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 perform-ance 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。
Objective To establish a stable method for the determination of various amino acid neurotransmitters in brain tissue by high performance liquid chromatography-fluorescence detection. METHODS: A variety of amino acid neurotransmitters were collected from mouse brain tissue using microdialysis technique. The content of neurotransmitters in mouse dialysate was determined by high performance-liquid chromatography (HPLC) with OPA. Asp, Glu, Gly and GABA were measured. Results Excitatory Asp, Glu and inhibitory Gly and GABA were completely separated within 18 min. There was a good linear relationship between peak area and concentration of Asp, Glu in the range of 0.625 ~ 5μmol / L. Conclusion HPLC method is rapid, accurate and sensitive and suitable for the determination of amino acid neurotransmitters in brain tissue.