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

Simultaneously determining 4 amino acid neurotransmitters in mice brain by high-performance liquid chromatography with fluorescence detector

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 of HPLC-FL to determine muhi-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 de- tection after o-Phthaldehyde （OPA） prc-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 rain. A good linear response was obtained for the 4 standards over the range of 0. 625 to 5 μmol/L with correlation coefficients. Conclusion Our method is quick, reliable and sensitive to measure the amino acid neurotransmitters in mice brain.