高效液相色谱-电化学法测定大鼠纹状体细胞外液中单胺类神经递质含量

目的: 建立应用常规高效液相色谱-电化学法(HPLC-ECD)测定大鼠脑微透析液中单胺类神经递质及其代谢产物。 方法: 将探针插入大鼠右侧纹状体,在清醒自由活动状态下,用Ringer氏液以1.5 μL·min^-1 的速度灌流,每20 min收集l管透析液,将其注入HPLC-ECD仪,考察本方法的专属性、线性范围、精密度和准确度等,并对其中所含的NE、DA、DOPAC 、HIAA、HVA 和 5-HT 进行含 量测定。 结果: 各递质在0.025~0.5 ng·μL^-1 范围内线性关系良好,方法加样回收完全,日间日内测定均RSD<3.8％。 结论: 本实验选用了高效液相色谱与电化学检测器连用的方法测定神经递质的含量,该方法简单准确、灵敏、特异性好。

Objective: To develop simple,rapid and sensitive HPLC-ECD method of monoamine neurotransmitter and its metabolites in rat cerebral microdialysates. Methods: Microdialysis probes were placed into the right stratums of SD rat brains and perfused with Ringers solution at a rate of 1.5 μL·min^-1.A reverse phase HPLC with electrochemical detection(ECD)were used to assay NE,DA,DOPAC,HIAA,HVA and 5-HT after the cerebral microdialysates were collected every
[20] minutes from awake,freely moving rats.In order to know the reliability of this method,selectivity,linear range,precision and accuracy were tested and the contents of NE,DA,DOPAC,HIAA,HVA and 5-HT in rat microdialysates were determined. Results: These six components were all found to be linear over a concentration range of 0.025-0.5 ng·μL^-1,relative recovery was more than 96.34％,and intra-day and inter-day precision(RSD)were less than 3.8%. Conclusion: The HPLC-ECD method is rapid,sensitive and specific for quantitative determination of monoamine neurotransmitter and its metabolites