大鼠脑组织单胺类递质及其代谢产物的检测方法研究

Research on testing method for monomine transmitters and metabolite in rat brain tissue

目的 :研究大鼠脑组织中单胺类递质及其代谢产物的高效液相反相离子对色谱测定法。方法 :采用LiChrosorbC18,10 μm色谱柱 ,流动相为甲醇 :水 (4 0 :60 ) ,含 0 .0 2 8g LEDTANa2 ,0 .15g LSDS ,0 .2ml LH2 SO4(pH 2 .5～ 3 ) ,荧光检测波长 :λEX=2 85 ,λEM=3 3 3。结果 :对 87只大鼠脑组织中 4种单胺类递质及其代谢产物的含量进行了同时测定 ,高香草酸 (HVA) 2 .5 0～ 40 .0 μg ml、去甲肾上腺素 (NE) 0 .0 1～ 0 .5 0 μg ml、多巴胺 (DA)0 .0 5～ 1.0 0 μg ml、5 羟色胺 (5 HT) 0 .0 2 5～ 0 .5 0 μg ml,峰面积与其含量呈良好的线性关系。 结论 :该法操作简便、快速、准确 ,为组织中单胺类递质及其代谢产物检测的一种理想方法 ,并适用于临床相关研究。

Objective:To establish an ion pair RP HPLC method for testing monomine transmitters and metabolite concentration in the brain tissues of rats.Methods:The tissue samples were dissolved with a solution (0.1?mol HClO 4 , 0.05% EDTANa 2) and Lichrosorb Column (C 18 ,10?μm,25?cm×4.6?mm) was employed.Results:The monomine transmitters and metabolite of four components can be simultaneously determined in 89 rats brain tissue. The calibration curves were linear in the range of 2.50～40.0?μg.ml -1 for HVA , 0.01～0.50?μg. ml -1 for NE , 0.05～1.00?μg.ml -1 for DA ,and 0.025～0.50?μg.ml -1 for 5 HT respectively .Conclusion:This method is simple , fast , accurate , and can be used to determine simultaneously the concentrations of monomine transmitters and metabolite in brain tissue of rats and is applicabte to relevant clinical studies .