大鼠脑微透析样品中左旋多巴、多巴胺及其代谢产物、羟自由基的测定

目的: 建立快速、准确、同时测定大鼠脑微透析样品中左旋多巴(L-DOPA)、多巴胺(DA)及其代谢产物、羟自由基指标的高效液相色谱-电化学检测方法。 方法: 应用玻璃碳工作电极、银/氯化银(Ag/AgCl)参比电极、Antec Leyden BV C₁₈分析柱,工作电压为0.52 V,单一流动相,检测大鼠脑微透析样品。 结果: 清醒自由活动大鼠纹状体微透析液中L-DOPA、DA及其代谢产物、羟自由基指标等10种物质在15 min内分离。标准曲线在一定浓度内线性良好。 结论: 所建方法可用于大鼠脑微透析样品中L-DOPA、DA及其代谢产物等的同时测定,为脑内L-DOPA药物浓度-药物效应-药物副作用同步研究提供可用的方法。

Objective: To establish a rapid,accurate HPLC-ECD method to simultaneously detect the L-dihydroxyphenylalanine(L-DOPA),dopamine( DA) and its metabolites,hydroxyl free radical.Method: The analytes were separated on a Antec Leyden BV C₁₈ column(3 μm,2.1 mm×100 mm) under the same mobile phase.The electrochemical detector was equipped with a glass carbon working electrode and a Ag/AgCl reference electrode.The working voltage was 0.52 V.Result: The extracellular L-dihydroxyphenylalanine(L-DOPA),DA and its metabolites,hydroxyl free radical in striatum of free-moving rats can be separated in 15 min.The calibration curves were linear within the limited concentration ranges.Conclusion: This method can be used to determine simultaneously the concentrations of L-DOPA,DA and its metabolites from the brain microdialysis of rats,providing the possible method of simultaneously measuring the concentration of L-DOPA in brain,target efficacy and studying the side effects of drugs.

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