高效液相色谱-电化学法测定大鼠脑微透析液_中多巴胺的方法建立及应用

目的: 建立高效液相色谱-电化学(HPLC-ECD)法测定大鼠脑脊液中多巴胺的含量。 方法: 高效液相色谱采用Waters Atlantis T3色谱柱,以乙腈-缓冲液 (75 mmol·L^-1三水合醋酸钠,24 mmol·L^-1无水柠檬酸溶液,1.2 mmol·L^-1十二烷基硫酸钠溶液,0.5 mmol·L^-1 EDTA-2Na溶液)(20: 80)为流动相,流动相pH 5.4,流速0.32 mL·min^-1。考察本方法的专属性、线性范围、精密度和回收率。进一步比较第一代(氟哌啶醇)及第二代(利培酮)抗精神病药对大鼠前额叶皮层(mPFC)和伏隔核(NAC)中多巴胺释放的影响。 结果: 多巴胺的保留时间为6.6 min,线性范围是0.2~20 nmol·L^-1(r=0.9998),日内与日间精密度均<7.3%,回收率在101.4~106.2%之间。给予氟哌啶醇后,mPFC中多巴胺含量基本无变化,NAC中多巴胺显著升高;给予利培酮后,mPFC和NAC中多巴胺均显著升高。 结论: 该方法准确、灵敏、稳定,适用于脑微透析样品中多巴胺的定量分析。第二代抗精神病药物(利培酮)可能通过升高前额叶皮层多巴胺水平改善阴性症状及认知功能。

Objective: To develop an HPLC-ECD method for determination of dopamine in rat cerebral microdialysates. Methods: A Waters Atlantis T3 column was adopted in the HPLC system.The mobile phase was acetonitrile-buffer solution(75 mmol·L^-1 sodium acetate trihydrate,24 mmol·L^-1 anhydrous citric acid, 1.2 mmol·L^-1 dodecyl sulfate sodium salt and 0.5 mmol·L^-1 EDTA-Na₂)with pH 5.4(20: 80).The flow rate was 0.32 mL·min^-1. Selectivity,linear range,precision and recovery were evaluated to investigate the reliability of this method.dopamine was determined with microdialysates in order to study the effects of haloperidol and risperidone(1 mg·kg^-1)s.c.on extracellular dopamine levels in the mPFC and the NAC. Results: The retention time of dopamine was 6.6 min;Good linearity(r=0.9998)was obtained over the range of 0.2 nmol·L^-1-20 nmol·L^-1;Intraday and interday precisions(RSDs)were less than 7.3%.Haloperidol increased extracellular dopamine levels in the NAC,significantly but not in the mPFC.Risperidone significantly increased extracellular dopamine levels in both NAC and mPFC. Conclusion: The HPLC-ECD method is rapid,sensitive and specific for quantitative determination of dopamine.