HPLC-ECD法同时测定海洛因急性中毒大鼠不同脑区中单胺类神经递质含量  

Simultaneous Determination of Monoamine Neurotransmitters in Different Brain Regions of Rats With Acute heroin Poisoning by HPLC-ECD

作　　者：陈园园 李锋 魏佳韵 李香豫 王丹[2,3] 燕瑾 王优美 徐鹏 狄斌 CHEN Yuan-yuan;LI Feng;WEI Jia-yun;LI Xiang-yu;WANG Dan;YAN Jin;WANG You-mei;XU Peng;DI Bin(China Pharm aceutical University,Nanjing 210009;Office of China National Narcotics Control Commission-China Pharm aceutical University Joint Laboratory on Key Technologies of Narcotics Control,Beijing 100193;Key Laboratory of Drug Monitoring and Control,Drug Intelligence and Forensic Center,Ministry of Public Security,Beijing 100193)

机构地区：[1]中国药科大学,南京210009 [2]国家禁毒委员会办公室中国药科大学禁毒关键技术联合实验室,北京100193 [3]毒品监测管控与禁毒关键技术公安部重点实验室,公安部禁毒情报技术中心,北京100193

出　　处：《中国药物滥用防治杂志》2021年第1期53-58,共6页Chinese Journal of Drug Abuse Prevention and Treatment

基　　金：十三五国家重点研发计划(2017YFC0803605,2018YFC0807402)支持。

摘　　要：目的:建立高效液相色谱-电化学检测器(HPLC-ECD)法同时测定大鼠伏隔核和纹状体两个不同脑区中单胺类神经递质多巴胺(DA)和5-羟色胺(5-HT)含量的方法,并对海洛因急性中毒大鼠不同脑区中单胺类神经递质的含量变化进行研究。方法:高效液相色谱采用QUATTRO C18色谱柱,以磷酸二氢钠-柠檬酸-辛烷磺酸钠缓冲液∶乙腈(94∶6)为流动相,流速为0.3 ml/min,进样量为20μl,考察本方法的专属性、线性范围、精密度、回收率和稳定性。测定海洛因急性中毒大鼠伏隔核和纹状体两个脑区中DA和5-HT的含量变化。结果:DA和5-HT的检测限均为0.2 ng/ml,定量限均为0.5 ng/ml;日内与日间精密度RSD均<5%;回收率范围87.8%~115.5%;4℃条件下,在脑组织匀浆液中4天内稳定性良好。给予急性中毒剂量海洛因后,大鼠两个脑区中的DA含量基本无变化,但5-HT含量均呈剂量依赖性显著升高。结论:该方法准确、灵敏、稳定,适用于大鼠不同脑区中DA和5-HT含量的测定。首次发现海洛因急性毒性作用可能与伏隔核和纹状体中5-HT含量显著升高有关,具体机制有待于进一步研究验证。Objective:To establish a HPLC-ECD method to simultaneously detect the contents of monoamine neurotransmitters dopamine(DA)and serotonin(5-HT)in the nucleus accumbens and striatum of rats and apply it to study the content changes of DA and 5-HT in different brain areas of rats with heroin acute poisoning.Methods:HPLC adopted QUATTRO C18 column,and used sodium dihydrogen phosphate-citric acid-sodium octane sulfonate buffer∶acetonitrile(94∶6)as the mobile phase,the flow rate was 0.3 ml/min,and the injection volume was 20μl.The specificity,linear range,precision,recovery and stability of this method were investigated and the contents of DA and 5-HT in the nucleus accumbens and striatum of rats with acute heroin poisoning were measured.Results:The detection limit and quantitative limit of DA and 5-HT were both 0.2 ng/ml and 0.5 ng/ml,respectively.Intraday and interday precisions(RSDs)were less than 5%;The recovery range was 87.8% to 115.5%;The brain tissue samples were stable after being placed in 4℃ for 4 d.After administration of acute toxic doses of heroin,the DA contents in the two brain regions of the rats were basically unchanged,but the 5-HT contents increased significantly in a dose-dependent manner.Conclusion:The method is accurate,sensitive and stable,and is suitable for the determination of DA and 5-HT contents in different brain regions of rats.It was first discovered that the acute toxicity of heroin may be related to the significant increase of 5-HT contents in the nucleus accumbens and striatum.The specific mechanism needs to be further studied and verified.

关 键 词：HPLC-ECD 伏隔核 纹状体 单胺类神经递质 海洛因 毒性 

分 类 号：R749.64[医药卫生—神经病学与精神病学]