微透析-UPLC-MS/MS法同时测定活体大鼠脑内烟碱及其代谢物  

Simultaneous Determination of Nicotine and its Metabolites in Rat Brain Using Microdialysis Coupled with Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry

作　　者：毛健[1] 卢剑清[2] 徐妍[2] 洪广峰[1] 卢斌斌[1] 孙世豪[1] 宗永立[1] 张建勋[1] 

机构地区：[1]中国烟草总公司郑州烟草研究院 [2]北京理工大学生命学院

出　　处：《烟草科技》2014年第5期37-41,共5页Tobacco Science & Technology

基　　金：国家自然科学基金项目"烟碱在大鼠脑内代谢的活体动态分析及其神经递质化学效应的实时评价"(21307163);郑州烟草研究院院长科技发展基金项目"微透析-LC/MS联用活体动态分析动物体内烟碱及其主要代谢物"(412011CA0280);"MIPs-SPE-HILIC-MSn在线分离鉴定平台的搭建及其在TSNAs分析中的应用"(602011CA0310)

摘　　要：建立了同时测定活体大鼠脑内烟碱（Nic）及其5种代谢物可替宁（Cot）、降烟碱（NNic）、烟碱氮氧化物（NNO）、降可替宁（NCot）和反-3＇-羟基可替宁（3-HC）的微透析-超高效液相色谱-串联质谱（UPLC—MS／MS）联用方法。采用微透析系统采集大鼠脑组织样品，加入内标Nic—d3，Cot—d3，NNic-d。和3-HC—d，后直接进行亲水相互作用色谱（HILIC）分离和多反应监测（MRM）分析。结果表明：Nic及其5种代谢物的日内和日间精密度及准确度均小于11％，检出限和探针回收率分别为0．012～0．029ng／mL和4．7％～9．3％。该方法快速简单、灵敏稳定，适合用于活体分析大鼠脑内的Nic及其代谢物。A method of microdialysis coupled with ultra-performance liquid chromatography tandem mass spectrometry （UPLC-MS/MS） was developed to simultaneously determine the nicotine （Nic） and its metabolites [cotinine （Cot） , nornicotine （NNic） , nicotine-N-oxide （NNO） , norcotinine （NCot） and trans-3-hydroxycotinine （3-HC）] in rat brain. After the addition of internal standards, Nic-d3, Cot-d3, NNic-d4 and 3-HC-d3, the microdialysed sample of rat brain tissue was directly separated by an HILIC column, and determined by mass spectrometry under multi-reaction monitoring （MRM） mode. The results showed that the inter- and intra-day precision and accuracy of Nic, Cot, NNic, NNO, NCot and 3-HC were all less than 11%. The limits of detection ranged from 0.012 to 0.029 ng/mL, and the recoveries of microdialysates ranged from 4.7% to 9.3%. This method is simple, rapid, sensitive, stable, and can be used to quantify nicotine and its metabolites in rat brain in vivo.

关 键 词：烟碱 代谢物 微透析 探针 UPLC—MS MS 鼠脑 活体分析 

分 类 号：TS412[农业科学—烟草工业]