HPLC-MS/MS同时测定大鼠血浆中烟碱、可替宁对映体及其药代动力学研究  

Simultaneous Determination Enantioseparation of Nicotine and Cotinine in Rat Plasma by Chiral HPLC-MS/MS and Pharmacokinetic Study

作　　者：金光祥 韩书磊[1] 陈欢[1] 付亚宁[1] 王红娟[1] 刘彤[1] 陈建[1] 史智浩 侯宏卫[1] 胡清源[1] JIN Guang-xiang;HAN Shu-lei;CHEN Huan;FU Ya-ning;WANG Hong-juan;LIU Tong;CHEN Jian;SHI Zhi-hao;HOU Hong-wei;HU Qing-yuan(China National Tobacco Quality Supervision and Test Center,Zhengzhou 450001,China)

机构地区：[1]国家烟草质量监督检验中心,河南郑州450001

出　　处：《质谱学报》2021年第2期172-180,共9页Journal of Chinese Mass Spectrometry Society

基　　金：国家烟草质量监督检验中心科技创新项目(552017CA0030);中国烟草总公司重大专项项目(110202001006(XX-02),110202001007(XX-03));国家烟草质量监督检验中心烟草控制框架公约科技项目(552018AQ0090)资助。

摘　　要：建立了高效液相色谱-串联质谱法(HPLC-MS/MS)同时测定大鼠血浆中烟碱、可替宁对映异构体,并开展药代动力学研究。样品用甲醇进行蛋白沉淀,Chiralpak IG-3手性色谱柱(250 mm×4.6 mm×3μm)分离,流动相为0.2%甲酸铵-甲醇溶液,流速1.2 mL/min,内标法定量。结果表明,烟碱、可替宁对映异构体能同时实现基线分离,其定量限为0.5μg/L,该方法的线性范围、选择性、准确性、精密度、基质效应、稳定性等均符合方法学验证要求。药代动力学研究结果表明,在大鼠皮下注射烟碱消旋体条件下,烟碱对映异构体的代谢行为无显著性差异,S-可替宁和R-可替宁的生成速率无显著性差异,但S-可替宁的清除速度显著慢于R-可替宁。A high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)method for simultaneous quantification of nicotine and cotinine enantiomers in rat plasma was developed and applied in a stereoselective pharmacokinetic study.The analytes in the rat plasma were extracted by one-step protein precipitation with methanol,then separated on a Chiralpak IG-3 column(250 mm×4.6 mm×3μm)using a mobile phase of 0.2%ammonium formate in methanol at a flow rate of 1.2 mL/min,and finally quantified with isotope internal standard method.The detection was performed on a triple quadrupole tandem mass spectrometer by multiple reaction monitoring mode(MRM)at positive electrospray ionization interface.The results showed that nicotine and cotinine enantiomers were separated by baseline with the limit of quantitation(LOQ)of 0.5μg/L.Parameters of method validation including the linearity,selectivity,accuracy,precision,matrix effect and stability were evaluated.The intra-day and inter-day precision and accuracy at low,medium and high concentration levels exhibited relative standard deviations(RSD)less than 12%and the accuracy ranged from 91.3%to 110.4%.This method is convenient,rapid and suitable for pharmacokinetic study of nicotine enantiomers.The main pharmacokinetic parameters including T max,C max,t 1/2 and AUC were calculated by noncompartment model.After subcutaneous injection of(R,S)-nicotine in rats,pharmacokinetic results suggested that no difference between nicotine enantiomers existed.Metabolic rate of cotinine enantiomers from nicotine was similar,however,the clearance rate of S-cotinine was lower than its antipodes.This study will provide valuable information for further stereoselective pharmacokinetic study and application of nicotine enantiomers.

关 键 词：烟碱 可替宁 对映异构体拆分 高效液相色谱-串联质谱(HPLC-MS/MS) 药代动力学 

分 类 号：O657.63[理学—分析化学]