亲水作用液相色谱串联质谱法测定头发中尼古丁和可替宁

Detecting Nicotine and Cortinine in Hair by Hydrophilic Interaction Liquid Chromatography Tandem Mass Spectrometry

目的建立亲水作用色谱串联质谱法测定头发中的尼古丁和可替宁的方法。方法样品在氢氧化钠溶液中水解,二氯甲烷萃取,氮气吹干,流动相复融后注入色谱-质谱系统分析。采用亲水作用色谱柱分离,甲醇-0.1%氨水为流动相,电子喷雾电离-三重四极杆质谱完成定量检测。采用建立的方法测定602例孕妇头发样品、31例志愿者头发和尿样中尼古丁和可替宁含量。结果采用本实验建立的亲水作用液相色谱串联质谱法检测样品,尼古丁和可替宁在0.030~100.000μg/L范围内线性良好,方法检出限分别为0.007 6μg/g和0.004 4μg/g。日间和日内精密度小于10%。样品加标回收率为81.0%~102.0%。602例孕妇头发样品中非暴露二手烟孕妇头发中的尼古丁和可替宁质量浓度为0.020~0.260μg/g和0.004 8~0.069 0μg/g,二手烟暴露孕妇头发中的尼古丁和可替宁质量浓度为0.029~0.350μg/g和0.005 6~0.085 0μg/g。志愿者报告的吸烟状态与样品中的尼古丁和可替宁含量均具有相关性(P<0.05);暴露的吸烟支数与头发中尼古丁含量关系密切,两者具有线性关系。结论本研究建立的方法准确灵敏,适用于头发中尼古丁和可替宁检测。头发中的尼古丁水平可能是评估烟草暴露剂量的特定生物标志物。

Objective To develop a method for detecting nicotine and cotinine in hair by hydrophilic interaction chromatography tandem mass spectrometry.Methods Hair samples were hydrolyzed in sodium hydroxide solution before extraction with dichloromethane.The samples were blown to dry with nitrogen and dissolved with mobile phase.The filtrate of the samples was injected into a chromatographic-mass spectrometry system for analysis.The separation was performed by a hydrophilic column,with which methanol-0.1%ammonia was used as the mobile phase.The quantitative detection of Nicotine and Cortinine was carried out with electron spray ionization-triple quadrupole mass spectrometry.The established method was used for detecting nicotine and cotinine in 602 hair samples of pregnant women and 31 hair and urine samples of volunteers.Results A standard curve was drawn for the established method of hydrophilic liquid chromatography tandem mass spectrometry.Good linearity was obtained for detecting nicotine and cotinine in the range of 0.030-100.000μg/L,with a detection limit(MDL)of 0.007 6μg/g and 0.004 4μg/g,respectively.The inter-day and intra-day precisions reached a level of less than 10%.The recoveries of the spiked samples ranged from 81.0%to 102.0%.About 0.020-0.260μg/g nicotine and 0.004 8-0.069 0μg/g cotinine were detected in the pregnant women without exposure to secondhand smoking(SHS),compared with 0.029-0.350μg/g nicotine and 0.005 6-0.085 0μg/g cotinine in those exposed to SHS.Nicotine and cotinine were also found in the hair and urine samples of volunteers,which were correlated with smoking(P<0.05).A dose-response relationship were found between smoking and hair nicotine.Conclusion The proposed method is accurate and sensitive for detecting nicotine and cotinine in hair samples.Hair nicotine can be a specific biomarker for assessing exposure to tobacco smoking.