Center for Human Toxicology, Department of Pharmacology and Toxicology, University of Utah, Salt Lake City, USA.
J Anal Toxicol. 2011 Jul;35(6):321-32. doi: 10.1093/anatox/35.6.321.
The aim of this exploratory study was to develop and validate a liquid chromatography-tandem mass spectrometry (LC-MS-MS) method for the quantification of nicotine, eight nicotine metabolites, and two minor tobacco alkaloids in fortified analyte-free hair and subsequently apply this method to hair samples collected from active smokers. An additional aim of the study was to include an evaluation of different wash procedures for the effective removal of environmentally deposited nicotine from tobacco smoke. An apparatus was designed for the purpose of exposing analyte-free hair to environmental tobacco smoke in order to deposit nicotine onto the hair surface. A shampoo/water wash procedure was identified as the most effective means of removing nicotine. This wash procedure was utilized for a comparison of washed and unwashed heavy smoker hair samples. Analytes and corresponding deuterated internal standards were extracted using a cation-exchange solid-phase cartridge. LC-MS-MS was carried out using an Acquity™ UPLC(®) system (Waters) and a Quattro Premier XE™ triple quadrupole MS (Waters) operated in electrospray positive ionization mode, with multiple reaction monitoring data acquisition. The developed method was applied to hair samples collected from heavy smokers (n = 3) and low-level smokers (n = 3) collected through IRB-approved protocols. Nicotine, cotinine, and nornicotine were quantified in both the washed and unwashed hair samples collected from three heavy smokers, whereas 3-hydroxycotinine was quantified in only one unwashed sample and nicotine-1'-oxide in the washed and unwashed hair samples from two heavy smokers. In contrast, nicotine-1'-oxide was quantified in one of the three low-level smoker samples; nicotine was quantified in the other two low-level smoker samples. No other analytes were detected in the hair of the three low-level smokers.
本探索性研究旨在开发和验证一种用于定量分析加标无分析物头发中尼古丁、八种尼古丁代谢物和两种次要烟草生物碱的液相色谱-串联质谱（LC-MS-MS）方法，并将该方法应用于从主动吸烟人群中采集的头发样本。该研究的另一个目的是评估不同的洗涤程序，以有效去除环境中烟草烟雾沉积的尼古丁。设计了一种装置，用于使无分析物头发暴露于环境烟草烟雾中，以将尼古丁沉积在头发表面上。发现洗发水/水洗程序是去除尼古丁的最有效方法。该洗涤程序用于比较经过洗涤和未经洗涤的重度吸烟者头发样本。使用阳离子交换固相萃取小柱提取分析物和相应的氘代内标。采用 Acquity™UPLC（®）系统（Waters）和 Quattro Premier XE™三重四极杆 MS（Waters）进行 LC-MS-MS 分析，采用电喷雾正离子化模式，采用多重反应监测数据采集。该方法应用于通过 IRB 批准的方案采集的重度吸烟者（n=3）和低水平吸烟者（n=3）的头发样本。在来自三个重度吸烟者的经洗涤和未经洗涤的头发样本中均定量了尼古丁、可替宁和去甲烟碱，而 3-羟基可替宁仅在一个未经洗涤的样本中定量，尼古丁-1'-氧化物在两个重度吸烟者的经洗涤和未经洗涤的头发样本中定量。相比之下，尼古丁-1'-氧化物在三个低水平吸烟者样本中的一个中定量，尼古丁在其他两个低水平吸烟者样本中定量。在三个低水平吸烟者的头发中未检测到其他分析物。
