Habibagahi Arezoo, Siddique Shabana, Harris Shelley A, Alderman Nicholas, Aranda-Rodriguez Rocio, Farhat Imen, Chevrier Jonathan, Kubwabo Cariton
Exposure and Biomonitoring Division, Environmental Health Science and Research Bureau, Health Canada, Ottawa, ON, Canada.
Department of Epidemiology & Department of Occupational and Environmental Health, Dalla Lana School of Public Health, University of Toronto, Toronto, ON, Canada.
J Chromatogr B Analyt Technol Biomed Life Sci. 2021 Jan 1;1162:122490. doi: 10.1016/j.jchromb.2020.122490. Epub 2020 Dec 15.
Tobacco use, of which cigarette smoking is the most common, is a global health concern and is directly linked to over 7 million premature deaths annually. Measurement of the levels of tobacco-related biomarkers in biological matrices reflects human exposure to the chemicals in tobacco products. Nicotine, nicotine metabolites, anatabine, and anabasine are specific to tobacco and nicotine containing products. However, as nicotine and its metabolites are ubiquitous in the environment, background contamination during sample preparation can occur, making the quantification of target analytes challenging. The main purpose of the present study was to examine quality control measures needed in the determination of urinary nicotine, nicotine metabolites, anatabine, and anabasine. Urine samples (n = 75) and NIST standard reference materials SRM 3671 and SRM 3672 were analysed. A one-step extraction procedure using cold acetone was used in this study, which involved no additional clean up. The blank matrices investigated included synthetic urine prepared with HPLC-grade water, synthetic urine prepared with Milli-Q water, and bovine urine. By adopting strategies for minimizing the background levels, very low detection limits for all the target analytes ranging from 0.025 ng/mL for 3-hydroxycotinine to 0.634 ng/mL for nicotine, were achieved. Recoveries ranged between 67% and 118% with RSD values below 20%. Intra-day and inter-day precisions were in the range of 1.1-11.7% and 4.8-25.2%, respectively. The levels of all target analytes were higher in daily smokers than in non-smokers, with the largest difference observed for 3-hydroxycotinine. No difference was observed in the levels of target analytes between individuals who were former smokers, who never smoked or who were exposed to environmental tobacco smoke (ETS), except for total nicotine equivalents (TNE), which was significantly higher in non-smokers exposed to environmental tobacco smoke compared with study participants who never smoked. The results obtained from SRM 3671 and SRM 3672 could inform a potential certification of additional biomarkers of exposure to tobacco products in those standard reference materials.
烟草使用，其中吸烟最为常见，是一个全球健康问题，每年直接导致超过700万人过早死亡。测量生物基质中与烟草相关的生物标志物水平反映了人类对烟草制品中化学物质的接触情况。尼古丁、尼古丁代谢物、新烟草碱和假木贼碱是烟草及含尼古丁产品所特有的。然而，由于尼古丁及其代谢物在环境中普遍存在，样品制备过程中可能会发生背景污染，这使得目标分析物的定量具有挑战性。本研究的主要目的是检验测定尿中尼古丁、尼古丁代谢物、新烟草碱和假木贼碱所需的质量控制措施。分析了尿样（n = 75）以及美国国家标准与技术研究院（NIST）标准参考物质SRM 3671和SRM 3672。本研究采用冷丙酮一步萃取法，无需额外净化。所研究的空白基质包括用高效液相色谱（HPLC）级水配制的合成尿、用超纯水（Milli-Q水）配制的合成尿和牛尿。通过采取使背景水平最小化的策略，实现了所有目标分析物的极低检测限，范围从3-羟基可替宁的0.025 ng/mL到尼古丁的0.634 ng/mL。回收率在67%至118%之间，相对标准偏差（RSD）值低于20%。日内精密度和日间精密度分别在1.1 - 11.7%和4.8 - 25.2%范围内。每日吸烟者所有目标分析物的水平均高于非吸烟者，其中3-羟基可替宁的差异最大。除了总尼古丁当量（TNE）外，在曾经吸烟者、从不吸烟者或接触环境烟草烟雾（ETS）的个体之间，目标分析物水平未观察到差异，接触环境烟草烟雾的非吸烟者的总尼古丁当量显著高于从不吸烟的研究参与者。从SRM 3671和SRM 3672获得的结果可为这些标准参考物质中其他烟草制品接触生物标志物的潜在认证提供参考。
