Li Mingxin, Wang Qian, Zhu Jing, Ren Lin, Yong Li, Zou Xiaoli
Department of Laboratory Techonology and Science of Public Health, West China School of Public Health, Sichuan University, Chengdu 610041, China.
Center for Disease Control and Prevention of Sichuan Province, Chengdu 610041, China.
Se Pu. 2017 Aug 8;35(8):826-831. doi: 10.3724/SP.J.1123.2017.05019.
The analytical method of nicotine and cotinine in human urine with hydrophilic interaction chromatography-tandem mass spectrometry (HILIC-MS/MS) was established. After the urine sample containing nicotine-d and cotinine-d isotope internal standards being diluted with water, the filtrate was introduced into ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) for analysis. Separation was performed on an ACQUITY UPLC BEH HILIC column (50 mm×3.0 mm, 1.7 μ m), in which methanol and 0.1% (v/v) ammonia were used as the mobile phases with isocratic elution at 0.2 mL/min of flow rate. Positive ion scan mode was used for mass spectrometry measurement and calibration curves were plotted for quantification determination. A good linearity could be obtained in the range of 1.0-1000 μ g/L for nicotine and cotinine with the linear coefficients of 0.9949 and 0.9958, respectively. The limits of detection of nicotine and cotinine were 0.082 μ g/L and 0.077 μ g/L, and the limits of quantification were 0.27 μ g/L and 0.26 μ g/L, respectively. The recoveries of the spiked urine samples were 90.4%-103.5% and 93.0%-104.6%, and the relative standard deviations (RSDs) were 4.80%-6.21% and 4.22%-7.15% for nicotine and cotinine respectively. The established method was applied to the analyis of 200 urine samples. Based on the investigation information of the urine of the smoking people, the nicotine contents were 26.68-854.30 μ g/L, and the cotinine contents were 36.66-1191.18 μ g/L (=86, =76.00 μ g/L, =83.52 μ g/L, :median); of the nonsmoking people, the nicotine contents were 5.08-69.66 μ g/L, and the cotinine contents were 3.16-28.21 μ g/L (=114, =7.53 μ g/L, =3.79 μ g/L). The method is simple, sensitive and rapid. It is suitable for batch analysis of nicotine and cotinine in urine, and it can meet the requirement of evaluating the human tobacco exposure.
建立了亲水作用色谱-串联质谱法（HILIC-MS/MS）测定人尿液中尼古丁和可替宁的分析方法。将含有尼古丁-d和可替宁-d同位素内标的尿液样品用水稀释后，取滤液引入超高效液相色谱-串联质谱仪（UPLC-MS/MS）进行分析。采用ACQUITY UPLC BEH HILIC色谱柱（50 mm×3.0 mm，1.7μm）进行分离，以甲醇和0.1%（v/v）氨水为流动相，等度洗脱，流速为0.2 mL/min。质谱测定采用正离子扫描模式，并绘制校准曲线进行定量测定。尼古丁和可替宁在1.0 - 1000μg/L范围内线性良好，线性系数分别为0.9949和0.9958。尼古丁和可替宁的检测限分别为0.082μg/L和0.077μg/L，定量限分别为0.27μg/L和0.26μg/L。加标尿液样品的回收率分别为90.4% - 103.5%和93.0% - 104.6%，尼古丁和可替宁的相对标准偏差（RSD）分别为4.80% - 6.21%和4.22% - 7.15%。所建立的方法应用于200份尿液样品的分析。根据吸烟人群尿液的调查信息，尼古丁含量为26.68 - 854.30μg/L，可替宁含量为36.66 - 1191.18μg/L（=86，=76.00μg/L，=83.52μg/L，：中位数）；非吸烟人群中，尼古丁含量为5.08 - 69.66μg/L，可替宁含量为3.16 - 28.21μg/L（=114，=7.53μg/L，=3.79μg/L）。该方法简便、灵敏、快速。适用于尿液中尼古丁和可替宁的批量分析，能够满足评估人体烟草暴露的要求。
