亲水作用色谱-串联质谱测定尿液中尼古丁和可替宁  

Determination of nicotine and cotinine in urine by hydrophilic interaction chromatography-tandem mass spectrometry

作　　者：李明昕[1] 王倩[1] 朱婧[1] 任琳[2] 雍莉[2] 邹晓莉[1] 

机构地区：[1]四川大学华西公共卫生学院卫生检验与检疫系,四川成都610041 [2]四川省疾病预防控制中心,四川成都610041

出　　处：《色谱》2017年第8期826-831,共6页Chinese Journal of Chromatography

基　　金：四川大学国际合作评估吸烟者牙种植体存活率的前瞻性;非随机化临床研究资助项目~~

摘　　要：建立了测定人尿液中尼古丁和可替宁含量的亲水作用色谱-串联质谱(HILIC-MS/MS)方法。尿样加入尼古丁-d4和可替宁-d3同位素内标后,用水稀释10倍,经过滤后的滤液由超高效液相色谱-串联质谱(UPLC-MS/MS)进行分离分析。采用ACQUITY UPLC~BEH HILIC色谱柱(50 mm×3.0 mm,1.7μm),以甲醇和体积分数为0.1%的氨水为流动相,流速为0.2 mL/min,在电喷雾电离源正离子模式下测定尿液中尼古丁和可替宁的含量,用标准曲线法定量。尼古丁和可替宁在1.0~1 000μg/L范围内线性关系良好,相关系数分别为0.994 9和0.995 8;检出限分别为0.082μg/L和0.077μg/L;定量限分别为0.27μg/L和0.26μg/L;加标回收率分别为90.4%~103.5%和93.0%~104.6%;相对标准偏差分别为4.80%~6.21%和4.22%~7.15%。应用所建立的方法测定了200份尿样,结果表明,吸烟人群尿中尼古丁含量为26.68~854.30μg/L,可替宁含量为36.66~1 191.18μg/L(n=86,M_(nicotine)=76.00μg/L,M_(nicotine)=83.52μg/L,M为中位数);非吸烟人群尿中尼古丁含量为5.08~69.66μg/L,可替宁含量为3.16~28.21μg/L(n=114,Mnicotine=7.53μg/L,M_(nicotine)=3.79μg/L)。该方法快速灵敏,操作简单,适用于尿样中尼古丁和可替宁的批量测定,能满足烟草暴露评价的需要。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(4) and cotinine-d(3) 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 (R) BEH HILIC column (50 mmx3.0 mm, 1.7 mu 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 1.0-1 000 mu g/L for nicotine and cotinine with the linear coefficients of 0.994 9 and 0.995 8, respectively. The limits of detection of nicotine and cotinine were 0.082 mu g/L and 0.077 mu g/L, and the limits of quantification were 0.27 mu g/L and 0.26 mu 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 cotininerespectively. 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 mu g/L, and the cotinine contents were 36.66-1 191.18 mu g/L (n = 86, M-nicotine = 76.00 mu g/L, M-cotinine = 83.52 mu g/L, M: median); of the nonsmoking people, the nicotine con. tents were 5.08 -69.66 mu g/L, and the cotinine contents were 3.16 -28.21 mu g/L (n = 114, M = 7.53 mu g/L, M (cotinine) = 3.79 mu 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.

关 键 词：亲水作用色谱-串联质谱 同位素稀释 尼古丁 可替宁 尿液 

分 类 号：O658[理学—分析化学]