Fast and sample cleanup-free measurement of nicotine and cotinine by stable isotope dilution ultra-performance liquid chromatography-tandem mass spectrometry

详细信息	查看全文 | 推荐本文 |

• 作者：Joachim Kuhn^a ; ^{jkuhn@hdz-nrw.de} ; Tanja Vollmer^a ; Claudia Martin^b ; Doris Hendig^a ; Cornelius Knabbe^a
• 关键词：LOD ; Limits of Detection ; LLOQ ; Lower Limit of Quantification ; UPLC&ndash ; MS/MS ; ultra-performance liquid chromatography&ndash ; tandem mass spectrometry
• 刊名：Journal of Pharmaceutical and Biomedical Analysis
• 出版年：2012
• 期刊代码：65_07317085
• 类别：ch
• 出版时间：August-September, 2012
• 卷：67-68
• 期：Complete
• 页码：137-143
• 文件大小：389 K

摘要

We developed a stable isotope dilution ultra-performance liquid chromatography-electrospray ionization tandem mass spectrometry assay to measure nicotine and cotinine, the major oxidative and pharmacologically less active metabolite of nicotine, in human urine. A simple dilution step was used as sample preparation and the measurement of nicotine and cotinine was performed during a 1.5-min run-time using nicotine-d₄ and cotinine-d₄ as internal standards. Multiple calibration curves for the analysis of both nicotine and cotinine exhibited a consistent excellent linearity and reproducibility in the range of 5-35,000 渭g/L (r > 0.999). Limits of Detection were 0.7 渭g/L for nicotine and 0.4 渭g/L for cotinine, and Lower Limits of Quantification were 1.7 渭g/L for nicotine and 1.1 渭g/L for cotinine. The intraassay coefficients of variation (CVs) for nicotine and cotinine were <4%and <2%, respectively, the interassay CVs were <6%for nicotine and <4%for cotinine. The inaccuracy was <6%for both substances. The mean recovery was 103.2%(range 96.8-105.1%) for nicotine and 97.4%(range 94.3-99.2%) for cotinine. A method comparison showed that the values of nicotine metabolites in human urine samples (n = 98) measured by a commercially available chemiluminescent immunoassay tested on analyzer IMMULITE 2000 were much higher than the cotinine concentration in the same urine samples measured by our UPLC-MS/MS assay. The Passing-Bablok regression line was: immunoassay = 4.62 (UPLC-MS/MS) + 3.64 [渭g/L]; r = 0.75. This robust, sensitive and interference-free UPLC-MS/MS assay permits rapid and accurate determination of nicotine and cotinine in human urine.