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Fast and sample cleanup-free measurement of nicotine and cotinine by stable isotope dilution ultra-performance liquid chromatography-tandem mass spectrometry.
J Pharm Biomed Anal. 2012 Aug-Sep; 67-68:137-43.JP

Abstract

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 μg/L (r>0.999). Limits of Detection were 0.7 μg/L for nicotine and 0.4 μg/L for cotinine, and Lower Limits of Quantification were 1.7 μg/L for nicotine and 1.1 μg/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 [μg/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.

Authors+Show Affiliations

Institut für Laboratoriums- und Transfusionsmedizin, Herz- und Diabeteszentrum Nordrhein-Westfalen, Universitätsklinik der Ruhr-Universität Bochum, 32545 Bad Oeynhausen, Germany. jkuhn@hdz-nrw.deNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Validation Study

Language

eng

PubMed ID

22608097

Citation

Kuhn, Joachim, et al. "Fast and Sample Cleanup-free Measurement of Nicotine and Cotinine By Stable Isotope Dilution Ultra-performance Liquid Chromatography-tandem Mass Spectrometry." Journal of Pharmaceutical and Biomedical Analysis, vol. 67-68, 2012, pp. 137-43.
Kuhn J, Vollmer T, Martin C, et al. Fast and sample cleanup-free measurement of nicotine and cotinine by stable isotope dilution ultra-performance liquid chromatography-tandem mass spectrometry. J Pharm Biomed Anal. 2012;67-68:137-43.
Kuhn, J., Vollmer, T., Martin, C., Hendig, D., & Knabbe, C. (2012). Fast and sample cleanup-free measurement of nicotine and cotinine by stable isotope dilution ultra-performance liquid chromatography-tandem mass spectrometry. Journal of Pharmaceutical and Biomedical Analysis, 67-68, 137-43. https://doi.org/10.1016/j.jpba.2012.04.036
Kuhn J, et al. Fast and Sample Cleanup-free Measurement of Nicotine and Cotinine By Stable Isotope Dilution Ultra-performance Liquid Chromatography-tandem Mass Spectrometry. J Pharm Biomed Anal. 2012 Aug-Sep;67-68:137-43. PubMed PMID: 22608097.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Fast and sample cleanup-free measurement of nicotine and cotinine by stable isotope dilution ultra-performance liquid chromatography-tandem mass spectrometry. AU - Kuhn,Joachim, AU - Vollmer,Tanja, AU - Martin,Claudia, AU - Hendig,Doris, AU - Knabbe,Cornelius, Y1 - 2012/05/03/ PY - 2012/02/14/received PY - 2012/04/24/revised PY - 2012/04/25/accepted PY - 2012/5/22/entrez PY - 2012/5/23/pubmed PY - 2012/10/12/medline SP - 137 EP - 43 JF - Journal of pharmaceutical and biomedical analysis JO - J Pharm Biomed Anal VL - 67-68 N2 - 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 μg/L (r>0.999). Limits of Detection were 0.7 μg/L for nicotine and 0.4 μg/L for cotinine, and Lower Limits of Quantification were 1.7 μg/L for nicotine and 1.1 μg/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 [μg/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. SN - 1873-264X UR - https://www.unboundmedicine.com/medline/citation/22608097/Fast_and_sample_cleanup_free_measurement_of_nicotine_and_cotinine_by_stable_isotope_dilution_ultra_performance_liquid_chromatography_tandem_mass_spectrometry_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0731-7085(12)00233-6 DB - PRIME DP - Unbound Medicine ER -