Pramod Upadhyaya1, Stephen S Hecht2. 1. Masonic Cancer Center, University of Minnesota, Minneapolis, MN. 2. Masonic Cancer Center, University of Minnesota, Minneapolis, MN hecht002@umn.edu.
Abstract
INTRODUCTION: Based on previous metabolism studies carried out in patas monkeys, we hypothesized that urinary 3'-hydroxynorcotinine could be a specific biomarker for uptake and metabolism of the carcinogen N'-nitrosonornicotine in people who use tobacco products. METHODS: We developed a method for quantitation of 3'-hydroxynorcotinine in human urine. [Pyrrolidinone-(13)C4]3'-hydroxynorcotinine was added to urine as an internal standard, the samples were treated with β-glucuronidase, partially purified by solid supported liquid extraction and quantified by liquid chromatography-electrospray ionization-tandem mass spectrometry. RESULTS: The method was accurate (average accuracy = 102%) and precise (coefficient of variation = 5.6%) in the range of measurement. 3'-Hydroxynorcotinine was detected in 48 urine samples from smokers (mean 393±287 pmol/ml urine) and 12 samples from individuals who had stopped smoking and were using the nicotine patch (mean 658±491 pmol/ml urine), but not in any of 10 samples from nonsmokers. CONCLUSIONS: Since the amounts of 3'-hydroxynorcotinine found in smokers' urine were approximately 50 times greater than the anticipated daily dose of N'-nitrosonornicotine, we concluded that it is a metabolite of nicotine or one of its metabolites, comprising perhaps 1% of nicotine intake in smokers. Therefore, it would not be suitable as a specific biomarker for uptake and metabolism of N'-nitrosonornicotine. Since 3'-hydroxynorcotinine has never been previously reported as a constituent of human urine, further studies are required to determine its source and mode of formation.
INTRODUCTION: Based on previous metabolism studies carried out in patas monkeys, we hypothesized that urinary 3'-hydroxynorcotinine could be a specific biomarker for uptake and metabolism of the carcinogen N'-nitrosonornicotine in people who use tobacco products. METHODS: We developed a method for quantitation of 3'-hydroxynorcotinine in human urine. [Pyrrolidinone-(13)C4]3'-hydroxynorcotinine was added to urine as an internal standard, the samples were treated with β-glucuronidase, partially purified by solid supported liquid extraction and quantified by liquid chromatography-electrospray ionization-tandem mass spectrometry. RESULTS: The method was accurate (average accuracy = 102%) and precise (coefficient of variation = 5.6%) in the range of measurement. 3'-Hydroxynorcotinine was detected in 48 urine samples from smokers (mean 393±287 pmol/ml urine) and 12 samples from individuals who had stopped smoking and were using the nicotine patch (mean 658±491 pmol/ml urine), but not in any of 10 samples from nonsmokers. CONCLUSIONS: Since the amounts of 3'-hydroxynorcotinine found in smokers' urine were approximately 50 times greater than the anticipated daily dose of N'-nitrosonornicotine, we concluded that it is a metabolite of nicotine or one of its metabolites, comprising perhaps 1% of nicotine intake in smokers. Therefore, it would not be suitable as a specific biomarker for uptake and metabolism of N'-nitrosonornicotine. Since 3'-hydroxynorcotinine has never been previously reported as a constituent of human urine, further studies are required to determine its source and mode of formation.
Authors: Lijiao Zhao; Silvia Balbo; Mingyao Wang; Pramod Upadhyaya; Samir S Khariwala; Peter W Villalta; Stephen S Hecht Journal: Chem Res Toxicol Date: 2013-09-18 Impact factor: 3.739
Authors: Sharon E Murphy; Sung-Shim L Park; Elizabeth F Thompson; Lynne R Wilkens; Yesha Patel; Daniel O Stram; Loic Le Marchand Journal: Carcinogenesis Date: 2014-09-18 Impact factor: 4.944
Authors: Silvia Balbo; Sandra James-Yi; Charles S Johnson; Michael G O'Sullivan; Irina Stepanov; Mingyao Wang; Dipankar Bandyopadhyay; Fekadu Kassie; Steven Carmella; Pramod Upadhyaya; Stephen S Hecht Journal: Carcinogenesis Date: 2013-05-13 Impact factor: 4.944
Authors: Stephen S Hecht; J Bradley Hochalter; Steven G Carmella; Yan Zhang; Diane M Rauch; Naomi Fujioka; Joni Jensen; Dorothy K Hatsukami Journal: Biomarkers Date: 2013-01-22 Impact factor: 2.658
Authors: Irina Stepanov; Pramod Upadhyaya; Steven G Carmella; Rachel Feuer; Joni Jensen; Dorothy K Hatsukami; Stephen S Hecht Journal: Cancer Epidemiol Biomarkers Prev Date: 2008-07 Impact factor: 4.254