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Published on: July 14, 2017
Quantitative analysis of 3'-hydroxynorcotinine in human urine
Pramod Upadhyaya1, Stephen S Hecht2
1Masonic Cancer Center, University of Minnesota, Minneapolis, MN.
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.
