Abstract
Biomarkers of exposure to harmful tobacco constituents are key tools for identifying individuals at risk and developing interventions and tobacco control measures. However, tobacco biomarker studies are scarce in many parts of the world with high prevalence of tobacco use. Our goal was to establish a robust method for simultaneous analysis of urinary total 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL), N′-nitrosonornicotine (NNN), and cotinine at the Advanced Centre for Treatment, Research and Education in Cancer (ACTREC) in Mumbai, India. These biomarkers are validated measures of exposure to the carcinogenic tobacco nitrosamines 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and NNN and the addictive alkaloid nicotine, respectively. The established method is characterized by excellent accuracy, linearity, and precision, and was successfully applied to the analysis of 15 smokeless tobacco (SLT) users and 15 non-users of tobacco recruited in Mumbai. This is the first report of establishment of such procedure in a laboratory in India, which offers the first in-country capacity for research on tobacco carcinogenesis in Indian SLT users.
Highlights
Tobacco use remains the leading preventable cause of morbidity and mortality globally, accounting for 8 million annual deaths worldwide[1]
The liquid chromatography-tandem mass spectrometry (LC–MS/MS) method for the analysis of NNAL, NNN, and cotinine in the purified urine samples was essentially based on the previously reported m ethods[21,22,23,24,25], except that a non-capillary system and a QTRAP instrument were used in this study
Given the higher flow rate used in this system, we optimized the HPLC mobile phase gradient to allow for chromatographic resolution of analytes from the potential peaks of residual interfering metabolites that may be present in the complex urine matrix
Summary
Tobacco use remains the leading preventable cause of morbidity and mortality globally, accounting for 8 million annual deaths worldwide[1]. Research employing biomarkers of toxic and carcinogenic tobacco constituents can provide important insights for identifying individuals at risk and developing interventions and tobacco control measures. Exposure to NNK can be measured via its biomarker urinary total 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) and exposure. In the United States, studies assessing cotinine, total NNAL, and total NNN in tobacco users generated important evidence to support tobacco control measures such as clean air laws and the proposal by the Food and Drug Administration to regulate levels of NNN in SLT products to protect public health[17]. In our ongoing and future research efforts in India, we aim to analyze all three biomarkers in large numbers of urine samples in order to characterize the links between the chemical composition of SLT and other tobacco products and the exposures and health outcomes in users. Simultaneous analysis of these three important tobacco biomarkers offers a cost-effective solution for conducting tobacco control research in low-resource settings
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