RELATIONSHIP OF THE METABOLOMIC PROFILING IN BLOOD TO SMOKING BEHAVIOR
Better biomarkers of cancer risk are needed for smokers to enhance early detection and support the FDA as it considers cigarette performance standards and evaluate health claims for modified tobacco products. Metabolomics is an ideal tool to assess the impact of cigarette smoke on human exposure and health because of its lesser cost, higher sensitivity and high-throughput. However, little has been shown to validate the reproducibility of metabolomic profiling using LC/MS on human blood samples. In order to assess reproducibility and feasibility of this, a series of studies were designed for the assessment of smokers' blood and a pilot study was conducted to validate the use of metabolomics for the assessment of smoke exposure by Ultra Performance Liquid Chromatography coupled to Quadrupole with Time-of-Flight Mass Spectrometry (UPLC-QTOF-MS). Minimal measurement variation and sample preparation variation in all experiments was found, and the identification of nicotine metabololites along with a characteristic profile distinguishing smokers from non-smokers were identified. After establishing the reproducibility of LC-MS-based metabolomics data on blood samples, a large-scale study of 160 well-characterized smokers phlebotomized before and after smoking a cigarette, and several determinants of smoke exposure were assessed for the relationships on their metabolomic profiling. Using subset analysis, there was significant discrimination in the metabolome between gender, race, smoking status, metabolic status, and type of cigarette smoked. Paired analysis of menthol and non-menthol smokers showed not only a significant discrimination between metabolome of two cigarette types, but also identified cigarette brand with the lowest nicotine yield from the highest. Putative metabolites were identified signifying the importance of sex-specific differences, and revealing features related to carcinogenesis. Further validation is needed to identify and characterize those metabolites. The UPLC-QTOF-MS methodology for assessing metabolomics is a reliable laboratory method for the assessment of human blood. It has the potential to identify metabolic phenotypes and new metabolites relating to cigarette smoke exposure and toxicity. Several future studies can be conceived from the data derived herein in cohort studies of cancer risk and early detection of lung cancer. They also can be used in clinical studies by the FDA to support their decision making.
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