Feasibility of Identifying the Tobacco-related Global Metabolome in Blood by UPLC–QTOF-MS.pdf

Feasibility of Identifying the Tobacco-related Global Metabolome in Blood by UPLC?QTOF-MS Ping-Ching Hsu,?,? Bin Zhou,?,§ Yi Zhao,§ Habtom W. Ressom,§ Amrita K. Cheema,§ Wallace Pickworth,∥ and Peter G. Shields*,? ?The Ohio State University Comprehensive Cancer Center, Columbus, Ohio, United States §Lombardi Comprehensive Cancer Center, Georgetown University, Washington, D.C., United States ∥Battelle Centers for Public Health Research Evaluation, Baltimore, Maryland, United States *S Supporting Information ABSTRACT: Metabolomics is likely an ideal tool to assess tobacco smoke exposure and the impact of cigarette smoke on human exposure and health. To assess reproducibility and feasibility of this by UPLC?QTOF-MS, three experiments were designed for the assessment of smokers’ blood. Experiment I was an analysis of 8 smokers with 8 replicates. Experiment II was an analysis of 62 pooled quality control (QC) samples from 7 nonsmokers’ plasma placed as every tenth sample among a study of 613 samples from 160 smokers. Finally, to examine the feasibility of metabolomic study in assessing smoke exposure, Experiment III consisted of 9 smokers and 10 nonsmokers’ serum to evaluate differences in their global metabolome. There was minimal measurement and sample preparation variation in all experiments, although some caution is needed when analyzing specific parts of the chromatogram. When assessing QC samples in the large scale study, QC clustering indicated high stability, reproducibility, and consistency. Finally, in addition to the identification of nicotine metabolites as expected, there was a characteristic profile distinguishing smokers from nonsmokers. Metabolites selected from putative identifications were verified by MS/MS, showing the potential to identify metabolic phenotypes and new metabolites relating to cigarette smoke exposure and toxicity. KEYWORDS: metabolomics, metabolism, nicotine, validation, carcinogens ■ INTRODUCTION Tobacco smoking is a major cause of m