The long-term health effects associated with cigarette smoking have been shown to be more harmful compared to those associated with the consumption of non-combustible tobacco products, such as moist snuff. In order to investigate the long-term effects of tobacco exposure, we evaluated the biochemical profiles of 40 smokers, 40 moist snuff consumers (MSC), and 40 non-tobacco consumers (NTC) using UHPLC-mass-spectrometry based global metabolomics. Twenty-four hour urine samples and matching plasma samples were collected from adult male subjects who abstained overnight from both food and tobacco. Metabolomic profiling and data analyses were performed at Metabolon Inc., (Durham NC). In this global profiling study, a total of 511 biochemicals (290 known and 221 unknown metabolites) were detected in the plasma, whereas 972 biochemicals (396 known and 596 unknown) were found in urine. For example, biochemicals from amino acid, carbohydrate, fatty acid, lipid, nucleotide and xenobiotic metabolism were identified. These biochemicals fit into distinct metabolic pathways such as oxidative stress and inflammation, and cholesterol, glucose and amino acid metabolism. In addition, a large number of structurally unknown biochemicals were detected. Cigarette smoking, relative to moist snuff consumption, appears to lead to the most changes in biochemical profiles observed in this study. Biochemical changes which point to a hyperglycemic state and hyperlipidemia are among the key perturbations noted in the smoker cohort, compared to non-smoking cohorts. In summary, these data show changes in global biochemical profiles in generally healthy cigarette smokers and moist snuff consumers. These differences in the metabolite profiles may be useful in understanding the higher risks associated with smoking relative to consumption of smokeless tobacco prod