Several physiological assessments have been reported in the literature as measures for detecting, predicting and monitoring cardiovascular disease (CVD). These non-invasive techniques were investigated in three exclusive all male cohorts (cigarette smokers, moist snuff consumers, and non-consumers of tobacco) to evaluate potential differences in CVD status. Flow-mediated dilation (FMD), carotid intima-media thickness (CIMT), and ankle brachial index (ABI) were selected to assess CVD endpoints. Secondarily, spirometry and expired carbon monoxide (ECO) were measured to assess lung function.The three cohorts were age-stratified into four groups: 26-31; 32-37; 38-43; and 44-49. FMD and ABI were measured on Days 1 and 2; the change between days was calculated. CIMT was measured on Day 2 only. For CIMT, a significant age group main effect was observed, demonstrating a tendency toward higher CIMT with age. For FMD, no significant age or cohort main effects were observed at any time-point. For ABI, the only significant difference was observed on Day 1 between smokers and non-consumers of tobacco, with smokers having the lower ABI mean value.Day 2 spirometry measures (% predicted FVC and % predicted FEV1) were significantly lower in smokers compared to moist snuff consumers and non-consumers. ECO and derived COHb were significantly higher in smokers compared to snuff consumers and non-consumers. No differences were observed between snuff consumers and non-consumers for either spirometry or ECO.Although some cohort or age differences were observed in CVD endpoints, the results are consistent with "within normal ranges" for each physiological assessment. ECO measures replicate data reported in other tobacco studies. Spirometry measures support lung function changes expected in smokers compared to smokeless tobacco consumers or non-consumers of tobacco.