The disruption of glucose homeostasis associated with the use of nicotine delivery systems may be due to a shift to lipid metabolism. Indirect calorimetry was used to measure the respiratory exchange ratio (RER) in female (N = 21) and male (N = 21) C57BL/6J mice exposed to room air (control) or e-cigarette vapor in a 1L chamber to test the hypothesis that lipid metabolism predominates in vaped mice. Metabolism was quantified via RER using a GA-200 gas analyzer (iWorx, Inc) and LabScribe v.4 (iWorx, Inc.) software. Blood glucose levels were assessed from a subset of the population using an Accu-Check glucometer (Roche Diagnostics, Inc.). Statistical analyses were conducted using R v.4.0.3. Median RER for controls was lower in females. Older females showed a reduction in RER when exposure occurred in the afternoon (p < 0.001), and in males when exposure occurred in the morning (p = 0.007). Glucose concentrations (mg/dL) were higher after e-cigarette inhalation compared with controls, but this difference was not significant (p = 0.464). The reduction in the respiratory exchange ratio supports the hypothesis that e-cigarette inhalation promotes lipid metabolism, and the magnitude of the effect is influenced by gender, age and time of day.