Tobacco consumption affects human health, but no studies have investigated its effect on the bone metabolome, or if any changes are traceable after long postmortem intervals. Human osteoarchaeological remains preserve small molecules, making them valuable for studies that aim to examine past conditions. We test if there are molecular differences in the metabolome of cortical bone between archaeological individuals who used tobacco and those who did not, and if these differences are distinct enough to assign tobacco use status to individuals with unknown tobacco use. Cortical bone of 323 known and unknown tobacco users was studied by an untargeted metabolomics assay using a liquid chromatography-high-resolution mass spectrometry platform. We identified 45 discriminating molecular features that differed between tobacco consumers (15 up-regulated features) and nonconsumers (17 up-regulated features). Tobacco consumption leaves a metabolic record in human bone distinctive enough to identify its use in individuals of unknown tobacco consumption. Future work will validate molecular features relating to tobacco consumption.