Stable isotope analyses in skeletal remains of 18th and 19th century African Americans from the Mid-Atlantic region of North America allow us to explore the origins of this historically marginalized population. In 1808, the United States slave trade was officially banned, yet legal slavery continued until 1865. During the apex of North American slavery, thousands of Africans and African Americans were trafficked and born into this system. A critical component of understanding this population involves tracing individual origins, yet written and historic records are rare. Oxygen, carbon, and nitrogen isotopes in bones and teeth provide insight into geographic origins and diet. The isotope values from African Americans and Caucasian Americans were compared to individuals from the cosmopolitan site of Elmina, Ghana, the population therein representing stable isotope values typical of West African slave export sites. Results show oxygen isotope values from Elmina are higher than those from North American Mid-Atlantic sites. These oxygen isotope values track geographic differences in drinking water, differentiating Elmina from Mid-Atlantic sites and identifying outliers as potential recent arrivals from West Africa. Given that very few outliers were identified in the Mid-Atlantic sites, this data lends insight into the establishment of slavery as a self-perpetuating system in the early United States. Additional results from carbon and nitrogen isotopes facilitate inferences on food choices in West Africa and suggest African Americans retained certain cultural food preferences. Overall these results show great promise for our ability to use biochemical techniques as a means to investigate and understand the identity of enslaved individuals whose life stories historically have been underrepresented.