A burial ground located in the Town of Colonie, NY along the Hudson River revealed fourteen individuals dated from the 17th through the early 19th centuries. Bioarchaeological analysis suggested some of these individuals were of African ancestry who had worked and died on the property owned by the prominent Schuyler family. Mitochondrial DNA (mtDNA) analysis was carried out on skeletal remains of seven adults using restriction fragment length polymorphism typing and direct sequencing of the control region to infer their origins and relatedness. Results show that none of the individuals were maternally related, with four individuals identified as African haplogroup L, one identified as Native American haplogroup X, and two identified as haplogroup M and M7. Individuals of African ancestry correlate with published mtDNA data on African Americans and their geographical origins corroborate with the various exit points during the African slave trade to New York State. Individuals identified as haplogroup M7 and M resemble lineages found in Madagascar. Historical documents suggest several hundred people were imported from Madagascar through illegal trading to New York by the end of the 17th century. This study highlights the diverse origins of the enslaved labor force in colonial New York and contributes to our understanding of African American history in the northeast.