Minimally invasive compositional analyses of glass trade beads have revolutionised the study of these highly portable and socially significant items. Here, the authors interrogate new and legacy compositional data to investigate how Indigenous communities in eastern North America, particularly Wendat confederacy members, obtained beads from European traders and connected to broader interregional exchange systems c. AD 1600–1670. Diagnostic chemical elements in glass compositions reveal down-the-line exchange and population movement into the Western Great Lakes region prior to the arrival of European settlers, which highlights active Indigenous participation in transatlantic economic networks during a historical period of dynamic reorganisation and interaction.