Abstract The NC State Center for Environmental and Health Effects of PFAS focuses on understanding the implications of PFAS exposure in humans and animals in the Cape Fear River Basin of North Carolina. The Cape Fear River in North Carolina is characterized by high levels of a variety of PFAS (per-and poly-fluoroalkyl substances) throughout the watershed. The specific chemicals vary by location with the lower Cape Fear characterized by the presence of novel fluoroethers from chemical manufacturing near Fayetteville, NC, while upstream, the levels of legacy PFAS such as PFOA and PFOS are elevated and frequently exceed the drinking water advisory levels for these chemicals. To better characterize the diversity of PFAS exposures among impacted communities along the Cape Fear River, we have developed this supplement proposal to: 1) further characterize human exposure in communities upriver from Fayetteville, 2) expand analytical capabilities to measure total organic fluorine in biological and environmental samples, and 3) demonstrate feasibility for novel blood collection methods that can be employed in the field for both human and animal blood PFAS analysis.