Per- and polyfluoroalkyl substances (PFAS) are a recalcitrant suite of chemicals that are environmentally persistent and have been detected worldwide in numerous human and environmental matrices. Human exposure to PFAS has been associated with a myriad of human health impacts due to their carcinogenic nature. A better understanding of various transportation routes that instigate human and environmental exposure in the Midwest will assist communities in making management decisions regarding PFAS usage and treatment. However, current PFAS research is limited due to existing analytical constraints and limitations. This thesis focuses on assessing current extraction methodologies for PFAS for four environmental matrices (POCIS, non-potable water, solids, plant tissue) and a study looking at the transportation of PFAS within a wastewater treatment plant, adjacent surface waters, and agricultural ecosystem land applied with municipal biosolids for a better assessment of PFAS within the Midwest.