Per- and polyfluoroalkyl substances (PFAS) have been previously detected near suspected sources in Alabama, but the overall extent of contamination across the state is unknown. This study evaluated the spatial distribution of 17 PFAS within the ten major river basins in Alabama and provided insights into their transport and fate through a mass flux analysis. Six PFAS were identified in 65 out of the 74 riverine samples, with mean ∑6PFAS levels of 35.2 ng L-1. The highest ∑6PFAS concentration of 237 ng L-1 was detected in the Coosa River, a transboundary river that receives discharges from multiple sources in Alabama and Georgia. PFAS distribution was not observed to be uniform across the state: while the Coosa, Alabama, and Chattahoochee rivers presented relatively high mean ∑6PFAS concentrations of 191, 100 and 28.8 ng L-1, respectively, PFAS were not detected in the Conecuh, Escatawpa, and Yellow rivers. Remaining river systems presented mean ∑6PFAS concentrations between 7.94 and 24.7 ng L-1. Although the short-chain perfluoropentanoic acid (PFPeA) was the most detected analyte (88%), perfluorobutanesulfonic acid (PFBS) was the substance with the highest individual concentration of 79.4 ng L-1. Consistent increases in the mass fluxes of PFAS were observed as the rivers flowed through Alabama, reaching up to 63.3 mg s-1, indicating the presence of numerous sources across the state. Most of the mass inputs would not have been captured if only aqueous concentrations were evaluated, since concentration is usually heavily impacted by environmental conditions. Results of this study demonstrate that mass flux is a simple and powerful complementary approach that can be used to broadly understand trends in the transport and fate of PFAS in large river systems.