Endocrine-disrupting compounds (EDCs), specifically estrogenic endocrine-disrupting compounds, vary in concentration and composition in surface waters under the influence of different landscape sources and landcover gradients. Estrogenic activity in surface waters may lead to adverse effects in aquatic species at both individual and population levels, often observed through the presence of intersex and vitellogenin induction in male fish. In the Chesapeake Bay Watershed, located on the mid-Atlantic coast of the USA, intersex has been observed in several sub-watersheds where previous studies have identified specific landscape sources of EDCs in tandem with observed fish health effects. Previous work in the Potomac River Watershed (PRW), the largest basin within the Chesapeake Bay Watershed, was leveraged to build random forest regression models to predict estrogenic activity at unsampled reaches in both the Potomac River and larger Chesapeake Bay Watersheds (CBW). Model outputs including important variables, partial dependence plots, and predicted values of estrogenic activity at unsampled reaches provide insight into drivers of estrogenic activity at different seasons and scales. Using the US Environmental Protection Agency effects-based threshold of 1.0ng/L 17 β-estradiol equivalents, catchments predicted to exceed this value were categorized as at risk for adverse effects from exposure to estrogenic compounds and evaluated relative to healthy watersheds and recreation access locations throughout the PRW. Results show immediate catchment scale models are more reliable than upstream models, and the best predictive variables differ by season and scale. A small percentage of healthy watersheds (< 13%) and public access sites were classified as at risk using the "Total" (annual) model in the CBW. This study is the first Potomac River Watershed assessment of estrogenic activity, providing a new foundation for future risk assessment and management design efforts, with additional context provided for the entire Chesapeake Bay Watershed.