Filtered water samples were collected by the USGS National Water Quality Program (NWQP) Southeastern Stream Quality Assessment (SESQA) from 59 perennial, wadeable (less than 10 m width and 1 m depth at base-flow) headwater stream sites in watersheds with varying degrees of urban land use in four states. Dataset includes sample site locations and information, analytical method information, water sample pharmaceutical concentrations and summary statistics, and corresponding watershed land-use-land-cover data and data dictionary.