AbstractA wide range of chemicals was measured in different types of drinking water and urine samples through target and non-target screening (NTS) to estimate human exposure. Tap water samples collected from 42 locations in Barcelona (August–October/2020, May/2021), tap water filtered with domestic activated carbon filters (AC, N = 6) and reverse osmosis (RO, N = 5), commercial bottled water (N = 10), and urine (N = 39) samples were included. 35 per- and polyfluoroalkyl substances (PFAS), bisphenol A, and nonylphenol were analyzed using LC–MS/MS and GC–MS/MS, and NTS using LC–HRMS. 9 PFAS were detected in unfiltered tap water of first sampling (79% samples, median = 30 ng/L), 6 in the second (69%, median = 9.8 ng/L), and 5 in 13% urine samples. NTS tentatively identified pharmaceuticals and other industrial chemicals in drinking water. PFAS were removed by RO and not by AC filters. Findings provide valuable information for exposure science and water quality monitoring of emerging drinking water contaminants.