Surface water samples were collected from 264 sites across 46 U.S national parks during the period of 2009-2019. The number of sites within each park ranged from 1 to 31 and the number of samples collected within each park ranged from 1 to 201. Samples were analyzed for up to 340 trace organic contaminants (TrOCs), including pharmaceuticals, personal care products, pesticides, and various contaminants indicative of anthropogenic influence (e.g., fragrances, surfactants, flame retardants). A total of 155 TrOCs was detected in at least one sample with concentrations ranging from the reporting level of 10 ng/L (multiple contaminants) to 11,900 ng/L (p-cresol). Except for bisphenol A, DEET, theobromine, and gabapentin, TrOCs were detected in <20% of samples. Despite the relatively low detection frequencies, when TrOCs were detected, concentrations were similar to those reported from other regional or national studies. We compared detected concentrations to bioactivity concentrations and water quality benchmarks, when available, to identify occurrences of elevated concentrations and to estimate the potential for biological effects to aquatic biota. Elevated concentrations of 27 TrOCs, mostly pesticides, were detected throughout the study. To gain insight regarding potential sources, we related watershed characteristics (e.g., land cover, presence of point sources) to the number of TrOCs detected at each site. We found that the presence of wastewater treatment plants and the proportion of the watershed classified as agricultural land were the most influential variables for describing the number of pharmaceuticals and the number of pesticides present, respectively. This study represents the largest-scale study characterizing the presence and magnitude of TrOCs in U.S. national park surface waters, to date. These data provide a baseline that can be used to inform future monitoring within the parks and to assess changes in water quality.