Urine source separation has been proposed as a novel method to isolate pharmaceuticals at a higher concentration than wastewater for more effective removal, and thereby reduce the load of pharmaceuticals to the environment. A life cycle assessment (LCA) and was performed to evaluate the overall environmental impacts and economic costs of removing pharmaceuticals by ion-exchange from source separated urine generated annually at the University of Florida. Eight treatment scenarios were evaluated: including centralized wastewater treatment with and without an advanced ozonation process, centralized treatment of source separated urine collected by vacuum truck or vacuum sewer, decentralized treatment of source separation urine. Within source separation scenarios, landfill disposal or regeneration of spent resin was also considered. Urine source separation had 90% lower environmental impact compared with centralized wastewater treatment due to reduced potable water production for flush water, reduced electricity use at the wastewater treatment plant, and reduced nutrient discharge due to nutrient recovery by struvite precipitation. Centralized wastewater treatment upgraded with ozone had the greatest reduction in pharmaceutical toxicity, however it had the greatest total ecotoxicity due to ozone operation and infrastructure. Among source separation scenarios, decentralized treatment of urine and centralized treatment of urine collected by vacuum truck had negligible cost differences compared with centralized wastewater treatment. Whereas, urine collected by vacuum sewer and centralized treatment with ozone cost 30% more than centralized treatment without ozone. Implementation of urine source separation may provide significant environmental benefits but negligible cost savings compared with centralized wastewater treatment.