As the level of consumption of opioids continues to rise globally, there is increasing concern over the potential impacts of continuous opioid discharges into aquatic ecosystems. Opioids are psychoactive compounds that are not completely removed during wastewater treatment, and little is known about their stability and fate in the environment. In the present study, we evaluated the stability of four highly used opioids, buprenorphine, codeine, fentanyl, and tramadol, in river water via batch degradation experiments. The opioids were spiked at environmentally relevant concentrations into 150 mL of river microcosms designed to distinguish among hydrolysis, abiotic degradation, biodegradation, and sorption. All opioids exhibited relatively high stability in river water, with removal rates of only 15% (tramadol) to 26% (buprenorphine) after 6 days. Biodegradation was the most important attenuation pathway for all four opioids, with first-order biodegradation constants ranging from 0.011 d −1 (tramadol) to 0.018 d −1 (buprenorphine). Overall, degradation rates were 1−4 orders of magnitude lower compared to the reported rates for wastewater systems. These results offer insights into the stability of opioids in freshwater systems and raise questions about the potential effects of their pseudopresence in surface waters on aquatic organisms.