Wastewater from manholes near two gyms and two sports centers was sampled at three-hour intervals over 3 days (Oct 2022 to Feb 2023). Samples were hydrolyzed, concentrated via SPE, and analyzed using LC–MS/MS. Multiple performance and image-enhancing substances were detected, with gyms showing the greatest diversity, including oxandrolone and stanozolol.

ABSTRACT

In this study, we assess the feasibility of wastewater sampling from manholes as a potential monitoring tool for performance and image-enhancing drugs (PIEDs) at local gyms and general sports centers in Denmark. Wastewater samples from two gyms and two sports centers were analyzed using two LC–MS-based analytical methods (UPLC–MS/MS and UPLC-TOF-MS) to detect the presence of PIEDs. Samples were collected at 3-h intervals during the opening hours on three selected days of the week for each location. The study showed the presence of anabolic agents such as oxandrolone, stanozolol, and ostarine in wastewater sampled from the gyms. All three compounds are class C drugs in the United States, and likewise, illegal to sell and possess in Denmark. Other PIEDs, such as aromatase inhibitors and psychoactive drugs, were detected in several samples from both gyms and sports centers. However, gyms showed a higher occurrence of PIEDs, whereas general sports centers showed a lower occurrence. Although it cannot be used as a traditional prevalence measure, wastewater sampling from manholes is a useful tool for studying the occurrence of PIEDs among recreational athletes at specific locations or events. Wastewater analysis may be used as an easy and cost-effective tool to complement both preventive and control measures, such as information campaigns, surveys, or personal testing. However, not all locations are suitable for manhole sampling, and it is necessary to consider the legal, ethical, and practical aspects before using wastewater analysis to monitor communities or specific populations.