Emergence and occurrence of performance-enhancing substance use in Australia determined by wastewater analysis

Steroidal androgens, non-steroidal androgens and other non-steroidal performance- and image-enhancing drugs that are Schedule III substances under the US Controlled Substances Act as well as Schedule 4 and 10 substances under the Australian Poisons Standard are being used by athletes and non-athletes. Recent data suggest that they are growing in popularity among those communities. However, more information is necessary to understand the extent of their use by the general population. This study provides more information by assessing the emergence and occurrence (temporal and spatial trends) of performance- and image-enhancing drug use within the general community through wastewater analysis. For this, archived wastewater collected from 2009 to 2021 from one treatment plant and wastewater collected in August 2021 from 51 treatment plants covering 11.6 million Australian people (45% of the national population) was extracted and analysed for 52 performance- and image-enhancing drugs. These included steroidal and non-steroidal androgens, peroxisome proliferator-activated receptor delta agonist metabolites, ReV-ErbA agonists, β 2 -agonists, myostatin inhibitors and growth hormone secretagogues. Analysis of the samples from 2009 to 2021 showed the earliest detection of a non-steroidal androgen, enobosarm, in 2011, followed by cardarine metabolites, ibutamoren and ligandrol and testolone in 2014, 2016 and 2017, respectively. The concentrations of all identified substances increased until 2021 following their first detection. Steroidal androgens and non-steroidal performance-enhancing drugs were detected in samples from 49 of the 51 investigated wastewater treatment plants. A higher number of different analytes were detected in samples representing catchments with larger populations. Our study demonstrates that wastewater analysis can be a useful tool for providing information on performance-enhancing drug use in the general population.