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However, current studies of SHs have primarily focused on estrogenic substances, with androgen-related studies being quite limited. We optimized the solid-phase extraction (SPE) pretreatment method, as well as the enzymolysis conditions of five androgens (androstenedione, boldenone, methandienone, nandrolone, and testosterone), to simultaneously determine their concentrations in the effluent from wastewater treatment plants and surface water samples. Then we evaluated the ecological risks of the five androgens in the effluent and Pearl River basin of Guangzhou (PR China) using the risk quotient method. The recovery rates of the targets were 90% to 99% in water samples when digested with \u03b2-glucosidase for 90\u2009min before solid-phase extraction, extracted with a Poly-Sery HLB column, and washed with 15% methanol aqueous solution and 2% ammonia. The established instrument's limit of detection was between 0.02 and 0.39\u2009\u03bcg\/L, and the limit of quantification was between 0.05 and 1.29\u2009\u03bcg\/L. Androstenedione, boldenone, methandienone, nandrolone, and testosterone were detected in all samples from the 2018 and 2022 wastewater influent and the 2018 surface water, with concentrations of 3.06\u2009\u00d7\u2009101\u2009ng\/L to 1.33\u2009\u00d7\u2009103\u2009ng\/L, 1.03\u20138.15\u2009\u00d7\u2009102\u2009ng\/L, and 0.93\u2009\u00d7\u2009101\u2009ng\/L to 5.50\u2009\u00d7\u2009102\u2009ng\/L, respectively. The ecological risks of androgens in wastewater influent and surface water were medium to high and low to medium, respectively. Moreover, the biotoxicity of androgens was predicted by the Ecological Structure Activity Relationships model, with methandienone and androstenedione having the highest and lowest acute and chronic toxicities, respectively. These results suggest that the risk of environmental androgens should not be ignored and that further research should be carried out. Environ Toxicol Chem 2024;43:915\u2013925. \u00a9 2023 SETAC<\/jats:p>","DOI":"10.1002\/etc.5805","type":"journal-article","created":{"date-parts":[[2023,12,13]],"date-time":"2023-12-13T00:28:15Z","timestamp":1702427295000},"page":"915-925","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Trace Analysis of Five Androgens in Environmental Waters by Optimization of Enzymolysis and Solid-Phase Extraction Ultra-Performance Liquid Chromatography\u2013Mass Spectrometry and its Risk Assessment"],"prefix":"10.1093","volume":"43","author":[{"given":"Yufei","family":"Xie","sequence":"first","affiliation":[{"name":"School of Environment and Energy, Higher Education Mega Center, South China University of Technology, Guangzhou, PR China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhihan","family":"Gao","sequence":"additional","affiliation":[{"name":"School of Environment and Energy, Higher Education Mega 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