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To date, a range of impairments to aquatic organisms has been reported with exposure to pharmaceuticals; however, further comparisons of their impacts across different species on the molecular level are needed. In the present study, the crustacean Daphnia magna and the freshwater fish Japanese medaka, common model organisms in aquatic toxicity, were exposed for 48\u2009h to the common analgesics acetaminophen (ACT), diclofenac (DCF), and ibuprofen (IBU) at sublethal concentrations. A targeted metabolomic-based approach, using liquid chromatography\u2013tandem mass spectrometry to quantify polar metabolites from individual daphnids and fish was used. Multivariate analyses and metabolite changes identified differences in the metabolite profile for D. magna and medaka, with more metabolic perturbations for D. magna. Pathway analyses uncovered disruptions to pathways associated with protein synthesis and amino acid metabolism with D. magna exposure to all three analgesics. In contrast, medaka exposure resulted in disrupted pathways with DCF only and not ACT and IBU. Overall, the observed perturbations in the biochemistry of both organisms were different and consistent with assessments using other endpoints reporting that D. magna is more sensitive to pollutants than medaka in short-term studies. Our findings demonstrate that molecular-level responses to analgesic exposure can reflect observations of other endpoints, such as immobilization and mortality. Thus, environmental metabolomics can be a valuable tool for selecting sentinel species for the biomonitoring of freshwater ecosystems while also uncovering mechanistic information. Environ Toxicol Chem 2024;43:1339\u20131351. \u00a9 2024 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.<\/jats:p>","DOI":"10.1002\/etc.5876","type":"journal-article","created":{"date-parts":[[2024,4,25]],"date-time":"2024-04-25T13:34:49Z","timestamp":1714052089000},"page":"1339-1351","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Metabolomic-Based Comparison of <i>Daphnia magna<\/i> and Japanese Medaka Responses After Exposure to Acetaminophen, Diclofenac, and Ibuprofen"],"prefix":"10.1093","volume":"43","author":[{"given":"Erico A.","family":"Oliveira Pereira","sequence":"first","affiliation":[{"name":"Environmental Nuclear Magnetic Resonance Centre and Department of Physical and Environmental Sciences, University of Toronto Scarborough, Toronto, Ontario, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Theresa R.","family":"Warriner","sequence":"additional","affiliation":[{"name":"Faculty of Science, Ontario Tech University, Oshawa, Ontario, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9472-7192","authenticated-orcid":false,"given":"Denina B. 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