Abstract

Abstract Pharmaceuticals in aquatic environments pose threats to aquatic organisms because of their continuous release and potential accumulation. Monitoring methods for these contaminants are inadequate, with targeted analyses falling short in assessing water quality's impact on biota. The present study advocates for integrated strategies combining suspect and targeted chemical analyses with molecular biomarker approaches to better understand the risks posed by complex chemical mixtures to nontarget organisms. The research aimed to integrate chemical analysis and transcriptome changes in fathead minnows to prioritize contaminants, assess their effects, and apply this strategy in Wascana Creek, Canada. Analysis revealed higher pharmaceutical concentrations downstream of a wastewater‐treatment plant, with clozapine being the most abundant in fathead minnows, showing notable bioavailability from water and sediment sources. Considering the importance of bioaccumulation factor and biota–sediment accumulation factor in risk assessment, these coefficients were calculated based on field data collected during spring, summer, and fall seasons in 2021. Bioaccumulation was classified as very bioaccumulative with values >5000 L kg –1 , suggesting the ability of pharmaceuticals to accumulate in aquatic organisms. The study highlighted the intricate relationship between nutrient availability, water quality, and key pathways affected by pharmaceuticals, personal care products, and rubber components. Prioritization of these chemicals was done through suspect analysis, supported by identifying perturbed pathways (specifically signaling and cellular processes) using transcriptomic analysis in exposed fish. This strategy not only aids in environmental risk assessment but also serves as a practical model for other watersheds, streamlining risk‐assessment processes to identify environmental hazards and work toward reducing risks from contaminants of emerging concern. Environ Toxicol Chem 2024;00:1–22. © 2024 SETAC