Daily Monitoring at a Full-Scale Wastewater Treatment Plant Reveals Temporally Variable Micropollutant Biotransformations

Despite decades of micropollutant (MP) monitoring at wastewater treatment plants (WWTPs), we lack a fundamental understanding of the time-varying metabolic processes driving MP biotransformations. To address this knowledge gap, we collected 24-h composite samples from the influent and effluent of the conventional activated sludge (CAS) process at a WWTP over 14 consecutive days. We used liquid chromatography and high-resolution mass spectrometry to (i) quantify 184 MPs in the influent and effluent of the CAS process; (ii) characterize temporal dynamics of MP removal and biotransformation rate constants; and (iii) discover biotransformations linked to temporally variable MP biotransformation rate constants. We measured 120 MPs in at least one sample and 66 MPs in every sample. There were 24 MPs exhibiting temporally variable removal throughout the sampling campaign. We used hierarchical clustering analysis to reveal four temporal trends in biotransformation rate constants and found MPs with specific structural features co-located in the four clusters. We screened our HRMS acquisitions for evidence of specific biotransformations linked to structural features among the 24 MPs. Our analyses reveal that alcohol oxidations, monohydroxylations at secondary or tertiary aliphatic carbons, dihydroxylations of vic-unsubstituted rings, and monohydroxylations at unsubstituted rings are biotransformations that exhibit variability on daily timescales.