Comprehensive analysis of antagonistic endocrine activity during ozone treatment of hospital wastewater Fabian Itzel a,b,c , Kevin S. Jewell d , Juri Leonhardt a , Linda Gehrmann a , Ulf Nielsen e , Thomas A. Ternes d , Torsten C. Schmidt b,c,f , Jochen Tuerk a,c, ⁎ a Institut für Energie - und Umwelttechnik e. V., (IUTA, Institute of Energy and Environmental Technology), Bliersheimer Str. 58-60, 47229 Duisburg, Germany b University of Duisburg-Essen, Instrumental Analytical Chemistry, Universitätsstrasse 5, 45141 Essen, Germany c Centre for Water and Environmental Research (ZWU), University of Duisburg-Essen, Universitätsstrasse 2, 45141 Essen, Germany d Federal Institute of Hydrology (BfG), Am Mainzer Tor 1, 56068 Koblenz, Germany e DHI Urban Water, Agern Alle 5, 2970 Horsholm, Denmark f IWW Water Centre, Moritzstr. 26, 45476 Mülheim an der Ruhr, Germany HIGHLIGHTS • Reduction of estrogene and androgene activity during Hospital wastewater treatment using ozone • Behaviour of agonistic and antagonistic endocrine activities during ozonation using Arxula Adeninivorans based yeast assays • Fractionation in combination with effect based analysis reduces the amount of relevant features • Reduction of antagonistic activities by granular activated carbon filter as post- treatment GRAPHICAL ABSTRACT abstract article info Article history: Received 13 September 2017 Received in revised form 15 December 2017 Accepted 16 December 2017 Available online xxxx To reduce the discharge of micropollutants, advanced wastewater treatment methods were investigated in the last years. Estrogenic effects were found to be reduced by ozonation. These activities are usually measured using genetically modified cell-based tests. As these bioassays are representing a sum parameter, also inhibitory effects such as antagonistic effects need to be further investigated as they are potentially reducing the detected activities. Therefore, a direct comparison of chemical target analysis and biological equivalent concentrations measured by bioassays is often difficult. To investigate the fate of antagonistic activities and their role in mixtures with agonistic activities, two hospital wastewater treatment plants were studied after different treatment steps. Thereby highly enriched samples were analyzed by a combination of bioassays with chemical target and non- target analyses. In order to achieve an in-depth characterization of the antagonistic activities a fractionation of the enriched samples was performed. To identify relevant compounds an effect directed identification approach was used by combining high-resolution mass spectrometry and bioassays. The results showed a high reduction for estrogene and androgene activities. However, a constant antagonistic activity after membrane bioreactor and ozone treatment was observed. A reduction of the antagonistic activity was observed after passing an activat- ed carbon filter. The fractionation approach showed a specific finger-print of each sample of the different treat- ment steps. Hereby we could show that the composition of agonistic and antagonistic active compounds is changing after each treatment step while the overall measured activity stays the same. Using fractionation and Keywords: Hospital wastewater Ozone Anti-estrogenic Anti-androgenic Effect directed analysis Non-target screening Science of the Total Environment 624 (2018) 1443–1454 ⁎ Corresponding author at: Institut für Energie- und Umwelttechnik e. V. (IUTA, Institute of Energy and Environmental Technology), Bliersheimer Str. 58-60, 47229 Duisburg, Germany. E-mail address: tuerk@iuta.de (J. Tuerk). https://doi.org/10.1016/j.scitotenv.2017.12.181 0048-9697/© 2017 Elsevier B.V. All rights reserved. Contents lists available at ScienceDirect Science of the Total Environment journal homepage: www.elsevier.com/locate/scitotenv