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Investigation of Agonistic and Antagonistic Endocrine Activity During Full-Scale Ozonation of Waste Water

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Investigation of Agonistic and Antagonistic Endocrine Activity During Full-Scale Ozonation of Waste Water Investigation of agonistic and antagonistic endocrine activity during full-scale ozonation of waste water Dissertation zur Erlangung des akademischen Grades eines Doktors der Naturwissenschaften – Dr. rer. nat. – vorgelegt von Fabian Itzel geboren in Moers Fakultät für Chemie der Universität Duisburg-Essen 2018 Die vorliegende Arbeit wurde im Zeitraum von September 2014 bis September 2018 im Arbeitskreis von Prof. Dr. Torsten C. Schmidt in der Fakultät für Chemie im Bereich Instrumentelle Analytische Chemie der Universität Duisburg-Essen durchgeführt. Tag der Disputation: 21.02.2019 Gutachter: Prof. Dr. Torsten C. Schmidt Prof. Dr. Elke Dopp Vorsitzender: Prof. Dr. Bettina Siebers I Summary The use of a wide variety of chemicals in our society, such as industrial chemicals, pharmaceuticals, personal care products, etc., leads to pollution of surface waters. Especially in densely populated urban areas such as the Ruhr catchment, sustainable water management poses a major challenge. Despite intensive use through various types of discharges (effluents of direct dischargers, municipal waste water treatment plants, industry, etc.), good water quality has always to be guaranteed in accordance to the European Water Framewor Directive. Endocrine disrupting chemicals can have an effect on aquatic organisms even at very low concentrations (pg/L range). In order to reduce the emission, ozonation was investigated as advanced waste water treatment for the elimination of organic trace compounds. An elimination performance of ≥ 80% for selected substances at specific ozone doses in the range of zspec. = 0.3 - 0.7 mgO3/mgDOC was achieved. Since 2015, estrogens are listed on the watch-list of the European Water Framework Directive with required detection limits in the pg/L range. These detection limits cannot be achieved with current instrumental methods. Therefore, sensitive effect based methods based on modified yeast cells (Arxula adeninivorans), which carry the human estrogen or androgen receptor, were used as a screening tool for corresponding activities. The results showed high elimination rates of the effects already at low ozone doses (zspec. = 0.3 mgO3/mgDOC). Inhibitory effects (antagonists) were investigated in parallel. No significant reduction in antagonistic effects was observed. A comprehensive analysis method based on effect-based analysis and a non-target approach using high-resolution mass spectrometry was developed to identify unknown active compounds. Due to concerns about possible toxicity of the transformation products formed during the ozonation, biological post-treatments are recommended for further treatment. To characterize the efficiency of post-treatment, the assimilable organic carbon together with a trend analysis done by a non-target approach of unknown features formed during ozonation was established and demonstrated to be a useful tool. II Zusammenfassung Der Einsatz verschiedenster Chemikalien in unserer Gesellschaft wie z. B. Arzneimittelwirkstoffen, Industriechemikalien, Körperpflegeprodukte, etc. führen zu einer Belastung der Oberflächengewässer. Besonders in dicht besiedelten Metropolen wie dem Ruhrgebiet stellt eine nachhaltige Bewirtschaftung der Gewässer eine große Herausforderung dar. Denn hierbei muss trotz intensiver Nutzung durch Einleitungen verschiedenster Art (Direkteinleiter, kommunale Kläranlagen, Industrie, etc.) eine stetig gute Wasserqualität nach der EU Wasserrahmenrichtline gewährleistet werden. Bei endokrin wirkenden Chemikalien wird bei bereits sehr niedrigen Konzentrationen (pg/L-Bereich) ein Effekt auf aquatische Organismen beobachtet. Um die Einträge zu verringern, wurde im Rahmen dieser Arbeit die Ozonung als erweiterte Abwasserbehandlung zur Elimination organischer Spurenstoffe untersucht. Es konnte eine Eliminationsleistung von ≥ 80% für ausgewählte Substanzen bei spezifischen Ozondosen im Bereich von zspec. = 0,3 - 0,7 mgO3/mgDOC erreicht werden. Seit 2015 sind Östrogene auf der Beobachtungsliste der EU Wasserrahmenrichtlinie mit erforderlichen Nachweisgrenzen im pg/L-Bereich aufgeführt. Diese Nachweisgrenzen sind mit derzeitigen instrumentellen Methoden nicht erreichbar. Daher wurden sensitive effektbasierte Methoden auf Basis modifizierter Hefezelle (Arxula adeninivorans), welche den humanen Östrogen- bzw. Androgenrezeptor enthalten, als Screening-Tool für die Messung entsprechender Aktivitäten eingesetzt. Die Ergebnisse zeigten bereits bei niedrigen Ozondosen (zspec. = 0,3 mgO3/mgDOC) hohe Eliminationsraten der hormonellen Aktivitäten. Parallel wurden inhibierende Effekte (Antagonisten) untersucht. Es konnte keine signifikante Reduktion antagonistischer Effekte beobachtet werden. Um entsprechend unbekannte Wirkstoffe zu identifizieren, wurde eine umfassende Analysemethode entwickelt, die auf der effektbasierten Analytik sowie einem non-target Ansatz mittels hochauflösender Massenspektrometrie basiert. Aufgrund von Bedenken zur möglichen Toxizität der bei der Ozonung gebildeten Transformationsprodukte, werden biologische Nachbehandlungen zur weiteren Behandlung empfohlen. Zur Charakterisierung der Effizienz einer Nachbehandlung konnte der assimilierbare organische Kohlenstoff zusammen mit einer Trendanalyse der gebildeten Features während der Ozonung als nützliches Tool etabliert werden. III Table of content Table of Content List of abbreviations .............................................................................................................. X List of units and symbols .................................................................................................... XIII List of figures ...................................................................................................................... XIV List of tables ..................................................................................................................... XVIII 1. Introduction ............................................................................................................. 1 1.1 Water quality and waste water management ........................................................... 1 1.2 Micropollutants and effects ...................................................................................... 2 1.3 Advanced waste water treatment ............................................................................. 4 1.4 Analytical tools for evaluation of treatment efficiancy ............................................... 5 1.4.1 Chemical analysis .................................................................................................... 5 1.4.2 Effect based methods .............................................................................................. 5 1.5 Objectives and scope of the thesis .......................................................................... 8 1.6 References .............................................................................................................. 9 2. Recent developments and concepts of effect based methods for the detection of endocrine activity and the importance of antagonistic effects .... 17 2.1 Abstract ..................................................................................................................17 2.2 Keywords ................................................................................................................17 2.3 Introduction .............................................................................................................17 2.4 Instrumental analysis of endocrine active compounds ............................................22 2.5 Effect based analysis ..............................................................................................25 2.5.1 Agonistic effects .....................................................................................................25 2.5.1.1 Relevance of agonistic effects ................................................................................25 2.5.1.2 Effect based trigger values .....................................................................................27 2.5.2 Antagonistic effects .................................................................................................30 2.5.2.1 Relevance of antagonistic active compounds .........................................................30 2.5.2.2 Anti-estrogenic effects ............................................................................................31 IV Table of content 2.5.2.3 Anti-androgenic effects ...........................................................................................34 2.6 Conclusions ............................................................................................................38 2.7 References .............................................................................................................38 3. Investigation of a full-scale ozonation at a municipal waste water treatment plant by a toxicity-based evaluation concept .................................... 55 3.1 Abstract ..................................................................................................................55 3.2 Keywords ................................................................................................................56 3.3 Introduction .............................................................................................................56 3.4 Materials and Methods ...........................................................................................58 3.4.1 Chemicals and Standards
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