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Interpreting Sources and Endocrine Active Components of Trace Organic Contaminant Mixtures in Minnesota Lakes

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INTERPRETING SOURCES AND ENDOCRINE ACTIVE COMPONENTS OF TRACE ORGANIC CONTAMINANT MIXTURES IN MINNESOTA LAKES by Meaghan E. Guyader © Copyright by Meaghan E. Guyader, 2018 All Rights Reserved A thesis submitted to the Faculty and the Board of Trustees of the Colorado School of Mines in partial fulfillment of the requirements for the degree of Doctor of Philosophy (Civil and Environmental Engineering). Golden, Colorado Date _____________________________ Signed: _____________________________ Meaghan E. Guyader Signed: _____________________________ Dr. Christopher P. Higgins Thesis Advisor Golden, Colorado Date _____________________________ Signed: _____________________________ Dr. Terri S. Hogue Professor and Department Head Department of Civil and Environmental Engineering ii ABSTRACT On-site wastewater treatment systems (OWTSs) are a suspected source of widespread trace organic contaminant (TOrC) occurrence in Minnesota lakes. TOrCs are a diverse set of synthetic and natural chemicals regularly used as cleaning agents, personal care products, medicinal substances, herbicides and pesticides, and foods or flavorings. Wastewater streams are known to concentrate TOrC discharges to the environment, particularly accumulating these chemicals at outfalls from centralized wastewater treatment plants. Fish inhabiting these effluent dominated environments are also known to display intersex qualities. Concurrent evidence of this phenomenon, known as endocrine disruption, in Minnesota lake fish drives hypotheses that OWTSs, the primary form of wastewater treatment in shoreline residences, may contribute to TOrC occurrence and the endocrine activity in these water bodies. The causative agents specific to fish in this region remain poorly understood. The objective of this dissertation was to investigate OWTSs as sources of TOrCs in Minnesota lakes, and TOrCs as potential causative agents for endocrine disruption in resident fish. Three research efforts were executed to investigate these topics: examining chemical and biological signatures of OWTS proximity in Minnesota lakes (Chapter 2), prioritizing potential causative agents of endocrine disruption using liquid chromatography tandem mass spectrometry (LC-HRMS) (Chapter 3), and conducting a suspect search of OWTS-associated LC-HRMS features at an adjacent Minnesota lake (Chapter 4). In Chapter 2, traditional targeted aqueous analyses indicated higher concentrations of TOrCs at locations more proximal to residences with OWTSs. Residential proximity also corresponded to feminization of male sunfish. The particular contaminants detected at these locations are considered weak indicators of wastewater presence in environmental compartments and inactive with current metrics for endocrine activity. Unexpected features unique to sites with pronounced endocrine disruption were identified in Chapter 3; however, these components were still considered endocrine inactive by current toxicology metrics. This suggests that temporal resolution in sampling was too low for this chronic-toxicity endpoint, or current regulatory efforts underestimate the effects of these contaminants as environmental mixtures. Finally, Chapter 5 indicated that LC-HRMS analysis and passive sampling allowed for identification of a broader suite of OWTS-associated compounds in adjacent lake water, but that higher spatial resolution was required to refine lake-specific OWTS-compound interest lists. The iii results from this dissertation encourage further investigation of residential inputs of TOrCs to Minnesota lakes, particularly prioritizing locations with endocrine disruption, so that local regulatory agencies may effectively manage these highly valued state resources. iv TABLE OF CONTENTS ABSTRACT ............................................................................................................................... iii LIST OF FIGURES ..................................................................................................................... viii LIST OF TABLES .......................................................................................................................... x ACKNOWLEDGEMENTS ......................................................................................................... xiii CHAPTER 1 INTRODUCTION .................................................................................................... 1 1.1 Objectives and Hypothesis ...................................................................................... 5 1.2 Dissertation Organization ....................................................................................... 7 CHAPTER 2 TRACE ORGANIC CONTAMINANT (TORC) MIXTURES IN MINNESOTA LITTORAL ZONES: EFFECTS OF ON-SITE WASTEWATER TREATMENT SYSTEM (OWTS) PROXIMITY AND BIOLOGICAL IMPACTS ..................................................................................... 10 2.1 Abstract ................................................................................................................. 10 2.2 Introduction ........................................................................................................... 11 2.3 Materials and Methods .......................................................................................... 13 2.3.1 Site Selection ............................................................................................. 13 2.3.2 Sample Collection ..................................................................................... 15 2.3.3 Sample Analysis ........................................................................................ 16 2.3.4 Quality Assurance and Quality Control .................................................... 18 2.3.5 Statistical Analysis .................................................................................... 19 2.4 Results and Discussion ......................................................................................... 20 2.4.1 Site Distinction Analysis ........................................................................... 20 2.4.2 TOrC Detections and Non-Parametric Two Group Comparisons ............ 21 2.4.3 Patterns in Inorganic Non-point Source Indicators ................................... 25 2.4.4 MPD Effect on Total TOrC Concentration ............................................... 26 2.4.5 Biological Data Two Group Hypothesis Tests .......................................... 28 v 2.5 Conclusions and Implications ............................................................................... 29 2.6 Acknowledgements ............................................................................................... 29 CHAPTER 3 PRIORITIZING POTENTIAL ENDOCRINE ACTIVE HIGH RESOLUTION MASS SPECTROMETRY FEATURES IN MINNESOTA LAKEWATER ....... 30 3.1 Abstract ................................................................................................................. 30 3.2 Introduction ........................................................................................................... 31 3.3 Materials and Methods .......................................................................................... 34 3.3.1 Site Selection ............................................................................................. 34 3.3.2 Sample Selection ....................................................................................... 35 3.3.3 Sample Analysis ........................................................................................ 35 3.3.4 Data Processing Workflow........................................................................ 36 3.3.5 Investigating Estrogen Disrupting Potential of Identified Features .......... 41 3.3.6 Prioritized Feature Occurrence in Reference Lake ................................... 41 3.3.7 Quality Control .......................................................................................... 41 3.4 Results and Discussion ......................................................................................... 42 3.4.1 Peak Picking and Feature Alignment ........................................................ 42 3.4.2 Statistical Prioritization of LC-HRMS ...................................................... 43 3.4.3 Feature Reduction and Elimination of Redundancies ............................... 45 3.4.4 Evaluating EAC Potential ......................................................................... 51 3.4.5 Suspect Search of a Reference Lake ......................................................... 55 3.5 Implications and Conclusions ............................................................................... 56 3.6 Acknowledgements ............................................................................................... 56 CHAPTER 4 SUSPECT ANALYSIS OF ON-SITE WASTEWATER TREATMENT TRACE ORGANIC CONTMAINANTS IN PASSIVE SAMPLERS DEPLOYED IN AN ADJACENT MINNESOTA LAKE ..................................... 57 4.1 Abstract ................................................................................................................. 57 4.2 Introduction ........................................................................................................... 58 4.3 Materials and Methods .......................................................................................... 61 4.3.1 Field Sampling .......................................................................................... 61 4.3.2 Sample Preparation ..................................................................................
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