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The Development of Varying Methodologies to Speciate and Monitor the Interactions of Selenium and Environmental Contaminants in Plants

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The Development of Varying Methodologies to Speciate and Monitor the Interactions of Selenium and Environmental Contaminants in Plants The development of varying methodologies to speciate and monitor the interactions of selenium and environmental contaminants in plants A dissertation submitted to the Division of Research and Advanced Studies of the University of Cincinnati In partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY In the Department of Chemistry of the College of Arts and Sciences 2008 By Scott Ellington Afton B.S., Chemistry, Andrews University, 2004 Committee Chair: Dr. Joseph A. Caruso Abstract of dissertation There is a multitude of contaminated waste sites worldwide due to anthropogenic activities. When assessing the potential toxicological effects of environmental contaminants, prior concern has dealt with simply quantifying the total concentration of the particular contaminating element. With the increasing awareness of the often significant differences in toxicity between the varying environmental contaminants, elemental speciation and percent distribution must be determined. Conventional remediation efforts have been effective in contaminant removal, but are generally very costly. As a result, phytoremediation, which utilizes plants, has recently gained popularity for removing contaminants from soil. It is also known that a toxic concentration of selenium and arsenic/mercury, if administered simultaneously, produce a nontoxic metabolite in a mammal. The studies in this dissertation utilize novel methodologies to speciate and monitor the interactions of selenium and environmental contaminants, arsenic and mercury, in plants. Eight predominant selenium and arsenic species were simultaneously separated using ion-pairing reversed phase liquid chromatography (IPRP) coupled with inductively coupled plasma mass spectrometry (ICPMS) and electrospray ionization ion trap mass spectrometry within 18 minutes and applied to river water, plant extract and urine matrices. The differences metabolic pathways, including location and identity, of selenium and arsenic species were elucidated after single and simultaneous supplementation in the Chlorophytum comosum, spider plant via size exclusion chromatography (SEC) and IPRP coupled to ICPMS. Although total elemental analysis demonstrates a selenium and arsenic antagonism, a compound containing selenium and arsenic was not found in the general aqueous extract of the plant. iii Increasing the difference between the voltage on the extraction lens and the octopole, plus using a positive voltage between the octopole and quadrupole provided the lowest sulfur detection limits using xenon as the collision reaction cell (CRC) gas in ICPMS. Energy 32 + 32 + discrimination is the predominant mechanism for removing the O2 interference of S . Optimization parameters were also suggested for a standard solution comprised of 52 elements, including sulfur. Similar detection limits were acquired when comparing Xe to He or H2 CRC gas with a general trend comparable to detection levels for He. Capillary reversed phase chromatography (capRPLC) and SEC were coupled to ICPMS to investigate the metabolic fate of mercury in conjunction with or exclusion of selenium in the Allium fistulosum. The data suggests a possible selenium-mercury association in a proteinaceous macromolecule which is not readily translocated to the aerial plant regions. Data from x-ray fluorescence mapping of a freshly excised root and capRPLC-ICPMS of homogenized root extract suggest the formation of a mercury-selenium species and a similarly structured mercury- sulfur species predominantly residing in the cell wall of the epidermal root tissue. Utilizing x- ray absorption near edge structure analysis, the local environment of mercury and selenium qualitatively coincided with the mercury-selenium species formed in a mammal via a Hg-Se- S(GSH) moiety. The local environment of mercury also coincided with (GS)2Hg. iv v Acknowledgements This dissertation is dedicated to my parents, Rick and Anne Afton. Besides being kind enough to bring me into the world, they have continuously supported me throughout my many years of education financially, spiritually and educationally. I hope the accomplishment of this dissertation represents a small token of my appreciation. I also would like to thank my parents for the multitude of important life lessons that have been instilled in me such as the seven P’s: prior proper planning prevents pitiful poor performance. I would also like to acknowledge my sister, Danielle, for conversations we have had throughout my graduate career. I could not have come this far without my family’s constant love and support. I would like to thank my advisor Dr. Joseph Caruso “Doc” for his continued support over the past few years. Of the many benefits gained by joining the Caruso group, Dr. Caruso has granted me almost complete independence to decide the direction and projects of my research. This has allowed for multiple failures and successes that have made me into the scientist I am today. In addition, he was there to provide support and guidance when needed and entrusted responsibilities to me that allowed for external collaborations that otherwise would not have been possible. He has been a wonderful advisor and friend. I would also like to thank Judy Caruso for her kindness and support. She has always made me feel as though I was part of her extended family. In addition to my advisor, I would like to thank my committee members Dr. William Heineman, Dr. Bruce Ault and a previous member Dr. Theresa Reineke. They each have offered invaluable advice and support that aided in reaching my research goals and I greatly appreciate the time and effort they extended. I would also like to thank the rest of the analytical faculty: Dr. Patrick Limbach, Dr. Thomas Ridgway and Dr. Apryll Stalcup for their support and insightful vi questions. In addition, I would like to thank Dr. Peter Padolik and Dr. John Breiner for all the good times during my teaching assistantship. I would like to thank the many collaborators that I have had the privilege of working with that has resulted in multiple publications and still continues to date: Dr. David McNear from the University of Kentucky; Dr. Steve Sutton and Dr. Matt Newville from Argonne National Laboratories; Dr. Julio Figueroa, Dr. Katarzyna Wrobel and Dr. Kazimierz Wrobel from the Universidad de Guanajuato; Dr. Jeff Lehman from Otterbein College; Dr. Mary Beth Genter, Dr. Zhenyu Qin, Dr. Michael Craig, Elizabeth LaPensee, Dr. Nira Ben-Jonathan, Dr. Erin Haynes, Dr. Bin Wang, Scott Schneider and Jed Thorn from the University of Cincinnati. I would also like to thank Agilent Technologies and CEM Corporation for continued support through instrumentation, which has enabled the Caruso group the capability to investigate and answer interesting research questions. I would also like to thank the past and present members of the Caruso group for their support, advice and willingness to incorporate a family atmosphere in the office and laboratory: Dr. Oktay Cankur, Dr. Baki Sadi, Dr. Juris Meja, Dr. Katie DeNicola, Dr. Monica Shah, Dr. Santha Yathavakilla, Dr. Sarath Jayasinghe, Dr. Kevin Kubachka, Dr. Douglas Richardson, Allison Krentz, Heather Trenary, Dr. Jenny Ellis, Dr. Kirk Lokits, Qilin Chan, Yaofang Zhang, Karolin Kroening, Jen Siverling, Chris Tompson, Dean Stuart, Brittany Catron and Renee Easter. It would not be possible to list everyone outside the Caruso group that has made my graduate experience enjoyable; however, I would specifically like to thank a few good men, which include Phillip Durham, Michael Haven and Kevin Parker. I thank you for all of the good times, off-color jokes and the plethora of awkward moments that have culminated into my graduate experience, which will be greatly missed. vii Last but certainly not least, I would like to thank my best friend and my wife, Dr. Lisa Afton. I will try not to hold a grudge against you for finishing your doctorate in audiology before me for too long. My wife was a major influence on me when I considered the possibility of attending graduate school. During our simultaneous graduate careers, we have shared in good times and hardships that have brought us closer together and strengthened our marriage. My graduate journey would have been nearly as fruitful without your love and support. I look forward to the many adventures and experiences ahead. I love you sooooooooooooooooooooo much. -SEA viii Table of contents Abstract of dissertation iii Acknowledgments vi Figures 4 Tables 7 Chapter 1 - Methodologies used for biological contaminant remediation and speciation 1.1 Contaminant remediation 1.1.1 Ex situ methodologies 1.1.2 In situ methodologies 1.1.3 Phytoremediation 1.1.4 Selenium, arsenic and mercury remediation 1.2 Inductively coupled plasma mass spectrometry (ICPMS) 1.2.1 Instrument setup and theory 1.2.2 Theory of interference removal with the collision/reaction cell 1.3 High performance liquid chromatography (HPLC) 1.3.1 Size exclusion chromatography (SEC) 1.3.2 Ion-pairing reversed phase chromatography (IPRP) 1.4 X-ray absorption fine structure (XAFS) 1.4.1 Instrument setup and operation 1.4.2 X-ray absorption fine structure theory 1.5 References Chapter 2 – Simultaneous characterization of selenium and arsenic analytes via ion-pairing reversed phase chromatography with inductively coupled plasma and electrospray ionization ion trap mass spectrometry for detection; applications of river water, plant extract and urine matrices 2.1 Abstract 2.2 Introduction 2.3 Experimental 2.3.1 Instrumentation 2.3.2 Reagents and standards
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