Thesis Outline
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Identification of brominated organic compounds in aquatic biota and exploration of bromine isotope analysis for source apportionment Maria Unger Doctoral Thesis Department of Materials and Environmental Chemistry (MMK) Stockholm University Stockholm 2010 © Maria Unger, Stockholm 2010 ISBN 978-91-7447-070-3 pp 1-58 Printed in Sweden by US-AB, Stockholm 2010 Distributors: Department of Materials and Environmental Chemistry (MMK), Stockholm University Department of Applied Environmental Science (ITM), Stockholm University Cover photo by Sven-Erik Nilsson. Smedfjärden/Kvädöfjärden. Farleden mellan Huvududden på Stora Askö och Huvudskär. 2 What will the winter bring if not yet another spring Kamelot, Serenade, Black Halo 2005 3 Forward momentum! 4 Abstract Brominated organic compounds (BOCs) of both natural and anthropogenic origin are abundant in the environment. Most compounds are either clearly natural or clearly anthropogenic but some are of either mixed or uncertain origin. This thesis aims to identify some naturally produced BOCs and to develop a method for analysis of the bromine isotopic composition in BOCs found in the environment. Polybrominated dibenzo-p-dioxins (PBDDs) in the Baltic Sea are believed to be of natural origin although their source is unknown. Since marine sponges are major producers of brominated natural products in tropical waters, BOCs were quantified in a sponge (Ephydatia fluviatilis) from the Baltic Sea (Paper I). The results showed that the sponge does not seem to be a major producer of PBDDs in the Baltic Sea. In this study, mixed brominated/chlorinated dibenzo-p-dioxins were however discovered for the first time in a background environment without an apparent anthropogenic source. The use of nuclear magnetic resonance spectroscopy (NMR) is unusual in analytical environmental chemistry due to its sample requirements. Preparative capillary gas chromatography was used to isolate a sufficient amount of an unidentified BOC from northern bottlenose whale (Hyperoodon ampullatus) blubber (Paper II) to enable NMR analysis for identification of the compound. The bromine isotopic composition of BOCs may give information on the origin and environmental fate of these compounds. The first steps in this process are the development of a method to determine the bromine isotope ratio in environmentally relevant BOCs (Paper III) and measuring the bromine isotope ratio of several standard substances to establish an anthropogenic endpoint (Paper IV). 5 Table of contents Abstract ............................................................................ 5 Table of contents ................................................................ 6 Abbreviations ..................................................................... 8 List of papers ..................................................................... 9 Statement ....................................................................... 10 Thesis objectives .............................................................. 11 1. Brominated organic compounds in the marine environment 13 1.1 Naturally produced brominated organic compounds ............................................................................ 13 1.2 Natural production of brominated organic compounds ............................................................................ 17 2. Bromine isotopes .......................................................... 20 2.1 Kinetic isotope effect ......................................... 20 2.2 Isotope fractionation processes ........................... 20 3. Samples and purification methods .................................. 23 3.1 Samples .......................................................... 23 3.2 Extraction and lipid removal methods .................. 24 3.3 Chromatographic separation ............................... 27 4. Analysis ....................................................................... 29 4.1 GC-MS ............................................................. 29 4.2 NMR ................................................................ 30 4.3 GC-multiple-collector-ICP-MS ............................. 31 5. Summary of papers....................................................... 33 5.1 Paper I - Identification and quantification of PBDDs and Br/Cl-DDs ........................................................ 33 5.2 Paper II - NMR-identification of a novel brominated organic compound isolated from whale blubber .......... 34 5.3 Paper III - Development of a GC-MC-ICP-MS method for brominated diaromatic compounds ...................... 34 5.4 Paper IV - Measurements of δ81Br in anthropogenic and natural monoaromatic compounds ...................... 35 6 6. Results and Discussion .................................................. 36 6.1 Identification of natural brominated organic compounds ............................................................ 36 6.2 Development of CSIA-Br .................................... 39 7. Conclusions .................................................................. 42 8. Future perspectives ....................................................... 43 9. Sammanfattning på svenska .......................................... 44 10. Acknowledgements ..................................................... 45 11. References ................................................................. 48 7 Abbreviations ACN Acetonitrile BOCs Brominated organic compounds Br/Cl-DDs Mixed brominated/chlorinated dibenzo-p- dioxins CIS Cooled injection system CSIA Compound specific isotope analysis DDT 2,2-bis(4-chlorophenyl)-1,1,1-trichloroethane ECNI Electron capture negative ionization EI Electron ionization GC-MC-ICP-MS Gas chromatography multiple collector inductively coupled ion plasma mass spectrometry GC-MS Gas chromatography – mass spectrometry HRMS High resolution mass spectrometry ICP Inductively coupled plasma KIE Kinetic isotope effect MBB Monobromobenzene MeOMe-BDE42 6-Methoxy-5-methyl-2,2’3,4’-tetrabromo diphenyl ether MeO-PBDEs Methoxylated polybrominated diphenyl ethers NMR Nuclear magnetic resonance spectroscopy OH-PBDEs Hydroxylated polybrominated diphenyl ethers PBB Polybrominated biphenyl PBDDs Polybrominated dibenzo-p-dioxins PBDEs Polybrominated diphenyl ethers PBDFs Polybrominated dibenzofurans PCBs Polychlorinated biphenyls PCDDs Polychlorinated dibenzo-p-dioxins PCGC Preparative capillary gas chromatograph R Isotope ratio RF Radio frequency SIM Selective ion monitoring TCDD 2,3,7,8-Tetrachlorodibenzo-p-dioxin TriBDD s Tribromodibenzo-p-dioxins 8 List of papers Paper I Maria Unger, Lillemor Asplund, Peter Haglund, Anna Malmvärn, Kristina Arnoldsson, Örjan Gustafsson. Polybrominated and Mixed Brominated/Chlorinated Dibenzo-p-Dioxins in Sponge (Ephydatia fluviatilis) from the Baltic Sea. Environmental Science & Technology, 2009, 43, 8245-8250 Paper II Maria Unger, Lillemor Asplund, Göran Marsh, Örjan Gustafsson. Characterization of an abundant and novel methyl- and methoxy-substituted brominated diphenyl ether isolated from whale blubber. Chemosphere, 2010, 79, 408-413 Paper III Henry Holmstrand, Maria Unger, Daniel Carrizo, Per Andersson, Örjan Gustafsson. Compound-specific bromine isotope analysis of brominated diphenyl ethers using GC-multiple collector-ICP-MS. Submitted to Rapid Communications in Mass Spectrometry Paper IV Daniel Carrizo, Maria Unger, Henry Holmstrand, Per Andersson, Örjan Gustafsson, Sean P. Sylva, Christopher M. Reddy. Compound-specific bromine isotope composition of natural and industrially-synthesized organobromine substances. Manuscript Papers I and II are reproduced with permission from American Chemical Society and Elsevier, respectively. 9 Statement I, Maria Unger, have contributed with the following to the papers included in this thesis: In Paper I, I planned and performed the sampling and a major part of the purification. I was responsible for data analysis and interpretation of mass spectra. I had main responsibility for writing the paper. In Paper II, I planned the project. I did a major part of the sample purification and was responsible for the PCGC isolation of the compound, data analysis and interpretation of mass spectra. I took part in the NMR spectra interpretation and had main responsibility for writing the paper. In Paper III, I took part in planning the set up of the instrument and experiments. I took part in the measurements and data evaluation and assisted in writing the paper. In Paper IV, I took part in planning the set up of the instrument and experiments. I took part in the measurements, data evaluation and in writing the paper. Papers not included in the thesis Emma L. Teuten, Carl G. Johnson, Manolis Mandalakis, Lillemor Asplund, Örjan Gustafsson, Maria Unger, Göran Marsh, Christopher M. Reddy Spectral characterization of two bioaccumulated methoxylated polybrominated diphenyl ethers. Chemosphere, 2006, 62, 197-203 Maria Unger and Örjan Gustafsson PAHs in Stockholm window films: Evaluation of the utility of window film content as indicator of PAHs in urban air. Atmospheric Environment, 2008, 42, 5550-5557 10 Thesis objectives The aims of this thesis were to identify natural brominated compounds in the aquatic environment and to develop a method for source apportionment of natural and anthropogenic organobromine compounds based on their stable bromine isotopic composition. The specific objectives for the papers were: Paper I: Investigate the role of an abundant sponge in the production of polybrominated dibenzo-p-dioxins (PBDDs) in the Baltic Sea by identification