Environmental field analysis provides advantages that allow real-time decisions, interactive sampling and cost effective solutions to the problems faced at the time of investigation. Gas chromatography (GC), a widespread technique for the determination of organic pollutants in the environment, has also shown to be useful in environmental field analysis. Thermal desorption of solid environmental sample provides a technique for liberation of volatile analytes from the samples without the need for solvent extraction. Combining the thermal desorption technique with a field gas chromatograph (GC) thus provides the possibility of on-site determination of organic contaminants in soils. However, to better characterize trace level contaminants in complex sample matrices, laboratory analysis using analytical instrument with great separation and resolution power is required. Comprehensive two-dimensional gas chromatography (GC X GC) is such a powerful analytical tool that provides enhanced separation and resolution capacity for the task. The project presented here involves the development and validation of a field method for the analysis of total polychlorinated biphenyls (PCBs) in soils, and determination of individual PCB congeners in the same samples by further laboratory analysis. The field analytical system developed was a field portable GC interfaced with a thermal desorber. The identification of PCB congeners was realized by a GC X GC system with a time-of-flight mass spectrometer (TOF-MS) as a detector. The field method was developed by optimizing and characterizing the method using PCB standards, followed by the application of the developed method to environmental soil samples. Finally, analyses of PCB congeners in environmental soil samples were performed using the GC X GC system.

Environmental field analysis provides advantages that allow real-time decisions, interactive sampling and cost effective solutions to the problems faced at the time of investigation. Gas chromatography (GC), a widespread technique for the determination of organic pollutants in the environment, has also shown to be useful in environmental field analysis. Thermal desorption of solid environmental sample provides a technique for liberation of volatile analytes from the samples without the need for solvent extraction. Combining the thermal desorption technique with a field gas chromatograph (GC) thus provides the possibility of on-site determination of organic contaminants in soils. However, to better characterize trace level contaminants in complex sample matrices, laboratory analysis using analytical instrument with great separation and resolution power is required. Comprehensive two-dimensional gas chromatography (GC X GC) is such a powerful analytical tool that provides enhanced separation and resolution capacity for the task. The project presented here involves the development and validation of a field method for the analysis of total polychlorinated biphenyls (PCBs) in soils, and determination of individual PCB congeners in the same samples by further laboratory analysis. The field analytical system developed was a field portable GC interfaced with a thermal desorber. The identification of PCB congeners was realized by a GC X GC system with a time-of-flight mass spectrometer (TOF-MS) as a detector. The field method was developed by optimizing and characterizing the method using PCB standards, followed by the application of the developed method to environmental soil samples. Finally, analyses of PCB congeners in environmental soil samples were performed using the GC X GC system.

Ein genereller Ansatz zur Charakterisierung von biologischen Systemen bietet die Untersuchung des Metaboloms, dessen Analyse als „Metabolomics“ bezeichnet wird. “Omics”- Technologien haben das Ziel, ohne Selektionskriterien möglichst alle Bestandteile einer biologischen Probe zu detektieren (identifizieren und quantifizieren), um daraus Rückschlüsse auf nicht vorhersehbare und somit neuartige Korrelationen in biologischen Systemen zu ziehen. Ein zentrales Dogma in der Biologie besteht in der Kausalität zwischen Gen – Enzym – Metabolite. Perturbationen auf einer Ebene rufen systemische Antworten hervor, die in einem veränderten Phänotyp münden können. Metabolite sind die Endprodukte von zellulären regulatorischen Prozessen, deren Abundanz durch die Resonanz auf genetische Modifikationen oder Umwelteinflüsse zurückzuführen ist. Zudem repräsentieren Metabolite ultimativ den Phänotyp eines Organismus und haben die Fähigkeit als Biomarker zu fungieren. Die integrale Analyse verschiedenster Stoffwechselwegen wie Krebszyklus, Pentosephosphatzyklus oder Calvinzyklus offeriert die Identifikation von metabolischen Mustern. In dieser Arbeit wurden sowohl das targeted Profiling via GC-TOF-MS als auch das untargeted Profiling via GC-TOF-MS und LC-FT-MS als analytische Strategien genutzt, um biologische Systeme anhand ihrer Metabolite zu charakterisieren und um physiologische Muster als Resonanz auf endogene oder exogene Stimuli zu erkennen. Dabei standen die metabolische, phänotypische und genotypische Plastizität von Pflanzen im Fokus der Untersuchungen. Metabolische Varianzen eines Phänotyps reflektieren die genotyp-abhängige Resonanz des Organismus auf umweltbedingte Parameter (abiotischer und biotischer Stress, Entwicklung) und können mit sensitiven Metabolite Profiling Methoden determiniert werden. Diese Anwendungen haben unter anderem auch zum Begriff des „Environmental Metabolomics“ geführt. In Kapitel 2 wurde der Einfluss biotischer Interaktionen von endophytischen Bakterien auf den Metabolismus von Pappelklonen untersucht; Kapitel 3 betrachtet die metabolische Plastizität von Pflanzen im Freiland auf veränderte biotische Interaktionsmuster (Konkurrenz/Diversität/Artenzusammensetzung); Abschließend wurde in Kapitel 4 der Einfluss von spezifischen genetischen Modifikationen an Peroxisomen und den daraus resultierenden veränderten metabolischen Fluss der Photorespiration dargestellt. Aufgrund der sensitiven Analyse- Technik konnten metabolische Phänotypen, die nicht zwingend in einen morphologischen Phänotyp mündeten, in drei biologischen Systemen identifiziert und in einen stoffwechselphysiologischen Kontext gestellt werden. Die drei untersuchten biologischen Systeme – in vitro- Pappeln, Grünland- Arten (Arrhenatherion-Gesellschaft) und der Modellorganismus (Arabidopsis) – belegten anschaulich die Plastizität des Metabolismus der Arten, welche durch endogene oder exogene Faktoren erzeugt wurden. / A general approach to characterise biological systems offers the analysis of the metabolome, named “metabolomics”. “Omics”- technologies are untargeted approaches without any selection criteria which aim to detect every potential analyte in a sample in order to draw conclusions about new correlations in biological systems. A central dogma in biology is the causality between gene – enzyme – metabolite. Perturbations on one level are reflected in systemic response, which possibly result in a changed phenotype. Metabolites are end products of its gene expression and metabolism, whose abundance is determined as a resonance of genetic modifications or environmental disturbance. Furthermore metabolites represent the ultimate phenotype of an organism and are able to act as a biomarker. The integral analysis of distinct metabolic pathways like TCA, Pentose phosphate and Calvin cycle consequently leads to the identification of metabolic patterns. In this work targeted profiling via GC-TOF-MS as well as untargeted profiling via GC-TOF-MS and LC-FT-MS were used as analytical strategies to characterise biological systems on the basis of their metabolites and to identify physiological patterns as resonance of endogenic or exogenic stimuli. The focus of the investigations concentrates on the metabolic, phenotypic and genotypic plasticity of plants. Metabolic variance of a phenotype is reflected in the genotypic dependence response of an organism on environmental parameters which may be detected via sensitive metabolic profiling methods. In chapter 2 the influence of biotic interaction of endophytic bacteria on the metabolism of their poplar host was analyzed; chapter 3 explores the metabolic plasticity of field-grown grassland species as a consequence of biotic interaction pattern (competition / diversity / species composition); In conclusion, chapter 4 illustrates the influence of specific genetic modifications on peroxisomes and the consequent changed metabolic flux in the photorespiration pathway. Due to the sensitive analytic methods, metabolic phenotypes in all three biological systems could be identified and classified in a physiological context. The three biological systems – in vitro poplar plants, field-grown grassland species and the model organism Arabidopsis – demonstrate the plasticity of the metabolism of species in response to stimuli.

京都大学 / 0048 / 新制・論文博士 / 博士(農学) / 乙第12847号 / 論農博第2799号 / 新制||農||1027(附属図書館) / 学位論文||H26||N4864(農学部図書室) / 31430 / (主査)教授 宮川 恒, 教授 西田 律夫, 教授 植田 和光 / 学位規則第4条第2項該当 / Doctor of Agricultural Science / Kyoto University / DFAM

This diploma thesis deals with Bisphenol A, especially with its impact on the aquatic ecosystem. Information about its properties, production and current use are summarized here. Its harmful impact on the environment, especially on the aquatic ecosystem and the human body is characterized. Also the methods of its degradation within the aquatic environment have been described. A summary of the options of a determination of Bisphenol A in water samples is incorporated and the method of gas chromatography with mass spectrometry (GC/TOF MS) and comprehensive two-dimensional gas chromatography with mass spectrometry (GCxGC/TOF-MS) is compared in the experimental part. Analytical determination precedes the isolation of the analyte from the water samples by solid phase extraction (SPE) using SupelcleanTM ENVITM - 18 and derivatization using the silylation reagent, N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA). The result of this work is the analysis of a series of real samples from wastewater treatment plants Brno Modřice and Luhačovice by a two-dimensional gas chromatography with mass spectrometry (GCxGC/TOF-MS).