In jüngster Vergangenheit hat die potentielle Belastung von Lebens- und Futtermitteln mit PA wiederholt Aufmerksamkeit erregt. Eine Exposition des Menschen mit PA kann über den Genuss von Tees, Phytopharmaka, pflanzlichen Lebensmitteln (z.B. Salatmischungen) oder, im Fall einer Verfütterung von PA-Pflanzen an Tiere, als sekundäre Kontamination über tierische Lebensmittel erfolgen. Im ‚International Programme on Chemical Safety (IPCS)’ der WHO ist die grundsätzliche Gefährdung der menschlichen Gesundheit durch PA dokumentiert. Aus Gründen des vorbeugenden Verbraucherschutzes gibt es demzufolge Rechtsvorschriften zur Regulierung PA-haltiger Phytopharmaka. Für diese gilt in Deutschland seit 1992 ein Grenzwert von 1 µg PA/Tag für 1,2-ungesättigte PA und deren N-Oxide bei oraler Aufnahme und einer Anwendungsdauer von max. 6 Wochen. Geht die Anwendung darüber hinaus, beträgt der Grenzwert 0,1 µg PA/Tag. Im ersten Teil der Arbeit wurde eine robuste, reproduzierbare und selektive analytische Methode basierend auf Zink-Staub-Reduktion, Festphasenextraktion (SCX-SPE), LiAlH4 Reduktion mit anschließender Silylierung sowie Kapillargaschromatographie-Massenspektrometrie (HRGC-MS)-Analytik erarbeitet. Durch ein solches Vorgehen werden die PA-N-Oxide in tertiäre PA überführt, so dass alle PA in ihrer tertiären Form vorliegen. Durch die anschließende chemische Reduktion werden alle Mono- und Diester-PA in die jeweiligen Necinbasen überführt. Durch die anschließende Derivatisierung zum di-TMS-Derivat, konnten über den Summenparameter Retronecin PA-Kontaminationen mit 1,2-ungesättigten PA-Strukturen verlässlich detektiert und hochselektiv mittels HRGC-MS im SIM-Modus angezeigt werden. Die Validierung der Methode erfolgte durch die Verwendung von Senecio vernalis-Extrakt sowie authentischen PA-Standards und deren N-Oxiden. Unter Modifikationen der Probenaufarbeitung war diese Methode sowohl für Honig und Pollen, als auch für honighaltige Lebensmittel einsetzbar. Im weiteren Verlauf der Arbeit wurde die Methode durch die Synthese des deuterierten Standards di-Butyroyl-[9,9-2H2]-Retronecin zur Stabilisotopen-Verdünnungsanalyse (SIVA) erweitert und optimiert. Die entwickelte Methode erlaubt erstmals, anders als bei bereits vorliegenden Arbeiten zur Bestimmung von PA in Pflanzenteilen, eine exakte und selektive Bestimmung von PA im Spurenbereich, unabhängig von deren botanischem Ursprung oder chemischer Struktur (tertiäres PA, N-Oxide). In einem breit aufgestellten Screening von 216 Honighandelsproben und 35 Forschungshonigen – letztere umfassten 27 Senecio- und 8 Echium-Honige - konnten zum Teil erhebliche Mengen an PA nachgewiesen werden. Die Belastungsrate der einzelnen Probensets reichte von 9 bis zu 100%. Die hierbei ermittelten Gehalte lagen, berechnet als Retronecin-Äquivalente, zwischen 0,019 µg/g und 4,66 µg/g. Ergänzt wurden die analytischen Daten durch die Erhebung von mellisopalynologischen Daten. Hierbei zeigte sich, dass eine Bestimmung von PA-Pflanzenpollen über die relative Pollenhäufigkeit nach DIN 10760 nur eine geringe Aussagekraft bei der Riskioabschätzung besitzt. Zwar war die Anwesenheit von PA-Pflanzenpollen immer ein Indikator für das Vorkommen von PA, jedoch konnten über den relativen Pollengehalt keine Aussagen über die Höhe der PA-Belastung getroffen werden. In einer weiteren Studie zu PA-Gehalten in Pollen und Pollenerzeugnissen sind in den nativen Pollen die erwartet hohen PA-Gehalte bestätigt worden. Aber auch die in Vollsortimentsupermärkten und Reformhäusern häufig vertretenen Pollenprodukte wiesen PA-Gehalte auf, die im Mittel weit über den bei Honig festgestellten Werten lagen. So ergaben sich für die nativen Pollen aller bedeutenden, PA-produzierenden Pflanzenfamilien PA-Gehalte von 0,57-4,07 mg/g, während sich für die Pollenprodukte Gehalte von 1,08-16,35 µg/g feststellen ließen. Eine zusätzliche Erhebung von mellisopalynologischen Daten bestätigte deren bereits bei den Honigproben festgestellte, eingeschränkte Aussagekraft hinsichtlich des PA-Gehaltes. Durch ein Screening von 60 honighaltigen Lebensmitteln mit unterschiedlichen Honiganteilen konnte eine potentielle Downstream-Kontamination durch den Einsatz von hoch PA-belasteten Honigen im Herstellungsprozess nachgewiesen werden. Bei einer Belastungshäufigkeit von 13% lagen die hierbei ermittelten PA-Gehalte, berechnet als Retronecin-Äquivalente, bei 0,010-0,484 µg/g. Abschließend ist in modellhaft durchgeführten Filtrationsversuchen gezeigt worden, dass PA-Pflanzenpollen erheblichen Einfluss auf den PA-Gehalt des Honigs ausüben. Dennoch stellt eine Honigfiltration, wie sie in Anlage 1 der Honigverordnung zulässig ist, keine Möglichkeit dar, hoch mit PA belasteten Honig im PA-Gehalt zu senken. Vielmehr ließ sich mit den durchgeführten Versuchen eine Diffusion der PA aus Pollen in den Honig nachweisen. / Recently, the potentially PA contamination of food and feeding stuff attracted high attention. Humans are exposed to these toxins by comsumption of herbal medicine, herbal teas, dietary supplements or food containing PA plant material. In the ´International Programme on Chemical Safety (IPCS) the WHO states the potential threath to human health by PA. In pharmaceuticals, the use of these plants is regulated by the German Federal Health Bureau to a total PA intake of 1 µg per day for 1,2-unsaturated PA and their N-oxides within a six-week period per year, or, if six weeks are surpassed, the level is reduced to 0.1 µg total PA content per day. With regard to food the DFG-Senate Commission on Food Safety (SKLM) passed an opinion that […] The existing data base dealing with the content of PAs in honey collected from PA-containing plants […] as well as the data base dealing with the exposure of consumers to PAs are judged to be inadequate […]. Furthermore, they reminded that […] The main goal of future research should be the careful analytical determination of the PA content of honey and pollen […]. Initiated by this, the essential intention of this work was to develop a reliable tool for selective and quantitative determination of PA in the relevant range of concentration at ≤ 0,1 ppm in this food. Furthermore, the generation of a broad dataset should be the basis for the estimation of the potential risk of PA. First of all, a robust, reproducable and selective method consisting of Zn-dust reduction, strong cation exchance solidphase extraction (SCX-SPE), chemical reduction with LiAlH4, subsequent silylation and capillary gas chromatography-mass spectrometry (HRGC-MS) using SIM mode was established. This procedure converted PA-N-oxides into their basic tertiary form and, due to the subsequent chemical reduction, all mono- and diester PA resulted in their particular necine backbones. The newly developed method represented a reliable tool to detect potential PA contamination with toxic 1,2-unsaturated PA structures using retronecine as sum parameter by capillary gas chromatography-mass spectrometry (HRGC-MS) in SIM mode. This procedure was validated using extracts of Senecio vernalis as well as authentic standards of PA and their N-oxides. This method was applicable to honey and, after some modifications, to pollen and honey-containing foods, respectively. Furthermore, the analytical method was optimised by synthesis of a deuterium labelled standard, di-Butyroyl-[9,9-2H2]-retronecine, used for stable isotope dilution assays. For the first time, the present method allows the exact and selective determination of toxic PA by simply detecting a sum parameter which contains the toxic principle of the PA (1,2-double bond), independently from botanical origin or the oxidation state of the PA (tertiary PA, N-oxides). The method was applied to 216 commercially available flower honey samples and 35 exploratory harvested honeys, comprising 27 Senecio- and 8 Echium-honeys. Among these honeys the contamination degree was between 9 and 100% and the measured PA content was in a range from 0.019 to 4.66 µg g-1, calculated as retronecine equivalents. These results were completed by mellisopalynological data, performed according to DIN 10760. In this context, only a marginal significance between identification and relative frequency of PA plant pollen and the amount of PA contamination could be proven. The presence of PA plant pollen can rather be a precursor for a presence of PA. In a further study, the expected high amounts of PA in pure, floral pollen were verified. Furthermore, pollen products, available as food supplements in various supermarkets and health food shops, showed a much higher PA contamination compared to honey. Within the pure, floral pollen of the major PA producing plant families showed a PA contamination of 0.57 to 4.07 mg g-1. The analysed pollen products revealed PA contaminations in a range of 1.08 to 16.35 µg g-1. An additional data acquisition of mellisopalynological data showed similar results referring the PA levels as it did within the honey samples. Additionally, the modificated method was applied to 60 honey-containing foods with various contents of honey. Thereby a downstream contamination caused by the usage of honey highly contaminated with PA could be proven. Among these foods the degree of contamination was about 13% and the products revealed PA contents in a range of 0.010 to 0.484 µg g-1. Finally, it could be proven by filtration experiments in a bench-scale model, that pollen of PA producing plants have a direct effect on the PA content in honey. Nevertheless, it is not possible to reduce the PA contamination in honey by filtration of the contaminated honey as it is allowed by the German honey-concerning legislation. In fact, PA diffuse from pollen to honey.

Die zunehmende Entstehung von Resistenzen macht die Entwicklung neuer potenter Wirkstoffe zur Therapie von Infektionskrankheiten immer wichtiger. Dieser Aufgabe stellt sich auch der interdisziplinär aufgebaute SFB 630, in den sich die vorliegende Arbeit eingliedert. Innerhalb des SFBs wurden Isochinolinalkaloid-Derivate (IQs) synthetisiert, die aktiv gegen verschiedene Mikroorganismen sind. Bioinformatische Modellierungen bilden die für den jeweiligen Mikroorganismus spezifischen Stoffwechselwege ab. In Netzwerkanalysen können Änderungen metabolischer Flüsse durch pharmakologisch aktive Substanzen vorhergesagt werden. Gemeinsam mit bioinformatischen Modellen liefern die Metabolommessungen Hinweise auf mögliche Wirkmechanismen. Im Rahmen der vorliegenden Arbeit wurden verschiedene analytische Methoden etabliert, um antiinfektive Wirkungen dieser verheißungsvollen Leitstrukturen auf das Metabolom verschiedener Mikroorganismen zu untersuchen. Die aus den Metabolommessungen erhaltenen Daten fließen in diese Modelle ein und tragen zu deren Optimierung bei. Die Mikroorganismen wurden für die Metabolomanalysen mit aktiven IQs (für S. aureus und C. albicans GB-AP-143, für L. major GB-AP-304) inkubiert. Bei C. albicans erfolgte die Probennahme zu unterschiedlichen Zeitpunkten (lag-, log-, stationäre Phase), um auch die Zeitabhängigkeit der Effekte zu untersuchen. Zusätzlich dienten bei C. albicans als Kontrollen neben parallel angesetzten Zellkulturen ohne Inhibitor, auch Zellkulturen, denen das Lösungsmittel DMSO zugegeben wurde. Es wurden Extraktionsmethoden für die betreffenden Metabolite der hier untersuchten Mikroorganismen (S. aureus, C. albicans, L. major) etabliert. Dabei lag der Fokus auf polaren Metaboliten, da bioinformatische Modellierungen für die Effekte der IQs Änderungen vor allem im Purin- und Pyrimidinstoffwechsel der Mikroorganismen vorhersagten. Zur Analyse des Nukleotidstoffwechsels wurde eine ionenpaarchromatographische HPLC-Methode entwickelt und optimiert. Mit dieser Methode konnten Nicotinamidderivate und Nukleotide des Purin- und Pyrimidinstoffwechsels in Zellextrakten von S. aureus, C. albicans und L. major quantifiziert werden. Für eine Analyse des Wirkmechanismus von GB-AP-143 wurde die Zusammensetzung des Metaboloms von C. albicans mittels einer GC/MS-Methode bestimmt. Nach einer Derivatisierung des Extrakts mit Methoxyamin-HCl und MSTFA konnten in einem Lauf zugleich Target- und Fingerprintanalytik durchgeführt werden. Die Auswertung der Targetanalytik fand unter Anwendung der NIST-Datenbank und Vermessung von Standards statt. Hierbei konnten vor allem Aminosäuren quantitativ erfasst werden. Der Fingerprint wurde durch Einsatz multivariater statistischer Verfahren ausgewertet. Die Daten für die mit GB AP 143 behandelten S. aureus und die mit GB AP 304 behandelten L. major-Promastigoten liefern Hinweise auf eine Wirkung der IQs auf den Komplex-I der mitochondrialen Atmungskette. Für die Behandlung der C. albicans-Kulturen mit GB-AP-143 konnten komplexe Änderungen im Nukleotid- und Aminosäurestoffwechsel gemessen werden. So beeinflusste bereits der Zeitpunkt der Probennahme (lag-, log- oder stationäre Wachstumsphase) die Zusammensetzung des Metaboloms und auch das Lösungsmittel, das für die IQs verwendet wurde, verursachte komplexe Änderungen im Metabolom von C. albicans. Zusätzlich wurden Nukleotid- und Aminosäurekonzentrationen Fluconazol-resistenter C. albicans-Mutanten (TAC, UPC und MRR) untersucht. Im Nukleotidstoffwechsel waren sowohl Konzentrationssteigerungen als auch ein Absinken der Konzentrationen im Vergleich zum Wildtyp zu verzeichnen. Der Aminosäurestoffwechsel zeigte insgesamt einen verminderten Gehalt an Aminosäuren der Mutanten gegenüber dem Wildtyp. Da GB-AP-143 auch Aktivität gegen diese Mutanten zeigte, wurde exemplarisch die MRR-Mutante mit GB-AP-143 inkubiert, um zu untersuchen, ob die durch GB-AP-143 hervorgerufenen Änderungen im Nukleotid- und Aminosäurestoffwechsel ähnlich zu denen des Wildtyps sind. Es konnten im Nukleotidstoffwechsel gegenläufige Effekte für die Inkubation von GB-AP-143 des Wildtyps und der Mutante verzeichnet werden. Die Daten aus den HPLC/UV- und GC/MS-Messungen werden von der Bioinformatik zur Optimierung der verwendeten Modelle genutzt, um auf diese Weise die Wirkmechanismen der IQs besser modellieren zu können. Da das Cytochrom-P-450-Enzymsystem am Metabolismus von etwa 95 % aller Arzneistoffe beteiligt ist, wurden die Effekte ausgewählter IQs auf die sechs wichtigsten arzneistoffmetabolisierenden Enzyme (CYP1A2, 2C8, 2C9, 2C19, 2D6 und 3A4) mit Hilfe eines bereits etablierten CYP-Assays analysiert und näher charakterisiert. Im CYP-Assay zeigte sich für drei IQs eine CYP2D6-Hemmung. Die ausgeprägte CYP2D6-selektive Hemmung von GB-AP-110 ergab einen IC50-Wert von nur 109 nM. Die Charakterisierung der Hemmung ergab einen reversiblen, kompetitiven Inhibitionsmechanismus. / The increasing frequency of resistance towards antibiotics in the therapy of infectious diseases highlights the importance of the development of novel drugs against infectious diseases. This work is integrated in the Collaboration Research Center 630 (SFB 630), which has been formed to search for innovative solutions by joint interdisciplinary approaches. In the framework of this SFB, some new isoquinoline alkaloid derivatives (IQs) could be synthesized, which show distinct activities against various microorganisms. The present work focussed on the development of different analytical methods in order to determine the antiinfective properties of these promising lead structures on the level of the metabolome of different microorganisms. Metabolome measurements together with bioinformatic models provide information about the possible mode of action. By using bioinformatic models one can predict changes in metabolic fluxes caused by pharmacologically active substances. The integration of data from metabolic measurements can optimize the predictive power of these models. For these measurements the microorganisms were incubated with the active IQs (GB-AP-143 for S. aureus and C. albicans, GB-AP-304 for L. major). In the case of C. albicans sampling was carried out at different time points (lag-, log and stationary growth stage) in order to examine the time dependence. In addition cell cultures without inhibitor and with the addition of the solvent DMSO were used as controls for C. albicans. Initially, extraction methods for the respective metabolites of the microorganisms S. aureus, C. albicans and L. major were established. Since bioinformatic models predicted alterations especially in the purine and pyrimidine metabolism of these microorganisms as a consequence of treatment with IQs, priority was put on the extraction of polar compounds. To analyze the nucleotide metabolism an ion pair chromatography method was developed and optimized. By means of this method nicotinamide derivatives and nucleotides of the purine and pyrimidine metabolisms could be quantified in cell extracts of S. aureus, C. albicans and L. major. For a more holistic analysis of GB-AP-143´s effect on the composition of the C. albicans metabolome a GC/MS-method was developed. After a derivatisation step of the cell extract by using methoxyamine hydrochloride and MSTFA target and fingerprint analysis were conducted simultaneously. Target analysis was conducted by means of the NIST database and measuring reference standards. Particularly, amino acids could be quantified. Fingerprint analysis was interpreted using multivariate statistics. A detailed biological interpretation of metabolome data was not the focus of this study, because the main goal was the method development. However, data of S. aureus and L. major treated with the IQs gave hints towards an effect of the IQs on complex I of mitochondrial respiration. For the treatment of C. albicans with GB-AP-143 a complex alteration of the metabolites has been detected: already the time of sampling and the solvent, which was applied for GB-AP-143, have an impact on metabolome pattern of C. albicans. Additionally, the nucleotide and amino acid concentrations of fluconazol-resistant C. albicans mutants (TAC; UPC and MRR) have been investigated. Nucleotides showed increased as well as decreased concentrations compared to the C. albicans wild type. The overall concentration of amino acids was decreased in the mutants. Since GB-AP-143 also showed activity against these resistant mutants, exemplarily the MRR-mutant was incubated with this compound in order to determine, whether the effect of GB-AP-143 on the nucleotide and amino acid metabolism in the wild type also occurs in the mutant. Nucleotide metabolism showed antidromic effects for the incubation of wild type and mutant for GB-AP-143. This indicates that the metabolism of the mutant, which differs from the wild type already without treatment, also reacts differently on GB-AP-143-incubation. Data from HPLC/UV- and GC/MS-analysis are used by the bioinformatics to optimize the developed models in order to develop improved models for the mode of action of IQs. Since cytochrome-P-450-enzymes are involved in the metabolism of about 95 % of all drugs, the effects of selected IQs on the six main drug processing enzymes (CYP1A2, 2C8, 2C9, 2C19, 2D6 and 3A4) were investigated. Therefore, a previously developed and well-established in vitro test system was used. Three of the tested IQs showed an inhibition of CYP2D6. The distinct CYP2D6-selective inhibition of GB-AP-110 showed an IC50 value as low as 109 nM. The accurate characterization revealed a reversible and competitive mode of inhibition.

O glifosato é o herbicida mais usado em todo o mundo. Sendo assim, é necessário que se tenham programas de monitoramento do seu uso, para garantir o bem estar da lavoura e da população. O seu metabólito principal é o ácido aminometilfosfônico (AMPA) que apesar de possuir baixa toxicidade, é mais persistente que o glifosato. O presente trabalho teve como objetivo desenvolver uma metodologia de análise para o glifosato e o AMPA por cromatografia gasosa acoplada à espectrometria de massas (CG/EM), a fim de avaliar possíveis contaminações em amostras ambientais nas imediações do Rio Monjolinho, em São Carlos. Para a faixa estudada (1,0 ug L-1 a 500 ug L-1, os limites de detecção e quantificação para o AMPA foram de 0,15 e 0,45 ug L-1 e para o glifosato, 0,67 e 2,02 ug L-1. As recuperações em água variaram entre 96,2 e 121% e para solo 70,1 a 119%. O método proposto apresentou boa linearidade, exatidão, seletividade e sensibilidade. A robustez foi avaliada de acordo com o teste de Youden. O procedimento de extração foi baseado em reações ácido-base e realizou-se etapa de clean-up para água e sedimento. Para os pontos amostrados, houve resíduo de AMPA em dois pontos (4,19 e 6,22 ug L-1). Os resultados encontrados para DBO foram altos, estando acima do limite estabelecido para um corpo d\'água Classe 3, de acordo com a CONAMA 357. Isso pode ter ocorrido devido à grande quantidade de esgoto despejado no leito do rio. Os valores de nitrogênio e fósforo também estão elevados, o que indica uma alta eutrofização do leito do rio. Vale ressaltar a necessidade de se ter uma legislação que estabeleça um limite máximo permitido para o AMPA, visto que ele é mais persistente no ambiente do que o glifosato. / The glyphosate is the most widely used herbicide worldwide. Therefore, it is necessary to have programs for monitoring their use to ensure the welfare of the farming and population. Its main metabolite is the acid aminomethylphosphonic (AMPA) that despite having low toxicity, is more persistent than glyphosate. This study aimed to develop a methodology for analyzing glyphosate and AMPA by gas chromatography coupled to mass spectrometry (GC/MS) in order to assess possible contamination of samples environment in the vicinity of Monjolinho River in São Carlos. In the range studied (1.0 ug L-1 to 500 ug L-1, limits of detection and quantification were 0.15 and 0.45 ug L-1 for AMPA and 0.67 and 2.02 ug L-1 for glyphosate. The recoveries in water varied between 96.2 and 121% and for soil from 70.1 to 119%. The proposed method showed good linearity, accuracy, selectivity and sensitivity. The robustness was evaluated according to the Youden test. The extraction procedure was based on acid-base reactions and included a clean-up step for water and sediment. For the sampling sites, it was determined residual AMPA at two points (4.19 and 6.22 ug L-1). The results for BOD were high, being above the limit set for a waterbody Class 3, according to CONAMA 357. This may be due to large amount of sewage dumped into the river bed. The values of nitrogen and phosphorus are also high, which indicates a high eutrophication of the bed river. It is worth emphasizing the need of having a legislation that establishes a maximum allowed value for AMPA, whereas it is more persistent in environment than glyphosate.

An analysis of 6 ceramic shards from the Viking Age town Birka was performed by use of Gas Chromatography-Mass Spectrometry. A new interpretation of a previously analyzed material, was performed by analyzing the raw data from 18 ceramic shards. The total of 26 ceramic shards will together form a material base that is used to study the vascular use in an urban environment from the Viking Age. The result of the analysis is used to compare five different parts of Birka with each other, in order to investigate whether there are differences or similarities in pottery use. The results show that residues of animals are most common in all premises. It is also possible to note that fish have been cooked both in the Garnison area and at Stadsvallen.

A contaminação das águas subterrâneas e recursos hídricos superficiais nas últimas décadas representam uma ameaça para a saúde pública. No Brasil, as águas subterrâneas desempenham importante papel no abastecimento público e privado, suprindo as mais variadas necessidades de água em diversas cidades e comunidades. Com a finalidade de garantir a qualidade da água consumida, o ser humano vem desenvolvendo desde a década de 70 diversas técnicas analíticas para monitorar a presença de contaminantes. No entanto, algumas dessas técnicas podem gerar resíduos piores ao meio ambiente do que as já existentes. Desta maneira a partir da década de 90, surge uma nova tendência na forma como conduzir as análises químicas, com o intuito de reduzir o impacto ambiental. No presente trabalho, foi explorada uma técnica de extração que atende tais objetivos, denominada como microextração líquido-líquido dispersivo DLLME, desenvolvida por M.Rezaee e colaboradores. Onde foram validados os hidrocarbonetos policíclicos aromáticos, atingindo limites entre 0,04 a 1,56 μgL-1, também foram analisadas amostras de águas subterrâneas coletadas aleatoriamente, em 4 postos de gasolina localizados na cidade de São Paulo. / Contaminations of groundwater and surface water resources in recent decades represent a threat to public health. In Brazil, groundwater plays an important role in public and private provision, meeting the diverse needs of water in various cities and communities. In order to ensure the quality of water consumed, the human being has been developing since the 70`s analytical techniques to monitor the presence of contaminants. However, some of these techniques can generate worst waste to the environment than the existing ones. Thus from the 90\'s, a new trend in how to conduct chemical analysis, aiming to reduce the environmental impact arises. In the present study, we explored a technique for extraction that meets these goals, termed as dispersive liquid-liquid microextraction - DLLME, developed by M.Rezaee and collaborators. Where polycyclic aromatic hydrocarbons have been validated, reaching limits between 0.040 to 1.558 μgL-1, also groundwater samples randomly collected from four gas stations located in the city of São Paulo analyzed.

Waste plastic treatment was a global issue currently. A sustainable recycling process was required to recovery the monomer units from the polymer, thus avoiding environmental impacts due to disposal and enhancing the economic benefit from the recovered products. Pyrolysis was one of the promising process and previous MQP group has studied the pyrolysis with molten salts. In this work, a standardized and optimized pyrolysis reaction process of polystyrene was developed, based on the previous work on molten salt pyrolysis 5. The literature of pyrolysis mechanism and catalytic effects were reviewed as a guide process design. The orientation of the reactor was standardized to be consistent with literature record of preceding work. The positioning of the flow tube for the carrier gas and thermocouple were standardized in locations to provide mixing process of the reactant, the removal of products, and the accurate measurement of reaction temperature. The product collection system was also investigated and optimized to maximize collection efficiency while avoiding excessively low temperatures. The experiment results with standardized reaction configuration showed an improved styrene yield, 65%, compared with a previous yield of 44%. Then with the standardized reaction configuration, the catalytic effects of molten salt were studied at 400℃ pyrolysis temperature. Quantitative analysis indicated that the molten salt improved the styrene selectivity of the monomer compared to the dimer. Analysis of product composition and mass balance indicated formation of heavy non-GC detectable species in the liquid products. Gas phase secondary reaction during pyrolysis, and re-polymerization inside the liquid products, were discussed to explain the different styrene yield and the existence of heavy species. Based on the current results, recommendations for the pyrolysis temperature, initial reactants loading, and condenser temperature and analysis strategies were provided for further study of the molten salt pyrolysis of polystyrene.

Part 1: The emergence of environmentally friendly commercial products and their impact on fire debris analysis. Environmentally friendly products (i.e. green products) are environmentally preferable choices relative to comparable commercial products. They are readily available to the public, often highly flammable, and can be used by criminals as accelerants to facilitate the start and/or spread of fire. It is critical for analysts to have an understanding of their composition and chromatographic characteristics. Green products include paint thinners, solvents, removers, and cleaning and surface preparation products. As the composition of commercial products continually change over time, the fire debris community needs to be aware of the variety of environmentally friendly ignitable liquids that could be encountered during casework. Traditionally, when fire debris analysts have been trained, they are taught that most of the ignitable liquid residues they will encounter in casework are petroleum-based products. With the increasing emergence of non-petroleum based green products in the consumer marketplace, such products may be encountered more often than ever before in fire debris evidence submitted to forensic laboratories. Analysts should become familiar with the chromatographic features of these products as neat liquids as well as when present in fire debris samples. The purpose of this study is to introduce fire debris analysts to the prevalence of green products and increase knowledge regarding a variety of green product compositions and the characteristics they exhibit when analyzed as neat liquids and in "mock" fire debris samples. Several green products were analyzed as neat liquid samples and subsequently extracted from fire debris samples using typical fire debris extraction and analysis techniques in order to familiarize fire debris analysts with the chromatographic and mass spectral features of these products. General information about different types of green commercial products, their chromatographic and mass spectral characteristics, and their interpretation will be summarized. Analytical methods were developed for the analysis of environmentally friendly products and included considerations of gas chromatography oven temperature and ramp rate, hold times, and flow rate, as well as the scan rate and range of the mass spectrometer. Analyses involving common substrates were performed, including spiking green products onto various substrates with subsequent analysis and comparison of burned and unburned samples. Part 2: Application of fast GC/MS analysis for the identification of ignitable liquids in fire debris samples. Fire debris samples that contain ignitable liquid residues undergo a two-step process of extraction, most commonly via passive adsorption elution (PAE) onto an activated carbon strip, and instrumental analysis by gas chromatography/mass spectrometry. Upon completion of PAE, adsorbed compounds are eluted from the adsorbent with a suitable solvent and analyzed using (GC/MS) for the potential identification of ignitable liquid residues. A thorough evaluation of the literature revealed the average run time for gas chromatography of fire debris samples that contain hydrocarbon or petroleum based ignitable liquids to be 30 minutes. Additionally, a blank sample is run before an evidentiary sample to ensure solvent purity and to ensure any chromatographic carry over has not occurred between subsequent injections. The average run time, along with case volume, extraction times and case reviews contributes significantly to the backlog of samples to be analyzed in most crime laboratories around the country. Fast-GC/MS would significantly reduce analysis time, lower operating costs and would use less consumables. Based on a process known as pattern recognition, an initial goal of a fire debris analyst is to identify a pattern that is consistent with an ignitable liquid class. The standard method followed by most fire debris analysts use or base standard operating procedures (SOPs) on the American Society of Testing and Materials (ASTM) E1618, which defines the classes of commercial ignitable liquids based on chemical composition and boiling point range (or volatility). This study was conducted to optimize current methods of ignitable liquid detection and to optimize fast-GC/MS conditions for the identification of ignitable liquids in fire debris samples. Additionally, this study was conducted to determine if fast-GC/MS can reduce chromatographic separation times without sacrificing peak resolution and subsequently allow for ignitable liquid discrimination. Method development included considerations of flow rate, initial GC oven temperature, ramp rate, and mid and end temperature hold times. Fast-GC/MS conditions were tested on neat ignitable liquids from all nine ASTM E1618 classes. Optimizing fast-GC/MS method parameters led to an increase in sample throughput in comparison to traditional GC/MS methods. As a result, the GC/MS identification of ignitable liquids and their residues was performed in a quarter of the amount of time when compared to traditional methods.

Im Rahmen der Arbeit wurde eine Methode für die Quantifizierung von freiem 17β-Estradiol, Estron sowie der hydroxylierten und methylierten Metabolite im Brustgewebe entwickelt. Aufgrund der geringen Probengehalte erforderte dies eine gezielte Isolierung der Analyte aus der Probenmatrix sowie eine effektive Aufreinigung und Aufkonzentrierung, so dass eine Extraktion mit anschließender Festphasenextraktion durchgeführt wurde. Zudem wurde eine empfindliche Mess-Methode etabliert, welche auf Grundlage einer multi-reaction-monitoring-Methode, mittels Gaschromatographie und gekoppelten Triple-Quadrupol-Massenspektrometer, entwickelt wurde. Die Anwendbarkeit der Aufarbeitungs- und Mess-Methode wurde überprüft, indem diese auf 30 Realproben übertragen wurde. Dabei sind die ermittelten Gehalte mit den publizierten Daten der Gewebekonzentrationen von 17β-Estradiol, Estron und deren Metaboliten verglichen und Korrelationen mit ausgewählten Brustkrebs-begünstigenden Risikofaktoren betrachtet worden. Um ein quantitatives Metabolitenprofil von 17β-Estradiol, Estron und deren Metaboliten im Gewebe zu erstellen, wurden mit Hilfe einer multi-reaction-monitoring-Methode für alle Metabolite ein spezifischer Quanti- und Qualifier-Übergang etabliert. Durch die Optimierung der Ionisierungs- und Kollisionsenergien sowie der Initial-, Transferline- und Ionenquell-Temperatur beziehungsweise der dwell-time wurden Methoden- und Geräte-bedingte Empfindlichkeitsverluste so weit wie möglich reduziert, so dass maximale Signalintensitäten aller Quantifier-Übergänge gewährleistet waren. Zur gezielten Isolation sowie Aufreinigung und Anreicherung der Analyten,... ...so dass trotz der geringen Anzahl analysierter Gewebe-spenden der Einfluss des Body-Mass-Index und die Einnahme oraler Kontrazeptiva auf die Gehalte von 17β-Estradiol in der prämenopausalen Frau deutlich wurden. Die entwickelte Mess-Methode ermöglicht den routinemäßigen Einsatz für die Quantifizierung von freiem 17β-Estradiol, Estron und deren Methyl-Catecholen in humanem Brustgewebe. Beim Vergleich der berechneten Nachweisgrenzen von Catechol-Estrogenen mit Literaturangaben wurde herausgestellt, dass empfindlichere flüssigchromatographische Methoden als Methode der Wahl bei deren Analytik heranzuziehen sind. Die Übertragung der in Standardlösungen durchgeführten Versuche zur enzymatischen Hydrolyse von Glucuronid-und Sulfat-Konjugaten auf Gewebematrix stellt für weiterführende Arbeiten den entscheidenden Ansatzpunkt dar, um ein quantitatives Metabolitenprofil von freiem und gebundenem 17β-Estradiol, Estron und den Metaboliten in Brustgewebe erstellen zu können. / The aim of the present work was to develop a method for the quantification of free estradiol, estrone, catechol estrogens and methylation products in breast tissue. Due to trace amounts in the samples it was necessary to isolate the analytes from the sample matrix, purify and concentrate them prior to tissue extraction and solid phase extraction. Additionally, a sensitive method of measurement based on multi reaction monitoring using gas chromatography and triple quadrupole mass spectrometry was developed. The applicability of the sample preparation and measurement method was tested in 30 tissue samples by comparing the measured levels with published data concerning concentrations of estradiol, estrone and their metabolites in breast tissue. Also it was necessary to find possible correlations between the analytes concentrations and selected risk factors for breast cancer. In order to develop a quantitative profile of estradiol, estrone and metabolites in tissue, qualifier and quantifier transitions were established using the multi reaction monitoring mode. By optimising the ionisation and collision energy, the initial, transferline and ion source temperatures and the dwell-time, it was possible to reduce the loss of sensitivity caused by both method and equipment and maximize the signal intensity of all quantifier transitions. For the isolation, purification and concentration of the analytes... ...have some influence on the levels of estradiol in premenopausal women. The developed measurement method enabled a routine usage for the quantification of free estradiol, estrone and methylcatechols in human breast tissue. The comparison between the calculated detection levels of catechol estrogens and the published data led to the conclusion that liquid chromatographic methods are more sensitive. The transference of these methods and experiments conducted, such as for the enzymatic hydrolysis of glucuronide- and sulfate-conjugates on tissue matrix in standard solution, seem to offer a starting point for future research projects. Especially, for the purpose of establishing a quantitative metabolite profile of free and conjugated estradiol, estrone and their metabolites.

Nowadays, there is an increasing awareness on the importance of biomass waste as a renewable source of energy, materials and chemicals. In this context, the European project MOBILE FLIP aims at developing and demonstrating mobile conversion processes suitable with variousunderexploited agro- and forest based biomass resources in order to produce energy carriers, materials and chemicals. One of these processes is torrefaction, which consists in a mild thermal treatment, occurring typically between 200 and 300°C during a few tens of minutes in a defaultoxygen atmosphere. The solid product obtained has thermal and processing properties closer to coal, and thus is suitable as fuel for combustion or gasification. During torrefaction, condensable coproducts are released, that may also be source of “green” chemicals. It is therefore crucial to characterize them to optimize the torrefaction process and design industrial units. Up to now, only few works have been focused on characterizing and modelling both solid and condensable species during torrefaction versus operating conditions and feedstock type. Furthermore, these studies typically include a reduced number of biomasses. Cellulose, hemicellulose and lignin,which constitute biomass macromolecular composition, are determining properties to predict biomass behaviour during torrefaction. However, torrefaction tests on these constituents are rare and always based on commercial compounds, which were proved as little representative of the native biomass. The objective of this study is to analyse the influence of biomass characteristics, mainly represented by the macromolecular composition in cellulose, hemicellulose and lignin, on the global behaviour of biomass in torrefaction, both in terms of solid mass loss and of productionprofiles of the volatile species released, in function of the operating conditions.14 biomasses from the main biomass families (deciduouswood, coniferous wood, agricultural byproductsand herbaceous crops) were selected for this study. An optimized extraction procedure was proposed to recover cellulose, hemicellulose and lignin fractions from 5 reference biomasses. Experiments were performed on a thermogravimetric analyzer coupled to a gas chromatography mass spectrometer device through a heated storage loop system (TGA-GC/MS). Solid degradation kinetics and volatile release profiles were followed during torrefaction experiments combining non-isothermal (200 to 300°C at 3°C/min) and isothermal (300°C, 30 min) conditions, ensuring the chemical regime thanks to the appropriate operating conditions. The results obtained with the raw materials demonstrated that biomass macromolecular composition is a main factor influencing biomass behavior in torrefaction. Consequently, the heterogeneity of the resource results in a diverse behavior in torrefaction, particularly in the case of agricultural biomasses. The results with the extracted components evidenced their very different behavior compared to thecommercial compounds, particularly in the case of cellulose. This suggests that a limitation could be induced by the common use in literature of commercial components for torrefaction modelling. The impact on the characterization of macromolecular components was also shown to be prevailing in their behavior in torrefaction, especially in the case of hemicellulose sugar composition and cellulose crystallinity. Furthermore, differences in release kinetics of volatile species during torrefaction were observed, even for volatiles belonging to the same chemical family (acids, furans, ketones). Derived from these results, a torrefaction model based on the additive contribution of extracted cellulose, hemicelluloses and lignin to the global behavior of biomass in torrefaction was proposed, and this for the 5 representative biomasses.

Analytical instrumentation for quantitative in vivo stable isotope metabolic studies has included gas chromatography-mass spectrometry (GC-MS) and gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS). Limitations of existing MS techniques include excessive parent ion fragmentation, time-consuming sample preparation, and complex instrument operating parameters. In this thesis, procedures for set up and implementation of four GC-MS techniques are described. The first three replicate existing GC-MS single quadrupole methods on an ion trap, and are electron ionization (EI), external methane positive chemical ionization (PCI), and methane negative chemical ionization (NCI). The fourth method is a novel GC-MS mass-selected ion trap internal isobutane positive chemical ionization technique. Four groups of rats were administered isotopic linoleic acid, and liver tissue was collected for labelled linoleic and n-6 polyunsaturated fatty acids (PUFA) metabolites analyses. Qualitative utility of EI was confirmed, and its quantitative limitations exposed. Labelled 18:2n-6 and n-6 PUFA metabolites were detected using external methane PCI, though limited due to significant fragmentation. Methane NCI also detected labelled 18:2n-6, as well as minimizing fragmentation. However, time-consuming sample preparation and non-linear responses were major limitations. Internal isobutane PCI was useful in detecting isotopic 18:2n-6 and n-6 PUFA metabolites. Fragmentation was reduced compared to EI and external methane PCI. Limitations include fragmentation of HUFAs such as EPA and DHA. The novel internal isobutane PCI is more sensitive than external methane PCI and NCI, produces highly linear responses, is simpler and less expensive to operate than C-IRMS, offers reliable instrument operation, and sample preparation time is minimal. Regular quantitative analyses of HUFAs such as EPA and DHA may require further refinements such as using lower energy reagents than isobutane, including acetonitrile and ammonia.