A amostragem ambiental por esfregaço ou swipe samples é uma metodologia que vem sendo empregada rotineiramente pela Agência Internacional de Energia Atômica (AIEA) para verificar se os Estados signatários do Acordo de Salvaguardas estão realizando atividades nucleares não declaradas. A análise dessas amostras ambientais é complementar aos procedimentos de salvaguardas convencionais para a verificação das informações cedidas pelos Estados. Neste trabalho, foi descrita uma metodologia que visa a fortalecer os processos de salvaguarda nuclear e da ciência forense nuclear no país. A proposta é estudar e avaliar a técnica de ablação à laser acoplada a um espectrômetro de massa de alta resolução com fonte de plasma indutivamente acoplado (LA-HR-ICP-MS Laser Ablation High Resolution Inductively Coupled Plasma Mass Spectrometry), como técnica alternativa para a análise das amostras de esfregaço. Adicionalmente, foram empregadas outras técnicas complementares (atividade radioativa, microscopia eletrônica de varredura e espectroscopia de energia dispersiva) com o intuito de garantir maior segurança ao processo de inspeção de salvaguardas e investigações forenses nucleares. A precisão, medida através do desvio padrão relativo (DPR),dos resultados obtidos para as razões isotópicas 234U/238U, 235U/238U e 236U/238U do padrão analisado (CRM -125A) para a medida do fator de discriminação de massa foram, respectivamente, 1,3 %, 0,2 % e 7,6 %. Já as incertezas percentuais (u %), que também contemplam o DPR das medições, variaram de 3,5 % a 29,8 % para as medições da razão isotópica 235U/238U e de 16,6 % a 42,9 % para a razão isotópica 234U/238U. Esses valores mostraram-se compatíveis com outros estudos que utilizaram a mesma técnica para análise de amostras reais coletadas em uma instalação nuclear. As amostras coletadas apresentaram nível de enriquecimento que variou de (2,3 ± 0,7) % (amostra 3 corredor de acesso à instalação) a (17,3 ± 2,8) % (amostra 18 bancada de recuperação de UF4). A partir da coleta de amostras nesses diversos pontos, desde a entrada da instalação até a bancada em que se manuseia o urânio enriquecido, foi possível detectar diversos níveis de enriquecimento no isótopo 235U. / Environmental sampling performed by means of swipe samples is a methodology frequently employed by International Atomic Energy Agency (IAEA) to verify if the signatory States of the Safeguards Agreements are conducing unauthorized activities. Swipe samples analysis is complementary to the Safeguards ordinary procedures used to verify the information given by the States. In this work it was described a methodology intending to strengthen the nuclear safeguards and nuclear forensics procedures. The proposal is to study and evaluate the laser ablation high resolution inductively coupled plasma mass spectrometry (LA-HR-ICP-MS) technique as an alternative to analyze the real-life swipe samples. The precision achieved through the standard(CRM -125A)measurements, represented by the relative standard deviation (RSD), was respectively 1.3 %, 0.2 % e 7.6 %for the 234U/238U, 235U/238U e 236U/238Uisotopes ratios. The percent uncertainties (u %), which covers the RSD, ranged from 3.5 % to 29.8 % to the 235U/238U measurements and from 16.6 % to 42.9 % to the 234U/238U isotope ratio. These results were compatible with former studies performed by the LA-HR-ICP-MS that analyzed real-life swipe samples collected at a nuclear facility. Swipe samples collected from several points of the nuclear facility presented enrichment level ranging from (2.3 ± 0.7) % (sample 3) to (17.3 ± 2.8) % (sample18). They also allowed detecting different enrichment levels within the facility.

La faïence est une céramique caractérisée par son revêtement vitreux blanc opaque, grâce à l’adjonction d’oxyde d’étain ; sur ce fond on peut apposer les décors polychromes. Résultant d’une technologie abbaside (VIIIe siècle), elle connait son apogée en France au XVIIIe siècle. Les précédentes études menées sur la faïence moderne ont mis en évidence la nécessité de procéder à une approche pluridisciplinaire pour appréhender ce matériau complexe, ses techniques de fabrication, et cerner les enjeux de sa production. Ce travail de recherche s’inscrit dans la continuité pour s’élargir aux productions faïencières du Sud Ouest de la France quelques peu délaissées du point de vue archéométrique. La manufacture Babut, première faïencerie établie à Bergerac, fut sélectionnée dans ce but. La production perdure sur cinquante ans environ, jusqu’à la Révolution Française.En prenant en considération ces observations, cette thèse a permis tout d’abord d’identifier la production du point de vue des matériaux et des techniques, de mettre en place un référentiel géochimique, ainsi qu’établir un corpus stylistique de base pour cette manufacture. Ensuite l’étude a porté sur l’évaluation du potentiel de méthodes d’analyses non destructives pour l’attribution de provenance directe à partir de la surface des glaçures. Et enfin ce travail a permis d’examiner la place de la faïence bergeracoise et de sa production au sein du tissu industriel régional, et les replacer dans le contexte plus général de la production faïencière française à l’aube de la Révolution industrielle. / Faiences are ceramics defined by a white and opaque glaze, due to the addition of tin oxide decorated or not. This technology appeared during the Abbasid period (VIIIth) and was imported in France around the XIIIth century; they peaked in the XVIIIth century. The previous studies realized on the modern faïences, underline the importance of proceeding to a pluridisciplinarity approach to comprehend this complex material, in the frame of a more global research program on faiences, this work focuses on the production of the South West of France, which have only received limited archaeometrics studies over the last years. The manufacture selected is the fabric of Jean Babut established in the city of Bergerac. The production lasted fifty years until the French Revolution. By analysing all the different results, this work establishes the possibility to proceed to the identification of the production site from technical and material standpoints, to establish a reference group based on the chemical analysis and to design a stylistic reference corpus. A non-destructive methodological approach was developed to evaluate the potential of non-destructive methods to reattribute faiences to a center of production. In addition, this work also reestablishes the production of faiences in the local industrial context and in the french production before the Industrial Revolution.

Abstract Platinum group elements (PGEs) have a high commercial value and variety of applications in different fields of industry. One of the well-known applications is the use of palladium, platinum and rhodium in the catalytic converters of automobiles to reduce the amount of harmful gases emitted to the environment. Advanced analytical methods are needed to deal with issues related to development of new catalyst materials, recycling of PGEs from spent materials and for monitoring PGE emissions to the environment. In the first part of this study the emphasis was on the catalyst materials. Especially, reliable determination of ruthenium content in catalyst materials required further studies. Consequently, acid digestions in closed vessels using a microwave oven or high pressure asher were compared with a previously reported fusion method. Furthermore, the recovery of PGEs from spent materials is important due to many factors, for example, the high value of these metals, environmental aspects related to their production and possible availability issues in the future. Thus, utilization of microwave-assisted leaching and cloud point extraction (CPE) for the recovery of palladium, platinum, rhodium and ruthenium from catalyst materials was investigated. The second part of this study concentrated on the PGEs in environmental samples and the analytical challenges related to PGE determinations with inductively coupled plasma mass spectrometry (ICP-MS). Due to the use of PGEs in catalytic converters of automobiles, they are emitted to the roadside environment. The use of Pleurozium schreberi, a terrestrial moss, for active biomonitoring of these emissions was evaluated. Advanced analytical methods were needed to perform interference-free determinations of palladium, platinum and rhodium in these samples. Two alternative approaches for interference elimination were studied. Firstly, the interfering elements were removed using CPE as a chemical separation method. Secondly, interferences were eliminated using ammonia as a reaction gas with the novel ICP-MS/MS (inductively coupled plasma tandem mass spectrometry) technique. / Tiivistelmä Platinaryhmän alkuaineita hyödynnetään monissa teknisissä sovelluksissa. Yksi tunnetuimmista on autoliikenteen haitallisten päästöjen vähentäminen käyttäen palladiumia, platinaa ja rodiumia autojen pakokaasukatalysaattoreissa. Luotettavia analyysimenetelmiä tarvitaan esimerkiksi kehitettäessä uusia katalyyttimateriaaleja, kierrätettäessä platinaryhmän alkuaineita käytetyistä materiaaleista tai seurattaessa platinaryhmän alkuaineiden määrää ympäristössä. Tämän tutkimuksen ensimmäinen osa liittyi katalyyttien ruteniumpitoisuuksien määrittämiseen ja platinaryhmän alkuaineiden kierrätykseen katalyyttimateriaaleista. Erityisesti näytteenhajotusvaihe aiheuttaa usein ongelmia ruteniummäärityksissä. Tämän vuoksi tutkimuksessa verrattiin keskenään mikroaaltotekniikalla tai korkeapainetuhkistimella suoritettuja happohajotuksia ja aiemmin raportoidulla sulatemenetelmällä suoritettuja hajotuksia. Mikroaaltoavusteista liuotusta sovellettiin yhdessä samepisteuuton kanssa tutkittaessa katalyyttimateriaalien palladiumin, platinan, rodiumin ja ruteniumin kierrätysmahdollisuuksia. Aihe on ajankohtainen, kun huomioidaan platinaryhmän alkuaineiden korkea hinta, niiden tuotantoon liittyvät ympäristöasiat sekä saatavuuteen liittyvät epävarmuustekijät. Tutkimuksen toisessa osassa keskityttiin ympäristönäytteisiin ja erityisesti niiden ICP-MS -analytiikan (induktiiviplasmamassaspektrometria) haasteisiin. Autojen katalysaattoreista lähtöisin olevia platinaryhmän alkuaineita päätyy ympäristöön lähinnä teiden varsille. Näitä päästöjä arvioitiin aktiivista biomonitorointia käyttäen. Spektraaliset häiriöt vaikeuttivat kerättyjen sammalnäytteiden (Pleurozium schreberi) palladium-, platina- ja rodiumpitoisuuksien määrityksiä. Tämän vuoksi mittauksissa esiintyvien häiriöiden poistossa hyödynnettiin kahta eri lähestymistapaa. Näistä ensimmäisessä häiritsevät alkuaineet poistettiin kemiallisen erotusmenetelmän, samepisteuuton, avulla. Toisessa tavassa häiriöt poistettiin uudella ICP-MS/MS -tekniikalla (induktiiviplasmatandemmassaspektrometria) käyttäen ammoniakkia reaktiokaasuna.

[pt] Neste trabalho, métodos analíticos foram desenvolvidos para a determinação de elementos terras raras (REE, do inglês Rare Earth Elements), por espectrometria de massa e espectrometria de emissão óptica com plasma indutivamente acoplado (ICP-MS e ICP OES) numa amostra de asfalto. Foram avaliados os resultados analíticos através da análise de um material de referência certificado de rocha (GSP-1) e através da comparação dos resultados obtidos pelas duas técnicas. Dois diferentes métodos de lixiviação ácida foram empregados para o tratamento da amostra de asfalto, com HNO3 ou H2SO4, de maneira a simular os efeitos intensificados da chuva ácida. Também foi avaliada a extração da amostra com solvente orgânico e posterior lixiviação ácida, para identificar a origem (orgânica ou inorgânica) dos REE na amostra do asfalto, concluindo que eles estão presentes apenas na fração inorgânica da amostra. Os limites de detecção variaram entre 0,002 e 0,010 microgramas L elevado a menos 1 para a lixiviação com HNO3 e entre 0,001 e 0,014 microgramas L elevado a menos 1 com H2SO4, por ICP-MS, e entre 0,05 e 2,5 micrograma L elevado a menos 1 com HNO3 e entre 0,06 e 3,1 micrograma L elevado a menos 1 com H2SO4, por ICP OES. As determinações foram feitas com curvas analíticas com padrões aquosos e Rh como padrão interno. Além disso, a interferência espectral do BaO sobre Eu foi reduzida, através da precipitacao do Ba como BaSO4 quando se usou ácido sulfúrico para a lixiviacao da amostra de asfalto, reduzindo cerca de 94 porcento a concentracao de Ba em relação àquela medida no lixiviado com HNO3. As concentrações de REE variaram de 0,47 a 65,6 microgramas g menos 1, sendo que os terras raras leves (LREE) representaram 83 porcento do total de REE no asfalto investigado. O Ce foi o mais abundante dos REE no lixiviado do asfalto, seguido por Nd e La. A ordem das concentrações médias de REE no asfalto foi, em ordem decrescente: Ce maior Nd maior La maior Pr maior Sm maior Eu maior Gd maior Dy maior Er maior Yb maior Ho maior Tm maior Lu. / [en] In this study, analytical methods were developed for the determination of Rare Earth Elements by inductively coupled plasma -mass and -optical emission spectrometries (ICP-MS and ICP OES), in an asphalt sample. The analytical results were evaluated through the analysis of a rock certified reference material (GSP-1) and through the comparison of the results with the two techniques. Two different methods of acid leaching were employed for the asphalt sample treatment, with HNO3 or H2SO4, in order to simulate intensified acid rain effects. Additionally, the extraction of the sample with organic solvent and subsequent acid leaching was also evaluated to identify the origin (organic or inorganic) of REE in asphalt sample, concluding that they are present in the sample inorganic fraction. The obtained limits of detection ranged from 0.002 to 0.010 microgam L to the power of minus 1 for the leaching with HNO3 and from 0.001 to 0.014 microgram L to the power of 1 with H2SO4, by ICP-MS, and from 0.05 to 2.5 microgram L to the power of minus 1 with HNO3 and from 0.06 to 3.1 microgram L to the power of minus 1 with H2SO4, by ICP OES. The determinations were made with analytical curves with aqueous standards and Rh as internal standard. Moreover, the spectral interference by BaO on Eu was reduced, through the precipitation of Ba as BaSO4 when sulfuric acid was used for the asphalt sample leaching, decreasing about 94 percent the Ba concentration in relation to that measured in the leachate with HNO3. The concentrations of REE in the asphalt sample ranged from 0.47 to 65.6 microgran g to the power of minus 1, being the light rare earth elements (LREE) accounted for 83 percent of total REE on the asphalt investigated. Cerium was the most abundant REE in the asphalt leached, followed by Nd and La. The order of average concentrations of REE in the asphalt was, in descending order: Ce > Nd > La > Pr > Sm > Eu > Gd > Dy > Er > Yb > Ho > Tm > Lu.

Die Analyse von organischen Polymeren mittels Massenspektrometrie mit induktiv gekoppeltem Plasma nach Laserablation unterliegt starken Matrixeffekten. Mögliche Korrekturmodelle sollten basierend auf der tatsächlich ablatierten Elementmasse sowie der Signalintensität des Kohlenstoffisotops C-13 (13C-Korrektur) entworfen werden. Damit die 13C-Korrektur erfolgreich verlaufen kann, ist ein direkter Zusammenhang zwischen ablatiertem Kohlenstoff und gemessenem 13C-ICP-MS Signal erforderlich. Dies wurde überprüft, indem unter Anwendung der konfokalen Mikroskopie das Ablationskratervolumen bestimmt und der ablatierte Kohlenstoff berechnet wurde. Dieser Zusammenhang konnte sowohl für undotierte, wie auch für elementdotierte Polymere und die Verwendung verschiedener Ablationsgase (Helium, Argon und Sauerstoff) ermittelt werden. Eigens hergestellte polymere elementdotierte Standardmaterialien wurden hinsichtlich der Matrixeffekte charakterisiert. Als größter Einflussfaktor stellte sich die unterschiedliche Ablationsrate bei der Ablation verschiedener Polymere heraus. Unter Anwendung der tatsächlich ablatierten Elementmasse sowie unter Anwendung der 13C-Korrektur konnte die Matrixabhängigkeit deutlich verringert werden.

[pt] A utilização de drogas à base de platina é o tratamento de primeira linha para diversos tipos de câncer. Pacientes tratados com estas drogas apresentam resultados melhores que os obtidos com outros regimes de quimioterapia para os mesmos tipos de malignidades. As principais limitações para a utilização destes compostos são os efeitos colaterais severos e a resistência dos tumores ao tratamento. A cisplatina foi a primeira droga dessa categoria a ser empregada. Desde o final dos anos 70 até hoje, esta droga vem sendo amplamente utilizada e com sucesso substancial. Acredita-se que o principal mecanismo de ação desta classe de medicamentos seja a ligação de dois sítios ativos da platina com o DNA das células tumorais, impedindo sua multiplicação e finalmente induzindo a apoptose, o que provoca a redução, e em alguns casos a eliminação, dos tumores. Entretanto, devido à complexidade dos mecanismos envolvidos, uma descrição clara da atuação intracelular destas drogas ainda não foi estabelecida. A combinação de técnicas de separação como eletroforese ou cromatografia líquida de alta performance com técnicas de espectrometria atômica tem se apresentado como uma poderosa alternativa para investigação de fenômenos biológicos que envolvem, de alguma maneira, espécies metálicas. A hifenação destas técnicas permite a separação e detecção em linha de biomoléculas contendo metais, possibilitando a obtenção de informações únicas sobre os processos biológicos. O presente trabalho apresenta o desenvolvimento de métodos analíticos utilizando eletroforese em gel de agarose (GE), cromatografia líquida de alta eficiência (HPLC) e espectrometria de massa com plasma indutivamente acoplado (ICP-MS) para a determinação de biomoléculas contendo platina em materiais biológicos. O principal objetivo do trabalho é fornecer ferramentas analíticas para o estudo dos mecanismos de ação de drogas à base de platina em humanos. Foram utilizados diversos materiais biológicos, como sangue, urina e culturas de células. A cromatografia líquida em fase reversa foi usada na determinação das drogas intactas e de seus produtos de hidrólise; a cromatografia de exclusão por tamanho foi empregada para a avaliação de proteínas presentes nas amostras enquanto a cromatografia de par iônico para separação de fragmentos de DNA. A detecção de platina nos eluatos por ICP-MS permitiu a obtenção de cromatogramas limpos apresentando claramente as moléculas contendo platina. A evidência da aplicabilidade dos métodos desenvolvidos foi avaliada com a prospecção de biomarcadores de eficiência do tratamento com cisplatina. Diversas linhagens celulares foram expostas a diferentes tratamentos com cisplatina e tiveram seus comportamentos avaliados. A determinação de adutos de DNA contendo platina apresentou-se como uma interessante perspectiva para a obtenção de um biomarcador de resistência ao tratamento com cisplatina. / [en] The use of platinum-based drugs is the first line treatment for many cancers. Patients treated with these drugs present better outcome when compared with other chemotherapy regimens for the same types of malignancies. The major limitations to the use of these drugs are the severe side effects and resistance tumors present to the treatment. Cisplatin was the first platinum-based drug to be approved for human use. Since the late 1970 s until today, this drug has been widely used with great success. It is believed that the major mechanism of action of these drugs is the binding of two active sites of platinum complexes with the DNA of the tumor cells, preventing their multiplication and finally inducing apoptosis, that leads to a reduction, and in some cases eliminating, tumors. However, due to the complexity of the mechanisms involved, a clear description of the intracellular action of these drugs has not been established. The combination of separation techniques such as electrophoresis or high performance liquid chromatography with atomic spectrometric techniques has emerged as a powerful alternative for investigation metal-related biological phenomena. The so called hyphenation of these techniques allows the separation and detection of biomolecules containing metals, making possible to obtain unique information about biological processes. This work presents the development of analytical methodologies using agarose gel electrophoresis (GE), high performance liquid chromatography (HPLC) and inductively coupled plasma with mass spectrometry (ICP-MS) for the determination of platinum-containing biomolecules in biological materials. The main objective of this work is to provide analytical tools for the study of the mechanisms of action of platinum-based drugs in humans. Various biological materials such as blood, urine and cell cultures were used. Reverse phase liquid chromatography was used for the determination of intact drugs and its hydrolysis products; size exclusion chromatography was used to assess the protein profile in samples while the ion-pair chromatography for separation of DNA fragments. The detection of platinum in the eluates by ICP-MS allowed the obtention of clean chromatograms clearly presenting the platinum-containing molecules. The evidence of the applicability of the developed methods was assessed with the search for biomarkers of efficacy of treatment with cisplatin. Several cell lines were exposed to different treatments of cisplatin and their behavior were evaluated. The determination of DNA adducts containing platinum presented an interesting approach for obtaining a marker of resistance to cisplatin treatment.

Le comportement des actinides dans l’environnement (site miniers, sites de stockagedes déchets et sites contaminés) dépend de l’interaction de ces éléments avec des ligandsprésents dans les différents milieux (air, sol, eaux superficielles et souterraines). Lesinteractions des actinides avec des ligands organiques polyfonctionnels peuvent piéger ceséléments ou au contraire favoriser leur migration.Dans ce travail, le comportement d’actinides aux degrés d’oxydation III, V et VI ensolution aqueuse a été étudié en présence de deux ligands polyaminocarboxyliques : l’acidenitrilotriacétique (NTA) et l’acide iminodiacétique (IDA).L’étude de la complexation du protactinium pentavalent par NTA et IDA a été conduiteselon une double approche : une étude thermodynamique réalisée par extraction par solvant,avec le protactinium à l’échelle des traces (C233Pa < 10-10 M) et une étude structurale, avecle protactinium en quantité pondérable (C231Pa ≈ 10-3 M), réalisée par Spectroscopied’Absorption des rayons X (SAX) et complétée par des calculs de chimie quantique.Les variations du coefficient de distribution de Pa(V) dans le systèmeTTA/Toluène/ HClO4 / NaClO4 /Pa(V)/ligand en fonction de la concentration de ligands àplusieurs températures et/ou acidités ont permis de déterminer la stœchiométrie maximale et lacharge moyenne des complexes Pa(V)-NTA et Pa(V)-IDA ainsi que les constantes de formationassociées à chaque espèce. Une stœchiométrie maximale de 2 a été observée pour les deuxsystèmes. Pour le système Pa-NTA, les deux complexes successifs prédominants dans la phaseaqueuse sont une espèce neutre PaO(NTA) pour le complexe (1:1) et une espèce de charge -3pour le complexe (1:2) qui pourrait correspondre à PaO(NTA) -:R. Une étude complémentairepar Electrophorèse Capillaire couplé à un spectromètre de masse (EC-ICP-MS) a permis deconfirmer la charge (-3) du complexe Pa − (NTA)-. Pour le système Pa-IDA, les résultats ontmontré la formation successive de deux complexes positifs PaO(IDA)3+ et PaO(IDA)+2.Lagéométrie de coordination et les distances interatomiques des complexes de stœchiométriemaximale ont été déterminés à partir de calculs DFT ou de dynamique moléculaire.L’étude de la complexation d’actinides trivalents (Am et Cf) et hexavalents (Np et Pu)par IDA a été réalisée par EC-ICP-MS. Les variations de la mobilité électrophorétique globaleen fonction de la concentration en IDA-R ont permis de déterminer les constantes de formationdes actinides avec IDA. Les constantes de formation des complexes de stœchiométrie (1:1)sont rapportées pour Am (III) et Cf (III). Concernant les complexes Np (VI) et Pu (VI) avecl'acide iminodiacétique, les constantes de formation des complexes de stœchiométriques (1:1)et (1:2) ont été déterminées. / The behaviour of actinides in the environment (in the surroundings of uranium miningsites, waste storage or contaminated sites) depends on the interaction of these elements with theorganic ligands present in these different systems. The interactions of actinides withpolyfunctional organic ligands could trap the element or inversely, favour their migration.In this work, the behaviour of actinides (III, V and VI) in aqueous solution was studied in thepresence of two polyaminocarboxylic ligands: nitrilotriacetic acid (NTA) and iminodiaceticacid (IDA).The study of the complexation of pentavalent protactinium by NTA and IDA was conductedaccording to a dual approach. A thermodynamic study carried out by liquid-liquid extraction inthe system TTA/Toluene/HClO4/NaClO4/Pa(V)/ligand, with protactinium at tracer scale (C233Pa< 10−10 M), has been implemented in order to determine the stoichiometry, the mean charge ofcomplexes and the associated formation constants. A structural approach with protactiniumin weighable quantity (C231Pa≈ 10−3 M) was conducted in order to determine the coordination geometry of complexes and interatomic distances.The variations of the distribution coefficient D of Pa(V) as a function of nitrilotriacetateconcentration enabled to determine that the maximum stoichiometry for the Pa-NTA and Pa-IDA systems is 2. For the Pa-NTA system, the two successive complexes predominant in theaqueous phase are a neutral species for (1:1) complex PaO(NTA) and charge -3 for the (1:2)which may correspond to PaO(NTA) -:R. In addition, Capillary Electrophoresis InductivelyCoupled Plasma Mass Spectrometry experiments have been performed to confirm the charge (-3) for the maximum stoichiometry complex. For the Pa-IDA system, the results have shownthe successive formation of two positive complexes PaO(IDA)3+ and PaO(IDA)+2. Optimized structures of Pa-NTA and Pa-IDA complexes were established using DFT or molecular dynamics calculations.The complexation of trivalent (Am and Cf) and hexavalent (Np and Pu) actinides with IDA was performed by Capillary Electrophoresis coupled with Inductively Coupled Mass Spectrometry(CE-ICP-MS). The variations of overall electrophoresis mobility of actinides species as function of CIDA2 allow the determination of the formation constants of actinides IDAcomplexes. The formation constants of complexes with (1:1) stoichiometry are reported forAm(III) and Cf(III). Concerning the complexes Np(VI) and Pu(VI) with iminodiacetic acid,formation constants of (1:1) and (1:2) stoichiometry complexes were determined

Pour accéder à la caractérisation des matrices complexes pétrolières, la chromatographie en phase liquide bidimensionnelle couplée à une détection spécifique comme la spectrométrie de masse à couplage inductif (LCxLC-ICP-MS/MS) s’avère être une solution pertinente. Un tel couplage permet d’envisager la spéciation des contaminants soufrés ou encore métallés (vanadium et nickel). Ce couplage reste, à notre connaissance, inédit aujourd’hui et sa mise en place a nécessité en premier lieu d’évaluer différents systèmes d’introduction de l’échantillon en amont de la détection. La comparaison de ces systèmes, au regard de la dispersion qu’ils génèrent, a été effectué afin de conserver la qualité de séparation obtenue en sortie du système LCxLC. La seconde partie du développement instrumental a concerné l’optimisation de la partie LC×LC. Le choix des différents mécanismes de rétention dans les deux dimensions étant primordial au vu de la complexité des échantillons (polarité, solubilité, poids moléculaire…). De plus, l’introduction de matrices organiques dans les sources plasma reste un réel défi qu’il a fallu évaluer, celles-ci pouvant être la cause de nombreuses contraintes analytiques. Enfin, une fois la méthodologie off-line SECxRPLC-ICP-MS/MS développée, elle a été appliquée à différents échantillons montrant qu’elle peut être considérée comme une solution intéressante pour expliquer le comportement de certaines matrices au sein des unités de raffinage, par l’intermédiaire de la comparaison de cartographies 2D / Two-dimensional liquid chromatography coupled with a specific detection such as inductively coupled plasma mass spectrometry (LCxLC-ICP-MS/MS) proves to be a relevant technique for the characterization of petroleum complex matrices. Such coupling makes it possible to consider the speciation of sulfur or metal contaminants (vanadium and nickel). Firstly, the evaluation and the comparison of several sample introduction systems was performed, with regard to the dispersion induced in the system, in order to keep a high efficiency from the LCxLC system. The second part of the instrumental development concern the optimization of both dimensions. The choice of the different retention mechanisms is essential given the complexity of the samples (polarity, solubility, molecular weight...). Additionally, the introduction of organic matrices in the plasma remains a real challenge which could be the cause of many instrumental and analytical issue. Finally, once the off-line SECxRPLC-ICP-MS/MS method was developed, it was applied to different samples showing how it can be considered as an interesting tool to explain the behavior of matrices within the refining units, through the comparison of 2D-contour plots

Durch die Untersuchungen der vorliegenden Arbeit ist es erfolgreich gelungen die beiden gram-positiven Mikroorganismen Lysinibacillus sphaericus JG-A12 und Lysinibacillus sphaericus JG-B53 unter geregelten und idealen Kultivierungsbedingungen im Bioreaktor in hinreichenden Biomasseausbeuten zu kultivieren. Aus der Biomasse beider Stämme ist es anschließend gelungen, die primären Zellwandkomponenten bestehend aus Membranlipiden, Peptidoglykan mit sekundären Zellwandpolymeren und S-Layer-Proteinen in reiner Form und in guten Ausbeuten zu extrahieren. Diese Zellwandkomponenten wurden dann unter Verwendung von biochemischer und strukturanalytischer Methoden charakterisiert. Dabei ist es erstmals gelungen, die Membranlipide beider genutzter Mikroorganismen in Bezug auf deren Zusammensetzungen der enthalten hydrophoben Fettsäuren und der hydrophilen phosphathaltigen Kopfgruppen zu charakterisieren. Durch die vergleichend durchgeführten Metallbindungsversuche im Batch-Verfahren konnten Bindungspräferenzen intakter Zellen von Lysinibacillus sphaericus JG-A12 und Lysinibacillus sphaericus JG-B53 und deren isolierten Zellwandkomponenten mit den Metallen As, Au, Cd, Eu, Pb, Pd, Pt bzw. U untersucht werden. Dabei konnten sowohl in den Untersuchungen intakter Zellen und der primären Zellwandbestandteile deutlich höhere Metallsorptionsraten und Metallentfernungseffizienzen für Lysinibacillus sphaericus JG-B53 festgestellt werden als dies bei Lysinibacillus sphaericus JG-A12 nachzuweisen war. Dies macht diesen Stamm für potentielle technische Anwendungen als metallselektives biosorptives Material weitaus interessanter. Die Untersuchungen der Einzelkomponenten in Suspension lieferten jedoch nur begrenzt Informationen zur Interaktion der Metalle mit den Schichten wie sie unter natürlichen Bedingungen in der Zelle vorkommen. Daher wurden unter Verwendung der QCM-D erstmals die primären Zellwandkomponenten beider Mikroorganismen (S-Layer und Peptidoglykan) sowie von Referenzlipiden an Grenzflächen erfolgreich im nanoskaligen Bereich abgeschieden und online verfolgt. Dadurch war es möglich vereinfachte Einzelschichtsysteme der gram-positiven bakteriellen Zellwand nachzubilden. In den Untersuchungen konnten stabile Schichten generiert werden, welche vergleichbar zu dem Schichtsystem vitaler Zellen sind. Zusätzlich konnte bei den Abscheidungen der S-Layer-Proteine SlfB und Slp1 der positive Effekt von Polyelektrolytmodifizierungen auf das Rekristallisationsverhalten, die Schichtstabilität und den Bedeckungsgrad auf der technischen Oberfläche aufgezeigt werden. Zur Untersuchung der Metallinteraktion zellulärer Einzelschichtsysteme wurden in dieser Arbeit exemplarisch nach den erfolgreichen Untersuchungen zur Rekristallisation, die S-Layer-Proteine als erste Interaktionsschicht des Gesamtzellsystems mit der QCM-D untersucht. Diese stabilen und intakten Schichten konnten analog zu den Schichtuntersuchungen der reinen biologischen Komponenten und nach den QCM-D Metallinteraktionsstudien mit den S-Layer Strukturen mittels der Rasterkraftmikroskopie (AFM) untersucht und bildgebend dargestellt werden. In weiteren spektroskopischen Untersuchungen (TRLFS) der Zellwandkomponenten konnten die lumineszierenden Eigenschaften von Europium ausgenutzt werden, um das Metallbindungsverhalten der einzelnen Komponenten als auch des Gesamtsystems der Zellen beider Mikroorganismen zu bestimmen. Somit konnte Europium als spektroskopische Sonde eingesetzt werden um Rückschlüsse die Biomolekül-Metallwechselwirkungen zu ermöglichen. Dabei konnten vor allem mit den beiden oberflächennahen Zellschichten Lösung teilweise sehr starke Metall-Biomolekül-Wechselwirkungen beobachtet werden.

Die Analyse von organischen Polymeren mittels Massenspektrometrie mit induktiv gekoppeltem Plasma nach Laserablation unterliegt starken Matrixeffekten. Mögliche Korrekturmodelle sollten basierend auf der tatsächlich ablatierten Elementmasse sowie der Signalintensität des Kohlenstoffisotops C-13 (13C-Korrektur) entworfen werden. Damit die 13C-Korrektur erfolgreich verlaufen kann, ist ein direkter Zusammenhang zwischen ablatiertem Kohlenstoff und gemessenem 13C-ICP-MS Signal erforderlich. Dies wurde überprüft, indem unter Anwendung der konfokalen Mikroskopie das Ablationskratervolumen bestimmt und der ablatierte Kohlenstoff berechnet wurde. Dieser Zusammenhang konnte sowohl für undotierte, wie auch für elementdotierte Polymere und die Verwendung verschiedener Ablationsgase (Helium, Argon und Sauerstoff) ermittelt werden. Eigens hergestellte polymere elementdotierte Standardmaterialien wurden hinsichtlich der Matrixeffekte charakterisiert. Als größter Einflussfaktor stellte sich die unterschiedliche Ablationsrate bei der Ablation verschiedener Polymere heraus. Unter Anwendung der tatsächlich ablatierten Elementmasse sowie unter Anwendung der 13C-Korrektur konnte die Matrixabhängigkeit deutlich verringert werden.