[pt] O presente trabalho visou estudar a geocronologia da contaminação na Baía de Guanabara por compostos orgânicos de estanho e alguns metais de relevância ambiental. Foram coletados testemunhos sedimentares em cinco pontos da Baía de Guanabara em regiões não afetadas diretamente por fontes de contaminação conhecidas. Os trinta primeiros centímetros de cada testemunho foram seccionados em segmentos finos nas estações onde haviam dados de taxas de sedimentação. Para duas estações, onde as taxas de sedimentação não eram conhecidas, os testemunhos coletados foram seccionados em camadas de três centímetros e, em seguida, datados pelo método de Pb-210. As concentrações dos metais foram determinadas por ICP-MS após decomposição das amostras a quente com HNO3 e H2O2. A especiação dos compostos orgânicos de estanho foi realizada por cromatografia gasosa com detecção por fotometria de chama pulsante após extração por solvente, derivação, eliminação de enxofre e limpeza dos extratos. As maiores concentrações (em ng g-1 sedimento, como Sn) para os diferentes compostos organoestânicos encontrados foram: TBT (82), DBT (28) e MBT (126). As distribuições espacial e temporal das espécies orgânicas de estanho nos sedimentos apresentaram-se muito irregulares e há pouca evidência de degradação após a sedimentação. Foram feitas tentativas de estimar a constante de degradação do TBT e os fluxos foram calculados utilizando-se as taxas de sedimentação e as concentrações. As concentrações de organoestânicos são baixas em comparação com as reportadas para a Marina da Glória por Fernandez (2001) e confirmam a tendência à rápida deposição dos compostos orgânicos de estanho próximo às fontes dos mesmos. As concentrações dos metais-traço investigados (em ug g-1 sedimento), As (1,31-11,5), Ca (2,08x103-8,72x103), Cd (< L. D.-7,94x10-1), Cu(0,222x101-5,58x101), Fe (0,564x104-3,33x104), Hg (0,046-2,35), Mg (0,128x104-1,31x104), Mn (0,496x102-11,8x102), Ni (0,288x101-3,17x101), Pb (0,505x101-8,18x101), Sn (0,130x101122), Zn (0,162x10²-2,30x10²) quando normalizados em relação à concentração de alumínio, mostraram, em geral, um acréscimo ao longo do tempo, demonstrando que, desde a última avaliação geocronológica, realizada na década de 80 do século XX, não há redução visível no aporte de contaminantes. Não foram encontrados dados pretéritos de arsênio em sedimentos da Baía de Guanabara. Em algumas estações, os valores encontrados para este elemento são comparáveis às concentrações em estuários sob influência de atividades de mineração. Foram feitas correlações de Spearman entre as propriedades medidas, incluindo carbono total e orgânico, nitrogênio total, enxofre total, pH, potencial redox, umidade e porosidade, revelando dependências de fontes e de propriedades físico-químicas. / [en] The present work aimed at studying the geochronology of contamination in Guanabara Bay as related to organotin compounds and a number of metals of environmental relevance. For this, sediment cores were taken from five different stations in the bay selected as to represent average conditions not immediately affected by known sources. The cores were sliced in thin segments up to 30 cm below the top layer. For two stations where sedimentation rates were not known cores were collected and sliced in 3 cm segments and thereafter dated by using Pb-210. Metal concentrations were determined by ICP-MS after digestion of freeze dried sediments in HNO3 and H2O2 under heat. Organotin speciation was carried out in a GC-PFPD instrument after solvent extraction, derivatization, sulfur elimination and clean-up. The higher organotin concentrations (ng g-1 as tin) found were: TBT (82), DBT (28) and MBT (126). Space-time distribution of organotin species in the sediments were very irregular and there is little evidence that extensive degradation occurs after sedimentation. Attempts are presented to derive an estimate for TBT degradation constant. Fluxes were calculated using the sedimentation rates and concentrations. Organotin concentrations are low in comparison to those reported for Marina da Glória by Fernandez (2001), and they confirm the tendency of organotins to settle down rapidly in the vicinities of the sources. The concentrations (ug g-1) of the investigated metals (As (1.31-11.5), Ca (2.08x103-8.72x103), Cd (<dl-7.94x10-1), Cu (0.222x101-5.58x101), Fe (0.564x104-3.33x104), Hg (0.046-2.35), Mg (0.128x104- 1.31x104), Mn (0.496x102-11.8x102), Ni (0.288x101-3.17x101), Pb (0.505x101-8.18x101), Sn (0.130x101-1.66x101), Zn (0.162x102-2.30x102)) when normalized to the Al concentration show, in general, an increase over time, demonstrating that since the last geochronological work conducted in the early 1980s there was no visible reduction in the contaminants input. No data for arsenic was found prior to this date for Guanabara Bay sediments. In some stations arsenic concentrations are comparable to those found in estuaries under influence of mining activities. Spearman correlations among the several measured properties, that include also total and organic carbon, total nitrogen, total sulfur, pH, redox potential water content and porosity, were calculated revealing source and physico-chemical dependencies.

Die chemische Zusammensetzung pharmazeutisch verwendeter Gläser (Ampullen, Infusionsglas, Fertigspritzenbehältnisse) wurde mit Laser-Ablation-ICP-Massenspektrometrie (LA-ICP-MS) untersucht. Dazu kam ein Nd:YAG-Laser mit einer Frequenz von 10 Hz im q-switch mode mit der Grundwellenlänge sowie der 4. Harmonischen (1064 und 266 nm) zum Einsatz. Ziel war die Identifizierung verschiedener Arten pharmazeutisch eingesetzter Gläser. Folgende Isotope wurden zur Charakterisierung bestimmt: 7Li, 11B, 23Na, 24Mg, 27Al, 28Si, 29Si, 30Si, 39K, 42Ca, 47Ti, 57Fe, 90Zr, 121Sb, 137Ba. Die quantitativen Ergebnisse zeigten relative Standardabweichungen von 1.8 % bis 8.0 %. Die Normierung der Daten erfolgte unter Bezug auf 29Si. Mit Referenzmaterial verschiedener Zusammensetzung (Natron-Kalk-Glas, Borosilikatglas und Bleiglas) wurden externe Mehrpunktkalibrationen erstellt. Für alle gemessenen Isotope konnten lineare Kalibrierfunktionen festgestellt werden. Die Richtigkeit des Verfahrens wurde durch Aufschluß des Standardmaterials wie auch der untersuchten Proben mit Flußsäure/Salpetersäure und Messung der Aufschlußlösungen nach externer Kalibration durch Multielement-Standards gezeigt. Aufschluß- und Laserablation-Ergebnisse zeigten gute Übereinstimmung und wichen um bis zu 8 % voneinander ab. Zur Prüfung repräsentativer Ablation der Proben wurden die Isotopenverhältnisse bestimmt. Auch hier zeigte sich eine ausreichende Übereinstimmung mit den theoretischen Werten. Weiterhin wurden Präzision, Empfindlichkeit, Selektivität, Nachweisgrenze, Bestimmungsgrenze und Robustheit des Verfahrens bestimmt. Im Unterschied zu den Ergebnissen unter der Grundwellenlänge führte die Verwendung der Laserwellenlänge 266 nm zu keinen signifikanten Verbesserungen der Resultate. Die Aufnahme transienter Signale sollte zur Erkennung von Elementverteilungen in Glasoberflächen dienen. Am Beispiel eines Natron-Kalk-Glases wurden in einzelnen Schichten verschiedene Elementkonzentrationen festgestellt. Weiterhin konnten mit dieser Technik Glasampullen erkannt werden, deren Oberflächen durch ein Silikonisierungsverfahren vergütet waren. Zum Abschluß der Untersuchungen wurde versucht, auf Grundlage der unnormierten Intensitätsdaten der LA-Messungen Gläser zu klassifizieren. Hierzu wurde die lineare Diskriminanzanalyse eingesetzt, mit deren Hilfe nach Variablenreduktion (8 von 13 Isotopen) alle eingesetzten Glasarten korrekt identifiziert werden konnten. Durch das entwickelte und validierte LA-ICP-MS-Verfahren steht eine leistungsfähige Technik zur quantitativen Untersuchung der chemischen Zusammensetzung von Gläsern sowie zur Identifizierung der jeweiligen Glassorte zur Verfügung. / The chemical composition of pharmaceutical glasses (ampoules, infusion bottles, plunger) has been determined by laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS). A Nd:YAG laser with 10 Hz repetition rate in the q-switch mode at its fundamental wavelength and its 4th harmonic (1064 and 266 nm) was used to identify common types of pharmaceutical glasses. The following isotopes were used for measurements: 7Li, 11B, 23Na, 24Mg, 27Al, 28Si, 29Si, 30Si, 39K, 42Ca, 47Ti, 57Fe, 90Zr, 121Sb, 137Ba. Relative standard deviations between 1.8 % and 8.0 % of the quantitative results were obtained. 29Si was used as internal standard. Standard reference materials (soda-lime-, lead- and borosilicate glasses) were used for external calibration of laser sampling. Linear calibration functions for each isotope were found. The accuracy was determined by digestion of all samples and standard materials in a two-step-procedure by nitric/fluoric acid, measurement and external calibration by ICP-MS with multi-elemental standard solutions. Digestion and laser ablation results agreed within 8 % with the certified values. The proof of representative ablation was given by sufficient agreement of intensity ratios of most isotopes with the corresponding theoretical values. Further precision, sensititivity, selectivity, limit of detection, limit of quantitation and robustness were determined. In contrast to the fundamental wavelength the laserablation technique with 266 nm showed no significant improvement in the quality of the results. By measuring transient signals element concentrations of surface layers should be detected. Differing concentrations could be determined in soda-lime-glass samples. Also different surface-treated borosilicate-glass ampoules were examined by this method. Silicon-treated glass surfaces could be identified. Finally raw data of LA measurements were used for applying linear discriminant analysis. After reduction of the used variables (8 of 13 isotopes) all types of glasses could be distinguished only by their intensity data of LA measurements. The developed LA-ICP-MS method is a powerful technique to distinguish different types of pharmaceutical glasses and to examine their chemical composition.

Lo scopo della tesi è di caratterizzare i rischi per la salute umana, che derivano dall’inquinamento delle acque (acque sotterranee) e dal cibo (riso) tramite l’applicazione e lo sviluppo di nuove tecniche analitiche. La tecnica “diffusive gradients in thin films (DGT)” con resina a base di ferrihydrite, è stata studiata per l'accumulo delle forme inorganiche (As(III) e As(V)) e organiche (DMA(V) e MMA(V)) di Arsenico in matrici acquose. I DGT sono stati applicati su campioni di acque sotterranee, raccolti in sei diverse città del Nord Italia, dove la concentrazione di As è molto alta. Antimonio (Sb) è associato all’As in diversi studi perché le proprietà fisiche e chimiche di questi due elementi sono simili, ed esso è stato recentemente riconosciuto come contaminante dell'acqua. In questa tesi per la prima volta sono state riportate le prestazioni dettagliate dei DGT con ferrihydrite, impiegati in soluzioni acquose di Sb(III) e Sb(V). L’analisi delle forme chimiche di Sb(III) e Sb(V) in campioni acquosi è stata realizzata con l’applicazione della diluizione isotopica. In generale il riso, a differenza di altri prodotti alimentari di origine terrestre, contiene quantità significative di Arsenico inorganico. L’analisi della speciazione di As è stata realizzata in 70 campioni di riso italiano, rappresentativo di diversi tipi di coltivazione. Le forme più abbondanti nel riso erano As(III) e DMA(V). Inoltre è stato interessante studiare la localizzazione dell’As nel chicco di riso, sottoposto a diversi processi di lavorazione (risone, riso integrale e lavorato con o senza parboiling), perché sia la distribuzione che la presenza delle diverse forme chimiche nel chicco sono fattori chiave che controllano la biodisponibilità del contaminante. La distribuzione dell’ As nei chicchi di riso di due varietà (Gladio e Ronaldo) sottoposti a diversi processi, è stato determinato con Laser Ablation ICP-MS. In realtà il contenuto di As era più alto nei chicchi di riso non-parboiled rispetto a quello parboiled. La relazione tra intensità dell’As e le varie parti del chicco di riso ha rivelato che i livelli di As diminuisco dalla parte esterna verso la posizione centrale, mentre i valori di As sembrano essere simili tra la parte media e interna dei non parboiled. / The aim of this thesis was to explore new analytical techniques as well as to carry out further characterisations of human health risks, which derive from water pollution, in particular groundwater, and food, in particular rice. The prevention of water pollution is an environmental aspect, that includes monitoring of both natural enrichment and outside pollution with routine analysis but also with new techniques, e.g. the application of passive sampling techniques and advanced technologies. The diffusive gradients in thin films (DGT) technique with ferrihydrite adsorbent, has been investigated for the accumulation of different species of Arsenic (As), like Inorganic Species (arsenite and arsenate) and Organic Arsenic (dimethylarsinic and monomethylarsenate) in aqueous matrix. To evaluate the performance of DGT method for accumulation of arsenic species, after deployment in synthetic solutions, DGT devices were carried out on groundwaters collected in six different towns in the North of Italy, where the As concentration is very high. Recently, health effects at arsenic exposures have been observed in areas where levels of inorganic As in drinking water are not excessive. Antimony (Sb) is associated to As in several studies because the physical and chemical properties of these two elements are similar, and it has been recently recognized as water contaminant. In this thesis I reported for the first time detailed performance characteristics of the Fe-oxide gel associated to DGT devices deployed in known aqueous solutions of trivalent and pentavalent Sb. Speciation analysis of Sb(III) and Sb(V) in aqueous samples was performed through extraction and on-line determination of isotope dilution concentration after a chromatographic separation. Generally rice, unlike food products of terrestrial origin, contains significant amounts of inorganic arsenic. Recently some Government Organizations (e.g. EFSA) debated the possibility to set an upper limit for total and inorganic arsenic in rice. Arsenic speciation was realized in 70 Italian rice samples from different representative cultivation conditions. The most abundant forms in rice were As(III) and DMA(V). After that, it was fundamental to investigate the localization of As in rice grains in different processes (raw, brown and milled rice with or without parboiling technique), because both speciation and distribution throughout the grain are key factors controlling bioavailability of the contaminant. The As distribution in rice grains of two varieties (Gladio and Ronaldo) from different processes, was determined by LA-ICP-MS. The distribution of As varied between the various parts of the grains (exterior, medium and interior part). During parboiling, the partial boiling of food as the first step in the cooking process, arsenic might have released from the grain to the boiling water. Thus, parboiling of rice grain may reduce the magnitude of arsenic intake in human body. Actually the As content was higher in non-parboiled rice grain than in parboiled rice. The relationship between As intensities and the different parts of rice grain revealed that As levels decreased from the external part towards the middle position, and then the intensity values seem to be similar between medium and internal part in non parboiled products.

Submitted by Ana Hilda Fonseca (anahilda@ufba.br) on 2016-02-25T13:31:28Z No. of bitstreams: 1 Tese Uenderson Araujo Barbosa -final.pdf: 2006362 bytes, checksum: 7f8502d02bb06b7f75032fb7a79d79bc (MD5) / Approved for entry into archive by Ana Hilda Fonseca (anahilda@ufba.br) on 2016-05-10T17:54:24Z (GMT) No. of bitstreams: 1 Tese Uenderson Araujo Barbosa -final.pdf: 2006362 bytes, checksum: 7f8502d02bb06b7f75032fb7a79d79bc (MD5) / Made available in DSpace on 2016-05-10T17:54:24Z (GMT). No. of bitstreams: 1 Tese Uenderson Araujo Barbosa -final.pdf: 2006362 bytes, checksum: 7f8502d02bb06b7f75032fb7a79d79bc (MD5) / CAPES e CNPq / Neste trabalho foram propostas estratégias analíticas visando à avaliação toxicológica de metais e metaloides em medicamentos utilizados no tratamento da anemia ferropriva, empregando técnicas cromatográficas e não cromatográficas. Na primeira parte desta tese determinou-se chumbo nos medicamentos aplicando espectrometria de absorção atômica com atomização eletrotérmica (ETAAS). O preparo da amostra foi realizado fazendo uso de digestão ácida em bloco digestor com sistema de refluxo “dedo frio” usando 3 mL de HNO3 (69% v/v) e 1 mL de H2O2 (30% v/v). Foram otimizadas as temperaturas de pirólise e atomização usando uma mistura de paládio e magnésio como modificador químico convencional, obtendo-se 1100ºC e 2000ºC para pirólise e atomização, respectivamente. Com uma massa característica de 21,3 pg e 0,25 e 0,82 µg L-1, para os limites de detecção (LD) e de quantificação (LQ), respectivamente. A precisão, expressa pelo desvio padrão relativo de 5.92% para uma amostra com concentração de chumbo igual a 2,98 µg g-1. A exatidão foi estimada por comparação entre os resultados obtidos por ETAAS e espectrometria de massas com plasma indutivamente acoplado (ICP-MS). As concentrações de chumbo nas amostras variaram entre 0,096 e 7,068 µg g-1. Na segunda parte deste trabalho, foi efetuada a determinação de mercúrio através de amostragem em suspensão e espectrometria de absorção atômica com alta resolução e fonte contínua com geração de vapor frio (HR-CS CVAAS). As condições ótimas de preparo das suspensões (1,5 mol L-1 de HCl, 1,25% (m/v) de tioureia e 15 minutos de sonicação) foram otimizadas por planejamento fatorial completo. Os limites de detecção e quantificação foram 0,63 ng g-1 e 2,11 ng g-1, respectivamente. O desvio padrão relativo para avaliação da precisão foi calculado usando duas amostras com concentrações 4,82 e 9,61 ng g-1 com resultados 10,89 e 6,84%, respectivamente. Foi estimada a possível perda de mercúrio durante o procedimento proposto e a exatidão, mediante testes de adição e recuperação, variando entre 85,0 e 106,0%. As concentrações de mercúrio ficaram entre 3,17 e 34,86 ng g-1. A terceira parte deste trabalho descreve a determinação de macro e microelementos e o estudo in vitro da bioacessibilidade de ferro em medicamentos usados no tratamento da anemia ferropriva. Foi escolhida a espectrometria de emissão ótica com plasma indutivamente acoplado (ICP OES) como técnica analítica de medida. Foi efetuada digestão ácida para tratamento das amostras e o método foi validado, demonstrando boa sensibilidade, com limites de quantificação (mg L-1) 0,52 para Ca, 0,14 para K, 0,03 para Mg, 0,07 para Mn, 0,40 para Na, 0,36 para P e 0,24 para Zn. A precisão do método foi reconhecida mediante experimentos repetitivos intradia e interdia com resultados abaixo de 20%. Durante estudo da exatidão, testes de adição e recuperação foram executados em diferentes concentrações com resultados entre 81,85 e 114,97%. A robustez do plasma foi calculada mediante uma razão entre as linhas iônica e atômica de magnésio, Mg(II)/Mg(I) e os resultados, mediante análise componentes principais (PCA) e análise por agrupamentos hierárquicos (HCA). O estudo de bioacessibilidade in vitro de Fe demonstrou que alguns medicamentos são melhores bioacessíveis, sendo portanto, mais recomendados para o tratamento da anemia, comprovando dados da literatura farmacológica. Na quarta parte deste trabalho descreve-se o estudo da determinação e especiação simultânea de arsênio e cromo por 11 cromatografia líquida de alta eficiência acoplada à ICP-MS (HPLC-ICP-MS). A determinação total de arsênio e cromo foi realizada usando uma digestão ácida em forno de micro-ondas, uma mistura de 4 mL HNO3 (69% v/v) e 1 mL de H2O2 (30% v/v) e análise por ICP-MS. Para o estudo de especiação foi elaborado um estudo com três possíveis procedimentos para extração das espécies, sendo selecionado um procedimento com água a 80ºC por 30 minutos em energia de micro-ondas. Usando uma fase móvel contendo 1,0 mM de hidróxido de tetrabutilamônio (TBAH), 0,7 mM de ácido etilenodiaminotetracético (EDTA) e 5% de metanol (v/v) em pH 7,2 foi possível separar as espécies presentes com tempo de retenção de 5 minutos com separação de As(III), As(V), Cr(III) e Cr(VI), com limites de detecção de 0,002, 0,003, 0,140 e 0,042 µg g-1, respectivamente. A exatidão do método foi avaliada conforme testes de adição e recuperação, entre 80,64 e 109,36%. O método foi aplicado em amostras do Brasil e Espanha, com concentrações entre 0,1-1,3 e 0,18-62,27 µg g-1 para As e Cr, respectivamente. / In this thesis work analytical strategies have been proposed for toxicological assessment of metals and metalloids in medicines used to treat iron deficiency anemia, using chromatographic and non-chromatographic techniques. In the first part of this thesis, lead was determined in drugs using electrothermal atomic absorption spectrometry (ETAAS). Sample preparation was performed using acid digestion in digester block with reflux system "cold finger" using 3 mL of HNO3 (69% v / v) and 1 mL of H2O2 (30% v / v). Pyrolysis and atomization temperatures were optimized using a mixture of palladium and magnesium as conventional chemical modifier, yielding 1100°C and 2000°C for pyrolysis and atomization, respectively. With a characteristic mass of 21.3 pg and 0.25 and 0.82 µg g-1, for limit of detection (LOD) and limit of quantification (LOQ), respectively. The accuracy, expressed as relative standard deviation was 5.92% for a sample with lead concentration of 2.98 µg g-1. The accuracy was evaluated by comparing the results obtained by ETAAS and inductively coupled plasma mass spectrometry (ICP-MS). Lead concentrations found in samples ranged between 0.096 and 7.068 µg g-1. In the second part of this work, the determination of mercury using slurry sampling and high resolution continuous source cold vapor atomic absorption spectroscopy (HR-CS CVAAS) was performed. The optimum conditions for preparation of slurries (1.5 mol L-1 HCl, 1.25% (w / v) thiourea and 15 minutes sonication) were optimized using a two level full factorial design. The limits of detection and quantification were 0.63 ng g-1 and 2.11 ng g-1, respectively. The relative standard deviation for accuracy evaluation was calculated using two samples with concentrations 4.82 and 9.61 ng g-1 from which was obtained as results 10.89 and 6.84%, respectively. The possible loss of mercury was evaluated for the proposed procedure and the accuracy was studied by addition/recovery test, with recoveries ranging from 85.0 to 106.0%. Mercury concentrations ranged between 3.17 and 34.86 ng g-1. The third part of this work describes the determination of macro and micro elements and the in vitro study of iron bioaccessibility in medicines used to treat iron deficiency anemia. It was used inductively coupled plasma optical emission spectrometry (ICP OES) measured as an analytical technique. Acid digestion treatment was conducted to the samples and the method was fully validated by demonstrating good sensitivity with quantification limits (mg L-1) 0.52 for Ca, 0.14 for K, 0.03 for Mg, 0.07 for Mn, 0.40 for Na, 0.36 for P and 0.24 for Zn. The precision of the method was evaluated by intraday and interday repetitive experiments with results below 20%. During accuracy evaluation, addition/recovery tests were performed at different concentrations with results between 81.85 and 114.97%. The robustness of plasma was evaluated by a ratio between magnesium atomic and ionic lines, Mg(II)/Mg(I). The results were evaluated by multivariate analysis tools, principal component analysis (PCA) and hierarchical cluster analysis (HCA). The study of in vitro bioaccessibility of Fe showed that some drugs are better bioaccessible, being therefore more recommended for anemia treatment, confirming data of pharmacological literature. The fourth part of this work describes the simultaneous determination of arsenic and chromium speciation using high-performance liquid chromatography coupled to ICP-MS (HPLC-ICP- MS). The total arsenic and chromium determination was carried out using an acid digestion in microwave oven, using a mixture of 4 mL HNO3 (69% v / v) 9 and 1 mL of H2O2 (30% v / v) and analyzed by ICP-MS. For speciation, a study was performed with three possible procedures for extraction of species, being employed a procedure with water at 80 °C for 30 minutes in microwave energy. Using a mobile phase containing 1.0 mM tetrabutylammonium hydroxide (TBAH), 0.7 mM ethylenediaminetetraacetic acid (EDTA) and 5% methanol (v / v) under pH 7.2 was possible the separation of the species present with retention time of 5 minutes with separation of As(III) As(V), Cr(III) and Cr(VI), with a LOD of 0.002, 0.003, 0.140 and 0.042 µg g-1, respectively. The accuracy was assessed using addition/recovery tests, ranging between 80.64 and 109.36%. The method was applied in samples from Brazil and Spain.

L'utilisation de lasers est largement répandue dans le domaine de la microanalyse directe des solides. La matière vaporisée, en focalisant un faisceau laser de forte puissance sur la cible, peut être analysée soit par spectrométrie d'émission optique sur plasma induit (LIBS, acronyme anglais pour Laser Induced Breakdown Spectroscopy), soit par une source à plasma induit par haute fréquence couplée à la spectrométrie d'émission optique (ICP-AES) ou à la spectrométrie de masse (ICP-MS). Avec une résolution spatiale à l'échelle microscopique, les techniques d'ablation laser permettent ainsi d'accéder à la composition élémentaire locale de la surface d'un matériau. Néanmoins, les performances analytiques de ces techniques pourraient être améliorées par l'utilisation combinée des informations LIBS et ICP afin également de comprendre et maîtriser davantage l'interaction laser/matière. Dans ce but, ce travail a consisté à développer une technique de microanalyse par ablation laser couplée avec une détection en simultané par ICP et par LIBS afin d'étudier les potentialités analytiques de cet instrument pour cartographier la surface des matériaux. Les performances et les limitations de ce système ont été évaluées d'une part, en caractérisant les aérosols produits par ablation laser et d'autre part, en étudiant les signaux LIBS et ICP obtenus à partir d'un même prélèvement de matière. Le phénomène de fractionnement élémentaire rencontré sur des matrices critiques telles que le laiton a été mis en évidence en microablation malgré des caractéristiques de l'interaction laser/matière différente de la macroablation. Une méthode de correction, a posteriori, par l'efficacité d'extraction de la cellule d'ablation a été proposée afin de pallier ces effets limitatifs pour l'analyse quantitative. Une cellule d'ablation, optimisée à partir d'une étude de simulation numérique, a été développée afin de s'adapter aux applications de cartographies de surface. Les performances analytiques du système ont été évaluées en termes de stabilité (8-10%), de résolution spatiale (5 µm) et de limites de détection (de l'ordre de la ppm dans le solide avec un détecteur de masse). La complémentarité des mesures LIBS et ICP représente à la fois un outil de diagnostic de l'interaction laser/matière et un instrument d’analyse très complet grâce à la double détection qui permet de suivre simultanément des traces et des majeurs sur une large gamme d'éléments de la classification périodique / Laser ablation is widely spread for solid sample microanalysis. A tightly focused laser beam allows direct sampling of matter, the ablated mass can then be analysed either with LIBS (Laser Induced Breakdown Spectroscopy) or with an inductively coupled plasma source combined with an optical emission spectrometer (ICP-AES) or a mass spectrometer (ICP-MS). With spatial resolution down to the micron scale, laser ablation techniques permit local elemental analysis of sample surface. Nevertheless, analytical performances of such techniques could be improved by combining LIBS and ICP information to understand and control laser/matter interaction. For this purpose, this work aimed to develop a microanalytical technique based on laser ablation coupled to simultaneous detection with LIBS and ICP to study analytical potentialities of such technique for elemental mapping of material surface. Performances and limitations of the system were studied on one hand, by characterizing laser-induced aerosols and on the other hand, by studying simultaneous LIBS and ICP signals. Elemental fractionation on critical matrices such as brass was evidenced in microablation despite a different laser/matter interaction compared with macroablation. A correction procedure a posteriori using the total extraction efficiency of the ablation cell was proposed to overcome this problem for quantitative analysis. An ablation cell, optimized from a numerical simulation study, was developed for mapping applications. Analytical performances were evaluated in terms of stability (8-10 %), spatial resolution (5 µm) and detection limits (in the ppm range with ICP-MS). The LIBS and ICP complementarity makes the double detection system a diagnostic tool for laser/matter interaction and an analytical instrument allowing simultaneous monitoring of traces and majors from a large element range of the periodic classification

La présence d'hétéroatomes et métaux dans des fractions lourdes de pétrole brut peuvent entraîner la désactivation des catalyseurs d’hydrotraitement et d’hydrocraquage. Les composés contenant ces éléments sont connus pour être inclus dans des agrégats macromoléculaires d’asphaltènes, qui sont considérées comme les composants les plus problématiques des produits pétroliers. Ces travaux ont pour objectif d’apporter une meilleure compréhension de l’environnement chimique de ces espèces et des phénomènes d’agrégation pour optimiser les processus de raffinage. Des techniques de séparation comme la chromatographie de perméation de gel ou Gel Permeation Chromatography (GPC), Advanced Polymer chromatographie (APC) et la chromatographie sur couche mince d’haute performance ou High Performance Thin Layer Chromatography (HPTLC) ont été combinées avec des techniques d’analyse élémentaire comme la spectrométrie de masse à plasma à couplage inductif ou Inductively Coupled Plasma Mass Spectrometry (ICP-MS) et la spectrométrie de masse moléculaire d’haute résolution pour la caractérisation des fractions pétrolières lourdes. Les profils de distribution en taille d’espèces qui contiennent du vanadium, du nickel et du soufre, les hétéroatomes et métaux les plus abondants dans les produits pétroliers, ont été déterminés par GPC couplé à une détection élémentaire par ICP MS. Les paramètres ayant le plus d’impact sur la distribution en taille et l’évolution des agrégats dans les solutions de différentes coupes pétrolières ont été optimisés. Cela nous a permis de faire différentes comparaisons, entre de nombreuses charges et effluents de procédés, ainsi que d’autres séries d’échantillons ou de mélanges provenant d’unités-pilotes d’hydrotraitement. Les résultats obtenus ont pu être mis en regard de caractéristiques macroscopiques comme la viscosité. Le couplage du nouveau système APC avec l’ICP MS a été mis au point avec succès. Cette technique a permis de réaliser des cartographies d’échantillons pétroliers plus rapides qu’en GPC tout en consommant moins de solvant. Cependant, les différences des chromatogrammes APC obtenus par rapport aux profils GPC suggèrent des interactions indésirables avec la phase stationnaire, qui ont été l’objet d’une évaluation approfondie.Enfin, un dernier type de séparation a été évaluée. L’HPTLC, qui utilise des plaques à usage unique, peut être appliquée à la séparation de produits lourds par polarité, et ce sans étapes de déasphaltage préalable, contrairement aux colonnes de chromatographie liquide qui peuvent être détériorées à cause de l'adsorption irréversible et de la précipitation des asphaltènes. Différents types de plaques et éluants ont été testés afin de développer une méthode simple pour la séparation des fractions SAR (Saturés, Aromatiques et Résines) des échantillons de distillat sous vide ou Vacuum Gas Oil (VGO). Ce travail avait pour but des migrations en une seule étape et analyse UV qui pourrait être mis en œuvre pour l’analyse de routine dans les laboratoires de contrôle avec une adsorption irréversible réduite. Des résultats très prometteurs ont été obtenus avec l’utilisation de plaques de cellulose pour la séparation des familles de polarité différentes au sein des agrégats d’asphaltènes. Des tests sur l'analyse directe des plaques par Désorption-Ionisation par Électronébulisation (DESI) MS, l'analyse directe en temps réel ou Direct Analysis in Real Time (DART) MS et Ablation Laser (LA) ICP MS ont été effectués. / The presence of heteroatoms and metals in heavy fractions of crude oil can cause the deactivation of hydrotreatment and hydrocracking catalysts. Compounds containing these elements are known to be included in macromolecular asphaltene aggregates, which are considered the most problematic components of heavy oil. This thesis aimed for a better understanding of the chemical environment of these species and on the aggregation phenomenon to optimize refining processes. Separation techniques such as Gel Permeation Chromatography (GPC), Advanced Polymer Chromatography (APC) and High Performance Thin Layer Chromatography (HPTLC) were combined with elementary analysis techniques such as Inductively Coupled Plasma Mass Spectrometry (ICP MS) or high-resolution molecular mass spectrometry for the characterization of heavy fractions of crude oil. Here, the size profiles of species that contain vanadium, nickel and sulfur, the most abundant heteroatoms and metals present in petroleum products, were determined by GPC online with elemental detection by ICP MS. The parameters having the most significant impact on size distribution and the evolution over time of aggregates from solutions of different petroleum cuts were optimized. This allowed the comparison between various feeds and effluents and other series of samples and mixtures from pilot units of refining processes by GPC ICP MS. Results obtained could be related to macroscopic characteristics such as viscosity. Hyphenation of the new APC System to ICP MS has been successfully completed. It allowed the mapping of petroleum samples faster than GPC, therefore with less solvent consumption. However, the differences of the APC chromatograms obtained with respect to the GPC profiles suggested significant unwanted interactions of the highly polar compounds of the petroleum samples with the stationary phase, which have been thoroughly studied. Finally, another type of separation was evaluated. HPTLC, of single-use plates, can be applied to the analysis of heavy petroleum products by polarity without prior deasphalting steps while chromatography columns can be deteriorated because of the irreversible adsorption and asphaltene precipitation. To this end, various types of plates and eluents were tested to develop straightforward SAR (Saturates, Aromatics and Resins) separations of Vacuum Gas Oil (VGO) samples. This method, based on single-step migrations with reduced irreversible adsorption and UV monitoring, could be implemented for routine analysis in control laboratories. Very promising results were obtained by the use of cellulose plates for the separation of different families within asphaltene aggregates. Tests on Desorption ElectroSpray Ionization (DESI) MS, Direct Analysis in Real Time (DART) MS and Laser Ablation (LA) ICP MS analyses of the plates have been performed.

[pt] Método de identificação e quantificação de peptídeos, através da otimização de estratégias para a marcação de peptídeos com metais e subsequente separação por nano-HPLC-UV, MALDI MS. Primeiramente, peptídeos foram marcados com 3 diferentes metais lantanídeos usando um reagente funcional NHS-DOTA. Os resultados demonstraram que a reação de derivatização usando o reagente quelante DOTA foi eficiente para peptídeos simples e misturas dos mesmos, verificada através do MALDI MS a partir da relação m/z. Em paralelo, análises ambientais foram realizadas pela otimização de um procedimento de extração de metalotioneína em bílis de peixe, uma vez que esta matriz tem sido reportada como um biomarcador ambiental de contaminação por metal. Diferentes procedimentos e agentes de redução foram aplicadas para a extração de metalotioneína em bílis e fígado de peixe (Oreochromis niloticus. Análises espectrofotométricas foram realizadas a fim de quantificar os extratos de MT, e gel SDS-PAGE foi usado para avaliação qualitativa dos diferentes procedimentos usados. Cada procedimento foi avaliado estatisticamente. Metodologia de superfície de resposta foi aplicada para amostras de bílis, a fim de avaliar a resposta desta matriz. Em um contexto ambiental, concentrações de MT biliar foi mais baixa que MT do fígado, no entanto, a primeira mostrou-se mais adequada para um monitoramento ambiental. / [en] This work developed a new method for the identification and quantification of peptides, by optimizing some of the available strategies suitable for labeling peptides with lanthanide metals with subsequent separation by nano-HPLC with UV detection, matrix-assisted laser desorption ionization-mass spectrometry (MALDI MS). First, peptides were labeled with the three different lanthanide metals using a functional DOTA-based reagent. The results demonstrate that the derivatization reaction using the chelating reagent DOTA-NHS-ester was effective for single peptides and peptide mixtures, verified from the m/z relation obtained by MALDI MS. In parallel, environmental analyses were conducted, by performing the standardization of metalloprotein purification in fish bile, since this matrix has been reported as a biomarker for environmental metal contamination. Different procedures and reducing agents were applied to purify MT isolated from fish (Oreochromis niloticus) bile and liver. Spectrophotometrical analyses were used to quantify the resulting MT samples, and SDS-PAGE gels were used to qualitatively assess the different procedure results. A response surface methodology was applied for bile samples. In an environmental context, biliary MT was lower than liver MT, and, bile MT seems to be more adequate in environmental monitoring scopes. / [fr] Ce travail a développé une nouvelle méthode pour l identification et la quantification des peptides, par l optimisation de certaines stratégies disponibles appropriées pour le marquage des peptides avec des métaux lanthanide, une séparation par nano-HPLC et détection UV, et suivi par MALDI MS. Tout d abord, les peptides ont été marqués avec les trois métaux lanthanides différents et un réactif fonctionnel - DOTA. Les résultats montrent que la réaction de transformation en dérivé à l aide du réactif chélateur DOTA-NHS-ester a été efficace pour des peptides individuels et des mélanges de peptides, vérifiées à partir de la relation m/z obtenue par MALDI MS. En parallèle, nous avons effectué l optimisation pour la purification de métalloprotéine dans la bile de poisson, qui est signalée entant que biomarqueurs de contamination métallique de l environnement. Des procédures différentes et les agents réduisant ont été apliqués pour purifier les MT isolées de la bile et du foie des poissons (Oreochromis niloticus). Des analyses spectrophotométriques ont été utilisées pour quantifier les échantillons de MT, et le gel SDS-PAGE a été utilisé pour évaluer qualitativement les différents résultats de la procédure. Chaque procédure a en suíte été évaluée statistiquement, une méhtode des surfaces de réponse a été appliquée. Les MT de la bile semblent être plus adéquate pour la surveillance de l environnement en ce qui concerne l exposition récente à des xénobiotiques qui peuvent influer sur l expression protéomique et metalloproteomique de cette matrice biologique.

Die Strukturen der Erdalkalimetall-Subnitride (EA2N) von Calcium, Strontium und Barium ermöglichen mit ihrem schichtartigen Aufbau aus EA6N-Oktaedern Intercalationsreaktionen. Die Redox-Intercalation von Stickstoff in Sr2N wurde an Einkristallen untersucht. Nur durch eine drastische Erhöhung des Reaktionsgasdruckes im Vergleich zu den Reaktionen an mikrokristallinen Proben wurde die Intercalation der Diazenidionen in die Kristalle zu Sr4N3 und SrN möglich. Für eine analoge Intercalation von Wasserstoff in Sr2N konnten die Reaktionsbedingungen dahingehend optimiert werden, dass erstmals phasenreines Strontiumnitridhydrid (Sr2N)H bzw. deuterid (Sr2N)D erhalten wurde. Anhand von Intercalationsreaktionen mit Sr2N Kristallen konnte gezeigt werden, dass der Intercalationsprozess, erkennbar an der deutlichen Farbänderung von schwarz nach bersteinfarben, von außen nach innen fortschreitet. Als Methode zur räumlich aufgelösten Analyse wurde die Laserablation, in Kombination mit einem ICP - Massenspektrometer (LA-ICP-MS) verwendet. In der Literatur beschriebene Quantifizierungsstrategien wurden auf die Anwendbarkeit für die gegebene Fragestellung untersucht. Mit der ortsaufgelösten Analyse von Einkristallen konnte gezeigt werden, dass die Intercalation von Stickstoff in die Kristalle kontinuierlich von den Kanten zur Kristallmitte fortschreitet.

L’utilisation de plus en plus fréquente des agents de contraste à base de Gd (AC-Gd) au cours des examens d’Imagerie par Résonnance Magnétique (IRM), engendre le rejet de ces produits pharmaceutiques dans les eaux usées retraitées en STation d’EPuration (STEP). En l’absence de retraitement spécifique de ces AC-Gd en STEP, ils sont rejetés dans le milieu aquatique, où de nombreuses études ont relevé leur présence aussi bien dans les eaux de surface que dans les eaux souterraines et jusqu’à l’eau du robinet. Le manque de connaissances concernant les effets des AC-Gd suite à leur rejet a mis en évidence la nécessité d’étudier leurs impacts environnementaux sur les organismes vivants dans les milieux aquatiques. Dans ce contexte, nous avons choisi de déterminer les concentrations en Gd d’origine anthropique à proximité de rejets de STEP et de zones de captage en eau potable situés en région Lorraine. Nos mesures ont permis de montrer la présence Gd d’origine anthropique sur l’ensemble des échantillons prélevés, avec des concentrations mesurées comprises de quelques ng(Gd)/L à quelques dizaines de µg(Gd)/L. Ces concentrations de Gd anthropique seraient dues à la présence d’AC-Gd. Nous nous sommes plus particulièrement intéressés aux effets de l’AC-Gd le plus stable et l’un des plus fréquemment commercialisé : le Gd-DOTA (Dotarem®). Pour cela, nous avons choisis plusieurs espèces représentatives des taxons rencontrés dans les cours d’eau Lorrains. Des essais ont été menés en conditions contrôlées de laboratoire afin de mesurer l’accumulation du Gd-DOTA dans les tissus et les effets de l’AC-Gd ont été appréhendés au travers de mesures de croissance, de reproduction et de mortalité au niveau individuel chez les microalgues vertes unicellulaires (Chlorella vulgaris et Pseudokirchneriella subcapitata), chez un microcrustacé (Daphnia magna) et chez un vertébré aquatique (Danio rerio) exposés à des concentrations en Gd-DOTA réalistes d’un point de vue environnemental. L’accumulation du Gd-DOTA a aussi été mesurée chez les bivalves (Corbicula fluminea et Dresseina rostriformis bugensis) et comparée à des mesures d’accumulation du Gd in situ. Les réponses physiologiques des bivalves ont été évaluées à l’aide d’une batterie de 11 biomarqueurs dans leurs branchies et leur glande digestive. Les effets de l’AC-Gd ont également été étudiés in vitro sur des fibroblastes de D. rerio (cellules ZF4 - ATCC-2050). Nos travaux ont montré que les AC-Gd et le Gd-DOTA en particulier étaient responsables d’effets écotoxiques indirects à différents niveaux d’organisation biologiques. Seuls les bivalves accumulent le Gd-DOTA mais tous les individus semblent mettre en place des mécanismes de dépuration pour parer à la toxicité de l’AC-Gd. Les résultats obtenus au cours de cette recherche ont confirmé la nécessité d’un suivi des concentrations en AC-Gd dans le milieu aquatique et la nécessité d’approfondir les études de l’écotoxicité de ces produits pharmaceutiques. Ceci pourra aboutir à une évaluation pertinente de leur risque environnemental et de proposer des solutions pour la gestion environnementale de ces composés / The increasing use of Gadolinium-based Contrast Agents (Gd-CA) for Magnetic Resonance Imaging (MRI) results in their rejection in Waste Water Treatment Plants (WWTPs). Due to the lack of specific recycling process in European WWTPs, these pharmaceutical produces enter the aquatic environment from river to tap water. The effects of Gd-CA in aquatic media have been not studied yet. The lack of knowledge about these effects highlighted the need to study their environmental impacts on aquatic organisms. In this context, we decided to measure anthropogenic concentrations of Gd in the aquatic environment in the Lorraine region (France) closed to WWTPs outputs and catchment areas used for drinking water. Our measurements underlined the presence of anthropogenic Gd on all the collected samples at concentrations ranging from few ng/L to several dozen of µg/L. In this research we focused on the effects of the most frequently used Gd-CA, the gadoteric acid (Gd-DOTA - Dotarem®) which is also the most stable one. Several representative species of aquatic environment were selected for ecotoxicological assays: i.e. unicellular green microalgae (Chlorella vulgaris and Pseudokirchneriella subcapitata), microcrustacean (Daphnia magna) and aquatic vertebrate (Danio rerio). Assays were conducted in laboratory under controlled conditions as well as in situ. Gd-DOTA accumulation was measured in the tissues of the different organisms. Environmental realistic concentrations of Gd-CA were used to assess their effects at the individual level by means of growth, reproduction and mortality measurements. The Gd-DOTA accumulation was also measured in bivalves’ tissues (Corbicula fluminea and Dresseina rostriformis bugensis) and compared to Gd ones in situ in these organisms. Physiological responses were assessed based on a battery of 11 complementary biomarkers measured in the digestive gland and in the gills of both bivalve species. At cellular level, the effects of Gd-DOTA were studied in vitro on D. rerio fibroblasts (ZF4 – ATCC-2050). Indirect ecotoxicological effects of Gd-CA and of Gd-DOTA especially have been highlighted at all biological levels. Accumulation of Gd-DOTA was observed in bivalves only, but defense systems were mobilized in all organisms to limit toxicity. Our results demonstrated that following both research on ecotoxicological effects of the Gd-CA and evolution of their concentrations in aquatic ecosystem are necessary to assess more precisely their environmental risk and to propose solutions for their environmental management

Magma which intrudes into carbonate rich crust, interacts with the carbonate in several ways, for example by contact metamorphism and formation of marble or by metasomatism resulting in calc-silicate skarn. These processes release volatiles, such as CO2, from the carbonate and might thus cause climate change. One volcanic complex where the intrusions and their surrounding metamorphic aureole are well exposed and therefore convenient for investigation of magma-carbonate interaction is the Carlingford Igneous Centre in NE of Ireland. The complex is dominated by a gabbro lopolith and a microgranite ring dyke, which intruded into limestone and meta-siltstone at around 62-60 Ma. The purpose of this study is to investigate the extent of magma-carbonate interaction and assess the amount of CO2 which could have been released from the aureole at Carlingford. This is done by analysing major and trace elements as well as carbon and sulphur content in skarn and marble samples from a transect along the calc-silicate metamorphic aureole. The analytical methods used are SEM- EDX, XRF, LA-ICP-MS and carbon and sulphur analyses. The CO2 release is calculated by comparing the carbon and CO2 content in the skarn and marble samples with that in the unmetamorphosed limestone. The results show that the skarn has experienced a strong interaction with the magma, as it contains several high-grade minerals, such as wollastonite, vesuvianite and pyrope, and has lost almost all its carbon. The extent of interaction and amount of CO2 release is generally much lower in the marble, even though a few marble samples show a strong interaction and a high degree of degassing. On the other hand, the amount of marble in the aureole turns out to be significantly higher than the amount of skarn (70-90% compared to 10-30%). The total mass of CO2 release from the calc-silicate aureole is calculated to 0.64-9.06 Gt, where 1.30-2.67 Gt being the most realistic amount. This released CO2 has probably not caused any significant climate change on its own but may have had a small contribution to global warming together with other volcanoes that were active during the same period of time. / Magma som tränger in i karbonatrik jordskorpa, integrerar med karbonaten på flera olika sätt, t.ex. genom kontaktmetamorfos och bildande av marmor eller genom metasomatos, vilket resulterar i kalk-silikatisk skarn. Dessa processer släpper ut flyktiga ämnen, som exempelvis CO2, från karbonaten och kan därmed orsaka klimatförändring. Ett vulkaniskt komplex där intrusioner och deras omgivande metamorfa aureoler är välexponerade och därmed lämpliga för undersökning av magma-karbonat-interaktion är Carlingford Igneous Centre i nordöstra Irland. Komplexet domineras av en gabbro-lopolit och en mikrogranitisk ring-gång, som trängde in i kalksten och metasiltsten för ca 62-60 Ma. Syftet med studien är att undersöka graden av magma-karbonat-interaktion och bedöma mängden CO2 som kan ha släppts ut från aureolen i Carlingford. Detta utförs genom analysering av huvudelement och spårelement samt av kol- och svavelinnehåll i skarn- och marmorprover från en transekt genom den kalk-silikatiska metamorfa aureolen. De analytiska metoder som används är SEM-EDX, XRF, LA-ICP- MS samt kol- och svavelanalyser. Mängden CO2 beräknas genom att jämföra kol- och CO2-innehåll i skarn- och marmorprover med innehållet kol och CO2 i den ickemetamorfa kalkstenen. Resultaten visar att skarn har genomgått en stark interaktion med magman, då bergarten innehåller flera mineral av hög metamorf grad, som exempelvis wollastonit, vesuvianit och pyrop, samt har förlorat nästan allt sitt kol. Graden interaktion och mängden CO2-utsläpp är generellt mycket lägre i marmorn än i skarnen, även fast några marmorprover visar stark interaktion och en stor del avgasning. Å andra sidan är mängden marmor i aureolen signifikant högre än mängden skarn (70-90% jämfört med 10-30%). Den totala massan av CO2-utsläpp från den kalk-silikatiska delen av aureolen är beräknad till 0,64-9,06 Gt, där 1,30-2,67 Gt anses vara den mest realistiska mängden. Detta utsläpp av CO2 har troligtvis inte orsakat någon signifikant klimatpåverkan på egen hand, men kan ha haft en liten bidragande effekt till global uppvärmning tillsammans med andra vulkaner som var aktiva under samma geologiska tidsperiod.