Les spéléothèmes (concrétions carbonatées se formant dans les zones karstiques) sont des archives paleo-climatiques reconnues, dont l’intérêt majeur est de pouvoir être datées précisément par la méthode Uranium-Thorium. En revanche, les traceurs traditionnellement utilisés pour reconstruire les climats passés à partir de ces objets géologiques ne sont pas directement quantifiables en termes de paramètres climatiques comme la température moyenne, ou la quantité de précipitation. Les variations des concentrations en éléments traces contenus dans les spéléothèmes ont pu être relié dans certains sites aux changements climatiques passés, mais des doutes existent sur la robustesse de leur signal au sein d’une même grotte et entre différents sites.Nous nous sommes appliqués à déterminer les variations au cours des 50 000 dernières années de plusieurs catégories d’éléments (alcalino-terreux, uranium, et terres rares) dans des stalagmites de deux grottes situées dans le sud de la France (les grottes de Villars en Dordogne et de Chauvet en Ardèche), par spectrométrie ICP-MS. Les spéléothèmes sélectionnés ont déjà été datés et ont enregistré les variations paleo-environnementales à travers les isotopes stables de la calcite. Trois périodes d’étude caractérisées par des changements particuliers sont étudiées: le stade isotopique 3 de la dernière période glaciaire (~50-30 ka), la dernière déglaciation (~20-10 ka) et la fin de l’Holocène (~2-0 ka).Le signal des variations des alcalino-terreux à Villars pendant le stade isotopique 3 est significatif et robuste. La variabilité du strontium notamment, qui provient de processus hydrologiques intra-karst, suit les événements climatiques rapides enregistrés dans l’hémisphère Nord. D’autre part, le comportement de nombreux éléments traces pendant la déglaciation est similaire entre les grottes de Villars et de Chauvet. Enfin, des changements du couvert végétal sont probablement à l’origine des changements synchrones enregistrés par les éléments traces et les isotopes stables de la calcite au cours des deux derniers millénaires à Villars.Par ailleurs, l’étude des coefficients de partition des alcalino-terreux, de l’uranium et des terres rares dans des conditions variées montre l’importante de la variabilité inter-site de leur partitionnement. / Calcareous deposits forming within caves, also known as speleothems, have become acknowledged paleoclimatic archives. One of their main interests is that they can be absolutely dated by Uranium-Thorium methods. However, traditionally used speleothem climatic proxies cannot be directly translated into environmental variables such as the mean annual temperature or the amount of annual rainfall. In some contexts, the variations of trace element concentrations in speleothem calcite could be linked to past climatic changes, but the robustness of trace element signals between speleothems of the same or nearby caves is still questionable.We determined by ICP-MS the concentrations of several families of chemical elements (alkaline-earth metals, uranium, rare-earth elements) in stalagmites from two caves located in Southern France (Villars cave in Dordogne and Chauvet Cave in Ardèche) spanning the last 50,000 years. The selected speleothems had already been dated and their stable isotope profiles had proven to record paleoenvironmental fluctuations occurring during three separate periods: the Marine Isotopic Stage 3 or MIS 3 (~50-30 ka), the Last Deglaciation (~20-10 ka), and the end of the Holocene (~2-0 ka).Variations of alkaline-earth metals recorded in two stalagmites from Villars Cave during MIS 3 are significant and robust. Notably, Sr concentrations follow the rapid climatic changes recorded in the Northern Hemisphere. Furthermore, several trace elements behave similarly during the Last Deglaciation in the Villars and the Chauvet Cave. Finally, changes of the vegetation cover above the cave are likely to have caused the synchronous fluctuations of the trace element and stable isotope contents that happened during the last two thousand years in Villars speleothems.Lastly, the partition coefficients of alkaline-earth metals, uranium and rare-earth elements were measured in different sites and conditions and proved to be very site-dependent.

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.

[pt] O objetivo deste trabalho foi o desenvolvimento de um método analítico para determinação simultânea de elementos na faixa de concentração de traços em óleos através das técnicas de espectrometria de massas (ICPMS) e espectrometria de emissão ótica (ICPOES), ambas com plasma indutivamente acoplado, após procedimento de digestão em equipamento de mineralização que trabalha sob altas temperaturas (300 °C) e pressões (100 bar). A digestão dos petróleos foi realizada em frascos fechados de quartzo de volume de 90 mL utilizando-se uma mistura de ácido nítrico e peróxido de hidrogênio concentrado, de elevada pureza. Foram avaliadas as melhores condições de digestão dos petróleos, possíveis interferentes resultantes das matrizes formadas durante o processo de digestão e as condições operacionais para a determinação simultânea dos elementos de interesse. O teor de carbono residual foi determinado para avaliar a extensão do processo de digestão e possíveis interferências em ambas às técnicas utilizadas. O efeito da concentração residual de ácido nítrico na intensidade dos sinais analíticos foi estudado, bem como o desempenho de diferentes padrões internos para corrigi- lo. Curvas analíticas com lineariadades r2 > 0,999 foram obtidas para a maioria do elementos determinados e limites de detecção (LDM-3(sigma)): < 1 (mi) g kg-1 para Sb, Be, Cr, Co, Cu, Fe, Li, Mg, Mn, Hg, Ni, Ag, Na, Sr, Ti, V, Zn, ETR, EGP, e na faixa de 1 a 10 (mi)g kg-1, para Al, Ba, Cd, Pb, K, Se. O método foi validado através da quantificação de elementos certificados nas amostras de referência NIST 1634c e NIST 1084a, obtendo-se repetitividades e exatidões melhores que 5% para a maioria deles. / [en] An analytical procedure was developed for the determination of trace elements in crude oil by inductively coupled plasma mass spectrometry (ICPMS) and inductively coupled plasma optical emission spectrometry (ICPOES) after sample digestion in a high pressure asher system operating at 100 bar and a temperature of 300 °C. Wet digestion was performed in closed quartz vessels of 90 mL volume using high purity nitric acid and hydrogen peroxide. Best conditions for the digestion of petroleum samples were studied and possible interferences caused by the resulting matrix were evaluated, as well as best measuring conditions for the analytes of interest. Residual carbon content was determined in order to assess the completeness of digestion and interferences caused by this element in both techniques. The effect of residual acid concentration on the analytical signals were studied, as well as the performance of different internal standards for its correction. Linear analytical curves (r2 >0.999) were obtained and method detection limits (MDL - 3(sigma)) of 1 (mi)g kg-1 for Sb, Be, Cr, Co, Cu, Fe, Li, Mg, Mn, Hg, Ni, Ag, Na, Sr, Ti, V, Zn, ETR, EGP, and in the range of 1 to 10 (mi)g kg-1 for Al, Ba, Cd, Pb, K and Se were obtained. Method validation was performed by quantification of certified elements in two standard reference materials (NIST: 1634c; 1084a) resulting in repeatabilities and accuracies of better than 5 % for most of the elements studied.

Unter den Techniken der chemischen Proteomik hat Capture Compound – Massenspektrometrie (CCMS) den Vorteil, Interaktionen von Molekülen mit geringer Affinität zueinander effektiv untersuchen zu können. CCMS beruht auf kleinen molekularen Sonden (Capture Compounds, CCs), die aus drei funktionalen Bestandteilen bestehen: die Selektivitätsfunktion, ist ein kleines Molekül, das mit einem Zielprotein eine schwache Wechselwirkung eingeht. Die zweite Funktionalität erlaubt kovalente Anhaftung der molekularen Sonde an Proteine. Der dritte Anteil erlaubt Detektion mit sehr guten Sensitivität; allerdings ist die Quantifizierung weiterhin ein Schwachpunkt dieser Technik. Ziel dieses Projektes ist, eine in CCMS verwendbare Quantifizierungsmethode zu entwickeln. Heutzutage gibt es zahlreiche MS-basierte Quantifizierungsstrategien; unsere beruht auf der Einführung von Lanthanoid-haltigen Labels – Metal Coded Affinity Tagging (MeCAT). In dieser Arbeit wurde erstmalig die erfolgreiche Verwendung mit Metall- Markern chemoproteomischer Sonden (CCs) zur Detektion und absoluten Quantifizierung von Zielproteinen mit schwacher Wechselwirkung etabliert. Mit den Experimenten an isolierten Enzymen und an lebenden Zellen wurde nachgewiesen, dass Metall-Marker keinen negativen Einfluss auf andere funktionelle Teile chemoproteomischer Sonden haben. CCs, die mit Lanthanoid-Chelaten funktionalisiert sind, zeigen ähnliche Affinität zu ihren Zielproteinen wie die Referenz-Sonden. Zudem erlauben Metall-Marker, die für diese Art molekularer Sonden verwendet werden, die Entwicklung einer element-basierten Technik zur Bilderzeugung. Der herausragende Vorteil der Metall-funktionalisierten CCs kombiniert mit ICP-MS ist, dass diese eine absolute Quantifizierung der Ausbeute der Quervernetzungen ermöglichen. / Capture compound mass spectrometry (CCMS) is a chemical proteomics technique that has the advantage of addressing low abundant target proteins in lysates as well as in living cells. The CCMS is based on small molecule probes (capture compounds) that consist of three functionalities: a small molecule (quite often it is a drug), which interacts with the target protein; the moiety that allows covalent attachment of the molecular probe to the protein; the one that allows detection. The detection moiety utilized for CCMS can offer high sensitivity; however, the challenge of absolute quantification is still a bottleneck of this technique. Metal Coded Affinity Tagging (MeCAT) is a quantitative approach based on the chemical labeling with lanthanide; it allows obtaining both the structural and quantitative information. In this work for the first time the successful utilization of chemoproteomic probes functionalized with a metal tag for the detection and absolute quantification of target proteins was established. With the experiments both on isolated enzymes and living cells it was determined that MeCAT does not negatively influence other functional parts of the probes; therefore, capture compounds functionalized with lanthanide chelates demonstrate similar affinity to the target as the reference probes. Moreover, metal tags utilized for this type of molecular probes can offer a promising elemental imaging technique. However, to achieve the sufficient resolution multiple metal tags per molecular probe are needed. The striking advantage of the approach of utilization metal functionalized capture compound combined with ICP-MS detection is that it allows absolute quantification of crosslink yield, what cannot be performed with other detection methods applied for this technology.

No início dos anos 90, os primeiros indícios do tráfico ilícito de materiais nucleares e radioativos foram observados principalmente na região européia. Uma década marcada por inúmeros casos de apreensão desse tipo de material. Como resultado, esses atos passaram a ser alvo de investigações criminais forenses, desenvolvendo-se a partir daí, a ciência forense nuclear. No Brasil não há registros oficiais do tráfico ilícito de material nuclear, entretanto, é amplamente conhecida a extração e o transporte ilegal de materiais geológicos radioativos, assim como a apreensão de fragmentos de materiais utilizados como blindagem de fontes radioativas. Uma das principais ferramentas utilizadas na ciência forense nuclear consiste no estabelecimento de bancos de dados de materiais nucleares. Esses documentos devem conter o maior número possível de informações sobre as características físicas, químicas e nucleares do material apreendido, permitindo a identificação de sua origem, processo de fabricação ou mesmo a idade (age). Assim, se estabelecem padrões de composição característicos de cada material, denominados assinaturas químicas (chemical finger print). Nesse trabalho foi adotado o protocolo forense nuclear seguindo as três etapas de avaliação sugeridas pela Agência Internacional de Energia Atômica (AIEA) na identificação da origem de Siliceto de urânio (U3Si2). Realizaram-se ensaios de caracterização física, química e isotópica dos materiais em estudo e compararam-se os dados com aqueles obtidos para outros compostos de urânio (Tetrafluoreto de urânio, UF4; óxidos de urânio, UO2 e U3O8; tricarbonato de amônio e uranila, TCAU) estabelecendo-se uma assinatura característica para cada um. A partir dos resultados foi possível classificar os compostos por grupos de origem, uma vez que são provenientes de diferentes processos de fabricação e/ ou origem. Demonstrou-se também a importância da criação e manutenção de um banco de dados nuclear na investigação de um evento forense nuclear. / In the early 1990´s, the first illicit trafficking of nuclear and radioactive materials was observed mainly in Europe. A decade marked by numerous cases of seizures of these materials. As a result, these events have become the subject of criminal forensic investigations and develop from there, nuclear forensics. In Brazil there are no illicit trafficking official records of nuclear material, however, is widely known the extraction and illegal transportation of radioactive geological materials, and the materials pieces attachment used as shielding for radioactive sources. One of the main tools used in nuclear forensics is the nuclear materials databases establishment. These documents must contain the most information as possible about the physical, chemical and nuclear material seized, allowing the identification of their origin, manufacturing process or age. Thus, it sets characteristic composition standards of each material, called \"chemical signatures (chemical finger print). In this work nuclear forensic protocol was adopted as well as the three stages of assessment suggested by International Atomic Energy Agency (IAEA) in identifying the origin of uranium silicide (U3Si2). Assays were performed in order to make physical, chemical and isotopic characterization of the studied materials and compared the data with those obtained for other uranium compounds (Uranium tetrafluoride, UF4; uranium oxide, UO2 and U3O8; Yellow cake) by establishing a characteristic signature for each one. Through the assays the uranium compounds were classify by origin groups, as far as they are from different manufactured process and/ or origin. It was also possible to show the importance of a nuclear forensic database during an investigation of a nuclear forensic event.

Neste estudo foi determinado o conteúdo de metais em adoçantes dietéticos de mesa, consumidos por um número crescente de pessoas preocupadas em manter a forma física e restringir o nível calórico da dieta havendo ou não recomendação médica. Foram coletadas 26 amostras de adoçantes dietéticos distribuídos no mercado de São Paulo, em uma grande variedade de apresentações, produtos, composições e marcas. Como a composição é dependente da produção, esta análise é indispensável ao controle da qualidade e relevante à saúde do consumidor. A Espectrometria de Emissão Óptica com Fonte de Plasma Indutivamente Acoplado (ICP OES) com vista axial, mostrou-se adequada determinação dos macronutrientes Na, K, Ca, Mg e dos microelementos essenciais Fe, Cu, Zn, Cr, Mn e Se, dos potencialmente tóxicos AI, Ni, As, Cd e Pb em adoçantes sólidos e líquidos, atendendo as exigências da legislação brasileira. Assim, foi realizado um estudo comparativo entre as determinações utilizando as metodologias clássicas de pré-tratamento da amostra, como o resíduo de cinzas, a dissolução ácida com aquecimento por microondas com o procedimento sem tratamento prévio das amostras, visando facilitar a análise, uma vez que esta é a etapa que mais consome tempo e esforços. A determinação simultânea do espectro favoreceu a análise, uma vez que 74 comprimentos de onda puderam ser selecionados sem aumento no tempo ou no custo de análise, auxiliando na seleção de 36 linhas analíticas mais adequadas (livre de interferências) à determinação sem tratamento prévio da amostra. Nenhuma amostra de adoçante excedeu o valor recomendado para o somatório dos contaminantes inorgânicos (Aspartame < 10 &#181;g g-1 e Sacarina < 20 &#181;g g-1). No entanto, 6 marcas de adoçantes em pó excederam o valor limite da legislação para As (1,0 &#181;g g-1), excederam o valor para Cr total (0,1 &#181;g g-1) e 2 marcas de adoçante líquido excederam o valor para Ni (0,1 &#181;g g-1). Mostrando que os adoçantes em pó apresentam (&#8776;5 &#181;g g-1) níveis maiores de metais potencialmente tóxicos que os adoçantes líquidos (&#8776;2,O &#181;g mL-1). / The metal content in artificial sweetener usually consumed by a growing population concerned to keep body shape and to restrict the diet caloric content, by medical recommendation or not, was measured. The samples were 26 artificial sweeteners available at São Paulo\'s market, under different forms, and several suppliers, with various sweetener composition, and brands. Because the composition depends on manufacture factors, this analysis is mandatory for quality control and relevant for the user health. Inductively coupled plasma atomic emission with axial view has demonstrated to be an appropriated technique to determine the macronutrient content, such as Na, K, Ca, Mg and essential microelements such as Fe, Cu, Zn, Cr, Mn and Se, besides the potentially toxic elements such as AI, Ni, As, Cd and Pb in solid and liquid sweeteners in accordance with to the Brazilian regulation. It was also performed a comparative study of the sample preparation methodology, using classical sample pretreatment, such as burning to ashes, and acid dissolution with microwave heating and a procedure without prior sample treatment (dissolution and/or convenient dilution), aiming at facilitating the analysis of dietetic sweetener because this step is the most time and labor consuming. The analysis was simplified using the simultaneous spectra acquisition, once 74 wavelengths were selected without time or cost increase, and helping the identification of 36 appropriate analytical lines (free from interferences) to apply to the samples determination without prior treatment. None sweetener exceeded the recommended value of inorganic contaminant (for aspartame samples the value was below 10 &#181;g g-1 and for saccharine samples that was below 20 &#181;g g-1). However, 6 brands of artificial sweetener in powder exceeded the legislation limit value for As (1.0 &#181;g g-1), and other 7 brands exceeded the Cr recommended maximum value (0.1 &#181;g g-1). Other 2 brands of Iiquid sweetener exceeded the Ni recommended maximum value. Therefore, for ali samples, the artificial sweetener in powder presented higher levels of potentially toxic metais (&#8776;5&#181;g g-1) than those in a Iiquid presentation (&#8776;2.0 &#181;g mL-1).

A aplicação de fertilizantes aos solos é frequente devido à atual demanda mundial pela produção de alimentos e às projeções feitas para o futuro. Fertilizantes orgânicos têm sido uma opção amplamente explorada devido ao seu preço, possibilidade de reuso de resíduos agriculturais e urbanos, além do caráter sustentável. Podem ser provenientes de diversas origens como lodo de esgoto, estercos e restos de animais e vegetais submetidos a processos de compostagem. Neste trabalho, foi desenvolvido método para determinação elementar total em fertilizantes orgânicos, substratos e condicionadores de solo, e realizados estudos de fracionamento e análise de especiação em fertilizante orgânico. Assim, foi realizada a caracterização geral das amostras com fracionamento por tamanho de partícula, análises dos teores de C e N, difração de Raios-X e determinação elementar total. Para este último, foi desenvolvido método de digestão assistida por radiação micro-ondas, com análise por ICP OES. O procedimento foi otimizado para diferentes misturas ácidas, as quais foram avaliadas quanto aos erros relativos para diversos elementos em CRM de lodo de esgoto, utilizando PCA como ferramenta estatística. A mistura escolhida, constituída por 3 mL HNO3 + 1 mL HBF4 + 2 mL H2O, foi utilizada para digestão das amostras. Apenas a concentração total não traz informações a respeito da mobilidade e disponibilidade dos elementos, então estudos de fracionamento elementar também foram realizados. Na primeira abordagem foi executado fracionamento por tamanho de partícula, no qual foi observado que quanto menor o tamanho de partícula menor o teor de C e maior o teor de N, e maior a concentração de micronutrientes e contaminantes (Cr, Cu, Fe, Mn, Pb e Zn), indicando que estes elementos podem estar mais associados a frações com mais características inorgânicas e maior teor de humificação (dado pela razão C/N). A segunda abordagem refere-se à associação dos elementos com diferentes componentes da matriz seguindo o procedimento de extração sequencial do BCR e extrações em etapa única para avaliar a mobilidade, sendo os extratos analisados por ICP-MS. O elemento Cr mostrou baixa mobilidade e bioacessibilidade. Cu está associado, principalmente, à fração orgânica da matriz, porém também pouco móvel. Fe e Pb também apresentaram baixa mobilidade e associados, principalmente, a óxidos da amostra. Mn e Zn mostraram alta mobilidade, sendo também associados a matéria orgânica e aos óxidos. A análise de especiação dos extratos obtidos foi realizada por SEC-ICP-MS, de modo a investigar a associação dos elementos extraídos com ácidos húmicos e fúlvicos. Nenhum sinal de Cr estava relacionado com a matéria orgânica, enquanto que os elementos Cu, Fe e Pb mostraram associação com ácidos húmicos. Mn apresentou espécies associadas a ácidos fúlvicos, enquanto que o Zn, não está associado a espécies orgânicas. / Fertilizers are frequently applied to soils due to actual global food demand and its future projections. Organic fertilizers have been extensively employed due to its price, as a reuse possibility to agricultural and urban wastes, and its sustainable character. They can be prepared from various sources, such as sewage sludge, manure and plants and animal scraps, submitted to composting process. In this work, it was developed a total elements determination method, as well as fractionating studies and speciation analysis of organic fertilizer. Thus, organic fertilizer samples were generally characterized using particle size fractionating, C and N content analysis, X-Ray diffraction and total elements determination. To the last, it was developed a sample preparation method by microwave-assisted digestion and ICP OES analysis. Digestion procedure was optimized using various acid mixtures, which were evaluated due to its relative errors related to sewage sludge CRM values, using PCA as a statistical tool. The chosen acid mixture constituted of 3 mL HNO3 + 1 mL HBF4 + 2 mL H2O was applied to samples digestion and their total elements determination. As total concentration is not enough to predict elements mobility and bioavailability, fractionation studies were conducted. The first approach was the particle size fractionation, were it was observed that the smaller the particle size the higher the micronutrients and contaminants concentration (Cr, Cu, Fe, Mn and Zn), along with lower C contents and higher N contents, indicating that these elements are associated with inorganic fractions, or with highly humified compounds (due to the lower C/N ratio). A second approach refers to elements association with various matrix components, using BCR and single-step extraction procedures to investigate elements mobility and bioavailability, with ICP-MS analysis of extracts. Cr was poorly mobile and bioavailable. Cu was bound to organic fraction, but also poorly mobile and bioavailable. Fe and Pb were also weakly mobile, and were mostly associated with oxides on matrix. Mn and Zn were greatly mobile and bioavailable, and were also mainly associated with oxides on matrix. Speciation analysis of extract was performed by SEC-ICP-MS, investigating elements association with humic and fulvic acids on previous extracts. Cr could not be associated with any humic substance, as expected with previous extractions results. Cu, Fe and Pb had identifiable species associated with humic acids, while Mn was associated with fulvic acids. Zn, even though high mobile, was not associated with any organic species.

A aplicação de fertilizantes aos solos é frequente devido à atual demanda mundial pela produção de alimentos e às projeções feitas para o futuro. Fertilizantes orgânicos têm sido uma opção amplamente explorada devido ao seu preço, possibilidade de reuso de resíduos agriculturais e urbanos, além do caráter sustentável. Podem ser provenientes de diversas origens como lodo de esgoto, estercos e restos de animais e vegetais submetidos a processos de compostagem. Neste trabalho, foi desenvolvido método para determinação elementar total em fertilizantes orgânicos, substratos e condicionadores de solo, e realizados estudos de fracionamento e análise de especiação em fertilizante orgânico. Assim, foi realizada a caracterização geral das amostras com fracionamento por tamanho de partícula, análises dos teores de C e N, difração de Raios-X e determinação elementar total. Para este último, foi desenvolvido método de digestão assistida por radiação micro-ondas, com análise por ICP OES. O procedimento foi otimizado para diferentes misturas ácidas, as quais foram avaliadas quanto aos erros relativos para diversos elementos em CRM de lodo de esgoto, utilizando PCA como ferramenta estatística. A mistura escolhida, constituída por 3 mL HNO3 + 1 mL HBF4 + 2 mL H2O, foi utilizada para digestão das amostras. Apenas a concentração total não traz informações a respeito da mobilidade e disponibilidade dos elementos, então estudos de fracionamento elementar também foram realizados. Na primeira abordagem foi executado fracionamento por tamanho de partícula, no qual foi observado que quanto menor o tamanho de partícula menor o teor de C e maior o teor de N, e maior a concentração de micronutrientes e contaminantes (Cr, Cu, Fe, Mn, Pb e Zn), indicando que estes elementos podem estar mais associados a frações com mais características inorgânicas e maior teor de humificação (dado pela razão C/N). A segunda abordagem refere-se à associação dos elementos com diferentes componentes da matriz seguindo o procedimento de extração sequencial do BCR e extrações em etapa única para avaliar a mobilidade, sendo os extratos analisados por ICP-MS. O elemento Cr mostrou baixa mobilidade e bioacessibilidade. Cu está associado, principalmente, à fração orgânica da matriz, porém também pouco móvel. Fe e Pb também apresentaram baixa mobilidade e associados, principalmente, a óxidos da amostra. Mn e Zn mostraram alta mobilidade, sendo também associados a matéria orgânica e aos óxidos. A análise de especiação dos extratos obtidos foi realizada por SEC-ICP-MS, de modo a investigar a associação dos elementos extraídos com ácidos húmicos e fúlvicos. Nenhum sinal de Cr estava relacionado com a matéria orgânica, enquanto que os elementos Cu, Fe e Pb mostraram associação com ácidos húmicos. Mn apresentou espécies associadas a ácidos fúlvicos, enquanto que o Zn, não está associado a espécies orgânicas. / Fertilizers are frequently applied to soils due to actual global food demand and its future projections. Organic fertilizers have been extensively employed due to its price, as a reuse possibility to agricultural and urban wastes, and its sustainable character. They can be prepared from various sources, such as sewage sludge, manure and plants and animal scraps, submitted to composting process. In this work, it was developed a total elements determination method, as well as fractionating studies and speciation analysis of organic fertilizer. Thus, organic fertilizer samples were generally characterized using particle size fractionating, C and N content analysis, X-Ray diffraction and total elements determination. To the last, it was developed a sample preparation method by microwave-assisted digestion and ICP OES analysis. Digestion procedure was optimized using various acid mixtures, which were evaluated due to its relative errors related to sewage sludge CRM values, using PCA as a statistical tool. The chosen acid mixture constituted of 3 mL HNO3 + 1 mL HBF4 + 2 mL H2O was applied to samples digestion and their total elements determination. As total concentration is not enough to predict elements mobility and bioavailability, fractionation studies were conducted. The first approach was the particle size fractionation, were it was observed that the smaller the particle size the higher the micronutrients and contaminants concentration (Cr, Cu, Fe, Mn and Zn), along with lower C contents and higher N contents, indicating that these elements are associated with inorganic fractions, or with highly humified compounds (due to the lower C/N ratio). A second approach refers to elements association with various matrix components, using BCR and single-step extraction procedures to investigate elements mobility and bioavailability, with ICP-MS analysis of extracts. Cr was poorly mobile and bioavailable. Cu was bound to organic fraction, but also poorly mobile and bioavailable. Fe and Pb were also weakly mobile, and were mostly associated with oxides on matrix. Mn and Zn were greatly mobile and bioavailable, and were also mainly associated with oxides on matrix. Speciation analysis of extract was performed by SEC-ICP-MS, investigating elements association with humic and fulvic acids on previous extracts. Cr could not be associated with any humic substance, as expected with previous extractions results. Cu, Fe and Pb had identifiable species associated with humic acids, while Mn was associated with fulvic acids. Zn, even though high mobile, was not associated with any organic species.

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. L'application optimisée d’un complexe (Cytochrome C digest) a montré des résultats comparables à ceux obtenus avec des peptides modèles. 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 (différents moments de centrifugation et différentes températures de traitement thermique) et les agents (DTT, β-mercaptoéthanol et TCEP) 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. Une procédure d’exposition à des métaux dans le laboratoire a montré que les métaux étaient significativement importante pour l’évaluation de la contamination à partir de la quantification de MT, selon le traitement de données par une techinique de réseau neural. / 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. The application of the optimized method in a more complex matrix (Cytochrome C digest) showed results comparable to those obtained with model peptides. 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 (varying centrifugation times and heat-treatment temperatures) and reducing agents (DTT, β-mercaptoethanol and TCEP) 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. Each procedure was then statistically evaluated. A response surface methodology was applied for bile samples, in order to further evaluate the responses for this matrix. In an environmental context, biliary MT was lower than liver MT, and, bile MT seems to be more adequate in environmental monitoring scopes regarding recent exposure to xenobiotics that may affect the proteomic and metalloproteomic expression of this biological matrix. A procedure for exposure to metals in the laboratory showed that some metals are significantly important for the assessment of contamination from the quantification of MT, according to the data processing by atifical neural network (ANN).

Les pétroles utilisés en raffinage étant de plus en plus lourds et chargés en métaux et hétéroélements, il est d’une grande importance pour les activités de raffinage de connaître la spéciation de ces espèces au sein des produits pétroliers et leurs comportements durant les procédés de raffinage. Afin d’apporter des réponses à cette problématique des techniques de caractérisation élémentaire et moléculaire ont été utilisées notamment par ICP MS Haute Résolution et FT ICR MS respectivement. Ces techniques analytiques sont appliquées à différents échantillons pétroliers provenant parfois de procédés de raffinage et les informations obtenues au niveau élémentaire et moléculaire sont utilisées de manières complémentaires pour améliorer notre compréhension des mécanismes se produisant au sein de nos échantillons durant les activités de raffinage. / The crude oils available for the refining industry are heavier and heavier and more concentrated in metals and heteroelements. It is thus of great importance to study the speciation of these species within the petroleum products and their behavior during the refining processes. To answer that problematic, elemental and molecular characterization techniques have been used, mainly High Resolution ICP MS and FT ICR MS for the elemental and molecular characterization respectively. The analytical techniques quoted are used for the analysis of several petroleum products sometimes originating from refining processes and the information obtained at the elemental and molecular level are combined to improve our understanding of the mechanisms occurring within our samples during refining activities.