O Zn é essencial para o metabolismo de plantas, animais e do ser humano. Existe a necessidade de buscar soluções para a baixa disponibilidade de Zn, frequente em solos do Brasil e do mundo, e de melhorar o valor nutricional dos grãos de cereais com este nutriente. O tratamento de sementes pode apresentar-se como uma importante técnica para o fornecimento de Zn na cultura do milho (Zea mays L.). Este estudo foi dividido em quatro capítulos, no primeiro, objetivou-se avaliar o potencial fisiológico de sementes de milho tratadas com Zn durante o armazenamento; no segundo, avaliou-se a distribuição do Zn em sementes tratadas durante o armazenamento das sementes por 6 e 12 meses, bem como a absorção do Zn por plântulas; no terceiro capítulo, foram avaliados os reflexos do tratamento de sementes com Zn sobre a produção de biomassa, rendimento e valor nutricional dos grãos e, por fim, no quarto capítulo, determinou-se, pelo uso da marcação do óxido de Zn com o nuclídeo 68Zn, a absorção do Zn pela planta, utilizando o método isotópico. Os resultados demonstram que o tratamento de sementes pode incrementar o vigor das plântulas provenientes de sementes com baixo potencial fisiológico; entretanto, em condições de armazenamento com temperatura e umidade relativa do ar não controladas, plântulas provenientes de sementes maiores tratadas, armazenadas por 12 meses nessas condições, podem apresentar fitotoxicidade, em função do tratamento. O Zn disponibilizado em revestimento, durante o armazenamento das sementes tratadas por 6 e 12 meses, sob condições não controladas, pode penetrar no pericarpo das sementes e encontrar-se acumulado em pontos específicos da semente. O ponto de maior acúmulo de Zn nas sementes é o embrião, acima da camada negra. Quanto à absorção do Zn disponibilizado via tratamento por plântulas, após horas 72 horas de as sementes terem sido colocadas para germinar, são visíveis os pontos de acúmulo do Zn na raiz primária, evidenciando que tecidos meristemáticos possuem alta demanda pelo elemento. O tratamento de sementes com Zn não propiciou aumento da biomassa da planta, rendimento dos grãos, acúmulo de Zn nas plantas e concentração de Zn nos grãos. Apesar desse resultado, o Zn disponibilizado via tratamento de sementes contribui de forma significativa para a nutrição das plantas de milho, conforme se avaliou por meio da marcação isotópica. / Zn is essential to the metabolism of plants, animals and human being. There is the need for solutions regarding the low availability of Zn, frequent on soils of Brazil and of the world, and for improving the nutritional value of grains with this nutrient. Treatment of seeds may present as an important technique for the provision of Zn in the corn culture (Zea mays L.). This study is organized in four chapters, in the first one, the objective is to determine the physiological potential of corn seeds treated with Zn during storage; in the second, the Zn distribution in seeds treated during storage for 6 and 12 months was analyzed, as well as the Zn absorption by plantules; in the third chapter, reflexes of the treatment of seeds with Zn over the production of biomass were evaluated and, lastly, the fourth chapter aims at determining the absorption of Zn by the plant, using the isotope method and marking zinc oxide with the 68Zn nuclide. Results showed that the treatment of the seeds may improve the vigor of plantules coming from seeds with low physiological potential; however, treated plantules coming from bigger seeds, stored for 12 months in uncontrolled conditions of temperature and relative humidity may present phytotoxicity because of the treatment. Zn made available in form of coating, during storage of seeds treated for 6 and 12 months, in uncontrolled conditions, may penetrate the pericarp of seeds and accumulate on specific spots. The biggest Zn accumulation spot Zn in seeds is the embryo, above the black layer. Regarding the absorption of Zn by the plantules made available via treatment, after 72 hours seeds germinated, accumulation spots of Zn in the primary root are visible, showing that the meristematic tissues have a high demand for the element. The treatment of seeds with Zn did not favor increase in the biomass of the plant, grain yield, accumulation of Zn in plants and concentration of Zn in grains. Despite this result, Zn made available via treatment of seeds contributes significantly for the nutrition of corn plants, as evaluated using isotope marking.

Submitted by Biblioteca de Pós-Graduação em Geoquímica BGQ (bgq@ndc.uff.br) on 2016-09-05T17:15:32Z No. of bitstreams: 1 MARTINS G.S. 2016.pdf: 11805657 bytes, checksum: 112f32d3100030fafec5c3f6222a4b72 (MD5) / Made available in DSpace on 2016-09-05T17:15:32Z (GMT). No. of bitstreams: 1 MARTINS G.S. 2016.pdf: 11805657 bytes, checksum: 112f32d3100030fafec5c3f6222a4b72 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Universidade Federal Fluminense. Instituto de Química. Programa de Pós-Graduação em Geociências - Geoquímica. Niterói, RJ / Este trabalho consiste na reconstrução paleoambiental de dois lagos amazônicos através de dados sedimentares e geoquímicos com especial ênfase a geoquímica orgânica e isotópica. Os lagos estudados apresentam-se em duas área climáticas distintas da Amazônia, sendo o Lago do Saci localizado ao sul do estado do Pará, em área de clima tropical com estação seca definida; e a Lagoa da Pata, localizada no extremo norte do Amazonas em regime de clima sempre úmido. O registro do Lago Saci abrange o intervalo entre 35.000 cal AP até o último milênio e o registro da Lagoa da Pata abrange os últimos 7500 cal AP. Na lagoa da Pata observou-se a existência de condições secas durante o Holoceno Inferior (7.500 – 6.500 anos cal AP) e o Holoceno Médio (6.500 – 3.600 anos cal AP) com baixos valores de COT (Carbono orgânico Total) e altos valores de 13C, sugerindo baixo nível do lago e aumento na contribuição de plantas C4. Durante o Holoceno Superior (3.600 anos cal AP até o presente) a estabilização de um clima mais úmido é observado na Lagoa da Pata, suportado pelo aumento significativo nos valores de COT e redução nos valores de 13C, sugerindo um aumento no nível do lago e contribuição de plantas C3. O registro molecular da Lagoa da Pata sugere a existência de uma vegetação adaptada a condições de alto estresse hídrico em função do tipo de solo presente na região do Morro dos Seis Lagos. A composição isotópica de carbono do n-alcano apresentou um range de valores característicos de vegetação C3 (-33,6‰ e -36,9‰) durante os últimos 7.500 anos cal AP. No Pleistoceno Superior, o Lago do Saci é caracterizado pelos menores valores de COT e valores mais enriquecidos de 13C sedimentar, sugerindo um baixo nível lacustre e grandes contribuições de plantas C4. O ápice do clima seco acontece entre 22.000 – 21.000 anos cal AP, com ocorrências significativas de incêndios, indicado pelos valores de BC (Carbono Grafítico). A transição para o Holoceno é marcado pela ausência de sedimentação entre 18.200-9.200 anos cal AP, sugerindo a ocorrência de uma fase seca severa ou evento erosivo atuante no fim do período glacial. O Holoceno no Lago do Saci marca um aumento no nível do lago, refletido pelo aumento na produtividade e acumulação da matéria orgânica. Esse período tem como característica a ocorrência de eventos de transporte de material da bacia de drenagem, representado pelos pulsos de areia.Os dados de biomarcadores moleculares sugerem condições de estresse hídrico durante o Pleistoceno e o Holoceno Médio na Lago do Saci, em função dos altos valores de ACL (Comprimento médio da Cadeia), sendo esses períodos os com maiores contribuições de plantas C4 como indicado pela razão C33:(C27+C29). Os valores de 13C dos n-alcanos (- 33.2‰ e -35.7‰) sugerem que a região foi coberta por vegetação C3 durante todo o período e mostram uma variação significativa entre o Pleistoceno Superior e o Holoceno, associada a alterações fisiológicas da vegetação. O registro de D sugere um período de precipitação reduzida no Pleistoceno e um período de chuvas mais abundantes no Holoceno. Ambos os registros (Lago do Saci e na Lagoa da Pata) sugerem condições ligeiramente mais úmidas durante o Holoceno, com a existência de uma fase seca no Holoceno Médio, caracterizados por uma redução significativa no nível lacustre e maior contribuição de plantas C4. As chuvas se tornam mais abundantes na Lagoa da Pata e o Lago do Saci a partir de 4.000 anos cal AP onde ocorre a expansão da floresta. Em Ambos os sítios, o sinal isotópico de biomarcadores, apesar de indicar alterações na precipitação, apresentou um enriquecimento do sinal isotópico em função da evapotranspiração ou alteração na vegetação fonte (C3 ou C4), alterando o processo de fracionamento do deutério. / In this work we present the paleoenvironmental reconstruction of two Amazonian lakes based in sedimentary and geochemical data with focus on organic and isotopic geochemistry.The lakes are presented in two distinct climatic area of the Amazon, the Saci Lake are located in south of Para state, in tropical area with dry season; and Pata Lake, located on the northern of the Amazon in a humid climate regime. The Saci lake core covers the range between 35,000 cal BP until the last millennium and Pata Lake covers the last 7,500 cal BP. Pata lake record dry conditions during the Early Holocene (7,500-6,500 cal years BP) and the Middle Holocene (6,500-3,600 cal years BP) with low TOC values (Total Organic Carbon) and high 13C values, suggesting low lake level and an increase of C4 plants contribution. During the Late Holocene (3,600 years cal BP to the present) is observed a stabilization of humid conditions, supported by the increase in TOC values and reduction in 13C values, suggesting an increase in lake level and C3 plants. The molecular record of Pata Lake suggests the existence of a vegetation adapted to high water stress conditions due the soil characteristic in the Six Lakes Morro region. The isotopic composition of n-alkane carbon presented a range of C3 vegetation (-33.6 ‰ and -36.9 ‰) over the last 7,500 years AP. In the Late Pleistocene, Saci lake is characterized by lower TOC values and more enriched 13C values, suggesting a low lake level and more C4 plants contributions. The dry climate occurs between 22,000 to 21,000 cal years BP, with significant occurrences of fires, indicated by the BC (Black Carbon) values. The transition to the Holocene is marked by the absence of sedimentation between 18,200-9,200 cal years BP, suggesting the occurrence of a severe drought phase or active erosive event at the end of the glacial period. The Holocene in Saci Lake is marked by an increase in lake level, reflected by the increase of productivity and organic matter accumulation. This period is characterized by the occurrence of runoff events, represented by sand pulses. The molecular biomarker data suggest water stress during the Pleistocene and the Middle Holocene in Lake Saci, with high ACL values (Average Chain Length), and more C4 plants contributions as indicated by the reason C33: (C27 + C29). The 13C values of nalkanes (-33.2 and -35.7 ‰ ‰) suggest that the area was covered by C3 vegetation during the whole period and show a significant variation between the Pleistocene and Holoceno associated with physiological changes in the C3 vegetation. The D record suggests a reduced precipitation in the Pleistocene and a period of more abundant rainfall in the Holocene. Both records (Saci lake and Pata Lake) suggest wetter conditions during the Holocene, with the existence of a dry phase in the Middle Holocene, characterized by a significant reduction in lake levels and greater contribution of C4 plants. The rains become more abundant in Pata Lake and Saci Lake after 4,000 cal years BP, where the forest expansion occurs. In both sites, the isotopic biomarkers indicate changes in precipitation, however the D enrichment could be explained by the evapotranspiration processes or change in vegetation source (C3 or C4), which interfers the deuterium fractionation process.

Les études isotopiques couplées avec des informations géothermiques peuvent constituer des outils pertinents pour l’exploration des eaux souterraines en tant que ressources en eau potable ou géothermiques. Ce travail combine les deux approches, isotopes de l’environnement et radioactifs associés à des données de température sur des aquifères profonds, dans l’objectif d’enrichir et d’améliorer la connaissance des mécanismes de recharge (Bassin d’Aquitaine, France) ainsi que des mécanismes de recharge et du potentiel géothermique (Bassin Crétacé de Bohème, République Tchèque).Les isotopes stables (18O, 2H, 13C) utilisés conjointement avec des radioisotopes (14C, 3H) sont utilisés pour estimer l’époque de la recharge ainsi que les conditions climatiques qui prévalaient lors de l’infiltration depuis la fin de Pléistocène jusqu’à nos jours. Définir le type de recharge et les conditions d’écoulement est nécessaire pour parvenir à modéliser de façon satisfaisante et fiable les grands systèmes aquifères profonds. Trois types de recharge ont été définis en Europe - (i) continue, (ii) interrompue lors du dernier maximum glaciaire (LGM) – un troisième type (iii) correspond à des situations particulières de recharge.Les conditions géographiques et climatiques très différentes rencontrées en France et en République Tchèque ont engendrées une importante hétérogénéité des conditions et processus de recharge. Le sud de la France, avec un climat relativement doux depuis les derniers 40 ka BP, n’a pas enregistré d’interruption de la recharge. Le temps de séjour des eaux souterraines en Bohème est estimé à environ 11 ka BP au maximum. Cependant, l’appauvrissement des teneurs en isotopes stables enregistré suggère une recharge liée à la fonte de la calotte glaciaire Nord Européenne après le dernier maximum glaciaire (LGM), autour de 18-20 ka BP. Des investigations sur les isotopes du carbone minéral dissous des eaux souterraines du bassin de Bohème ont montrées d’importantes interactions avec différentes sources de carbone qui ont été identifiées.Pour le site d’étude tchèque, les informations apportées par la géochimie ont été complétées par des données géothermiques afin d’améliorer la connaissance des flux et de la dynamique des eaux souterraines. Plus d’une centaine d’enregistrements diagraphiques de température ont été utilisés pour estimer le gradient géothermique. Plusieurs phénomènes viennent perturber le gradient géothermique de la région. Les flux d’eau souterraine verticaux et les variations lithologiques et topographiques sont à l’origine d’une distribution complexe du flux de chaleur, étant majoritairement conditionné par les écoulements souterraines. Les discontinuités peu profondes et les nombreux pointements volcaniques exercent aussi une influence importante sur l’écoulement souterrain et donc aussi sur le potentiel géothermique du réservoir. Les investigations sur la géothermie ont ainsi fourni des informations fondamentales sur le potentiel géothermique mais aussi sur les conditions d’écoulement des eaux souterraines. La prise en compte de ces informations s’avère nécessaire afin de proposer des modèles mathématiques d’écoulement réalistes. / Isotopic investigations combined with geothermal applications represent powerful tools for the exploration of groundwater potential as a drinking or geothermal resource. This Ph.D. thesis combines both approaches, environmental and radioactive isotopes together with temperature data in deep aquifers, in order to enrich and update the knowledge concerning the aquifer recharge processes in the Aquitaine Basin (France) and the aquifer recharge processes and geothermal potential in the Bohemian Cretaceous Basin (Czech Republic).Stable isotopes (18O, 2H, 13C) combined with radioisotope data (14C, 3H) are used to estimate the recharge timing and climatic conditions prevailing during the infiltration from the Late Pleistocene up to modern time. The character of groundwater recharge and regime are necessary to generate relevant source data for the accurate modelling of complex groundwater systems. Three groups of groundwater recharge types can be distinguished throughout Europe – (i) continuous recharge and (ii) interrupted recharge during Last Glacial Maximum and (iii) a group corresponding to particular recharge conditions.The contrasted geographic and climate conditions at both study sites in France and Czech Republic have entailed a great heterogeneity of the recharge conditions and processes. Southern France, with generally mild climatic conditions during the last 40 ka BP, did not experienced considerable hiatus in groundwater recharge. The residence time of groundwater in the Bohemian aquifers is estimated about 11 ka BP at the maximum but the depletion in the stable isotopes suggests that this groundwater originates in the melting of the north European ice sheets after the Last Glacial Maximum period, i.e. 18-20 ka BP. Further investigations on both stable and radioactive carbon isotopes indicated numerous groundwater interactions within the reservoir that were used to delineate the carbon origin within the Bohemian aquifers.Information on groundwater geochemistry was supplemented in the Czech case study by geothermal data in order to improve our knowledge of groundwater flow and dynamics. More than a hundred of temperature records from well-logging measurements were used to assess the geothermal gradient in the Bohemian Cretaceous Basin which is the most promising heat accumulation within the country. Many phenomena can affect the thermal field in the region. Vertical groundwater flow and variations in the lithology and the topography lead to a complicated areal distribution of the geothermal gradient and the heat flux which is dominantly controlled by groundwater. Shallow tectonic structures and numerous volcanic rocks exercise an influence on groundwater flow and therefore exert a secondary effect on the thermal field. The geothermal investigation provided useful information on the geothermal resources within the region but also represents an important tool for understanding groundwater flow, and for constructing realistic hydrogeological models in such a complex geological, tectonic and geothermal context.

Les travaux s'inscrivent dans le développement de catalyseurs conducteurs ioniques à base de zircone stabilisée à l'oxyde d'yttrium (YSZ) pour l'oxydation des suies dans les filtres à particules (FAP) qui équipent les véhicules Diesel. L'oxydation de la suie sur YSZ a été observée à l'échelle nanométrique par microscopie électronique en transmission environnementale. La réaction d'oxydation se déroule à l'interface (20-40 nm) entre le catalyseur et la suie, l'activité étant liée au nombre de points de contact. Nous avons associé un vecteur d'oxygène mobile, l'argent, à YSZ. Les résultats montrent une synergie entre YSZ et les nanoparticules d'argent qui promeut l'activité notamment en cas de contact grossier avec la suie. Des expériences isotopiques d'échanges et d'oxydation ont démontré que l'argent active l'intégration de l'oxygène gazeux dans le réseau de YSZ. La caractérisation des propriétés physicochimiques des nanoparticules d'argent dispersées sur différents oxydes (une autre zircone yttriée aux propriétés différentes : YSZb, ZrO2, CeZrO2, SiO2) a permis de relier les performances catalytiques avec la teneur d'argent métallique en surface. Des tests de vieillissement hydrothermal ont montré que la formulation supportée sur YSZb présente une activité maintenue en contact médiocre avec la suie et une faible perte en argent. Des mini-FAPs imprégnés de catalyseur ont ensuite été testés sur un banc de gaz synthétiques capable de produire des particules de suie en suspension. L'activité des formulations catalytiques une fois mises en forme ainsi que leur capacité de filtration ont été évaluées. Un mini-FAP contenant la formulation Ag/YSZb oxyde les suies dès 350°C et s'auto-régénère (oxydation des suies plus rapide que leur collecte) dès 450°C / This study deals with the development of ionic conductive catalysts based on yttria-stabilized zirconia for soot oxidation in Diesel Particle Filter (DPF). Soot oxidation was observed at the nanoscale using Environmental Transmission Electron Microscopy. Reaction was shown to occur at the interface between YSZ and soot, ranging from 20 to 40 nm, and activity was related to the number of contact points between soot and catalyst. In order to promote activity, we associated to YSZ an oxygen carrier: silver, under the form of silver nanoparticles (AgNPs) scattered over YSZ surface. Results display a synergy between YSZ and AgNPs, allowing for the promotion of soot oxidation under poor contact conditions. Isotopic exchange and oxidation experiments demonstrated AgNPs promote oxygen integration into YSZ lattice. Characterizations of supported AgNPs over different oxides such as YSZb (displaying different properties than YSZ), ZrO2, SiO2, CeZrO2, indicate that the soot oxidation activity is related to the amount of metallic silver species on the oxide surface. Hydrothermal ageing tests indicate that Ag/YSZb formulation displays a stable activity in poor contact conditions and a low silver loss. Baby-DPF (1 inch diameter, 3 inches length) were impregnated with silver-supported catalysts and tested on a synthetic gas bench producing a gas stream containing suspended soot particles. Soot oxidation activity and filtration efficiency were measured. A baby-FAP containing the Ag/YSZb formulation was shown to oxide soot at a temperature as low as 350°C and to induce self-regeneration (soot oxidation overcoming the collection) at 450°C

Le tritium est l’isotope radioactif de l’hydrogène. Il est capable d’intégrer la plupart des moléculesbiologiques. Bien que sa radiotoxicité soit faible, les effets du tritium seraient augmentés s’il étaitcapable de se concentrer dans des compartiments critiques du vivant. Afin de mieux comprendre lacirculation du tritium dans l’environnement et mettre en évidence les constantes de transfert entrecompartiments, nous avons étudié la tritiation de différentes matrices agricoles exposéeschroniquement à du tritium. La première étape de notre démarche est la validation des techniquesmises en oeuvre dans la préparation des échantillons. Nous avons démontré la possibilité de stocker deséchantillons environnementaux à Valduc et avons mis en évidence différents biais liés à l’extraction del’eau. Des solutions ont été proposées. Une hypothèse est formulée sur l’origine physique du facteurde séparation isotopique original que nous avons mis en évidence lors de l’extraction de l’eau du lait.Dans l’étude environnementale, les mesures faites sur les végétaux confirment l’importance del’exposition atmosphérique dans la tritiation de la plante. L’absence de différence liée à l’espèce dansla tritiation de la matière sèche a été démontrée. La diversité des résultats obtenus sur la tritiation desdifférentes fractions de matrices végétales encouragent la poursuite de cette voie de recherche. Nousavons comparé la tritiation des principales fractions hydrogénées du lait, d’abord entre elles puis entreéchantillons afin de mettre en évidence les facteurs à l’origine des différences observées. Nous avonsainsi montré la très grande corrélation entre l’eau d’abreuvement et l’eau libre du lait ainsi qu’entre lesactivités tritium de la matière sèche du bol alimentaire et des fractions organiques du lait. Nous avonségalement mis en évidence l’importance des paramètres métaboliques dans l’excrétion du tritium dansle lait. La synthèse de ces résultats montre l’importance des effets de la dilution de l’hydrogène dansles compartiments considérés devant les mécanismes de discrimination isotopique, y compris au seind’une chaîne alimentaire exposée chroniquement dans son ensemble. / Tritium is the radioactive isotope of hydrogen. It can be integrated in most of the biological molecules.Even if its radiotoxicity is weak, effects of tritium could be increased if it can concentrate in somecritical compartments of beings. In order to better understand the tritium circulation in the environment and highlight constants of transfer between compartments, we have studied the tritiationof different agricultural matrices chronically exposed to tritium. The first step of our study is thevalidation of the different techniques used to prepare our sample. We have also demonstrated that itwas possible to store environmental samples in the Valduc centre and have underlined some biasesdue to the extraction of free water. Some ways of improvements are proposed. A hypothesis has alsobeen formulated on the origin of an original isotopic fractionation effect during the extraction of the free water of milk. In the environmental study, the specific activities measured on plants confirm the importance of the atmospheric exposure on their tritiation. No difference in the tritiation has beenmeasured between wheat, barley and colza. Some differences have been measured in the tritiation ofsome organic components of vegetal matrices. These results underline the interest of continuing thiskind of study. We have also compared the tritiation of the main hydrogenated components of milk,first, component to component, then, sample to sample. Some origins of the measured differences havebeen shown. We have demonstrated the correlation between the specific activities of drinking waterand the free water of milk as between the tritiations of dry matter of cattle’s food and of the mainorganic components of milk. Our results show also the importance of the metabolism on thedistribution of tritium in the different compartments. The overall synthesis of our results show the importance of the dilution of hydrogen in the considered environmental compartments, above theisotopic effects, even in a food chain chronically exposed to tritium.

Depuis de nombreuses années, une approche a été mise en œuvre reposant sur l'identification de l'origine géographique d'un produit agro-alimentaire via la connaissance de ses isotopes du Sr, notamment via l'analyse du rapport 87Sr/86Sr. L'objectif de cette thèse est d'appliquer cette stratégie de reconnaissance aux huiles d'olive, en s'appuyant sur la production des huiles AOP de Nîmes dans le Sud de la France (Gard). La viscosité, la charge organique élevée, la teneur faible en Sr de cette matrice, rendent néanmoins inadaptés les protocoles de purification du Sr existants. La première approche de l'étude correspond donc à la définition et à la validation d'une méthode de traitement des huiles d'olive en vue de la purification du Sr afin de procéder à l'analyse de sa composition isotopique (CI). La seconde partie de l'étude est consacrée à la définition des processus d'assimilation et de transfert du Sr dans l'olivier. Cette approche, menée grâce à une étude détaillée d'un des moulins AOP de Nîmes, a permis de démontrer que le transfert du Sr des sols vers l'olivier et au sein des oliviers se fait sans fractionnement isotopique.La dernière partie de l'étude s'appuie sur la mesure et la caractérisation des rapports isotopiques 87Sr/86Sr des dix huiles d'olive AOP de Nîmes, et une comparaison de cette signature avec celle des formations géologiques sur lesquelles reposent les parcelles de culture. L'identité de la CI huile-territoire de culture a permis de conclure, dans le contexte de production de l'huile d'olive AOP de Nîmes, que les isotopes du Sr constituent un très bon outil de traçage de l'origine géographique des produits oléicoles. / For many years, an approach based on the identification of the geographical origin for food product has been implemented through the knowledge of its Sr isotopes, particularly through the analysis of 87Sr / 86Sr ratio. The objective of this thesis is to develop a similar strategy for the olive oils, from the example of PDO Nimes (Gard), in southern France. A specific methodology is necessary since the existing purification protocols of Sr revealed themselves inadequate because of viscosity, high organic load, low Sr content of this matrix In order to proceed to the analysis of the isotopic composition (IC), the first part of the study highlights the definition and validation of a specific method for the treatment of olive oils and strontium purification. The second part of the study is dedicated to expose the processes of assimilation and transfer of strontium into the olive tree. This approach, conducted through a detailed study of one of Nîmes PDO mills, demonstrated that the transfer of strontium from soils to olive tree, and within the olive tree, occurs without isotope fractionation.The last part of the manuscript studies the isotopic ratios 87Sr / 86Sr of the ten PDO olive oils of Nimes and consists in a comparison of this signature with the geological formations underlying the cultivation soils. This approach allows discussing quality of Sr isotopic tool in the geographical recognition of the olive products. The observed similarity between th IC from oils and soil allow us to conclude that, in this context of production, the Sr isotopes are a efficient tool to trace and certify the geographical origin of the oil.

As proteínas são consideradas importantes alvos para os oxidantes, devido à abundância em sistemas biológicos e às altas constantes de reações com estas espécies. Adicionalmente, têmse demonstrado que o triptofano (W) é um aminoácido extremamente susceptível a oxidação, inclusive pelo oxigênio singlete (1O2). A reação do W com o 1O2 tem despertado o interesse de diversos pesquisadores. Recentemente, esta reação tem atraído mais atenção, uma vez que produtos de oxidação do W tais como N-formilquinurenina (FMK) e quinurenina (kn) têm sido associados com algumas condições patológicas, tais como o desenvolvimento de catarata e a formação de agregados covalentes da superóxido dismutase envolvidos na esclerose lateral amiotrófica. Entretanto, há poucos trabalhos enfocando detalhadamente as reações, com estudos de estabilidade, identificação de subprodutos e propostas mecanísticas. Desta forma, pretendemos com este trabalho contribuir no esclarecimento do mecanismo de oxidação do triptofano pelo 1O2, através da análise e caracterização de produtos de oxidação gerados. Com este intuito, dois hidroperóxidos de triptofano isômeros (WOOH cis e trans, em relação ao grupamento carboxila) foram completamente caracterizados por análises de HPLC/espectrometria de massa e RMN como os principais produtos de oxidação do W pelo 1O2. Estes hidroperóxidos demonstraram ser relativamente estáveis às temperaturas ambiente e fisiológica, decompondo lentamente para os alcoóis correspondentes. O aumento do pH e/ou o aquecimento das soluções contendo os WOOH leva a decomposição quimiluminescente dos WOOH à FMK. Utilizando hidroperóxidos isotopicamente marcados com [18O] (W18O18OH) foi possível confirmar que a FMK formada durante esta decomposição era marcada com dois átomos de oxigênio. Este resultado demonstra que os dois átomos da FMK são derivados do grupamento hidroperóxido. Em adição, estas reações são quimiluminescentes, sugerindo o envolvimento de um intermediário dioxetano. Este mecanismo foi confirmado uma vez que o espectro de quimiluminescência da decomposição dos WOOH pode ser sobreposto ao espectro de fluorescência da FMK, inequivocamente identificado a FMK como a espécie emissora. Dioxindoilalaninas diastereoisoméricas também foram caracterizadas como produtos de oxidação do triptofano pelo 102; uma possível via radicalar foi excluída. Em suma, este estudo contribuiu na elucidação das bases químicas envolvidas na oxidação do triptofano por 102, através da caracterização dos produtos formados e da investigação detalhada dos mecanismos de decomposição destes produtos. / Proteins have been considered important targets for reactive oxygen species. Indeed, tryptophan (W) has been shown to be a highly susceptible amino acid to many oxidizing agents, including singlet molecular oxygen (1O2). The reaction of 1O2 with W has long been a matter of concern, and has recently attracted considerably more attention because W-derived oxidation products such as N-formylkynurenine (FMK) and kynurenine (kn) have been associated with some pathological conditions such as the development of cataracts and the formation of covalent aggregates of superoxide dismutase, which has been implicated in amyotrophic lateral sclerosis. Despite the intense interest in the mechanism of W oxidation, there are a lot of gaps that remains to be elucidated. In this context, the current study was undertaken to investigate the chemical basis involved in W oxidation by 1O2. We are concerned about the chemistry of the initially formed hydroperoxides, their stability, further reactions and the mechanism leading to FMK conversion. With this purpose, two cis and trans tryptophan hydroperoxide (WOOH) isomers were completely characterized by HPLC/mass spectrometry and NMR analyses as the major W-oxidation photoproducts. Also, they were shown to be relatively stable at ambient and physiological temperatures, leading to a slow decomposition to the corresponding alcohols. Increasing the pH or heating the solutions gives rise to a luminescent decomposition of the WOOH to FMK. Using 18O-labeled hydroperoxides (W18O18OH), it was possible to confirm the formation of a two-oxygen-labeled FMK molecule derived from W18O18OH decomposition. This result shows that both oxygen atoms in FMK are derived from the hydroperoxide group. In addition, these reactions are chemiluminescent (CL), indicating a dioxetane cleavage pathway. This mechanism was confirmed since the CL spectrum of the WOOH decomposition matched the FMK fluorescence spectrum, unequivocally identifying FMK as the emitting species. Diastereoisomeric dioxindoyalanine were also characterized as oxidation products derived from the reaction of W with 1O2. The involvement of radicals in this reaction was excluded. In summary, this work offers further insights into the chemistry involved in W-oxidation, through the characterization of photoproducts and the detailed investigation about the decomposition mechanism of these products.

En complément des analyses de concentration et de spéciation, les analyses des isotopes stables du mercure par MC-ICP-MS sont à l’heure actuelle un outil majeur pour tracer les sources et les transformations de cet élément dans l’environnement. Cette thèse propose le développement de techniques de pré-concentrations couplées au MC-ICP-MS afin de réaliser des analyses isotopiques de mercure au niveau du ng.L-1 (Ultra-trace). La stratégie de pré-concentration en ligne va générer des signaux transitoires courts dont la gestion en MC-ICP-MS représente un réel défi. Afin d’y répondre, une méthode de traitement de données adaptée à ce type de signaux ainsi qu’une méthode de correction du phénomène de dérive isotopique ont été développées. Dans le but de déterminer des compositions isotopiques en mercure total à de faibles concentrations, le couplage d’une génération de vapeur froide, d’une double amalgamation sur or (CVG-DGA) et d’un MC-ICP-MS a été développé. Cette nouvelle technique donne des précisions externes de l’ordre de 0.20 à 0.30‰ (2SD) pour des concentrations en mercure de 5 ng.L-1 en solution. Cette thèse présente également une nouvelle méthode d’analyse isotopique par espèce chimique (CSIA) réalisée à l’aide du couplage d’une chromatographie en phase gazeuse équipée d’un injecteur PTV (GC-PTV) et d’un MC-ICP-MS. Celle-ci permet l’analyse isotopique de plusieurs espèces chimiques avec des précisions externes de l’ordre de 0.30 à 0.40‰ (2SD) pour des concentrations en mercure de 150 ng.L-1 dans des échantillons biologiques.Les développements analytiques de cette thèse ont permis de mesurer directement et de façon automatisée des compositions isotopiques de mercure à des niveaux d’ultra-trace (jusqu’à 5ng.L-1) tout en conservant des précisions compatibles avec les principales questions environnementales posées. Ceci va permettre d’analyser des compartiments environnementaux contenant de faibles quantités de mercure (eaux naturelles, planctons,…) et ouvre ainsi de nouvelles perspectives pour une meilleure compréhension du cycle bio-géochimique du mercure. / In addition to the quantitative and speciation analysis, the analysis of mercury stable isotopes by MC-ICP-MS are now a tool of choice to track sources and pathways of this element in the environment. This PhD thesis presents the development of hyphenation between pre-concentration techniques and MC-ICP-MS to measure isotopic composition of mercury at ng.L-1 levels (Ultra-trace). The on-line pre-concentration strategy will create short transient signals which represent a real challenge for MC-ICP-MS. In order to solve it, a data treatment strategy for this particular signals and a correction method for the isotopic drift were developed.The hyphenation between a cold vapor generation, a dual gold amalgamation (CVG-DGA) and a MC-ICP-MS was developed to determine total mercury isotopic composition. This new technique gives external precisions ranging from 0.20 to 0.30‰ (2SD) for Hg concentration in solution of 5 ng.L-1. This PhD thesis also reports a new method to perform mercury compound specific isotopic analysis (CSIA) using a gas chromatography fitted with a PTV injector (GC-PTV) coupled to the MC-ICP-MS. This allows the isotopic analysis of several species with external precisions ranging from 0.30 to 0.40‰ (2SD) for mercury concentration down to 150 ng.L-1 in biological samples.The analytical developments proposed in this PhD thesis allows to automatically and directly measure mercury isotopic compositions at ultra-trace levels (down to 5 ng.L-1) while keeping precision compatible with main environmental questions. This will allow to analyze environmental compartment containing very low amount of mercury (natural waters, planktons,…) and then open new perspectives for a better understanding of the bio-geochimical cycle of mercury.

The present PhD is focused on the development of new analytical methods based on ICP techniques to carry out the elemental and isotopic analysis of bioethanol samples and other specimens taken along the bioethanol production process. The total sample consumption system, so-called hTISIS, has been used as sample introduction system in ICP-OES for the quantification of major and minor metals, and ICP-MS for the determination of trace metals. The hTISIS has also been successfully set up to carry out the lead isotopic analysis of bioethanol samples. Additionally, metals found in bioethanol have been monitored along the production process of this biofuel to identify the origin of these metals. Finally, volatile organic compounds have been determined in different bioethanol samples to perform the characterization of the matrices and their effect on the accuracy of the methods based on ICP techniques, developed in the present PhD for the elemental and isotopic analysis of these kind of samples. / La presente Tesis Doctoral se centra en el desarrollo de nuevos métodos de análisis basados en técnicas de ICP para llevar a cabo el análisis elemental e isotópico de muestras de bioetanol y muestras tomadas a lo largo del proceso de obtención de dicho biocombustible. Se ha empleado un sistema de consumo total de muestra, conocido como hTISIS, como sistema de introducción de muestras en ICP-OES para la cuantificación de metales mayoritarios y minoritarios, y en ICP-MS para la determinación de metales traza en bioetanol. El sistema hTISIS también se ha empleado con éxito para llevar a cabo el análisis isotópico de plomo en dichas muestras. Además, los metales encontrados en las muestras de bioetanol han sido monitorizados a lo largo del proceso de obtención del mismo para identificar el origen de dichos metales. Finalmente, se han determinado los compuestos volátiles presentes en diferentes muestras de bioetanol, con el objetivo de caracterizar las matrices y como estas pueden afectar a la exactitud de los métodos desarrollados, que emplean técnicas ICP para el análisis elemental e isotópico de este tipo de muestras.

Unicellular algae (microalgae) are able to produce a number of substances such as starches, oils, proteins, carotenoids, polyphosphates, or crystalline purines directly from inorganic sources by photosynthesis. Different species of microalgae can be used for the economic production of various biomolecules. Due to their autotrophic nature, microalgae are also unique as they can synthesize complex isotopically labeled biomolecules from simple isotopically labeled inorganic substances. Analysis of the chemical composition of microalgae by means of chemical-analytical methods is relatively complex, time-consuming, and laborious. Confocal Raman microscopy represents one of the optical methods by which the chemical composition of microalgae can be determined in situ, i.e. directly within intact cells. This technique combining confocal optical microscopy with Raman spectroscopy enables fast and non- destructive analysis of the chemical composition of substances in the investigated objects, including the effect of isotopic labeling. The chemical composition of the investigated objects is reflected by their Raman spectra, in the case of Raman mapping of microscopic objects by their chemical maps. In this work, a specific case of isotopic labeling was studied, namely the effect of heavy water (D2O) on the deuteration...