Le germanium est un métalloïde « stratégique » dans l’industrie high-tech, notamment pour la transition énergétique et le secteur des communications. Étant distinctement sidérophile, lithophile, chalcophile et organophile, le Ge possède un fort potentiel comme traceur géochimique. Ces travaux de thèse visent à améliorer la compréhension de la géochimie du cycle du Ge et des facteurs qui contrôlent son incorporation dans les minéraux et les gisements métalliques. Les cibles de cette étude concernent le gisement filonien à Zn de Saint-Salvy (Massif Central, France), le gisement filonien à Cu de Barrigão (Ceinture pyriteuse ibérique, Portugal), et le gisement à Zn–Cu de Kipushi (R.D. Congo). Les porteurs de Ge les plus importants sont respectivement la sphalérite (jusqu’à 2580 ppm), la chalcopyrite (jusqu’à 5750 ppm) et la réniérite (5,0–9,1 %). Les résultats montrent qu’il existe une relation de premier ordre entre la concentration en Ge dans la sphalérite et le type de gisement. La spectroscopie XANES par rayonnement synchrotron met en évidence la présence de Ge4+ en site tétraédrique dans les sulfures analysés. Les corrélations élémentaires observées dans la sphalérite et la chalcopyrite suggèrent une incorporation du Ge par co-substitutions, e.g., 3Zn2+ ↔ Ge4+ + 2(Cu,Ag)+ et 3Fe3+ ↔ 2Ge4+ + Cu+, ou via la création de lacunes cristallographiques, e.g., 2Zn2+ ↔ Ge4+ + ?. La corrélation positive δ74Ge–[Ge]ZnS des sphalérites de Saint-Salvy indiquerait que coefficient de partage (KdGe) augmenterai avec T. Les compositions isotopiques δ74Ge des sulfures étudiés varient de –5,72‰ à +3,67‰. Les compositions légères mesurées dans les gisements de Saint-Salvy et Barrigão semblent liées à des variations de température des fluides (basse à moyennes T) lors de l’incorporation de Ge en système ouvert, alors que la tendance marquée vers les compositions isotopiques lourdes à Kipushi indiquerait un fractionnement de Rayleigh. / Germanium is a critical metalloid in many high-tech industries, especially for the energy transition and the communication sector. Being distinctly siderophile, lithophile, chalcophile and organophile, Ge can be a particularly useful geochemical tracer. This thesis aims at understanding the Ge geochemistry and the factors controlling its concentration in Ge-bearing minerals and ore deposits. Three contrasted Ge-bearing deposits were studied, the Saint-Salvy Zn vein-type deposit, French Massif Central, the Barrigão Cu vein-type deposit, Iberian pyrite belt, Portugal, and the Kipushi Zn–Cu carbonate-hosted deposit, Central African copper-belt, D.R. Congo. The most important Ge-bearing minerals are sphalerite (up to 2580 ppm Ge), chalcopyrite (up to 5750 ppm Ge), and renierite (5.0–9.1 wt.% Ge). The results show a first order relation between the Ge content and the deposition temperature. Synchrotron-based XANES spectroscopy showed that Ge4+ occur in tetrahedral sites in the studied sulphides. Element correlations suggest that Ge is chiefly incorporated in sphalerite and chalcopyrite through coupled substitutions, e.g., 3Zn2+ ↔ Ge4+ + 2(Cu,Ag)+ and 3Fe3+ ↔ 2Ge4+ + Cu+, respectively, or via the creation of lattice vacancies, e.g., 2Zn2+ ↔ Ge4+ + ?. The positive δ74Ge–Ge content correlation observed in sphalerite from Saint-Salvy could indicate that Ge partition coefficient (KdGe) increases with temperature. Ge isotopes in sulphides yield δ74Ge values spanning from –5.72‰ to +3.67‰. The light δ74Ge compositions of Saint-Salvy and Barrigão ores appear to be related to variations in low to moderate fluid temperatures during Ge uptake in open system (e.g., fluid cooling), while the trend towards heavy δ74Ge compositions observed at Kipushi likely translates a Rayleigh fractionation effect during ore formation in closed system, associated with significant fluid modification.

A exposição à aflatoxina B1 (AFB1) ocorre predominantemente através da ingestão de alimentos contaminados. A AFB1 é biotransformada por enzimas hepáticas, levando à formação de adutos AFB1-N7-guanina e AFB1-lisina (AFB1-Lys), entre outros compostos que podem permanecer como resíduos no fígado, tais como aflatoxina M1 (AFM1) e aflatoxicol (AFL). Os produtos de biotransformação da AFB1 podem ser utilizados como biomarcadores de exposição às aflatoxinas através da dieta. O presente trabalho teve por objetivo verificar a aplicabilidade da determinação de biomarcadores de AFB1 na avaliação da eficiência de adsorventes em suínos. Foram utilizados 24 suínos machos castrados com 28 dias de idade, que após 21 dias de adaptação foram divididos em 4 tratamentos experimentais, em arranjo fatorial 2 x 2, correspondendo à dois níveis de incorporação de AFB1 às rações (0 e 1,1 mg/kg) e dois níveis de incorporação (0 e 0,5%) de Aluminossilicato de Cálcio e Sódio Hidratado (HSCAS) por 42 dias. Semanalmente foram analisados parâmetros de desempenho, coletadas amostras de soro e plasma para análises bioquímicas, hematológicas e dosagem de AFB1-Lys. Nos primeiros dez dias de intoxicação foram coletadas amostras diárias de urina para análise de produtos da biotransformação de AFB1 incluindo AFB1-N7-guanina. Após este período, as coletas de urina foram semanais. Ao final do experimento, foram coletadas amostras de soro, plasma heparinizado e com EDTA para avaliação de AFB1-Lys, e os animais foram abatidos para avaliação da carcaça e coleta de órgãos para avaliação do peso relativo, histopatologia e análise de resíduos. O HSCAS foi efetivo na adsorção in vitro de AFB1, cujos percentuais de ligação variaram de 36,83 a 100%, para concentrações do adsorvente entre 0,005 e 10 mg/10 mL. A adição de HSCAS restaurou significativamente (P<0,05) os efeitos deletérios sobre a maioria dos parâmetros avaliados. Em fígado, os níveis de AFB1, AFB2 e AFG1 foram menores (P<0,05) no tratamento com AFB1 e HSCAS, indicando que houve proteção do adsorvente nos animais deste grupo. Em urina, a AFM1 consistiu em ~93% do total de aflatoxinas excretadas quantificada. A inclusão de adsorvente nas dietas reduziu (P<0,05) os níveis urinários de AFB1, AFQ1 e AFB1-N7-guanina durante todo o experimento. Nos níveis de AFM1 houve redução significativa entre os dias 1 e 28 de intoxicação. Comparando-se com os resultados obtidos em soro e plasma heparinizado, os níveis de AFB1-Lys foram fortemente suprimidos em amostras com EDTA devido à interferência deste composto no processo de digestão enzimática. Os níveis semanais de AFB1-Lys séricos nos animais tratados com AFB1 e HSCAS foram menores (P<0,05) do que nos animais alimentados somente com AFB1 durante todo o período de intoxicação. Os dados indicam que níveis de AFB1, AFM1, AFQ1 e AFB1-N7-guanina urinária bem como AFB1-Lys sérica podem ser utilizados como biomarcadores de exposição para aflatoxinas em suínos. Além disso, as determinações de AFB1 e AFB1-N7-guanina urinária e AFB1-Lys sérica podem ser utilizadas para avaliar a capacidade de proteção dos adsorvente como biomarcadores de dose interna de AFB1, sendo capazes de demonstrar à severidade da intoxicação em cada animal. Outros estudos são necessários para esclarecer a toxicocinéticados biomarcadores avaliados em suínos. / Exposure to aflatoxin B1 (AFB1) occurs primarily through ingestion of contaminated food. AFB1 is biotransformed by liver enzymes leading to the formation of adducts AFB1-N7-guanine and AFB1-lysine (AFB1-lys), among other compounds which may remain as residue in the liver, such as Aflatoxin M1 (AFM1) and aflatoxicol (AFL). The products of AFB1 biotransformation can be used as exposure biomarkers to aflatoxins through diets. This study aims to verify the applicability of the determination of AFB1 biomarkers in the evaluation of adsorbents efficiency in swine. Twenty four barrows with 28 days of age were used. After 21 days of adaptation they were divided into 4 experimental treatments in a factorial 2 x 2, corresponding to two AFB1 incorporation levels to feed (0 to 1.1 mg / kg) and incorporating two levels (0 and 0.5%) of sodium calcium aluminosilicate hydrate (HSCAS) for 42 days. Performance parameters were weekly analyzed, as well the biochemical and hematological, and AFB1-Lys dosage. Urine samples were collected daily in the first ten days of intoxication to assess biotransformation products of AFB1, including AFB1-N7-guanine. After this period, the collection of urine samples occurs weekly. At the end of the experiment, serum, heparinized and EDTA plasma samples were collected for AFB1-Lys evaluation, and the animals were slaughtered to evaluate carcass and collection agencies to assess the relative weight, histopathology and residue analysis. The HSCAS was effective in in vitro adsorption of AFB1 binding percentages that ranged from 36.83 to 100% for the adsorbent concentrations between 0.005 and 10 mg/10 ml. Adding HSCAS restored significantly (P<0.05) the deleterious effects on the majority of the parameters evaluated. In liver, levels of AFB1, AFB2 and AFG1 were lower (P<0.05) in the treatment AFB1 and HSCAS, indicating that there was protection of the adsorbent in the animals of this group. In urine, AFM1 consisted of ~ 93% of total aflatoxins excreted and quantified. The inclusion of adsorbent in diets reduced (P<0.05) urinary levels of AFB1, AFQ1 and AFB1-N7-guanine throughout the experiment. In AFM1 levels decreased significantly between days 1 and 28 of poisoning. Compared with the results obtained in serum or heparinized plasma, AFB1-Lys levels were strongly suppressed in the samples with EDTA interference due to this compound in the enzyme digestion process. Weekly serum levels of AFB1-Lys in animals treated with AFB1 and HSCAS were lower (P<0.05) than animals treated with AFB1 alone during all the poisoning period. The data indicate that the urinary levels of AFB1, AFM1, AFQ1 and AFB1-N7-guanine and serum AFB1-Lys can be used as biomarkers of exposure to aflatoxins in pigs. Furthermore, the determinations of AFB1 and AFB1-N7-guanine urinary and serum AFB1-Lys can be used to evaluate the protective ability of the adsorbent as internal dose biomarkers, being able to demonstrate the severity of toxicity in each animal. Further studies are necessary to provide physiologically based toxicokinetics of the evaluated biomarkers in pigs.

O presente estudo avaliou o perfil farmacogenômico de 338 pacientes, sob terapia antiagregante. Os pacientes foram submetidos a tratamento prévio com AAS (100mg/dia) e clopidogrel (75mg/dia) por no mínimo cinco dias antes da angioplastia coronária. Os indivíduos com resposta considerada indesejada <30% de inibição de PRU (do inglês, P2RY12 Reaction Unit) para clopidogrel e >550 ARU (do inglês, Aspirin Reaction Unit), foram considerados como não respondedores. As concentrações plasmáticas dos antiagregantes foram determinadas por cromatografia líquida acoplada à espectrometria de massa do tipo triploquadrupolo (LC-MS/MS). A taxa da inibição da agregação plaquetária foi medida utilizando-se o sistema VerifyNow®. A expressão gênica global das células totais do sangue periférico foi avaliada pela tecnologia de microarranjos de DNA Human Exon ST 1.0 Array. Características genotípicas dos pacientes também foram avaliadas pelo sistema Sequenom®. Assim, foi possível obter como resultados a identificação de 64% e 10% para pacientes não respondedores ao clopidogrel e AAS respectivamente, sendo que para o primeiro foi possível identificar a associação desta não resposta a variáveis clínicas como diabetes (p = 0,003), hipertensão (p = 0,011) e hábito de fumar (p = 0,041) e sexo (p = 0,022) e idade dos pacientes (p = 0,004) em relação à resposta ao AAS. O método de quantificação simultânea do clopidogrel, seu metabólito majoritário e do AS (metabólito do AAS), apresentou limites de quantificação entre de 2 a 500 ng/mL, 2 a 2000 ng/mL e de 20 a 2000 ng/mL, respectivamente. O estudo de associação encontrou uma relação significante da presença dos SNPs presentes nos genes CYP5A1 (rs2299890) e CYP2C19 (rs4244285 e rs3758580), com a variação na resposta ao clopidogrel, obtendo um valor de p corrigido pelo teste de permutação inferior a 0,001. Como também, uma fraca associação da variação na resposta do AAS com o SNP rs9605030 do gene COMT (p = 0,009). Os resultados do microarranjos relacionaram a resposta terapêutica ao clopidogrel com os genes CA2, MKRN1, ABCC3 e MBP seguido dos genes NFIA e IGF1R para a resposta ao AAS. Concluindo que o estudo farmacogenômico apresentou todo o seu potencial para relacionar variáveis como resposta, concentração farmacológica plasmática, SNPs e expressão global de RNAm, possibilitando assim compreender melhor a variação no tratamento antiagregante. / This study investigated the pharmacogenomics profile of 338 patients under antiplatelet therapy. Patients undergoing pretreatment with ASA (100 mg/day) and clopidogrel (75mg/day) for at least five days prior to coronary angioplasty. Individuals with response <30% of PRU (P2RY12 reaction unit) were considering non responder for clopidogrel and >550 of ARU (aspirin reaction unit), were considered as non responders for ASA. Plasma concentrations of the antiagregation drugs were determined by liquid chromatography followed mass spectrometry of triple quadrupole detection (LC-MS/MS). The rate of inhibition of platelet aggregation was measured using the VerifyNow® system. The global gene expression of total cells in blood was assessed by DNA microarray technology Human Exon 1.0 ST Array. Genotypic characteristics of the patients were also evaluated by the Sequenom® system. Thus it was possible to obtain results such as identification of 64% and 10% for patients non responders to clopidogrel and aspirin respectively, and for the first could identify the association of this response to variables such as diabetes (p = 0.003), hypertension (p = 0.011) and smoking (p = 0.041) for clopidogrel and sex and age in relation to response to ASA (p = 0.022 and p = 0.004, respectively). The method of simultaneous quantification of clopidogrel and its major metabolite of AS (metabolite of ASA), had quantification limits between 200 to 500 ng/mL 2000-2000 ng/mL and 20 to 2000 ng/mL, respectively. The association study found a significant grating presence of SNPs present in genes CYP5A1 (rs2299890) and CYP2C19 (rs4244285 and rs3758580), with the variation in the response to clopidogrel, obtaining a corrected p value by permutation test below 0.001. As well, a weak association of variation in the response of ASA with the SNP rs9605030 of the gene COMT (p = 0.009). The results of microarray related therapeutic response to clopidogrel with genes CA2, MKRN1, ABCC3 and MBP followed by NFIA and IGF1R genes for response to ASA. Concluding that the pharmacogenomics study showed its potential to relate variables such as response, plasma drug concentration, SNPs and global expression of mRNA, thus enabling better understand the variation in antiplatelet treatment.

A contaminação de corpos d\'água por fármacos é um tema de extrema relevância, tendo em vista problemas como a escassez de água, florações de cianobactérias tóxicas e lançamentos clandestinos de efluentes domésticos. Sendo assim, este trabalho teve como objetivo determinar a presença de cafeína (CAF), fluoxetina (FLX), levotiroxina (LVX) e bezafibrato (BZF) em mananciais do estado de São Paulo, bem como avaliar a toxicidade desses compostos à cianobactéria Microcystis aeruginosa LTPNA 08. Um método por LC-MS/MS foi desenvolvido e validado, de acordo com a RDC nº 166 da ANVISA, para a detecção de CAF, FLX, LVX e BZF em amostras ambientais. As represas Guarapiranga e Billings, bem como os rios Taiçupeba, Sorocaba, Baixo Cotia, Grande e Paraíba foram monitorados de abril a setembro de 2017. A toxicidade dos fármacos foi avaliada por meio do monitoramento do crescimento, produção de microcistinas e viabilidade celular da cianobactéria M. aeruginosa LTPNA 08. CAF foi detectada em todas as amostras analisadas, com concentrações que variaram de 6,6 ng.L-1 a 16,47 &#181;g.L-1. No Rio Cotia foram verificadas as maiores concentrações de CAF, FLX e BZF (16,47 &#181;g.L-1; 3,5 ng.L-1 e 322 ng.L-1, respectivamente). A LVX, cujos produtos de biotransformação não foram monitorados, não foi detectada em nenhuma amostra analisada. A concentração de 50 &#181;g.L-1 de FLX inibiu o crescimento da cianobactéria em 82,3% (CE50: 31,4 &#181;g.L-1). Em relação à produção de microcistinas totais, os fármacos inibiram a liberação da fração extracelular para a maior concentração testada ao longo do tempo de monitoramento, embora não tenham demonstrado efeito sobre a viabilidade celular. Sendo assim, considerando-se que fármacos estão presentes nos mananciais monitorados no estado de São Paulo e que a FLX pode causar efeito sobre a M. aeruginosa, os efeitos decorrentes da exposição a concentrações ambientais contínuas e cumulativas de fármacos em corpos d\'água devem ser estudados. Além disso, uma vez que a ocorrência destas substâncias e outros contaminantes antropogênicos no ambiente aquático natural é uma questão emergente devido aos efeitos adversos potenciais que estes compostos representam para a vida aquática e os seres humanos, os tipos e níveis destes compostos, que têm um impacto maior na qualidade da água, deve ser constantemente monitorada. Práticas de gestão que investem em saneamento e na redução da descarga de efluentes não tratados, e um plano de proteção de recursos hídricos com o objetivo de garantir a segurança da água seriam medidas essenciais para reduzir o aporte de contaminantes nos corpos d\'água do estado de São Paulo. / Contamination of water bodies by drugs is a subject of extreme relevance considering related problems such as water scarcity, harmful cyanobacterial blooms and discharge of untreated domestic effluents. Therefore, the aim of this work was to determine the presence of caffeine (CAF), fluoxetine (FLX), levothyroxine (LVX) and bezafibrate (BZF) in springs in the State of São Paulo, and to evaluate the toxicity of these compounds in cyanobacteria Microcystis aeruginosa LTPNA 08. A LC-MS/MS method was developed and validated according to RDC nº 166 of ANVISA to assess the concentration of CAF, FLX, LVX and BZF in environmental samples. Guarapiranga and Billings reservoirs, as well as the Taiçupeba, Sorocaba, Baixo Cotia, Grande and Paraíba rivers were monitored from April to September 2017.The drugs toxicity in M. aeruginosa LTPNA 08 was assessed by monitoring their effects on cyanobacterial growth, microcystins production and cell viabilityby flow cytometry. CAF was detected in all analyzed samples at concentrations ranging from 6.6 ng to 16.47 &#181;g.L-1.Among studied sites, Cotia river showed the highest concentrations of CAF, FLX and BZF (16.47 &#181;g.L-1, 3.5 ng.L-1 and 322 ng.L-1, respectively). LVX, which biotransformation products were not monitored, was not detected in any of the analyzed samples. Regarding the drugs toxicity, 50 &#181;g.L-1 of FLX inhibited the cyanobacterial grow thin 82.3% (EC50 of 31.4 &#181;g.L-1). Although no effect on cell viability was seen by flow cytometry, the highest concentrations of all compounds tested were able to inhibit the release of microcystins. Therefore, considering that some of the drugs monitored showed to be present in water sources in São Paulo State and that FLX affects cyanobacteria M. aeruginosa growth, the effects of continuous and cumulative exposure at environmental drug concentrations of in water bodies should be evaluated. Also, since the occurrence of these substances and other anthropogenic contaminants in the natural aquatic environment is an emerging issue due to the potential adverse effects these compounds pose to aquatic life and humans, thet ypes and levels of these compounds, which have a greater impact on water quality, should be constantly monitored. Management practices investing in sanitation and in reducing discharge of untreated effluents, as well as a plan for water resources protection with the goal of ensuring water security would be essential measures in reducing drugs loading into water bodies situated in São Paulo State.

As micotoxinas s&atilde;o compostos produzidos por fungos, sendo potencialmente perigosos &agrave; sa&uacute;de humana e animal. A Ocratoxina A (OTA) &eacute; uma das micotoxinas mais amplamente estudadas, sendo encontrada em v&aacute;rias matrizes aliment&iacute;cias. A concentra&ccedil;&atilde;o dessa micotoxina nos alimentos &eacute; geralmente muito baixa (da ordem de ng g-1), sendo, portanto, necess&aacute;rio o emprego de t&eacute;cnicas de preparo de amostras que realizem a purifica&ccedil;&atilde;o e pr&eacute;-concentra&ccedil;&atilde;o no analito. Os m&eacute;todos de separa&ccedil;&atilde;o e detec&ccedil;&atilde;o empregados na an&aacute;lise de OTA tamb&eacute;m devem oferecer sensibilidade adequada para a quantifica&ccedil;&atilde;o do analito. Logo, os objetivos do trabalho foram desenvolver uma metodologia utilizando a t&eacute;cnica in-tube SPME no modo de extra&ccedil;&atilde;o &uacute;nica (flow through extraction) com part&iacute;culas de C18 como fase extratora e separa&ccedil;&atilde;o e detec&ccedil;&atilde;o por HPLC-MS/MS. Al&eacute;m disso, otimizar e validar a metodologia proposta e detectar e quantificar a OTA em amostras de vinho. Para isso, a t&eacute;cnica in-tube SPME foi configurada e um tubo de PEEK foi empacotado com part&iacute;culas de C18, o qual foi utilizado na extra&ccedil;&atilde;o. A otimiza&ccedil;&atilde;o do m&eacute;todo foi realizada utilizando-se um planejamento experimental composto central 22 + 3 pontos centrais, tendo como fatores a porcentagem de ACN e o tempo durante o carregamento da amostra. A valida&ccedil;&atilde;o da metodologia empregada foi realizada conforme o guia de valida&ccedil;&atilde;o da ANVISA e, posteriormente, amostras de vinho tinto seco e vinho branco seco foram analisadas. O m&eacute;todo proposto foi desenvolvido, tendo sua funcionalidade atestada e suas condi&ccedil;&otilde;es de an&aacute;lise melhoradas, sendo utilizado 22 % de ACN e 6 minutos no carregamento da amostra. O m&eacute;todo foi validado, mostrando sensibilidade adequada, com limites de detec&ccedil;&atilde;o e quantifica&ccedil;&atilde;o iguais &agrave; 0,02 e 0,05 &micro;g L-1, respectivamente. A linearidade e precis&atilde;o da metodologia foram avaliadas, apresentando coeficiente de correla&ccedil;&atilde;o igual a 0,996 e DPR menor que 6%, respectivamente. O m&eacute;todo mostrou-se exato nos n&iacute;veis de concentra&ccedil;&atilde;o m&eacute;dio e alto e a recupera&ccedil;&atilde;o m&aacute;xima obtida para o n&iacute;vel alto foi pr&oacute;ximo 73%. Amostras brasileiras e estrangeiras de vinho tinto seco e vinho tinto seco branco foram analisadas e a OTA n&atilde;o foi detectada em nenhuma delas. Entretanto, a OTA pode essa estar presente nas amostras analisadas em concentra&ccedil;&otilde;es mais baixas que as determinadas pelos limites de detec&ccedil;&atilde;o e quantifica&ccedil;&atilde;o, n&atilde;o sendo potencialmente perigosos &agrave; sa&uacute;de. / Mycotoxins are compounds produced by fungus, being a potential danger to human and animal health. Ochratoxin A (OTA) is a mycotoxin widely studied, being found in a varied of food matrices. OTA concentration in food is generally low (the order of ng g-1), being, therefore, necessary the use of sample preparation techniques to perform the analyte purification and preconcentration in the sample. The separation and detection methods used in OTA analysis also should offer proper sensibility in order to quantify the analyte adequately. In short, the goals of the present work were to develop a methodology using in-tube SPME in flow through extraction mode with C18 particles as extraction phase and separation and detection by HPLC-MS/MS. Further, optimize and validate the proposed methodology and detect and quantify OTA in wine samples. For this, in-tube SPME was configured and a PEEK tube was packed with C18 particles, which was used in the extraction step. The method optimization was achieved using a central composite 22 + 3 central points experimental design, having as factors the percentage of ACN and time during sample loading step. Validation method was done following ANVISA validation guide after which red dry wine and white dry wine samples were analyzed. The proposed method was developed, having its functionalities attested and its analysis conditions enhanced, using 22% of ACN and 6 minutes in sample loading. The method was validated, demonstrating proper sensitivity, with detection and quantification limits equal to 0.02 and 0.05 &micro;g L-1, respectively. Linearity and precision were evaluated, exhibiting correlation coefficient equal to 0,996 e RSD under 6%, respectively. The method was accurate in medium and high concentration levels and maximum recovery was 73% in high concentration level. Brazilian and foreign red dry wine and white dry wine samples were analyzed and OTA was not detected in any of them. However, OTA may be present in samples analyzed in lower concentrations than that ones determined by detection and quantification limits, not being a potential danger to human health.

La surveillance radiologique de l’environnement est un enjeu majeur pour étudier l Impact des activités industrielles et humaines mettant en œuvre des substances radioactives. Umalyse de l:tImet Pu présents à l’échelle des ultra-traces dans les sols et les sédiments (- mBq.kg-1) est souvent menée par spectrométrie (après séparation chimique. Ces méthodes souffrent de plusieurs inconvénients. Les objectifs de cette thèse sont par conséquent de simplifier la méthode de séparation et d'obtenir une limite de détection basse et un temps d’analyse relativement court en utilisant lICPMS-HR couplé au système d’introduction Apex-Q/ACM. Sur la base des données de la littérature, une méthode chimique utilisant une seule résine d’extraction chromatographique (DGA) est proposée pour isoler les concentrations ultra-traces d’Am et de Pu des différents types d'éléments interférents. La méthodologie proposée a été évaluée avec des solutions synthétiques avant l'utilisation de solutions de lixiviation d’échantillons préalablement analysés par spectrométrie alpha. La méthode retenue demande plusieurs procédure d’élution pour pouvoir séparer les majeurs, les interférents isobariques, les lanthanides, l’uranium et le thorium. Des rendements de chimie supérieurs à 90% pour Am et 60% pour Pu ont pu être mesurés. La robustesse de la méthode d’analyse a été validée avec un standard de référence certifié (AIEA-38S) .Enfin, le protocole analytique d’isolation et de dosage des ultra-traces d241Am et de 239Pu par ICP-MS-HR solution a été appliqué à des échantillons de sédiments échantillonnés dans l’estuaire de la Loire pour caractériser L’état radiologique du fleuve. / Radiological monitoring of the environment is a key issue for studying the impact of industrial and human activities using radioactive substances. The analysis of Am and Pu present at ultra-trace concentrations in soils and sediments (-mBq.kg-l) is often carried out by spectrometry after chemical separation. This procedure suffers from several drawbacks (multi-step separation process and long coun.tingtime). The objectives of this PhD are consequently simplify separation process and to achieve a low limit of detection with a relatively short analysis time using a high-resolution plasma source mass spectrometer HR-ICP-MScoupled to the introduction system Apex-Q/ACM. Base on literature data, a chemical methodology using a single column extraction chromatography resin (DGA) is proposed for isolating ultra-trace concentrations of Am and Pu from different types of interfering elements. The proposed methodology is assessed with synthetic solutions prior to the use of real leaching solutions previously analyzed by alpha spectrometry. The chosen method requires several elution procedures in order to separate the majors element, isobaric interferes, lanthanides, uranium and thorium. Recovery yields greater than 90% for Am and 60% for Pu were determined. The robustness of the analytical method was validated with a certified reference standard sample (IAEA-385). Finally, the analytical process for isolation and analysis of 241Am and 239Pu at ultra-trace concentration by ICP-MS-HRsolution was applied to samples of sediment taken from Loire estuary to characterize the radiological status of this river.

As micotoxinas s&atilde;o compostos produzidos por fungos, sendo potencialmente perigosos &agrave; sa&uacute;de humana e animal. A Ocratoxina A (OTA) &eacute; uma das micotoxinas mais amplamente estudadas, sendo encontrada em v&aacute;rias matrizes aliment&iacute;cias. A concentra&ccedil;&atilde;o dessa micotoxina nos alimentos &eacute; geralmente muito baixa (da ordem de ng g-1), sendo, portanto, necess&aacute;rio o emprego de t&eacute;cnicas de preparo de amostras que realizem a purifica&ccedil;&atilde;o e pr&eacute;-concentra&ccedil;&atilde;o no analito. Os m&eacute;todos de separa&ccedil;&atilde;o e detec&ccedil;&atilde;o empregados na an&aacute;lise de OTA tamb&eacute;m devem oferecer sensibilidade adequada para a quantifica&ccedil;&atilde;o do analito. Logo, os objetivos do trabalho foram desenvolver uma metodologia utilizando a t&eacute;cnica in-tube SPME no modo de extra&ccedil;&atilde;o &uacute;nica (flow through extraction) com part&iacute;culas de C18 como fase extratora e separa&ccedil;&atilde;o e detec&ccedil;&atilde;o por HPLC-MS/MS. Al&eacute;m disso, otimizar e validar a metodologia proposta e detectar e quantificar a OTA em amostras de vinho. Para isso, a t&eacute;cnica in-tube SPME foi configurada e um tubo de PEEK foi empacotado com part&iacute;culas de C18, o qual foi utilizado na extra&ccedil;&atilde;o. A otimiza&ccedil;&atilde;o do m&eacute;todo foi realizada utilizando-se um planejamento experimental composto central 22 + 3 pontos centrais, tendo como fatores a porcentagem de ACN e o tempo durante o carregamento da amostra. A valida&ccedil;&atilde;o da metodologia empregada foi realizada conforme o guia de valida&ccedil;&atilde;o da ANVISA e, posteriormente, amostras de vinho tinto seco e vinho branco seco foram analisadas. O m&eacute;todo proposto foi desenvolvido, tendo sua funcionalidade atestada e suas condi&ccedil;&otilde;es de an&aacute;lise melhoradas, sendo utilizado 22 % de ACN e 6 minutos no carregamento da amostra. O m&eacute;todo foi validado, mostrando sensibilidade adequada, com limites de detec&ccedil;&atilde;o e quantifica&ccedil;&atilde;o iguais &agrave; 0,02 e 0,05 &micro;g L-1, respectivamente. A linearidade e precis&atilde;o da metodologia foram avaliadas, apresentando coeficiente de correla&ccedil;&atilde;o igual a 0,996 e DPR menor que 6%, respectivamente. O m&eacute;todo mostrou-se exato nos n&iacute;veis de concentra&ccedil;&atilde;o m&eacute;dio e alto e a recupera&ccedil;&atilde;o m&aacute;xima obtida para o n&iacute;vel alto foi pr&oacute;ximo 73%. Amostras brasileiras e estrangeiras de vinho tinto seco e vinho tinto seco branco foram analisadas e a OTA n&atilde;o foi detectada em nenhuma delas. Entretanto, a OTA pode essa estar presente nas amostras analisadas em concentra&ccedil;&otilde;es mais baixas que as determinadas pelos limites de detec&ccedil;&atilde;o e quantifica&ccedil;&atilde;o, n&atilde;o sendo potencialmente perigosos &agrave; sa&uacute;de. / Mycotoxins are compounds produced by fungus, being a potential danger to human and animal health. Ochratoxin A (OTA) is a mycotoxin widely studied, being found in a varied of food matrices. OTA concentration in food is generally low (the order of ng g-1), being, therefore, necessary the use of sample preparation techniques to perform the analyte purification and preconcentration in the sample. The separation and detection methods used in OTA analysis also should offer proper sensibility in order to quantify the analyte adequately. In short, the goals of the present work were to develop a methodology using in-tube SPME in flow through extraction mode with C18 particles as extraction phase and separation and detection by HPLC-MS/MS. Further, optimize and validate the proposed methodology and detect and quantify OTA in wine samples. For this, in-tube SPME was configured and a PEEK tube was packed with C18 particles, which was used in the extraction step. The method optimization was achieved using a central composite 22 + 3 central points experimental design, having as factors the percentage of ACN and time during sample loading step. Validation method was done following ANVISA validation guide after which red dry wine and white dry wine samples were analyzed. The proposed method was developed, having its functionalities attested and its analysis conditions enhanced, using 22% of ACN and 6 minutes in sample loading. The method was validated, demonstrating proper sensitivity, with detection and quantification limits equal to 0.02 and 0.05 &micro;g L-1, respectively. Linearity and precision were evaluated, exhibiting correlation coefficient equal to 0,996 e RSD under 6%, respectively. The method was accurate in medium and high concentration levels and maximum recovery was 73% in high concentration level. Brazilian and foreign red dry wine and white dry wine samples were analyzed and OTA was not detected in any of them. However, OTA may be present in samples analyzed in lower concentrations than that ones determined by detection and quantification limits, not being a potential danger to human health.

Würzburg, Universiẗat, Diss., 2007.

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