The use of chemicals, which generates a lot off benefits, is responsible for the contamination of soil, water and foods. The concern about contamination of surface and ground water systems by pesticides has grown in scientific circles, especially by the presence of pesticides and other compounds in water sources. Thus, rapid analytical methods, sensitive and efficient must be developed in order to verify the concentration levels of pesticides and other organic compounds in water samples agree with those established by law to protect human health and the environment media. This study aimed to optimize, validate and implement an extraction method to determine residues of mononitro, ketones, trifluralin, oxirane, lindane, 2,4-D ester, chlorpyrifos, bromoketal, α-endosulfan, β-endosulfan and endosulfan sulfate in samples of tap water, industrial wastewater and groundwater by Dispersive Liquid-Liquid Microextraction (DLLME) modified and Gas Chromatography with Mass Spectrometry Tandem using Triple Quadrupole type (GC-(TQ)MS/MS). The method is based on the rapid injection, with a syringe, of a mixture of an extractor solvent (50 μL of carbon tetrachloride) and an dispersor solvent (2.0 mL acetone) previously cooled (ice batch, 0 - 4 °C) in an aqueous sample (5.0 mL), contained in a 10 mL graduated glass tube with conical bottom, with no pH correction and cooled (ice batch, 0 - 4 °C). It is not necessary to stir the sample in this step. Remove 25 μL of the sedimented phase separated by simple sedimentation for 10 min. from the bottom of conical tube with a microsyringe and transferred to a vial of 200 μL for GC- (TQ)MS/MS determination. The preconcentration factor obtained was 100 times. The method was validated by determining the limits of detection and quantification limits (LOD and LOQ), linearity, precision and accuracy. The calibration curves showed adequate linearity between 0.05 e 10 μg L-1 with coefficients of determination higher than 0.993. The method showed good recovery values between 70 and 115%, RSD(%) ranged from 10.9 to 17.2%, LOQs between 0.02 and 0.09 μg L-1, and was considered adequate for the analysis of pesticide residues and other organic compounds evaluated in water samples. After validation, the method was applied to samples of drinking water, industrial wastewater and groundwater. No pesticide residues and other organic compounds evaluated were found in the water samples tested. / A utilização de substâncias químicas, que por um lado gera benefícios, por outro é responsável pela contaminação do solo, água e alimentos.A preocupação com a contaminação de sistemas aquáticos superficiais e subterrâneos por pesticidas tem crescido no meio científico, em especial pela presença de pesticidas e outras susbtâncias químicas em mananciais de água. Dessa forma, métodos analíticos rápidos, sensíveis e eficientes têm de ser desenvolvidos para verificar se os níveis de concentração dos pesticidas e outras substâncias químicas em amostras de água estão de acordo com os estabelecidos pela legislação, a fim de resguardar a saúde humana e o meio ambiente. Este estudo teve como finalidade otimizar, validar e aplicar um método de extração e determinação de resíduos de mononitro, ketona, trifluralina, oxirano, lindano, 2,4-D-éster, clorpirifós, bromoketal, α-endosulfan, β-endosulfan e endosulfan sulfato em amostras água de torneira, efluente industrial e água subterrânea, aplicando a técnica DLLME modificada e quantificação por Cromatografia Gasosa acoplada à Espectrometria de Massas Sequencial tipo Triplo Quadrupolo (GC-(TQ)MS/MS). A técnica de extração DLLME está baseada na injeção rápida, com auxílio de uma seringa, de uma mistura dos solventes extrator (50 μL de tetracloreto de carbono) e dispersor (2,0 mL de acetona) previamente resfriada em banho de gelo a uma temperatura entre 0 e 4 °C, em uma amostra aquosa (5,0 mL) contida em um tubo de vidro, com fundo cônico, graduados com volume de 10 mL, sem correção de pH e mantida resfriada em banho de gelo entre 0 e 4 °C. O procedimento não requer agitação da amostra. Retira-se a fase sedimentada separada por sedimentação simples após 10 min., do fundo do tubo cônico com auxílio de uma microseringa de 25 μL e transfere-se para um frasco de 200 μL, para determinação no sistema GC-(TQ)MS/MS. O fator de pré-concentração obtido é de 100 vezes. Na validação do método determinaram-se os limites de detecção e limites de quantificação (LOD e LOQ), linearidade, precisão e exatidão. As curvas analíticas apresentaram linearidade adequada entre 0,05 e 10 μg L-1 com valores de coeficiente de determinação superior a 0,993. O método apresentou bons valores de recuperação, entre 70 e 115% e RSD(%) entre 10,9 e 17,2%, com LOQs entre 0,02 e 0,09 μg L-1. O método foi considerado adequado para a análise de resíduos dos pesticidas em estudo em amostras de água. Após a validação, o método foi aplicado em amostras de água de potável, efluente industrial e água subterrânea. Não foram encontrados resíduos dos pesticidas e outras substâncias químicas estudados nas amostras analisadas.

Conselho Nacional de Desenvolvimento Científico e Tecnológico / The use of pesticides always has been associated with the effective control of pests or invasive weeds to ensure an increase in the food production. However, the indiscriminate use of these substances has caused the degradation of water resources. In Brazil, the Ministry of Health through Ordinance 2914 defines several parameters of potability, among them, the maximum limits allowed for same pesticides. In this work it was developed and validated a method for the determination of residues of 70 pesticides in drinking water using (SPE) for sample preparation and determination by Gas and Liquid Chromatography coupled to tandem mass spectrometry, triple quadrupole analyzer (GC-(TQ)MS/MS and LC- (TQ)MS/MS). It was evaluated different sample volume, sorbents and solvent of elution. The best results were obtained using 100 mL sample acidified at pH 2.5, Oasis® SPE cartridge HLB 60 mg/3 mL and dichloromethane/methanol as eluent. Analytical curves were linear between 10 and 250 Sg L-1, with r2 values greater than 0.99 for all compounds. The values of method LOQ were 0.02 Sg L-1 for aldrin, dieldrin and chlordane and 0.5 Sg L-1 for the other compounds. To evaluate accuracy the blank samples ware fortified at 0.5, 1.5 and 4.0 Sg L- 1 and an extra level at 0.02 Sg L-1 for aldrin, dieldrin and chlordane. The method showed good precision, with RSD values below to 20% and good accuracy, with recoveries between 70 and 120%. Only the compounds methamidophos, aldicarb, benfuracarb, terbufos, benomyl and thiophanate methyl were not recovered adequately. The matrix effect was evaluated, showing upper 10% for the most compounds. In order to compensate this effect, analytical curves were obtained with standarts prepared in blank extracts of the matrix. The validated method was applied to 12 samples of drinking water of different characteristics (river, shed, well and treated water), and just one of the river samples presented residues of lambda-cyhalothrin. The results indicate that the proposed method is suitable for analysis of pesticides residues in drinking water, since all the validation parameters met the suggested limits and parameters for validation of chromatographic methods. / O uso de agrotóxicos sempre esteve associado à efetividade no controle de pragas ou plantas invasoras para aumentar a produção de alimentos. No entanto, o uso indiscriminado dessas substâncias vem causando a degradação dos recursos hídricos. No Brasil, o Ministério da Saúde através da Portaria 2914 define diversos parâmetros de potabilidade, entre eles, os limites máximos permitidos para alguns agrotóxicos. No presente trabalho foi desenvolvido e validado um método para a determinação de resíduos de 70 agrotóxicos em água potável, utilizando Extração em Fase Sólida (SPE) para o preparo de amostra e determinação por Cromatografia Gasosa e Líquida, acopladas à Espectrometria de Massas sequencial, com analisador triplo quadrupolo (GC-(TQ)MS/MS e LC- (TQ)MS/MS). Foram avaliados diferentes volumes de amostra, sorventes e solventes de eluição. Os melhores resultados foram obtidos com 100 mL de amostra acidificada em pH 2,5, cartucho SPE Oasis® HLB 60 mg/3 mL e diclorometano/metanol como eluente. As curvas analíticas apresentaram linearidade entre 10 e 250 Sg L-1, com valores de r2 maiores que 0,99 para todos os compostos. Os valores de LOQ do método foram 0,02 Sg L-1 para aldrin, dieldrin e clordano e de 0,5 Sg L-1 para os demais compostos. Para avaliação da exatidão as amostras branco foram fortificadas em 0,5, 1,5 e 4,0 Sg L-1 e um nível extra em 0,02 Sg L-1 para os compostos aldrin, dieldrin e clordano. O método apresentou boa precisão, com valores de RSD inferiores a 20% e boa exatidão, com recuperações entre 70 e 120%. Apenas os compostos metamidofós, aldicarbe, benfuracarbe, terbufós, benomil e tiofanato metílico não foram recuperados de forma adequada. O efeito matriz foi avaliado, mostrando-se superior a 10% para a maioria dos compostos. A fim de compensar este efeito, utilizou-se curvas analíticas preparadas no extrato branco da matriz. O método validado foi aplicado em 12 amostras de água potável de diferentes características (rio, vertente, poço e água tratada), sendo que apenas uma das amostras de água de rio apresentou resíduos de lambda-cialotrina. Os resultados indicam que o método proposto é adequado à análise de resíduos de agrotóxicos em água potável, visto que todos os parâmetros de validação atenderam os limites e parâmetros sugeridos para validação de métodos cromatográficos.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Due to the complexity of its matrix, more efficient methods have been studied for the determination of pesticide residues in maize. The use of QuEChERS method coupled to low temperature clean up technique has showed advantages to reduce lipids during sample preparation in matrix with high fat content. It was optimized and validated in this study a multiresidue method for the determination of pesticides in maize employing QuEChERS and GC-MS/MS. The extraction procedure was optimized though the factorial design, and consisted in weighing 2.5 g of sample and adding 10 mL of ultrapurified water followed by vortex shaking for 1 min. After this, It was added 10 mL of acetonitrile and the tube was stirred for 1 h in a platform shaker at 200 rpm. Partition step was performed using 1 g of NaCl and 6 g of MgSO4, followed by manual shaking for 1 min and centrifugation at 3400 rpm for 8 min, at 10 ºC. An aliquot of 4 mL of the extract was transferred to a 15 mL tube for clean up step at low temperature for 12 h around -20 ºC. Finally, another clean up stage was realized applying dispersive SPE, 1 mL of the extract was transferred to a tube containing 150 mg of MgSO4, 50 mg of C18 and 50 mg of PSA followed by shaking and centrifugation. The final extract was filtered and analyzed by GC-MS/MS. Analytical parameters evaluated in this work were linearity of the analytical curve, limit of detection (LOD), limit of quantification (LOQ), precision, accuracy and matrix effect for the 12 compounds studied. The method showed efficient, with recovery results in a range of 55.3 to 110.1% in the spiked levels of concentration of 40, 100 and 200 μg kg-1 and RSD ≤ 20%. Values of instrument LOD and LOQ were 1.5 and 5.0 μg L-1, respectively. Method LOQ was 100 μg kg-1 for most compounds. Thereby, It was concluded that the proposed method demonstrated to be adequate for the analysis of most pesticide residues studied in maize, since the parameters evaluated were in according to the limits required for chromatographic methods validation applied in residues analysis. / Devido à complexidade da sua matriz, métodos mais eficientes para a determinação de resíduos de agrotóxicos em milho têm sido estudados. O uso do método QuEChERS, juntamente com a técnica de precipitação a baixa temperatura tem apresentado vantagens para a redução de lipídios durante o preparo de amostra de matrizes com alto teor de gordura. Neste estudo, otimizou-se e validou-se um método multirresíduo para determinação de agrotóxicos em milho empregando QuEChERS e GC-MS/MS. O procedimento de extração, otimizado através de planejamento fatorial, constituiu na pesagem de 2,5 g da amostra, adição de 10 mL de água purificada, seguida de agitação em vórtex por 1 min e posterior adição de 10 mL de acetonitrila seguindo de agitação em agitador pendular por 1 h a 200 rpm. A partição utilizando sais foi realizada com 1 g de NaCl e 6 g de MgSO4 e agitação manual por 1 min, seguida de centrifugação a 3400 rpm por 8 min a 10 ⁰C. Uma alíquota de 4 mL do extrato foi retirada e transferida para um tubo de 15 mL para a etapa de limpeza do extrato a baixa temperatura por 12 h a aproximadamente -20 ⁰C. Após, efetuou-se a etapa final de limpeza do extrato com SPE dispersiva onde retirou-se 1 mL deste extrato e transferiu-se para um tubo contendo 150 mg de MgSO4, 50 mg de C18 e 50 mg de PSA, repetindo a agitação por 1 min e centrifugação a 3400 rpm por 8 min. O extrato final foi filtrado e analisado por GC-MS/MS. Os parâmetros analíticos avaliados neste trabalho foram a linearidade da curva analítica, limite de detecção (LOD), limite de quantificação (LOQ), precisão, exatidão e efeito matriz para os 12 compostos estudados. O método demonstrou-se eficiente, com resultados de recuperação na faixa de 55,3 a 110,1% nos níveis de fortificação 40, 100 e 200 μg kg-1 e com RSD ≤ 20%. Os valores de LOD e LOQ do instrumento foram de 1,5 e 5,0 μg L-1, respectivamente, e o valor de LOQ do método foi de 100 μg kg-1para a maioria dos compostos. Desta forma, conclui-se que o método otimizado mostrou-se adequado para análise da maioria dos resíduos de agrotóxicos estudados em milho, pois os parâmetros avaliados encontram-se dentro dos limites exigidos para validação de métodos cromatográficos aplicados a análise de resíduos.

Les extraits naturels tels que les huiles essentielles, les absolues ou les résinoïdes utilisés en parfumerie, arômes et cosmétique sont produits à partir de matières premières naturelles. La culture de ces matières premières est souvent basée sur la monoculture et implique l'utilisation de pesticides. La réglementation pour les matières premières destinées à ces industries devient de plus en plus stricte en termes de niveaux résiduels autorisés de pesticides, en raison de leur impact sur la santé publique et sur l'environnement. Dans ce manuscrit, un état de l’art montrant la problématique pour l’analyse des pesticides dans les extraits naturels et les travaux dédiés à cette étude, est présenté. Une méthode d’analyse multi-résidus de pesticides a ensuite été mise au point dans les roses et les extraits (concrètes et absolues) issus de la rose. La préparation d’échantillon s’appuie sur l’approche QuEChERS utilisant l’extraction et la purification par SPE-dispersive. Les échantillons ont ensuite été analysés par GC-MS/MS. Les méthodes ainsi développées ont permis d’étudier les devenir des pesticides au cours du procédé d’extraction des roses. Le travail a été fait dans un premier temps sur des roses issues d’un champ bio puis dopées avec 57 pesticides. Chaque intermédiaire de transformation, excepté les cires, a été analysé mettant en évidence la répartition des pesticides. Il y est montré que les pesticides présents dans les roses sont co-extraits durant le procédé d’extraction et sont concentrés entre 100 et 300 fois dans l’absolue. Cette étude a ensuite été réalisée sur des roses et leurs extraits issus de champs avec des traitements phytosanitaires « classiques ». / Natural extracts in form of essential oils, concretes, resinoids, as well as absolutes used in perfume, aroma, and cosmetic industries are produced from natural raw materials. The cultivation of natural raw materials is often based on monoculture and involves the use of pesticides. Legal regulations for raw materials intended for these industries become increasingly stricter in terms of the allowed residual levels of chemicals used for treatment because of their impact on public health and on the environment. In this manuscript, a review showing pesticide analysis problems in natural extracts and works dedicated to this study, is presented. Next, a method for multi-residue pesticides analysis was developed in roses and extracts (concrete and absolute) from roses. The sample preparation is based on the QuEChERS approach using extraction and purification by SPE-dispersive. The samples were then analyzed by GC-MS / MS. Thus the developed methods were used to study pesticides behavior during the extraction process of the roses. In a first time, the work was done on roses from organic field and spiked with 57 pesticides. Each transformation intermediate, except waxes, was analyzed showing the distribution of pesticides. It showed that pesticides which were present in rose flowers were coextracted during extraction process and concentrated by factor between 100 and 300 in absolute. This study was then performed on roses and extracts from fields with pesticide "classic" phytosanitary treatments. The trend observed for these samples was the same as for the model: pesticides were concentrated in the absolute reaching concentrations of several milligrams per kilogram.

O tramadol é um analgésico de ação central amplamente utilizado em répteis. Seu efeito ocorre por meio da ativação de receptores opioides e do bloqueio da recaptação de serotonina e norepinefrina. Este estudo objetivou avaliar a atividade antinociceptiva e a farmacocinética do tramadol em jabutis-piranga. Oito jabutis receberam quatro tratamentos, com intervalo mínimo de 21 dias: solução salina (grupo controle), tramadol nas doses de 5 mg/kg (TIM5) e 10 mg/kg (TIM10) por via intramuscular e 5 mg/kg (TIV5) por via intravenosa. Um estímulo nociceptivo térmico foi aplicado na superfície plantar dos animais e a avaliação da latência do reflexo de retirada do membro foi realizada nos tempos 0, 30 minutos, 1, 2, 4, 6, 8, 10, 24, 48, 72 e 96 horas. Foi realizada também colheita de sangue e análise do plasma por meio de cromatografia gasosa acoplada à espectrometria de massas triplo quadrupolo. A farmacocinética do tramadol foi calculada por abordagem não-compartimental. A concentração máxima para TIM5 e TIM 10 foi de 128,98 ± 59,46 ng/mL e 613,87 ± 524,26 ng/mL, respectivamente. A meia-vida de eliminação foi de 21,22 ± 8,74 horas para TIV5, 32,84 ± 6,41 horas para TIM5 e 22,5 ± 8,02 horas para TIM10. As doses de tramadol 10 mg/kg por via intramuscular e 5 mg/kg por via intravenosa não apresentaram alteração do tempo de latência para o reflexo de retirada do membro. O grupo que recebeu 5 mg/kg por via intramuscular apresentou diferença nos tempos 1 hora e 24 horas após a administração. O tramadol não apresentou efeito antinociceptivo consistente por via intramuscular nas doses de 5 e 10 mg/kg e por via intravenosa na dose de 5 mg/kg em jabutis-piranga. Independente da via de administração, o tramadol apresentou meia-vida de eliminação longa. / Tramadol is a centrally acting analgesic widely used in reptiles. It promotes activation of opioid receptors and inhibits serotonin and norepinephrine reuptake. This study aimed to evaluate the antinociceptive effects and pharmacokinetics of tramadol in red-footed tortoises. Eight animals were treated with four different protocols, with a 3-week washout period: saline IM (control group), tramadol 5 mg/kg IM (TIM5) and 10 mg/ kg IM (TIM10) and 5 mg/kg IV (TIV5). Infrared heat stimuli was applied to the plantar surface of the animals and thermal withdrawal latencies were measured at 0 and 30 minutes, 1, 2, 4, 6, 8, 10, 24, 48, 72 and 96 hours after drug treatment. Blood samples were obtained and plasma analysis performed by gas chromatography - triple quadrupole tandem mass spectrometry. Pharmacokinetics parameters were determined using non-compartimental equations. The average peak drug concentration was 128.98 ± 59.46 ng/mL (TIM5) and 613.87 ± 524.26 ng/mL (TIM10). The elimination half-life was 21.22 ± 8.74 hours (TIV5), 32.84 ± 6.41 hours (TIM5) and 22.5 ± 8.02 hours (TIM10). TIM10 and TIV5 showed no changes in thermal withdrawal latencies. TIM5 showed a significant increase in thermal withdrawal latencies at 1 hour and 24 hours after administration. Tramadol did not induce a consistent antinociceptive effect at 5 mg/kg and 10 mg/kg IM and ar 5 mg/kg IV in red-footed tortoises. Regardless of the route of administration, tramadol had a long half-life and plasma concentrations close to those reported in other species.

Die Rückstandsanalytik tierischer Lebensmittel ist eine anspruchsvolle Aufgabe aufgrund des hohen Lipidanteils der Proben sowie des sich stetig vergrößernden Wirkstoffspektrums. Heutzutage werden für die Probenaufarbeitung die DFG S 19 Methode, mit der vorrangig unpolare Analyten nachgewiesen werden und zunehmend die QuEChERS Methode eingesetzt, die insbesondere auf die Erfassung polarer Pestizide abzielt. In dieser Arbeit wurde eine moderne Multirückstandsmethode für Hühnereier entwickelt, um sowohl gas- als auch flüssigkeitschromatographisch (GC, LC) erfassbare Wirkstoffe zu analysieren. Dazu gehören unpolare PCBs, Pyrethroide und Organochlorpestizide, aber auch polarere Organophosphate, Triazole und Carbamate. Das Verfahren basiert auf der Extraktion mittels Matrix Solid Phase Dispersion, der Reinigung auf Grundlage einer modifizierten Gelpermeationschromatographie (GPC) und zwei verschiedenen Festphasenextraktionen (SPEs) für GC- und LC-erfassbare Pestizide sowie der Quantifizierung mittels GC- und LC-MS/MS. Dünnschichtchromatographisch wurde die effektive Entfernung hochmolekularer Lipide durch die modifizierte GPC und niedrigmolekularer Fette durch die SPEs belegt. Laut der für Ei durchgeführten Validierung erfüllten 164 der 172 untersuchten Pestizide und alle sechs PCBs die Leistungskriterien für die amtliche Rückstandskontrolle - zumeist am niedrigsten validierten Level (5 µg/kg bzw. 0,5 µg/kg). Ausnahmen bildeten sehr polare LC-Pestizide (z.B. Aminopyralid, Clopyralid, MCPA, Quinmerac) und pH-Wert-abhängige GC-Analyten (Nicotin, Tolylfluanid, Dichlofluanid), die auch mit den etablierten Verfahren schwierig zu analysieren sind. Weiterhin verdeutlichte die erfolgreiche Untersuchung von verschiedenen Ringversuchsmaterialien, dass die ursprünglich für Eier entwickelte Methode auch für mageres Geflügelfleisch und Sahne genutzt werden kann. Gegenüber den etablierten Verfahren wies die neue Methode deutliche Vorzüge auf. So belegte die Dünnschichtchromatographie, dass mit der neuen Methode Cholesterin, aber auch freie Fettsäuren besser abgetrennt werden als mit den etablierten Verfahren. Die neue Methode verbrauchte im Vergleich zur DFG S 19 Methode 46 % weniger Lösungsmittel und ermöglichte eine Verdopplung des Probendurchsatzes innerhalb von 8 h. Zudem eignete sich das entwickelte Verfahren laut den Validierungsdaten für GC-Analyten deutlich besser als die QuEChERS Methode und etwas besser als die DFG S 19 Methode (v.a. für Pyrethroide). Hinsichtlich der LC-Analyten unterschieden sich die neue und die QuEChERS Methode nur bei wenigen Analyten. Mit dem neuen Verfahren konnten folglich im Gegensatz zu den etablierten Methoden sowohl unpolare GC- als auch polare LC-Analyten sicher erfasst werden.

Malgré une sensible baisse des ventes depuis quelques années, la France reste le premier utilisateur européen de phytosanitaires. Ceci n’est pas sans conséquences sur l’environnement et les organismes non visés par ces produits. De nombreuses études ont permis d’une part de mieux comprendre les phénomènes de transfert des pesticides dans les compartiments eau-air sols et d’autre part de mettre en évidence leur contamination. Les données concernant l’eau sont plus nombreuses du fait de son statut de réceptacle ultime des pollutions.et des normes établies par les textes réglementaires. La Directive Cadre européenne sur l’Eau (DCE) adoptée en 2000 vise à atteindre le bon état chimique et écologique des eaux d’ici 2015 avec une obligation de résultats. Le bon état chimique dépend du respect des normes de qualité environnementales (NQE) établies notamment pour 24 pesticides. En 2012, 5% des points de mesure en cours d’eau et 19% des points de mesure en eaux souterraines ne respectent pas ces NQE.Le Languedoc Roussillon est depuis de nombreuses années la première région viticole de France et souffre d’une pression phytosanitaire importante. En 2014, un rapport de l’Agence Régionale de Santé (ARS) fait même état de l’utilisation encore récente de pesticides interdits (simazine, terbuthylazine…) depuis le début des années 2000. L’ARS a de plus affirmé que la plupart des cas de non-conformité avérés pouvaient être rattachés à la présence d’activité agricole dans le bassin versant du captage.Remédier à ces contaminations passe par le suivi de la qualité des eaux et la caractérisation de la pollution. Les outils actuels s’avèrent ne pas être toujours représentatifs de cette pollution ;de nouveaux outils sont donc mis au point. L’étude présentée ici porte sur l’utilisation de l’échantillonnage passif de type POCIS (Polar Organic Chemical Integrative Sampler) pour le suivi de la contamination en pesticides de trois bassins versants languedociens : Paulhan,Puissalicon, Roujan.Dans un premier temps, l’étude des limites possibles du dispositif POCIS a été faite pour évaluer son applicabilité dans les conditions d’exposition imposées par le terrain. Le comportement des molécules au sein de l’outil a été étudié pour une meilleure compréhension des phénomènes de diffusion qui s’y produisent. Ces études ont mené à la mise en place de calibrations en laboratoire en conditions contrôlées. [...] La mise en place des dispositifs POCIS sur les trois sites d’études a mis en évidence la prédominance de pesticides utilisés en viticulture et a montré tout l’intérêt de l’outil d’un point de vue qualitatif. La comparaison de l’outil avec l’échantillonnage automatisé montre des disparités entre concentration réelle et concentration estimée via le POCIS. Ce dernier reste à améliorer d’un point de vue quantitatif pour obtenir un échantillonneur fiable et robuste pour ce type de milieu d’exposition. / Despite a noticeable sales decrease since few years, France remains the first european user of phytosanitary products. This is not without consequences on the environment and the untargeted organisms. Several studies allowed on one hand to better understand phenomena of pesticides transfer in water-air and soils compartments and on the other hand to highlight their contamination. Data concerning the water are more numerous cause of its status of ultimate receptacle of pollution and standards established by regulatory texts. The european Water Framework Directive (WFD) adopted in 2000 aims to achieve good chemical and ecological status of water bodies by 2015 with an obligation of results. The good chemical status depends on respect for the Environmental Quality Standards (EQS) established, for instance, for 24 pesticides. In 2012, 5% of river measurement points and 19%of groundwater measurement points do not respect these EQS. For many years, Languedoc Roussillon is the first wine-producing region of France and suffers from a significant pesticide pressure. In 2014, a report from the Regional Health Agency (RHA) has mentioned a relatively recent use of banned pesticides since the early 2000’s like simazin and terbutylazin. Moreover, RHA stated that most of recognized cases of non-compliance could be linked to the presence of agricultural activity in the catchment basin.[...] The establishment of POCIS devices on the three study sites showed the predominance ofpesticides used in viticulture and showed all the interest of the tool on a qualitative point ofview. The comparison of the tool with the automated sampling shows disparities between real concentration and concentration estimated with the POCIS. However, it remains to improvethe tool on a quantitative way to obtain a reliable and robust sampler for this type of exposure medium.

La consommation excessive d'alcool présente des risques élevés pour l'individu et pour la société ; elle est fréquemment associée à une augmentation du risque d'accidents, d'actes de violence, et peut également conduire à court et/ou à long terme à de graves maladies et à des problèmes sociaux. Dès lors, l'utilisation de marqueurs fiables permettant de détecter une consommation excessive d'alcool, ponctuelle ou chronique, s'avère nécessaire pour prévenir des conséquences néfastes de l'abus d'alcool. L'éthylglucuronide (EtG) est un marqueur d'alcoolisation utilisé en toxicologie clinique (alcoologie) et médicolégale. Par rapport aux marqueurs indirects d'alcoolisation (CDT, γ-GT), ce métabolite mineur de l'éthanol est très spécifique et est quantifiable dans diverses matrices biologiques. La production d'EtG est catalysée par des enzymes de la famille des UDP-glucuronosyl-transférases (UGT). Cependant, les UGT impliquées dans la glucuronoconjugaison de l'éthanol, ainsi que les sources potentielles de variabilité interindividuelle de la production d'EtG, sont encore mal connues. Nos travaux ont ainsi consisté à (1) développer et valider une méthode de dosage de l'EtG dans différentes matrices biologiques par chromatographie en phase gazeuse couplée à la spectrométrie de masse en tandem, (2) identifier les UGT humaines impliquées dans la glucuronoconjugaison de l'éthanol et étudier la contribution relative de chaque isoforme active au niveau hépatique, (3) étudier l'impact de substances fréquemment utilisées par les consommateurs d'alcool sur la production d'EtG in vitro, (4) étudier l'impact de polymorphismes génétiques fonctionnels des UGT sur la production hépatique d'EtG, et enfin (5) évaluer l'impact de la consommation de cannabis et d'autres drogues sur la production d'EtG à l'aide de prélèvements post-mortem. Ces travaux ont notamment permis de montrer que (1) l'éthanol est glucuronoconjugué principalement par le foie, puis dans une moindre mesure par les reins et par l'intestin, (2) les UGT1A9 et 2B7 sont les deux enzymes majoritairement impliquées dans la glucuronoconjugaison de l'éthanol, quel que soit l'organe considéré, (3) la morphine, la codéine, la nicotine et la cotinine n'entraînent aucune modification des taux de production d'EtG in vitro ; le lorazépam et l'oxazépam augmentent légèrement cette production (p = 0,2 et 0,065, respectivement) ; le cannabidiol inhibe la glucuronoconjugaison de l'éthanol par un mécanisme non-compétitif (CI50 = 1,17 mg/L; Ki = 3,1 mg/L), alors que le cannabinol augmente cette glucuroconjugaison de manière concentration-dépendante (p <0,05), (4) les SNP c.-900G>A affectant l'UGT2B7 et IVS1+399T>C affectant l'UGT1A9 augmentent légèrement la production d'EtG in vitro. Enfin (5) le rapport des concentrations sanguines EtG/éthanol apparaît significativement plus élevé chez des co-consommateurs de cannabis et/ou d'autres drogues que chez des consommateurs d'alcool seul. L'ensemble de ces résultats démontre l'existence de plusieurs facteurs pouvant potentiellement influencer la production d'EtG et devraient donc être pris en considération lors de l'interprétation de sa concentration in vivo.

Modern agriculture started using different techniques and materials in order to minimize losses in production and meet the growing demand for food, combining productivity and profitability. However, the indiscriminate use of these different inputs (pesticides and fertilizers) can lead to several environmental damage. Study conducted by the National Agency for Sanitary Vigilance regarding the pesticide market in Brazil reveals that there pace of expansion in consumption, putting the country on alert in conferring the enhancement of environmental contamination arising from the use of pesticides. The present work aims to study multiresidue method for determination of pesticides using solid phase extraction and quantification by gas chromatography coupled to mass spectrometry in tandem GC-(TQ) -MS/MS, as well as evaluating the storage of compounds directly extraction cartridges in different time periods. To optimize the extraction procedure was used factorial planning, assessing three kinds of sorbents, Oasis ® HLB, C18 Strata Strata-X ® and varying proportions of methanol in methylene chloride as well as different pH values. The extraction procedure is optimized for use with the sorbent cartridge SPE Oasis ® HLB polymeric mg/3mL 60, the sample filtered with 0.22 nylon membrane micrometers. Conditioning the cartridge with 3 mL of methanol followed by 3 mL of ultrapure water and 3 ml of ultrapure water at pH (6.2). Percolation 100 mL sample previously acidified to pH 6.2. For elution was used a mixture of MeOH: DCM 45:55 (v, v) with two additions of 1 mL, leaving two minutes of contact with the sorbent before elution. The assessment of figures of merit for the proposed method were satisfactory, with linear calibration curves with r2 greater than 0.99 in the range 5-200 mg L-1, with values of LODm LOQm and 0.03 and 0.10 mg L-1, respectively. When evaluated recoveries found values between 70 and 120% for 39 of the 43 pesticides evaluated with precision values RSD ≤ 20%. The matrix effect was evaluated and proved to be over 10% for most compounds. The application of the method was performed on 10 samples of dams in different cities of Colonial Northwest region of Rio Grande do Sul, found pesticide residues in eight. The method proved to be effective for the determination of pesticide residues by SPE and GC-(TQ) -MS/MS, being able to be used in routine analysis. / A agricultura moderna passou a utilizar diferentes técnicas e insumos a fim de minimizar as perdas na produção e atender a demanda crescente por alimentos, aliando produtividade e rentabilidade. Entretanto, o uso indiscriminado desses diferentes insumos (agrotóxicos e fertilizantes) pode acarretar diversos prejuízos ambientais. Estudo desenvolvido pela Agência Nacional de Vigilância Sanitária referente ao mercado de agrotóxicos revela que no Brasil há ritmo de expansão no consumo, colocando o país em alerta no que confere a potencialização da contaminação ambiental oriunda do uso de agrotóxicos. O presente trabalho visa o estudo de método multirresíduo para determinação de agrotóxicos utilizando extração por fase sólida e quantificação por cromatografia em fase gasosa acoplada a espectrometria de massas em série GC-(TQ)-MS/MS, bem como avaliar a armazenagem dos compostos diretamente nos cartuchos de extração em diferentes períodos de tempo. Para otimização do procedimento de extração, foi utilizado planejamento fatorial em estrela, avaliando três tipos de sorventes, Oasis® HLB, Strata C18 e Strata-X®, variando proporções de metanol em diclorometano , bem como diferentes valores de pH. O procedimento de extração otimizado consiste em utilização de cartucho SPE com o sorvente polimérico Oasis® HLB 60 mg/3mL, filtração da amostra com membrana de nylon de 0,22 μm. Condicionamento do cartucho com 3 mL de metanol, seguido de 3 mL de água ultrapura e 3 mL de água ultrapura em pH (6,2). Percolação de 100 mL de amostra previamente acidificada em pH 6,2. Para eluição foi utilizada uma mistura de MeOH:DCM 45:55 (v,v), com duas adições de 1 mL, deixando dois minutos de contato com o sorvente antes da eluição. A avaliação das figuras de mérito para o método proposto foram satisfatórias, apresentando curvas analíticas lineares com r2 superiores a 0,99 na faixa de 5 a 200 μg L-1, apresentando valores de LODm e LOQm de 0,03 e 0,10 μg L- 1, respectivamente. Quando avaliada as recuperações foi encontrado valores entre 70 e 120% para 39 dos 43 agrotóxicos avaliados, com valores de precisão RSD ≤ 20%. O efeito matriz foi avaliado, mostrando-se superior a 10% para a maioria dos compostos. A aplicação do método foi realizada em 10 amostras de açudes de diferentes municípios da região Noroeste Colonial do Rio grande do Sul, sendo encontrado resíduo de agrotóxicos em oito delas. O método proposto mostrou ser eficaz para a determinação de resíduos de agrotóxicos por SPE e GC-(TQ)-MS/MS, sendo passível de utilização em análises de rotina.

Although Lake Victoria is threatened by pollution there is a lack of knowledge about pharmaceuticals contaminants drained into the lake from large cities bordering the lake. Hence, the purpose of this project was to develop, validate and apply a method for analysis of pharmaceutical compounds accumulating in the Lake Victoria sediments. A simple quantitative method for 11 pharmaceuticals combining accelerated solvent extraction, solid phase extraction, trimethylsilylation derivatization, and gas chromatography mass spectrometry was developed, partly validated, and applied to 18 surface sediments and a sediment core dated using the 210Pb method. The results showed the presence of the pharmaceuticals estriol, gemfibrozil, metoprolol, ketoprofen, naproxen, 17α-ethinylestradiol and estrone concentrated around the regions Napoleon Gulf and Thurston Bay with accumulation rates decreasing towards the top of the sediment core. Nonetheless, a randomness in the distribution of these compounds behooves a systematic assessment investigating not only the provenance of these compounds but also further investigations to errors meaning that this study should be treated as a preliminary investigation.