Este estudo apresenta o desenvolvimento da metodologia de extração em fase sólida e separação em cromatográfica líquida acoplada a espectrometria de massas em sequencia, SPE-LC-MS/MS, para a determinação de 21 (vinte e um) fármacos pertencentes a diferentes classes terapeuticas, 1 (uma) droga de abuso e seu principal metabólito, em amostras de água superficial. A separação cromatográfica foi otimizada estudando o desempenho de fases estacionárias e fases móvies. A quantificação dos compostos selecionados foi realizada com a ionização por eletronebulização (electrospray ionization- ESI) e o espectrômetro de massas operando no modo de Monitoramento de Múltiplas Reações (Multiplas Reaction Monitoring- MRM). A validação da metodologia proposta foi realizada utilizando os parâmetros de seletividade, efeito de matriz, faixa de trabalho, linearidade, limites de detecção (LD) e quantificação (LQ), precisão, exatidão, recuperação e robustez. A validação da metodologia permitiu a sua aplicação na avaliação da distribuição dos 23 compostos selecionados, nas águas da represa Guarapiranga, um dos principais sistemas produtor de água potável da Região Metropolitana de São Paulo (RMSP). A presença desses poluentes nos ambientes aquáticos é proveniente da liberação direta do esgoto urbano das habitações do seu entorno, como consequência do precário sistema de saneamento básico. As águas da represa Guarapiranga foram avaliadas em 14 (quatorze) locais estrategicamente escolhidos e amostradas durante 3 (três) campanhas de coleta de amostra (agosto de 2011, setembro de 2012 e abril de 2013). Nessas amostras foram quantificados acetaminofeno (9,6 - 254 ng L-1), atenolol (8,5 177 ng L-1), benzoilegonina (7,9 139 ng L-1), cafeína (27 27386 ng L-1), carbamazepina (12 358 ng L-1), clortalidona (9,4 35 ng L-1), cocaína (12,8 2650 ng L-1), diclofenaco (8 35 ng L-1), enalapril (20 ng L-1), losartana (6,7 114 ng L-1) e valsartana (9,7 - 47 ng L-1). O ponto de coleta denominado de GU103-12 (23°4188.5S 46°4467.3W) foi a região que apresentou os valores mais elevados quanto ao nível de concentração dos compostos avaliados e ao índice de risco integrado de poluição química aquática (Integrated Risk Index of Chemical Aquatic Pollution IRICAP). O estudo também foi realizado em amostras de água de reservatórios das Unidades de Gerenciamento de Recursos Hídricos (UGRHI) 5 e 6 do Estado de São Paulo. Os resultados demonstraram que o uso e a ocupação do solo influenciam diretamente na qualidade da água dos reservatórios, evidenciando a necessidade de implementar melhorias no sistema de coleta de esgoto e de ocupação irregular para evitar a contaminação e o descarte inadequado em ambientes aquáticos. / This study presents the development of the methodology of solid phase extraction and liquid chromatography - tandem mass spectrometry, SPE-LC-MS/MS, for the determination of 21 (twenty one) pharmaceuticals belonging to different therapeutic groups, 1 (one) illicit drug and its major metabolite, in surface water samples. The chromatographic separation was optimized by studying the performance of different stationary and mobile phases. Quantitation of selected compounds was performed by electrospray ionization (ESI) and the mass spectrometer operating in a multiple reaction monitoring (MRM) mode. The validation of the proposed methodology was performed using the parameters of selectivity, matrix effect, dynamic range, linearity, limit of detection (LOD), limit of quantification (LOQ), precision, accuracy, recovery and robustness. The validation of methodology allowed to apply the methodology in the evaluation of the distribution of the 23 (twenty one) selected compounds, in Guarapiranga Dam waters, an of the major producer system of drinking water of the Metropolitan Region of São Paulo (MRSP). The presence of these pollutants in aquatic environments is from the direct release of urban sewage from the homes of your surroundings, as a result of poor sanitation system. The waters of Guarapiranga dam were evaluated in 14 (fourteen) locations strategically chosen and sampled in 3 (three) campaigns of sample collection (August 2011, September 2012 and April 2013). In these samples were quantified acetaminophen (9.6 - 254 ng L-1), atenolol (8.5 - 177 ng L-1), benzoylegonine (7.9 - 139 ng L-1), caffeine (27 - 27386 ng L-1) carbamazepine (12 - 358 ng L-1), chlorthalidone (9.4 - 35 ng L-1), cocaine (12.8 - 2560 ng L-1), diclofenac (8 - 36 ng L-1), enalapril (20 ng L-1), losartan (6.7 - 114 ng L-1) and valsartan (9.7 - 47 ng L-1). The sample siting GU103-12 (23°4188.5S 46°4467.3W) was the region with the highest values in the level of concentration of the target compounds and the integrated risk index of chemical aquatic pollution (IRICAP). The study was also conducted on water samples from reservoirs of the UGRHI (Unit of Water Resources Management) 5 and 6, State o São Paulo. The results showed that the use and occupation of land directly influence the reservoir water quality highlighting the need to implement improvements in sewage collection system and illegal occupation to prevent contamination and the improper disposal in aquatic environments.

Made available in DSpace on 2016-08-19T12:56:32Z (GMT). No. of bitstreams: 1 VERONICA ALVES GONCALVES DA SILVA.pdf: 6289705 bytes, checksum: 1c73081d0d3faa6d1cdc03eef3a8bc0c (MD5) Previous issue date: 2008-06-16 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Honey is a natural substance produced by bees and by your nature it is considered a healthy product. Presence of additives and/or preserve agents is not permitted. On apiculture, sulfonamydes are largely used on prevention and control of beehive disease, such as American foulbrood caused by Paenibacillus larvae. A consequence of this practice is the presence of these residues in honey, that should cause allergy reactions on consumers. In the last years, some publicated works is discussing the presence of antibiotic and sulfonamides residues in honey. In Brazil, the presence of antibiotics residues in food is controled by the Agência Nacional de Vigilância Sanitária (ANVISA) an by the Ministério da Agricultura, Pecuária e do Abastecimento (MAPA). The realization of monitoring contaminants residues in food programs, as well as the Plano Nacional de Controle de Resíduos (PNCR) from MAPA are essentials to the sanitary vigilance actions development, preventing and controlling possible hazard to the consumer health. This work has the objective the development and validation of multiresidue analytical method to the sulfadiazine, sulfathiazole, sulfadimedine and sulfadimethoxine determination in bee honey using high performance liquid chromatography (HPLC) with diode array detector (DAD). The established methodology consisted on such conditions: solid phase extraction with Nexus sorbent, elution with acetonitrile 5 mL, and reconstitution with sodium acetate buffer (0,05 mol L-1) pH 4,5, with 1% methanol addition, separation on analytical column C18, acetonitrile and water mobile phase, on gradient elution mode and detection on 278 nm. The sample preparation consisted on the acid hydrolysis with hydrochloric acid 2 mol L-1 of a honey rate and adjust to pH 4,5 with sodium acetate buffer solution 0,05 mol L-1 pH 4,5. The method was validated and the results demonstrated accuracy and precision between the limits established by the current legislation. Good linearity range was encountered, obtaining recovery values varying from 41 to 89% and relative standard deviation estimative varying between 2,36 and 14% to the four studied compounds. The four sulfonamides detection limits in honey sample varying from 20 to 30 ^g kg-1 and the quantification limits varied from 60 to 90 ^g kg-1. Considering 100 ^g kg-1 the residue maximum limit established by the Brazilian legislation to the SDZ, STZ, SMZ and SDM total concentration, the described method permit suitable determination of the four sulfonamides in bee honey. / O mel é uma substância natural, produzida pelas abelhas e por sua natureza é considerado um produto saudável, não sendo permitido a presença de aditivos e/ou agentes conservadores. Na apicultura as sulfonamidas são muito utilizadas na prevenção e controle de doenças em colméias, como a cria pútrida americana causada pelo Paenibacillus larvae. Uma conseqüência desta prática é a presença destes resíduos no mel, o que pode causar reações alérgicas nos consumidores. Nos últimos anos, alguns trabalhos na literatura vêm discutindo a presença de resíduos de antibióticos e sulfonamidas no mel. No Brasil, a presença de resíduos de antibióticos nos alimentos é controlada pela Agência Nacional de Vigilância Sanitária (ANVISA) e pelo Ministério da Agricultura, Pecuária e do Abastecimento (MAPA). A realização de programas de monitoramentos de resíduos de contaminantes em alimentos, assim como o Plano Nacional de Controle de Resíduos (PNCR) do MAPA são essenciais para que ações de vigilância sanitária possam ser desenvolvidas, prevenindo e controlando possíveis danos à saúde do consumidor. Este trabalho tem por objetivo desenvolver e validar um método analítico multiresidual para a determinação de sulfadiazina, sulfatiazol, sulfadimidina e sulfadimetoxina em mel de abelhas usando cromatografia líquida de alta eficiência (CLAE) com detector de arranjo de fotodiodos (DAD). A metodologia estabelecida consistiu nas seguintes condições: extração em fase sólida com sorvente Nexus, eluição com 5 mL de acetonitrila e reconstituição com solução-tampão acetato de sódio (0,05 mol L-1) pH 4,5, com adição de 1% de metanol, separação em coluna analítica C18, fase móvel acetonitrila e água, no modo de eluição em gradiente e detecção a 278 nm. O preparo de amostra consistiu na hidrólise ácida com ácido clorídrico 2 mol L-1 de uma alíquota de mel e ajuste para pH 4,5 com solução tampão acetato de sódio 0,05 mol L-1 pH 4,5. O método foi validado e os resultados demonstraram exatidão e precisão dentro dos limites estabelecidos pela legislação vigente. Foi encontrada boa faixa de linearidade, obtendo-se valores de recuperação que variaram de 41 a 89% e estimativa do desvio padrão relativo variando entre 2,36 e 14% para os quatro compostos estudados. Os limites de detecção das quatro sulfas em amostra de mel variaram de 20 a 30 ^g kg-1 e os limites de quantificação variaram de 60 a 90 ^g kg-1. Considerando o limite máximo de resíduo estabelecido pela legislação brasileira de 100 ^g kg-1 para as concentrações totais de SDZ, STZ, SMZ e SDM, o método descrito permite adequada determinação das quatro sulfas em amostras de mel de abelhas.

Made available in DSpace on 2016-08-19T12:56:39Z (GMT). No. of bitstreams: 1 Dissertacao Marconiel.pdf: 1507450 bytes, checksum: 7ab99051017cb605155d971d0a8bc70b (MD5) Previous issue date: 2009-01-30 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Honey is a product of proven therapeutic and nutritional properties, and which may act as an excellent bioindicator, given the way the bees gather and process the raw material used in its preparation. The monitoring of pesticide residues in honey is important both to control product quality in domestic and international markets, and in assessing the potential for contamination in areas near agricultural crops. Considering these facts, we developed a method for analyzing carbamate insecticides (carbaryl and carbofuran), organochlorine (aldrin and dieldrin), organophosphates (dimethoate and parathion methyl) and pyrethroid (permethrin), and the group of anilides herbicide (propanil) and fenoxiácidos (2,4-D) in honey. Hence, in this work the analytical conditions were established for the proper determination of residues of pesticides cited using the technique of solid phase extraction and liquid chromatography with UV detection with a diode array. The study aimed to compare different analytical conditions for the determination of pesticides, looking for better separation, sensitivity, precision and accuracy in the determination of pesticide residues in samples of honey bees. Standard solutions at concentrations ranging from 1.0 to 20.0 μ g mL-1 were evaluated in different mobile phases, using a C18 column (250 x 4.6 mm id, 5 μ m). The mobile phase composition more suitable was acetonitrile: water at pH 3.0 (acidified with acetic acid) in gradient mode, for monitoring the compounds at a wavelength of 220 nm. In the established conditions, except for aldrin and dieldrin and dimeotate, it was observed recoveries between 75.26% and 113.58%, with variation Coefficients Between 2.16% and 11.31% for concentration levels raging from 0:10 to 1.00 μ gg-1 (correlation coefficients between 0.9978 and 0.9998). Recoveries lower than 50% to the cited insecticides were observed when using ethyl acetate solvent extractor and the solvent elution by ethyl acetate, in such a way new studies can be done to obtain better recoveries for these compounds. / O mel é um produto de reconhecidas propriedades nutritivas e terapêuticas e, o qual pode atuar como um excelente bioindicador, dada à forma como as abelhas recolhem e processam a matéria-prima utilizada na sua elaboração. O monitoramento de resíduos de pesticidas no mel é importante, tanto para o controle de qualidade do produto nos mercados nacional e internacional, quanto na avaliação do potencial de contaminação em áreas próximas a cultivos agrícolas. Considerando tais fatos, foi desenvolvido um método para análise dos inseticidas carbamatos (carbaril e carbofurano), organoclorados (aldrin e dieldrin), organofosforados (dimetoato e parationa metílica) e piretróide (permetrina), além do herbicida do grupo das anilidas (propanil) e fenoxiácidos (2,4-D) em mel. Para tanto, neste trabalho foram estabelecidas condições analíticas para a adequada determinação dos resíduos dos citados pesticidas utilizando a técnica de extração em fase solida e cromatografia líquida com detecção espectrofotométrica com arranjo de diodos. O trabalho teve como objetivo comparar diferentes condições analíticas para a determinação dos pesticidas, buscando-se melhor separação, sensibilidade, precisão e exatidão na determinação de resíduos dos pesticidas em amostras de mel de abelhas. Soluções-padrão em concentrações variando de 1,0 a 20,0 μg mL-1 foram avaliadas em diferentes fases móveis, utilizando-se coluna C18 (250 x 4.6 mm d.i., 5 μm). A composição da fase móvel mais adequada foi acetonitrila:água em pH 3,0 (acidificada com ácido acético), em modo gradiente, monitorando-se os compostos em comprimento de onda de 220 nm. Nas condições estabelecidas, excetuando-se aldrin, dieldrin e dimetoato, foram observadas recuperações entre 75,26% a 113,58%, com coeficientes de variação entre 2,16% a 11,31%, para níveis de concentração entre 0,10 a 1,00 μg.g-1 (coeficientes de correlação entre 0,9978 e 0,9998). Recuperações inferiores a 50 % foram, entretanto, observadas para os citados inseticidas, utilizando-se solvente extrator acetato de etila e eluição com acetato de etila, sendo necessários novos estudos para obtenção de melhores recuperações para tais compostos.

No presente estudo, duas espécies de fungos isoladas do ambiente marinho tiveram seus extratos brutos ativos contra Staphylococcus aureus e Candida albicans para uma espécie de P. citrinum e, atividade citotóxica e contra Mycobacterium tuberculosis para uma espécie de P. oxalicum. Estas foram estudadas com a finalidade de otimizar suas condições de crescimento para produção de metabólitos secundários. Foram realizadas análises multivariadas utilizando planejamento fatorial fracionário. Foram obtidas duas condições ótimas de crescimento para as duas linhagens de Penicillium. O crescimento em maior volume em condições de cultura otimizadas para P. oxalicum permitiu observar a presença de três metabólitos secundários no extrato do meio de cultura. Dois puderam ser isolados e identificados: a meleagrina 52 e a oxalina 26. A metodologia utilizada para se obter uma maior quantidade de metabólitos secundários proporcionou, ainda, o incremento em cerca de 150% na área do pico da oxalina. A partir do crescimento em maior volume em condições de cultura otimizadas para P. citrinum foi possível observar presença de pelo menos onze diferentes metabólitos na análise dos extratos obtidos do meio de cultura. Foi possível identificar e isolar quatro compostos já conhecidos: a (8E)-1-(2,3-diidropirrol-1-il)-2-metildec-8-eno-1,3-diona 56; a 1-(2,3-diidropirrol-1-il)-2-metildecano-1,3-diona 58; a 2-((E)-hept-5-enil)-6,7,8,8a-tetraidro-3-metilpirrolo[2,1-b][1,3]oxazin-4-ona 59 e a citrinina 31. Também foram isolados dois novos alcalóides indólicos contendo um grupo nitro na molécula que foram nomeadas de citrinalinas A 60 e B 62. Foi realizado, também, um estudo com P. citrinum visando a maior produção das citrinalinas, que possibilitou o incremento na produção da citrinalina B. / In this study, two species of fungi isolated from the marine environment had their active extracts against Staphylococcus aureus and Candida albicans to a strain of P. citrinum, and activity cytotoxic and against Mycobacterium tuberculosis to a strain of P. oxalicum. This studied aims the optimization their growth conditions for the production of secondary metabolites. Multivariate analysis using a fractional factorial design were used to establish optimal growth conditions for both Penicillium strains. Two optimal growth conditions were obtained for both Penicillium strains. A largest growth volume of P. oxalicum using optimized conditions enabled the detection of three secondary metabolites in the culture media crude extract. Two of such compounds were isolated and identified: meleagrin 52 and oxaline 26. The methodology used to increase the production of secondary metabolites by P. citrinum enabled an increase of 150% in the peak area of oxaline. A largest growth volume of P. citrinum led to the detection of eleven different metabolites in the culture media. Four of these compounds were isolated and identified as the known (8E)-1-(2,3-dihydropyrrol-1-yl)-2-methyldec-8-ene-1,3-dione 56; the 1-(2,3-dihydropyrrol-1-yl)-2-methyldecane-1,3-dione 58; the 2-((E)-hept-5-enyl)-6,7,8,8a-tetrahydro-3-methylpyrrolo[2,1-b][1,3]oxazin-4-one 59 and citrinin 31. Two new indole alkaloids containing a nitro group were also isolated and identified, named citrinalins A 60 and B 62. A further study with P. citrinum aiming an enhanced production of citrinalins allowed a significant increase in the production of citrinalin B.

Submitted by Biblioteca da Faculdade de Farmácia (bff@ndc.uff.br) on 2017-03-23T19:00:13Z No. of bitstreams: 1 Peçanha, Bruna Rachel de Britto [Dissertação, 2012].pdf: 3907379 bytes, checksum: f2acabc3c1c39363b86f0d651d6b5936 (MD5) / Made available in DSpace on 2017-03-23T19:00:13Z (GMT). No. of bitstreams: 1 Peçanha, Bruna Rachel de Britto [Dissertação, 2012].pdf: 3907379 bytes, checksum: f2acabc3c1c39363b86f0d651d6b5936 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Polímeros de impressão molecular (MIPs) foram sintetizados e aplicados como adsorventes na técnica de extração em fase sólida (EFS). O método de polimerização por precipitação foi utilizado para a síntese dos polímeros, devido à simplicidade de preparo, altos rendimentos e obtenção de partículas mais uniformes, devido a não trituração do polímero. O MIP foi sintetizado com ácido metacrílico (MAA) como monômero funcional, trimetacrilato de trimetilolpropano (TRIM) e dimetacrilato de etilenoglicol (EDMA) como agentes de reticulação e o cloridrato de amilorida (AMI) foi escolhido como molécula-molde. Diferentes proporções de MAA, TRIM, EDMA, volume e tipo de solvente foram utilizadas para ajuste das condições ideais de síntese. Os MIP foram avaliados quanto à capacidade de adsorção comparando-se a polímeros sintetizados na ausência da molécula-molde (NIP, polímeros não impressos). O solvente de elevada polaridade empregado na síntese (THF:MeOH:H2O) permitiu o emprego da técnica para moléculas polares como AMI. O controle no volume de solvente permitiu a obtenção de partículas maiores, de modo que a EFS foi realizada em condições usuais, o que confere um potencial para aplicação dessa técnica de polimerização na preparação de adsorventes para EFS. O polímero que apresentou maior capacidade adsortiva no ensaio realizado em tampão citrato-acetato pH 6,5 foi o MIP/NIP 12 (AMI:MAA:TRIM 1:8:10), com uma taxa média de adsorção de 83 e 88% para NIP e MIP, respectivamente. A adsorção foi elevada devido a interação iônica entre MAA e AMI promovida pelo controle de pH, porém foi não específica. O polímero MIP/NIP 12 foi aplicado como adsorvente na EFS, onde a recuperação de AMI foi avaliada nos resíduos de carregamento e eluição com solventes. O carregamento com tampão citrato-acetato pH 6,5 foi o ideal, favorecendo a interação iônica do polímero com o analito. A eluição total de AMI do cartucho somente ocorre após lavagem com o solvente na presença de ácido, que protona os grupos carboxila do polímero, rompendo assim a interação iônica com o analito / Molecularly imprinted polymers (MIPs) were synthesized and applied as adsorbents in solid-phase extraction technique (SPE). The polymers have been synthesized by precipitation polymerization method because of its simplicity, high yields and good control of final size and shape of particles. MIP was synthesized using methacrylic acid (MAA) as functional monomer, trimethylolpropane trimethacrylate (TRIM) and ethyleneglycol dimethacrylate (EDMA) as cross-linker and amiloride hydrochloride (AMI) was chosen as template. Different ratios of MAA, TRIM and EDMA, volume and type of solvent were used to adjust the optimal synthesis conditions. The MIP were tested for adsorption capacity compared to the polymers synthesized in the absence of template molecule (NIP, non-imprinted polymers). The polar solvent mixture used (THF:MeOH:H2O) allowed the synthesis of MIP of polar molecules as AMI. The solvent volume control afforded the larger particles so the SPE was performed in the usual conditions, giving a potential application for this polymerization technique in the preparation of adsorbents for SPE. The polymers with higher adsorption capacity at the test performed in citrateacetate buffer pH 6,5 was MIP/NIP 12 (AMI:MAA:TRIM 1:8:10) with adsorption rate of 83 and 88% for NIP and MIP, respectively. The recognition of MIP was due to ionic interaction between MAA and AMI promoted by pH control, but was not specific. The polymer MIP/NIP 12 was used as a solid-phase extraction sorbent and the recoveries of AMI was evaluated using different loading and elution conditions. The loading with buffer citrate-acetate pH 6,5 was optimal, due to ionic interaction of the polymer with the analyte. Total elution of AMI bound to the polymers only occurs after washing with a acid-containing solvent, because of protonation of the carboxyl groups of the polymer and disrupting the ionic interaction with the analyte

This work is focused on the occurrence of surface-active substances in the environment. It deals with the development and optimization of methods for the determination of selected surfactants in samples from rivers and from influent and effluent of wastewater treatment plants. Representatives of anionic, nonionic and zwitterionic surfactants were selected as target compounds. Based on optimization of analytical methods and their pilot testing abroad on samples of surface water, the selection of analytes for the conditions in the Czech Republic were corrected and operatively expanded. For the final analysis liquid chromatography with mass spectrometry was used. Analysis of cationic surfactants was realized using liquid chromatograph with UV-VIS detector. This optimized method for analysis of anionic surfactants was applied to real samples. Samples of surface running water were from the River Thurso in Scotland and then from watercourses in the catchment area of Moravia river were collected. In addition to the grab sampling also continuous weekly sampling of water from Tvaroženský potok and Litava was realized. Waste water was collected at inflow and outflow of three wastewater treatment plants (WWTP) in South Moravia (Brno – Modřice, Břeclav and Hodonín).

Dans cette thèse, nous avons démontré la faisabilité de la préparation de polymères à empreinte moléculaires (MIP) hautement sélectifs pour la reconnaissance dans des matrices aqueuses complexes avec des applications dans les cosmétiques et en technologie alimentaire. Les MIP (de l'anglais molecularly imprinted polymers) sont des récepteurs synthétiques comparables aux anticorps, qui sont synthétisés par co-polymérisation de monomères fonctionnels et réticulants en présence d'un gabarit moléculaire. Leurs propriétés de reconnaissance moléculaire, associées à leur grande stabilité, robustesse mécanique, faible coût et leur synthèse facile les rendent extrêmement intéressants comme matériaux de capture sélective, avec des applications dans les séparations analytiques, la détection et la vectorisation des médicaments. Cependant, leur reconnaissance sélective dans des milieux aqueux reste toujours problématique et c'est l'une des raisons de leur expansion commerciale restreinte. Dans une première partie, nous avons développé un MIP fonctionnant en milieu aqueux pour son application comme ingrédient actif dans un déodorant. Les odeurs corporelles sont principalement dues à des acides gras volatils générés à partir de leurs précurseurs, des conjugués de glutamine par des enzymes hydrolytiques produites à partir de bactéries présentes sur la peau. La plupart des anti-transpirants et des déodorants actuellement commercialisés contiennent des sels d'aluminium et des agents antibactériens non spécifiques, respectivement. Cependant, l'utilisation extrêmement étendue de ces produits nécessite des solutions alternatives en ce qui concerne divers problèmes (environnement, respect de l'écosystème de la peau, toxicité, etc.). Pour cette raison, un MIP a été synthétisé pour capturer les précurseurs conjugués de glutamine afin qu'ils ne soient plus disponibles aux bactéries, empêchant ainsi leur transformation en composés malodorants. Afin de générer des liaisons sélectifs dans des environnements aqueux, un monomère à base d'amidinium qui peut former une interaction électrostatique stoechiométrique forte avec les groupes carboxyle sur le gabarit moléculaire a été synthétisé. Le MIP, mélangé dans une formulation dermo-cosmétique, pourrait capter sélectivement les précurseurs conjugués de glutamine, au milieu d'une multitude d'autres molécules présentes dans la sueur humaine. En outre, le MIP n’affecte pas les bactéries de la peau, ouvrant la voie à des déodorants innovateurs de nouvelle génération, moins problématiques pour la santé. Dans une deuxième partie, nous avons développé une procédure rapide et efficace basée sur l'extraction en phase solide à empreinte moléculaire (MISPE) pour la purification sélective de la bétanine et de son stéréoisomère l’isobétanine à partir d'extraits de betterave. La bétanine est un pigment naturel ayant un fort pouvoir antioxydant et dont les propriétés pharmacologiques sont de plus en plus étudiées. Ce pigment est actuellement utilisé comme simple colorant alimentaire. Dans notre étude, l'acide dipicolinique a été utilisé comme gabarit moléculaire pour la synthèse de MIP, en raison de sa similarité structurelle avec le groupe chromophore de la bétanine. Les procédures MISPE ont été optimisées permettant l'élimination presque complète des glucides issus de la matrice végétale ainsi que la majorité des protéines, ce qui permet d'obtenir un rendement élevé d'extraction de la bétanine / isobétanine en une seule étape. De plus, toute la procédure d'extraction a été réalisée dans des solvants respectueux de l'environnement, tels que l'éthanol ou l'eau. Pour conclure, nous sommes convaincus que ce travail pave le chemin au développement d'une nouvelle génération des MIP fonctionnant en milieu aqueux avec des propriétés de reconnaissance améliorées dans des environnements complexes, qui pourra s'appliquer également à d'autres domaines biotechnologiques et biomédicaux. / In this thesis, we have demonstrated the feasibility of preparing highly selective molecularly imprinted polymers (MIPs) for recognition in complex aqueous matrices with applications in cosmetics and food technology. MIPs are synthetic tailor-made receptors, with binding affinities and specificities comparable to those of natural antibodies. Their molecular recognition properties, combined with their high stability, mechanical robustness, low cost and easy synthesis make them extremely attractive as selective capture materials with applications in analytical and preparative separations, sensing and drug delivery, among others. However, their selective recognition in aqueous samples still remains problematic and is one of the reasons for their so far lilited commercial expansion. In the first part, we developed a water compatible MIP for its application as an active ingredient in a deodorant. Body odors are mainly due to volatile fatty acids generated from their glutamine conjugate precursors by hydrolytic enzymes from bacteria present on the skin. Most currently marketed anti-perspirants and deodorants contain, respectively aluminum salts and unspecific antibacterials. However, the extremely wide use of these products requires alternative solutions with regard to various problems (environmental, respect of skin ecosystem, toxicity, etc.). For this reason, a MIP was developed to capture the glutamine conjugate precursors so that they are no longer available to the bacteria, thus preventing their transformation to malodorous compounds. In order to generate binding selectivity in aqueous environments, an amidinium-based monomer which can form a strong stoichiometric electrostatic interaction with the carboxyl groups on the template, was synthesized. The MIP, blended in a dermo-cosmetic formulation, could capture selectively the glutamine precursors, amidst a multitude of other molecules present in human sweat. Furthermore, the MIP did not affect the skin bacteria, paving the way to an innovative and 'safer ' future-generation deodorant. In the second part, we developed a fast and efficient procedure based on molecularly imprinted solid­ phase extraction (MISPE) for the selective clean-up of betanin and its stereoisomer isobetanin from red beetroot extracts. Betanin is a natural pigment with significant antioxidant and biological activities currently used as food colorant. Dipicolinic acid was used as template for the MIP synthesis, because of its structural similarity to the chromophore group of betanin The MISPE procedures were optimized allowing the almost complete removal of carbohydrates and the majority of proteins, resulting in high extraction recovery of betanin / isobetanin in a single step. Moreover, the whole extraction procedure was performed in environmentally friendly solvents with either ethanol or water. To conclude, we believe that this study paves the way towards the development of a new generation of water compatible MIPs with improved recognition properties in highly complex aqueous environments, and should be applicable to other biotechnological and biomedical areas as well.

La pollution des eaux est particulièrement problématique pour les pays industrialisés et les pays en développement. Des suivis environnementaux se sont alors avérés nécessaires afin de progresser dans la compréhension des origines et des conséquences de la présence de ces polluants. Parmi les polluants organiques les plus dangereux, les Hydrocarbures Aromatiques Polycycliques (HAP) et les pesticides qui peuvent se retrouver à l’état de traces dans les milieux aquatiques. Leur analyse nécessite des méthodes analytiques à la fois spécifiques et sensibles. L’Extraction sur Phase Solide (SPE) suivi d’une analyse par la Chromatographie Gazeuse couplée à la Spectrométrie de Masse (GC-MS) est la méthode la plus couramment employée. Les travaux de thèse s’inscrivent ainsi dans l’optique généralevisant à développer des nouvelles méthodes analytiques pour l’extraction et l’analyse de cesdeux familles de polluants organiques présents dans l’eau. Afin de pallier les contraintes classiques de la SPE, une nouvelle approche a été développée qui consiste à introduire latechnique de centrifugation dans certaines étapes de la procédure SPE. La nouvelle méthodeélaborée a permis de réduire considérablement le temps de préparation et d’économiserl’utilisation et le rejet des solvants organiques, en réduisant presque 10 fois les volumes nécessaires à l’obtention des rendements de récupération satisfaisants. Une méthoded’analyse par injection à température programmable (PTV) a été optimisée afin d’améliorerles seuils de détection des HAP. Cette étude a permis également d’avoir les informations surla qualité des eaux de pluie, eaux souterraines et eaux de surface au Liban quant au niveau deleur contamination pour les HAP et les pesticides. / Water pollution presents a very critical problem facing industrial and developping countries. The environmental monitoring of the contaminants seems necessary to understand their sources and impacts. Among a wide variety of organic pollutants present in water,polycyclic aromatic hydrocarbons (PAHs) and pesticides are of particular importance as widespread, persistent, and toxic contaminants. They are usually present at trace levels in theacquatic surfaces; therefore their detection and control require selective and sensitiveanalytical procedures. The Solid-Phase Extraction (SPE) followed by the Gas-Chromatography coupled to Mass Spectrometry (GC-MS) are the most commonly usedtechniques for their analysis in water. Thesis objectives are focused on the development of new analytical methods for the extraction and analysis of these two families of pollutants present in water. To overcone the contraints of the traditional SPE, a new approach was developed consisting on the introduction of the centrifugation in several steps of the procedure. The new method showed practical environmental and economical advantages interms of sample preparation time, simplicity, reduction in solvent use, and cost and isparticularly suitable for routine applications requiring a high sample throughput. Aprogrammed temeperature vaporizing (PTV) injection method was also optimized and validated in order to improve the detection limits for the GC-MS analysis of PAHs. The evaluation of the quality of different water systems in Lebanon including rainwater,groundwater, drinking water and surface water was accomplished in this study.

Indiana University-Purdue University Indianapolis (IUPUI) / Paper spray - mass spectrometry (PS-MS) has been developed as a rapid and direct ionization method for qualitative and quantitative analysis of complex samples at trace levels. In this work, different sampling devices for PS-MS were investigated to improve the assay’s simplicity and sensitivity over traditional approaches. In particular, chapter two characterizes an alternate paper substrate to enhance drug detection on surfaces like asphalt, cloth, concrete, aluminum, and glass. Analysis occurs on a single spray ticket coated with pressure-sensitive adhesive (PSA), also known as Post-it notes to detect and quantify drug residues. A PS-MS method utilizing PSA paper was developed to detect a mixture of ten drugs off of various surfaces to evaluate the qualitative and quantitative capabilities of the aforementioned substrate. After the method development on a conventional linear ion trap mass spectrometer, the assay was translated for use on a portable mass spectrometer to evaluate the suitability of the pressure-sensitive adhesive paper substrate in the field in chapter three. Chapter four introduces a sampling device combined with a snap-in solid-phase extraction (SPE) column. The new cartridge design not only inherits the functions from the first iteration SPE cartridge, including extraction and preconcentration from complex samples, but also exhibits greater flexibility in volume control and ease of use for on-site sample collection.

Currently there is a spate of interest in the presence of pharmacologically active substances in the environment. These substances are excreted in active or metabolized form and with wastewater pass the wastewater treatment plant. Current treatment technologies do not always eliminate all pharmaceuticals effectively and therefore they enter the environment. One of these active groups is the group of steroid hormones. Steroid hormones belong to the group of endocrine disruptors and they are considered to be dangerous for the ecosystems. Due to the hydrophobic character of steroid hormones they undergo partial or total sorption from wastewater to sludge. Sewage sludge is the by product of wastewater treatment and contains heavy metals, organic contaminants and pathogenic bacteria. A part of produced sewage sludge is used as a fertilizer in the Czech Republic. This issue follows the regulation 347/2016 – conditions of agricultural use of sewage sludge. Waste policy of EU will alter the conditions of sludge disposal, so it is necessary to obtain data about the concentration levels, fate and behaviour of those pollutants. This thesis was focused on five natural female hormones and four synthetic, which are used as a part of contraceptive pills and substitutional hormonal therapy. Determination of hormones was performed in four steps including ultrasonic assisted extraction, clean up by solid phase extraction, derivatization and final analysis by gas chromatography with mass spectrometry on triple quadrupole in MS/MS mode.