Développés pour améliorer la santé et le bien-être, certains composés pharmaceutiques sont désormais sous haute surveillance car considérés comme des contaminants émergents des eaux. Pour répondre à ce nouvel enjeu, nous visons à développer un microsystème d'analyse permettant l'identification et la quantification de ces contaminants dans des échantillons d'eau. L'aspect original de ce microsystème repose sur l'intégration au sein du canal d'une zone de préconcentration dans laquelle sont immobilisés des ligands (aptamères dans notre étude), permettant l'extraction sélective de la cible et sa concentration. Pour développer ce microsystème, deux types de fonctionnalisation de surface doivent être mis en œuvre : (1) globale, sur toute la surface des canaux, pour contrôler leurs propriétés de surface et ainsi éviter l'adsorption et contrôler les écoulements de liquides, et (2) locale, pour greffer les ligands sélectifs dans une zone confinée du canal. Les polymères COC et THV, prometteurs pour la conception de puces microfluidiques grâce à leur transparence dans le domaine UV-visible et leur excellente résistance aux solvants, ont été sélectionnés pour la microfabrication du système. Cependant, leur inertie chimique rend difficile la fonctionnalisation de leur surface, et de nouvelles méthodes de traitement de surface ont été développées. Nous présentons ainsi plusieurs méthodes innovantes pour la fonctionnalisation de ces matériaux, basées sur un dépôt plasma, une modification électrochimique et/ou une réaction chimique. La possibilité d'encapsuler les ligands dans une phase monolithique à l'intérieur d'un microcanal grâce à un procédé sol-gel a également été évaluée. / Developed to promote human health and well-being, certain pharmaceuticals are now attracting attention as crucial emerging water contaminants. To deal with this concern, we aim at developing an analytical microsystem for the identification and quantitation of these contaminants in water samples. The original aspect of this lab-on-a-chip relies on the integration inside the channel of a preconcentration zone in which ligands (aptamer in our study) are immobilized, in order to concentrate the target and extract it from the rest sample matrix. Development of this microsystem requires surface treatments to modify the microchannel surface at two scales: (1) globally (on the entire channel walls) to control surface properties and thus avoid adsorption as well as control fluid flows, or (2) locally to immobilize selective ligands in restricted areas for selective target extraction and preconcentration. Polymers COC and THV, attractive for the conception of microfluidic chips thanks to their UV-visible transparency and high resistance to aggressive solvents, were selected as the microchip material. However due their chemical inertness new functionalization techniques have to be developed to modify their surface. In this work, innovative surface treatment strategies have been developed for both materials, based on plasma, electrochemical and chemical approaches. The possibility of encapsulating aptamers in a monolithic phase inside microchannel by sol-gel process was also explored.

Made available in DSpace on 2015-05-14T13:21:51Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 3041446 bytes, checksum: ababcafc4806728e34bf8d0bd8ea021e (MD5) Previous issue date: 2010-09-10 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / This study proposed a miniaturized flow-batch system for chemical analysis. The technique used microfabricated urethane-acrylate, a commercial polymer. The microsystem was evaluated and optimized by analysis of synthetic dye samples. Afterwards, it was employed for the determination of tannins in tea samples. The tannins are a group of polyphenols of significant relevance in the food industry and pharmaceuticals. The samples used were green and black tea, obtained from the local market. The determinations were performed by turbidimetric methods using copper (II) in an acetate medium, with photometric methods and ferrous tartrate as a reference. Miniaturization in urethane-acrylate implies low cost and low maintenance, rapid prototyping and includes the satisfactory physicochemical properties of polymer. These characteristics combined with the general advantages of miniaturization in analytical devices, such as high frequency analysis and low waste generation, make the system a great source in academic research. For analysis of tannins in tea samples, the system had precise and accurate results, and high speeds. This flow-batch microsystem was able to perform up to 300 tests per hour, for the photometric method of reference and up to 200 tests per hour for the turbidimetric method. Each analysis performed generated waste volumes lower than 70 μL. Data validation of statistical models obtained have proved very satisfactory and promising for new optical applications. / Neste trabalho de pesquisa foi proposto o uso da técnica de microfabricação em polímero comercial uretana-acrilato para a miniaturização de um sistema automático de análises químicas em fluxobatelada, o flow-batch. O microssistema desenvolvido foi avaliado e otimizado pela análise de amostras sintéticas de corantes. Posteriormente, o microssistema, foi empregado para a determinação de taninos, grupo de polifenóis de expressiva relevância industrial, em amostras de chá verde e preto, por dois métodos ópticos distintos, fotométrico e turbidimétrico. A técnica de microfabricação em uretana-acrilato se caracteriza pelos baixos custos de implementação e manutenção, satisfatórias propriedades físicoquímicas do polímero e a rápida prototipagem de sistemas microfluídicos. Tais características aliadas às vantagens inerentes da miniaturização de dispositivos analíticos, como a elevada frequência de análise e a baixa geração de resíduos, conferem a esse sistema de análise uma ótima fonte de pesquisa acadêmica. Para análise dos taninos em amostras de chá, o sistema apresentou resultados precisos e exatos, além de uma alta velocidade analítica para ambos os métodos ópticos, sendo capaz de executar até 300 análises por hora, no método fotométrico e 200 análises por hora pelo método turbidimétrico. Cada análise efetuada gerou resíduos cujos volumes foram inferiores a 70 μL. Os dados de validação estatística dos modelos obtidos se mostraram bastante satisfatórios e promissores para novas aplicações ópticas.