Ces travaux de recherches concernent l'association de la technologie des polymères à empreintes moléculaires aux dispositifs acoustiques à onde de Love afin de réaliser un biocapteur dans le cadre d'un projet ANR Tecsan. La première partie de ces travaux de thèse a dressé plus spécifiquement la mise au point d'un protocole de dépôt localisé de polymères imprimés (MIP) et non imprimés (NIP) en films minces compatibles avec la propagation de l'onde élastique. La seconde aprtie des travaux a visé une caractérisation des films et des capteurs ainsi réalisés, par microscopie à balayage et apr mesures de détection sous gaz. Des éléments relatifs aux propriétés mécaniques (porosité, surfaces spécifiques) des MIPs et NIPs, avant extraction de la molécule cible, puis après extraction et après recapture, ont permis de valider le principe du capteur, ouvrant la voie à l'application en milieu liquide. / This work deals with the association of molecularly imprinted polymers technology and Love wave devices in order to develop a biosensor in the frame of ANR project, Tecsan. The first part of this work concerned the development and the validation of a protocol for localized deposition of imprinted (MIP) and non-imprinted (NIP) polymeric thin film compatible with the propagation of acoustic wave. The second part exhibited the cahracterization of the coated sensors, by scanning microscopy and deep gas characterization. The proposed characterization technique gives information on surface morphology and porosity of thin MIP films before extraction of the target molecule, then after extraction and after rebinding. It allowed hence, the validation of the sensor principle. These results constitute a god background for future application in medium liquid.

京都大学 / 新制・課程博士 / 博士(工学) / 甲第23513号 / 工博第4925号 / 新制||工||1769(附属図書館) / 京都大学大学院工学研究科材料化学専攻 / (主査)教授 大塚 浩二, 教授 松原 誠二郎, 教授 秋吉 一成 / 学位規則第4条第1項該当 / Doctor of Philosophy (Engineering) / Kyoto University / DFAM

Le vétérinaire aide le producteur laitier à garder son troupeau en santé. Lorsqu’une vache est malade, il peut prescrire des antibiotiques. Le cas échéant, le lait de la vache traitée aux antibiotiques est jeté. Il n’est donc pas vendu pour consommation. Tout le lait produit est analysé avant d’être pasteurisé et transformé afin de s’assurer que les produits laitiers ne contiennent pas de résidus d’antibiotiques. Si les analyses indiquent que le lait renferme des traces d’antibiotiques, il est jeté et le producteur en cause doit assumer la perte. Les épreuves de dépistage actuelles de ces résidus médicamenteux sont onéreuses et inapplicables sur le terrain. Pour résoudre cette problématique aux pieds de la vache, la solution proposée dans ce projet est la fabrication d’un kit de détection basé sur les polymères à empreinte moléculaire. Il s’agit de polymères dont la conformation moléculaire est complémentaire à celle des antibiotiques. Dans ce projet, il est question d’améliorer l’efficacité des épreuves de dépistages des résidus de tétracyclines en augmentant le nombre de sites d’interaction entre l’antibiotique et des polyesters. Trois polymères sont utilisés portant respectivement des groupements aromatiques, carboxyliques et hydroxyles. Une étude antérieure dans notre laboratoire avait déjà donné un pourcentage de rétention de tétracycline de 50% pour une composition de 1/3 PLAOH- 1/3 PLACOOH- 1/3 PLAOBn. Avec des ajustements, le pourcentage passe à 38.93 % pour une composition de 1/4 PLAOH- 1/2 PLACOOH- 1/4 PLAOPh, de 44.81 % pour une composition de 1/4 PLACOOH- 1/2 PLAOH- 1/4 PLAOPh, de 66.34 %pour une composition de 1/4 PLAOH- 1/4 PLACOOH- 1/2 PLAOPh et de 78.07 % pour une composition de 1/6 PLAOH- 1/6 PLACOOH- 2/3 PLAOPh. Notre hypothèse était que la présence accrue du groupement phényle augmenterait le nombre de sites d’interaction spécifique avec l’antibiotique augmentant ainsi le pourcentage de rétention de l’antibiotique à travers les MIP. Les résultats ont confirmé cette hypothèse. / The veterinarian is helping the dairy farmer to keep his flock healthy. When a cow is sick, he may prescribe antibiotics. But in that case, the cow milk treated with antibiotics is discarded, and not sold for consumption. All milk products are analyzed before being pasteurized and processed to ensure that dairy products do not contain residues of antibiotics. If the analyses show that milk contains traces of antibiotics, it is discarded and the producer must bear the loss. On place, the current screening tests for drug residues are expensive and inapplicable. To resolve this problem at the feet of the cow, the solution proposed in this project is the production of a detection kit based on molecular imprinted polymers. These are polymers whose molecular conformation is complementary to that of antibiotics. This project is about improving the effectiveness of screening tests for residues of tetracyclines by increasing the number of interaction sites between the antibiotic and polyester. Three polymers are used respectively, aromatic, carboxyl and hydroxyl groups. A previous study in our laboratory had already shown a tetracycline retention percentage of 50% for a composition of 1 / 3 PLAOH-1 / 3 PLACOOH-1 / 3 PLAOBn. After variation in the polymers ratio, the percentage obtained is, 38.93 % for a composition of 1 / 4 PLAOH-1 / 2 PLACOOH-1 / 4 PLAOPh, 44.81 % for a composition of 1 / 4 PLACOOH- 1 / 2 PLAOH- 1 / 4 PLAOPh, 66.34 % for a composition of 1/4 PLAOH- 1/4 PLACOOH- 1/4 PLAOPh and 78.07 % for a composition of 1/6 PLAOH- 1 / 6 PLACOOH- 2 / 3 PLAPh. Our hypothesis was that the increase of the amount of the phenyl group, increases the number of specific interaction sites with the antibiotic, and so increases the percentage of retention of the antibiotic through the MIP. The results confirmed this hypothesis.

Le vétérinaire aide le producteur laitier à garder son troupeau en santé. Lorsqu’une vache est malade, il peut prescrire des antibiotiques. Le cas échéant, le lait de la vache traitée aux antibiotiques est jeté. Il n’est donc pas vendu pour consommation. Tout le lait produit est analysé avant d’être pasteurisé et transformé afin de s’assurer que les produits laitiers ne contiennent pas de résidus d’antibiotiques. Si les analyses indiquent que le lait renferme des traces d’antibiotiques, il est jeté et le producteur en cause doit assumer la perte. Les épreuves de dépistage actuelles de ces résidus médicamenteux sont onéreuses et inapplicables sur le terrain. Pour résoudre cette problématique aux pieds de la vache, la solution proposée dans ce projet est la fabrication d’un kit de détection basé sur les polymères à empreinte moléculaire. Il s’agit de polymères dont la conformation moléculaire est complémentaire à celle des antibiotiques. Dans ce projet, il est question d’améliorer l’efficacité des épreuves de dépistages des résidus de tétracyclines en augmentant le nombre de sites d’interaction entre l’antibiotique et des polyesters. Trois polymères sont utilisés portant respectivement des groupements aromatiques, carboxyliques et hydroxyles. Une étude antérieure dans notre laboratoire avait déjà donné un pourcentage de rétention de tétracycline de 50% pour une composition de 1/3 PLAOH- 1/3 PLACOOH- 1/3 PLAOBn. Avec des ajustements, le pourcentage passe à 38.93 % pour une composition de 1/4 PLAOH- 1/2 PLACOOH- 1/4 PLAOPh, de 44.81 % pour une composition de 1/4 PLACOOH- 1/2 PLAOH- 1/4 PLAOPh, de 66.34 %pour une composition de 1/4 PLAOH- 1/4 PLACOOH- 1/2 PLAOPh et de 78.07 % pour une composition de 1/6 PLAOH- 1/6 PLACOOH- 2/3 PLAOPh. Notre hypothèse était que la présence accrue du groupement phényle augmenterait le nombre de sites d’interaction spécifique avec l’antibiotique augmentant ainsi le pourcentage de rétention de l’antibiotique à travers les MIP. Les résultats ont confirmé cette hypothèse. / The veterinarian is helping the dairy farmer to keep his flock healthy. When a cow is sick, he may prescribe antibiotics. But in that case, the cow milk treated with antibiotics is discarded, and not sold for consumption. All milk products are analyzed before being pasteurized and processed to ensure that dairy products do not contain residues of antibiotics. If the analyses show that milk contains traces of antibiotics, it is discarded and the producer must bear the loss. On place, the current screening tests for drug residues are expensive and inapplicable. To resolve this problem at the feet of the cow, the solution proposed in this project is the production of a detection kit based on molecular imprinted polymers. These are polymers whose molecular conformation is complementary to that of antibiotics. This project is about improving the effectiveness of screening tests for residues of tetracyclines by increasing the number of interaction sites between the antibiotic and polyester. Three polymers are used respectively, aromatic, carboxyl and hydroxyl groups. A previous study in our laboratory had already shown a tetracycline retention percentage of 50% for a composition of 1 / 3 PLAOH-1 / 3 PLACOOH-1 / 3 PLAOBn. After variation in the polymers ratio, the percentage obtained is, 38.93 % for a composition of 1 / 4 PLAOH-1 / 2 PLACOOH-1 / 4 PLAOPh, 44.81 % for a composition of 1 / 4 PLACOOH- 1 / 2 PLAOH- 1 / 4 PLAOPh, 66.34 % for a composition of 1/4 PLAOH- 1/4 PLACOOH- 1/4 PLAOPh and 78.07 % for a composition of 1/6 PLAOH- 1 / 6 PLACOOH- 2 / 3 PLAPh. Our hypothesis was that the increase of the amount of the phenyl group, increases the number of specific interaction sites with the antibiotic, and so increases the percentage of retention of the antibiotic through the MIP. The results confirmed this hypothesis.

Submitted by Liliane Ferreira (ljuvencia30@gmail.com) on 2018-11-27T10:54:13Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Tese - Thays Colletes de Carvalho - 2018.pdf: 10167238 bytes, checksum: e63cc8b9621f34c961582121789dd84c (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2018-11-27T11:56:05Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Tese - Thays Colletes de Carvalho - 2018.pdf: 10167238 bytes, checksum: e63cc8b9621f34c961582121789dd84c (MD5) / Made available in DSpace on 2018-11-27T11:56:05Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Tese - Thays Colletes de Carvalho - 2018.pdf: 10167238 bytes, checksum: e63cc8b9621f34c961582121789dd84c (MD5) Previous issue date: 2018-10-05 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Ionization is a crucial step in mass spectrometry (MS) and governs the versatility of this important analytical technique. Ionization is responsible for transferring atoms and molecules, in their respective ionized forms, into the high vacuum environment of mass spectrometers, where they are discriminated as a function of their m/z ratio. Among the several techniques of ionization of MS, the technique of ionization by Paper spray (PS) is one of the simplest, versatile and can be implemented easily in MS system. In this work, several PS-MS applications in the forensic area were developed, demonstrating their ability to screen new synthetic drugs as a complementary tool to the thin layer chromatography (TLC) method and as a tool to monitor Date-rape Drugs in blood. In the first application, PS-MS was shown to be an effective and rapid method for the identification of synthetic drugs lysergic acid diethylamide (LSD), 2,5-dimethoxy-4-chloroamphetamine (DOC), 2,5-dimethoxy-4-bromoamphetamine (DOB), 25C-NBOMe, 25B-NBOMe and 25I-NBOMe) in bottles, both by the isotopic profile of the substances and by the fragmentation presented in tandem mass spectrometry. In the second application, the TLC was coupled to the PS-MS in order to achieve greater reliability in the CCD results. In this way, a CCD method for the analysis of cocaine and adulterants (caffeine, benzocaine, lidocaine and phenacetin) was optimized by evaluating its sensitivity and selectivity. In order to improve the Detection and Quantification Limits for cocaine and adulterants, the spots were analyzed by PS-MS, obtaining improvements in the. Finally, in order to solve the low PS-MS selectivity, a new substrate was developed, especially when the analytical target is in complex mixtures, such as blood. It is a membrane coated with a molecularly printed polymer (MIP) with dual function: simultaneous extraction and ionization of targets in the same device. The developed membrane was applied in the determination of benzodiazepines in blood samples from alleged date rape- drugs victims. Blood is a complex sample containing several compounds that can suppress the analyte signal. With this modification, any suppression is avoided, obtaining excellent results, both qualitative and quantitative. In conclusion, PS-MS is a fast and low-cost technique that can replace or complement conventional analyzes in a laboratory of expertise, increasing the productivity of Brazilian justice. / A ionização é uma etapa crucial em espectrometria de massas (MS) e rege a versatilidade desta importante técnica analítica. A ionização é a responsável por transferir átomos e moléculas, em suas respectivas formas ionizadas, para o ambiente de alto vácuo dos espectrômetros de massas, onde são discriminados em função de sua razão massa sobre carga (m/z). Dentre as diversas técnicas de ionização de MS, a técnica de ionização por Paper spray (PS) é uma das mais simples, versátil e pode ser implementada facilmente em sistema de MS. Nesse trabalho diversas aplicações do PS-MS na área forense foram desenvolvidas, demonstrando sua capacidade para screening de novas drogas sintéticas, como ferramenta complementar ao método de cromatografia de camada delgada (TLC) e como ferramenta para monitorar “boa noite cinderela” em sangue. Na primeira aplicação, o PS-MS mostrou ser um método eficaz e rápido para identificação de drogas sintéticas dietilamida do ácido lisérgico (LSD), 2,5-dimetoxi-4-cloroanfetamina (DOC), 2,5-dimetoxi-4- bromoanfetamina (DOB), 25C-NBOMe, 25B- NBOMe e 25I-NBOMe) em selos, tanto pelo perfil isotópico das substâncias, quanto pela fragmentação apresentada na espectrometria de massas tandem. Já na segunda aplicação, acoplou-se TLC ao PS-MS visando alcançar maior confiabilidade nos resultados de TLC. Desta maneira, otimizou-se um método de TLC para análise de cocaína e adulterantes (cafeína, benzocaína, lidocaína e fenacetina) avaliando sua sensibilidade e seletividade. Com intuito de melhorar os Limites de Detecção e Quantificação para a cocaína e adulterantes, os spots foram analisados por PS-MS, obtendo melhorias nos resultados. Por fim, com o intuito de resolver a baixa seletividade do PS-MS, um novo substrato foi desenvolvido, principalmente quando o alvo analítico está em misturas complexas, como sangue. Trata-se de uma membrana recoberta com um polímero molecularmente impresso (MIP) com dupla função: extração e ionização simultânea de alvos em um mesmo dispositivo. A membrana desenvolvida foi aplicada na determinação de benzodiazepínicos em amostras de sangue de supostas vítimas de benzodiazepínicos. O sangue é uma amostra complexa que contém vários compostos que podem suprimir o sinal do analito. Com essa modificação qualquer supressão é evitada, conseguindo-se ótimos resultados, tanto qualitativos quanto quantitativos. Sendo assim, o PS-MS é uma técnica rápida e de baixo custo, que pode substituir ou complementar as análises convencionais em um laboratório de perícia, aumentando a produtividade da justiça brasileira.