Profenofós é um organofosforado empregado como inseticida e acaricida amplamente utilizado no Brasil para o controle de pragas de cebolas, milho, soja, café, tomate, algodão, feijão, batata e outros. A irradiação é um processo empregado em todo o mundo e recomendada por diversos órgãos de saúde para a conservação de alimentos. A radiação ionizante utiliza raios gama, raios X ou aceleradores de elétrons e tem sido aplicada para eliminar ou reduzir a ação de agentes patogênicos e contribuir para aumentar o tempo de estocagem de vários alimentos. Os objetivos desse trabalho foram: (a) avaliar a degradação de soluções aquosas de profenofós submetidas à radiação ionizante, identificar e quantificar a formação de novos produtos por GC-MS; (b) analisar o efeito de feixe de elétrons em ervilhas inoculadas com soluções aquosas de profenofós; (c) sintetizar Polímeros Molecularmente Impressos (MIP) e Sílica Impressa Molecularmente (MIS), posteriormente, caracterizar os adsorventes em fase sólida e verificar sua seletividade para profenofós. O tratamento com aceleradores de elétrons com dose 31,6 kGy promoveu a formação de um novo produto de degradação e redução de 93,40 % de profenofós em soluções aquosas. Em ervilhas inoculadas com 1 μg de profenofós submetidas à radiação ionizante de 30,4 kGy promoveu uma redução na concentração de profenofós em 57,46 %. Além disso, foram realizadas sínteses de MIP e MIS para a extração em fase sólida de profenofós. Os MIS sintetizados por sol-gel mostraram-se eficazes para o reconhecimento molecular e extração seletiva de profenofós. / Profenofos is an organophosphate widely used in Brazil as insecticide and acaricide in the control of pests in onions, corn, soybeans, coffee, tomato, cotton, beans, potatoes among others. Irradiation is a process used worldwide and recommended by many health agencies for food preservation. Food irradiation preserving process uses accelerated electrons, gamma rays or X-rays. Ionizing radiation treatment is applied to eliminate or to reduce the action of pathogens and to increase the shelf life of some foods. The objective of this study were (a) to evaluate the degradation of aqueous solutions of Profenofos by ionizing radiation, identify and quantify the formation of new products by GC-MS; (b) to analyze the effects of electron beam in peas inoculated with aqueous solutions of Profenofos; (c) to synthesize Molecularly Imprinted Polymer (MIP) and Molecularly Imprinted Silica (MIS), subsequently characterize the adsorbents in solid phase and check its selectivity for profenofos. The treatment with electron accelerators with 31.6 kGy dose promoted the formation of a new by-product and 93.40 % reduction of profenofos in aqueous solutions. In peas inoculated with 1 μg of profenofos by ionizing radiation of 30.4 kGy promoted a reduction of 57.46 % in the concentration of profenofos. Furthermore, the MIP and MIS were performed for solid phase extraction of profenofos. The MIS synthesized by sol-gel proved to be effective for the recognition molecular and selective extraction of profenofos.

Telomerase is expressed in the majority (>85%) of tumours but not in differentiated normal tissue. This enzyme catalyses the elongation of telomeres – a process critical for continued cell proliferation. Telomerase is a potential novel target for molecularly-targeted radiotherapy (mRT), due to its nuclear localization and expression profile. The radiolabelling of telomerase inhibitors may accelerate and enhance the cytotoxicity of such molecules, as a result of irradiation of the DNA. An oligonucleotide targeting telomerase RNA (hTR), shown to inhibit enzyme activity in vitro, was selected for study. Complementary and non-targeting control oligonucleotides were conjugated to a metal chelator (DTPA) to allow radiolabelling with indium-111. The radioiodination of MST-312, BIBR-1532 and flavonoid-derived small molecule inhibitors of telomerase was also pursued. The inhibitory activity of the candidate molecules was analysed using the telomeric repeat amplification protocol (TRAP). The internalization of inhibitors was assessed by gamma-counting following cell lysis. The clonogenic assay was employed to measure the effect of modified inhibitors on cell survival. Small molecule telomerase inhibitors were modified for labelling with iodine-123, which led to a modest decrease in inhibitory potency, compared to the parent molecules. Radiolabelled small molecules exhibited poor stability and internalization into cancer cells, so were unsuitable for mRT. Modified oligonucleotides potently inhibited telomerase activity, whereas a non-targeting oligonucleotide exhibited no inhibitory activity. Indium-111 radiolabelled oligonucleotides decreased the clonogenic survival of telomerase-positive breast cancer cells but not telomerase-negative cells, in a sequence-specific manner. Accordingly, complementary radiolabelled oligonucleotides were found to induce the DNA damage marker γH2AX. Oligonucleotides localized to nuclear Cajal bodies, the sites of telomerase assembly, in a proportion of cancer cells. Telomerase inhibitors of different classes were radiolabelled with Auger electron-emitting radionuclides, and delivered to cells. Radiolabelled oligonucleotides targeting telomerase significantly reduced the clonogenicity of cancer cells in vitro. This study represents a novel approach for the mRT of telomerase-positive cancers.

A ocorrência de N-nitrosaminas em produtos cosméticos e de higiene pessoal está relacionada com os ingredientes utilizados na formulação de tais artefatos. A N-nitrosodietanolamina (NDELA) é formada na presença da dietanolamina associada a outros componentes, tais como íons nitrito empregados como conservantes em matrizes cosméticas. São cancerígenas e, portanto, as condições em que tais itens de consumo são fabricados é um fator que deve ser mensurado. Assim, é aconselhável a realização de estudos que mostrem a ocorrência de NDELA em cosméticos, seus principais métodos de determinação, bem como os aspectos legais que indicam os níveis de concentração deste contaminante presente em diversas formulações. Foi avaliado neste trabalho o potencial de algumas técnicas de preparo de amostras na concentração de NDELA, tais como microextração líquido-líquido dispersiva em fase reversa (DLLME-RP), extração líquido-líquido com salting out e clean up no modo on-line. Os resultados obtidos foram confrontados com os dados presentes na literatura. Adicionalmente, utilizando os conceitos de polimerização via sol-gel e via precipitação para a síntese de polímeros molecularmente impressos (MIP), propõem-se uma possível rota sintética para a confecção de materiais seletivos para a pré-concentração da nitrosamina. / The occurrence of N-nitrosamines in cosmetic and personal hygiene are related to the ingredients used in the formulation of such artefacts. N-nitrosodiethanolamine (NDELA) is formed in the presence of diethanolamine associated with other components such as nitrite ions employed as preservatives in cosmetic matrices. They are carcinogenic and, therefore, the conditions under which such consumer items are produced is a factor that must be measured. It is therefore advisable to carry out studies that show the occurrence of NDELA in cosmetics, their main methods of determination as well as the legal aspects that indicate the levels of concentration of this contaminant present in various formulations. It was evaluated in this study the potential for some sample preparation techniques in the concentration of NDELA such as liquid-liquid microextraction dispersive reverse phase (DLLME-RP) liquid-liquid extraction with salting out and clean up online. The results were compared with data in the literature. Additionally, using the concepts of polymerization via sol-gel and precipitation route for the synthesis of molecularly imprinted polymers (MIP) are proposed as a possible synthetic route for the preparation of materials for selective pre-concentration of nitrosamine.

A ocorrência de N-nitrosaminas em produtos cosméticos e de higiene pessoal está relacionada com os ingredientes utilizados na formulação de tais artefatos. A N-nitrosodietanolamina (NDELA) é formada na presença da dietanolamina associada a outros componentes, tais como íons nitrito empregados como conservantes em matrizes cosméticas. São cancerígenas e, portanto, as condições em que tais itens de consumo são fabricados é um fator que deve ser mensurado. Assim, é aconselhável a realização de estudos que mostrem a ocorrência de NDELA em cosméticos, seus principais métodos de determinação, bem como os aspectos legais que indicam os níveis de concentração deste contaminante presente em diversas formulações. Foi avaliado neste trabalho o potencial de algumas técnicas de preparo de amostras na concentração de NDELA, tais como microextração líquido-líquido dispersiva em fase reversa (DLLME-RP), extração líquido-líquido com salting out e clean up no modo on-line. Os resultados obtidos foram confrontados com os dados presentes na literatura. Adicionalmente, utilizando os conceitos de polimerização via sol-gel e via precipitação para a síntese de polímeros molecularmente impressos (MIP), propõem-se uma possível rota sintética para a confecção de materiais seletivos para a pré-concentração da nitrosamina. / The occurrence of N-nitrosamines in cosmetic and personal hygiene are related to the ingredients used in the formulation of such artefacts. N-nitrosodiethanolamine (NDELA) is formed in the presence of diethanolamine associated with other components such as nitrite ions employed as preservatives in cosmetic matrices. They are carcinogenic and, therefore, the conditions under which such consumer items are produced is a factor that must be measured. It is therefore advisable to carry out studies that show the occurrence of NDELA in cosmetics, their main methods of determination as well as the legal aspects that indicate the levels of concentration of this contaminant present in various formulations. It was evaluated in this study the potential for some sample preparation techniques in the concentration of NDELA such as liquid-liquid microextraction dispersive reverse phase (DLLME-RP) liquid-liquid extraction with salting out and clean up online. The results were compared with data in the literature. Additionally, using the concepts of polymerization via sol-gel and precipitation route for the synthesis of molecularly imprinted polymers (MIP) are proposed as a possible synthetic route for the preparation of materials for selective pre-concentration of nitrosamine.

Submitted by JOSIANE SANTOS DE OLIVEIRA (josianeso) on 2017-12-06T13:59:20Z No. of bitstreams: 1 Tainã Coelho Quevedo_.pdf: 1779262 bytes, checksum: 1c805c11de74d708f12d1e1a98e63886 (MD5) / Made available in DSpace on 2017-12-06T13:59:20Z (GMT). No. of bitstreams: 1 Tainã Coelho Quevedo_.pdf: 1779262 bytes, checksum: 1c805c11de74d708f12d1e1a98e63886 (MD5) Previous issue date: 2017-08-21 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / PROSUP - Programa de Suporte à Pós-Gradução de Instituições de Ensino Particulares / Os cachalotes, Physeter macrocephalus Linnaeus, 1758, são amplamente distribuídos por todos os oceanos do mundo. Todavia, os machos e fêmeas têm diferenças marcadas em seu padrão de distribuição. Globalmente, estudos genéticos, ecológicos e de comportamento vocal comparando populações sugerem a existência de uma forte estrutura social matrilinear com alta filopatria das fêmeas nas regiões tropicais, sendo o fluxo gênico mediado pelos machos. As populações do oceano Atlântico Sul Ocidental (ASO) ainda são tidas como as menos pesquisadas e conhecidas. A fim de identificar possíveis unidades de manejo da espécie na região, o presente estudo apresentou a primeira análise da diversidade genética e a avaliação da potencial estruturação das populações de cachalotes ao longo do ASO, utilizando sequências da região controladora do DNA mitocondrial (DNAmt) de 565 pb obtidas de amostras coletadas na região. Além disso, objetivou-se também estabelecer as relações filogeográficas entre estas populações do ASO e as do resto do mundo. Para a realização deste estudo foram analisadas 58 amostras de cachalotes de três áreas geográficas na costa brasileira (nordeste =15, sudeste =3, sul = 36) e uma da costa argentina (n = 4), e comparadas com 1.577 sequências de outros oceanos, disponíveis publicamente no GenBank (Atlântico = 362, Índico = 159 e Pacífico = 1.056). Além disso, o sexo de 39 espécimes foi determinado molecularmente, a fim de se testar a existência de mais de uma população de cachalotes no ASO, a partir da detecção de fêmeas em áreas além do nordeste brasileiro. A análise das sequências do DNAmt revelou a existência de quatro grupos genéticos e sete haplótipos no ASO. As diversidades haplotípica (Hd) e nucleotídica (π) observadas para a espécie como um todo no ASO foram Hd = 0,6824 e π = 0,002296, respectivamente. Os testes de neutralidade seletiva não sugeriram mudanças significativas no tamanho efetivo e nem expansão populacional recente da espécie no ASO. Os resultados das análises de variância molecular (AMOVA) revelaram que entre 9,14% (ΦST) e 12,31% (FST) da variação genética observada deve-se a diferenças entre as populações. Contudo, essa diferenciação geográfica só foi significativa entre as populações do nordeste e sudeste-sul do Brasil, as quais possuíam altos índices de fixação entre si (nordeste e sudeste-sul do Brasil: FST = 0,1089; ΦST = 0,1378; nordeste do Brasil e Argentina: FST = -0,1076; ΦST = -0,0512; sudeste-sul do Brasil e Argentina: FST = 0,0742; ΦST = 0,1206; P<0,00001). A população do nordeste do Brasil apresentou a diversidade genética mais baixa do estudo (Hd = 0,5128 e π = 0,001796), quando comparada com as populações do sudeste-sul (Hd = 0,6659 e π = 0,002482), e Argentina (Hd = 0,8333 e π = 0,002043). A costa sudeste-sul do Brasil apresentou três haplótipos exclusivos num total de sete, enquanto a costa da Argentina apresentou um haplótipo particular. Apenas a região sudeste-sul apresentou um grupo genético exclusivo, sugerindo que a distribuição da variação genética é melhor compreendida com a existência de três grupos ao longo do ASO (nordeste, sudeste-sul e Argentina), os quais seriam considerados diferentes unidades de manejo. Em relação a comparação mundial entres as populações de cachalotes, foram recuperados 39 haplótipos e quatro grupos genéticos, sendo que apenas o oceano Pacífico apresentou um grupo genético exclusivo. A AMOVA revelou que entre 8,89% (FST) e 16,39% (ΦST) da variação genética observada deve-se a diferenças entre as populações do mundo. A AMOVA indicou que há diferenciação geográfica entre o ASO e os demais oceanos, os quais possuíam altos índices de fixação entre si (ASO e Atlântico Norte: FST = 0,1837; ΦST = 0,3142; Atlântico Sul Ocidental e Índico: FST = 0,0898; ΦST = 0,1661; Atlântico Sul Ocidental e Pacífico: FST = 0,1140; ΦST = 0,0757; P<0,00001). A amostra unificada do oceano ASO não apresentou haplótipos exclusivos. Entretanto, os indivíduos da região sudeste-sul do Brasil compartilharam seis dos seus sete haplótipos com a população do oceano Pacífico. Dos 39 cachalotes com sexo determinado molecularmente nove eram machos e 30 fêmeas, tendo sido encontradas 18 fêmeas na região sul, fora da região tropical, sugerindo que há pelo menos mais de uma população reprodutiva no ASO. Estes resultados, aliados as diferenças de variabilidade genética e ao pouco compartilhamento de haplótipos entre as populações estudadas, sugere que os cachalotes do sudeste-sul do Brasil seriam uma unidade de manejo, potencialmente isolada em termos reprodutivos das demais. Contudo, o DNAmt é um marcador matrilinear e apresenta exclusivamente a história evolutiva das fêmeas. Desta forma, a continuidade destes estudos, incluindo novas amostras e marcadores nucleares, será fundamental para a identificação de reais unidades de manejo da espécie no ASO e principalmente em águas brasileiras. / Sperm whales, Physeter macrocephalus Linnaeus, 1758, are widely distributed throughout the world's oceans. However, males and females have marked differences in their distribution pattern. Worldwide studies on genetics, ecology and vocal behavior comparing populations suggested the existence of a strong matrilineal social structure with female high philopatry in tropical regions, and gene flow mediated by males. The populations of the southwestern Atlantic Ocean (SWA) are still considered the least known and studied. In order to identify possible management units of the species in the SWA, the present study presented the first analysis of the genetic diversity and the evaluation of the potential structuring of the sperm whale populations along the SWA, using sequences from the mitochondrial DNA control region (mtDNA) of 565 bp obtained from samples collected in the region. In addition, it was also aimed to establish the phylogeographic relationships among these SWA populations and those of the rest of the world. Fifty-eight sperm whales from three geographic areas on the Brazilian coast (northeast = 15, southeast = 3, south = 36) and one from the coast of Argentina (n = 4) were analyzed and compared with 1577 sequences from other oceans and available in GenBank (Atlantic = 362, Indian = 159 and Pacific = 1056). In addition, the sex of 39 specimens was molecularly determined in order find females outside of the northeastern coast of Brazil, which would confirm the existence of more than one sperm whale population in the SWA. The analysis of mtDNA sequences revealed the existence of four genetic groups and seven haplotypes in SWA. The haplotype (Hd) and nucleotide (π) diversities observed for the species as a whole in SWA were Hd = 0.6824 e π = 0.002296, respectively. The selective neutrality tests did not suggest significant changes in the effective population size or recent population expansion for the species in SWA. The results of the molecular analysis of variance (AMOVA) revealed that between 9.14% (ΦST) and 12.31% (FST) of the genetic diversity variation observed are due to differences between populations. However, this geographical differentiation was only significant between the populations of northeast and southeast-south Brazil, which had the highest fixation index between them (northeast and southeast-south Brazil: FST = 0.1089, ΦST = 0.1378; northeast Brazil and Argentina: FST = - 0.1076; ΦST = - 0.0512; southeast-south of Brazil and Argentina: FST=0.0742; ΦST=0.1206; P<0.00001). The population of northeastern Brazil had the lowest genetic diversity of the study (Hd = 0.5128 and π = 0.001796), when compared to the southeast-south populations (Hd = 0.6659 and π = 0.002482), and Argentina (Hd = 0.8333 and π = 0.002043). The southeast-south coast of Brazil presented three exclusive haplotypes in a total of seven, while the coast of Argentina presented a particular haplotype. Only the samples from the southeast-south region of Brazil presented an exclusive genetic group, suggesting that the distribution of the genetic variation is better understand with the existence of three groups of sperm whales along the SWA (northeast of Brazil, southeast-south of Brazil and Argentina), which could be considered as different management units. Regarding the worldwide genetic structure of sperm whales, 39 haplotypes and four genetic groups were recovered, and only the Pacific Ocean presented an exclusive genetic group. AMOVA revealed that between 8.89% (FST) and 16.39% (ΦST) of the observed genetic variation was due to differences among the populations of the world. AMOVA indicated that there is a geographical differentiation between the SWA and the other oceans, which had high indexes of fixation between them (SWA and North Atlantic: FST = 0.1837, ΦST = 0.3142, SWA and Indian Ocean: FST = 0.0898; ΦST = 0.1661; SWA and Pacific: FST = 0.11140; ΦST = 0.0757; P<0.00001). The unified sample of the SWA did not present exclusive haplotypes. However, individuals from the southeast-south of Brazil only shared six of seven haplotypes with the population of the Pacific Ocean. From 39 sperm whales with sex molecularly determined, nine were males and 30 females, 18 females were found in the southern region, outside the tropical region, suggesting that there are at least more than one reproductive population in WSA. These results, together with the differences in genetic variability and the lack of sharing of haplotypes among the studied populations, suggest that sperm whales from southeast-south of Brazil would be a management unit, potentially isolated, in reproductive terms, from the others. However, mtDNA is a matrilineal marker and exclusively presents the evolutionary history of females. In this context, the continuity of these studies, including new samples and nuclear markers, will be fundamental for the identification of real units of management of the species in the SWA and particularly in Brazilian waters.

Orientadora: Profa. Dra. Káthia Maria Honório / Dissertação (mestrado) - Universidade Federal do ABC, Programa de Pós-Graduação em Ciência & Tecnologia - Química, 2014. / Polimeros de impressao molecular (MIPs) sao materiais sinteticos que possuem sitios de reconhecimento molecular especificos para determinado analito de interesse (molecula molde), e tem sido empregados para a pre-concentracao e extracao em amostras complexas, considerando a maior seletividade em relacao aos sorventes convencionais empregados na extracao em fase solida. A escolha dos reagentes utilizados na sintese de um MIP deve ser criteriosa, a fim de que sejam criados sitios de ligacoes especificos. Metodos computacionais de Quimica Teorica podem ser empregados para racionalizar o planejamento de MIPs, por intermedio do estudo de parametros fisico-quimicos das moleculas envolvidas e as interacoes que ocorrem no sistema de pre-polimerizacao. Com a utilizacao de metodos de quimica quantica, foi objetivo desse trabalho o estudo previo das interacoes entre as moleculas moldes e os possiveis reagentes empregados na sintese de MIPs, como forma de predicao das melhores interacoes que promovam um MIP de desempenho ideal. O estudo teorico foi direcionado para a molecula de ¿¢9-tetrahidrocanabinol (¿¢9-THC), substancia presente na planta Cannabis sativa e que e a substancia ilicita mais produzida e consumida mundialmente, e seu principal metabolito de excrecao urinario, o 11-nor-9-carboxi-¿¢9-THC (THC-COOH). A simulacao computacional foi realizada utilizando a Teoria do Funcional da Densidade (DFT), e a metodologia envolveu a comparacao das energias de interacao (¿¢E) dos adutos de pre-polimerizacao entre a molecula molde e os reagentes de sintese. Com os resultados das simulacoes, verificou-se que as melhores interacoes ocorrem quando o monomero utilizado tem caracteristica acida, como o acido acrilico e o acido metacrilico; e que o solvente mais adequado para a sintese sao os que apresentam baixa constante dieletrica, como o cloroformio e o tolueno, por interferirem menos na formacao do complexo de interacao entre a molecula molde e o monomero na etapa de pre-polimerizacao. Detalhes das interacoes existentes entre as moleculas do meio reacional tambem foram analisadas e forneceram informacoes sobre a possivel competicao entre as moleculas no acesso aos sitios de interacao das moleculas moldes, informacao crucial para obtencao de MIPs seletivos e de grande afinidade a molecula molde desejada. A partir das predicoes do estudo computacional, os MIPs de THC-COOH foram sintetizados com os monomeros indicados, juntamente com o polimero nao impresso (NIP), em alguns dos solventes simulados (agua, acetonitrila, metanol e agua), para avaliacao da interferencia do solvente na eficiencia do MIP obtido. Foi realizada a analise dos MIPs por espectroscopia vibracional na regiao do infravermelho, e avaliada as bandas de absorcao nas regioes dos grupos C=O e O-H, para analise da formacao de interacoes por ligacao de hidrogenio. Concluiu-se, portanto, que sem substituir os ensaios experimentais, a abordagem computacional pode ser utilizada como uma ferramenta preliminar e complementar a selecao experimental. / Molecularly imprinted polymers (MIPs) are synthetic materials with specific sites of molecular recognition for a given analyte of interest (template molecule), and have been used for the extraction and pre-concentration in complex samples, considering the higher selectivity presented in relation to the sorbent employed in conventional solid phase extraction. The choice of the reagents used in the synthesis of a MIP must be chosen carefully, so that site-specific linkages are created. Computational methods in Theoretical Chemistry can be used to rationalize the planning of MIPs through the study of physico-chemical parameters of the molecules involved and the interactions that occur in the pre-polymerization system. Using quantum chemical methods, the aim of this study was a preliminary investigation of the interactions between the template molecules and the possible reagents used in the synthesis of MIPs, as a way to predict the best interactions that promote MIPs with optimal performance. The theoretical study was directed to the molecule of Ä9-tetrahydrocannabinol (Ä9-THC), present in the plant Cannabis sativa, and that is the illicit substance most widely produced and consumed globally, as well as its major metabolite urinary excretion, 11 -nor-9-carboxy-Ä9-THC (THC-COOH). The computational simulation was performed using the Density Functional Theory (DFT), and the methodology involved the comparison of the interaction energies (ÄE) of adducts in pre-polymerization between the template molecule and reagents for synthesis. From the results of the simulations, it was found that the best interactions occur when the monomer has acid characteristic, such as acrylic acid and methacrylic acid; and that the most suitable solvent for synthesis are those with low dielectric constant such as toluene and chloroform, by interfering less in the formation of complex interaction between the template molecule and monomer in the pre-polymerization step. Details of the existing interactions between the molecules of the reactional medium were analyzed and provided information about the possible competition between the molecules in the access to the interaction sites of the template molecules, which is a crucial information for obtaining selective MIPs and with high affinity to the template molecule desired. From the computational predictions obtainde in this study, the MIPs of THC-COOH was synthesized with the indicated monomers, and also a non-imprinted polymer (NIP), considering some of simulated solvents (water, acetonitrile, methanol and water) for evaluation of the interference of solvent in the efficiency of the MIP obtained. Analyses on the formed MIPs were performed by spectroscopy in the infrared region, and the absorption bands in the regions of the groups C=O and OH were evaluated with the aim of analysing the formation of hydrogen bond interactions. Therefore, it is possible to conclude that, without replacing the experimental trials, the computational approach can be used as a preliminary and complementary tool to experimental selection.

Biomakromoleküle sind in der Natur für viele Abläufe in lebenden Organismen verantwortlich. Dies reicht vom Aufbau der extrazellulären Matrix und dem Cytoskelett über die Erkennung von Botenstoffen durch Rezeptoren bis hin zur Katalyse der verschiedensten Reaktionen in den Zellen selbst. Diese Aufgaben werden zum größten Teil von Proteinen übernommen, und besonders das spezifische Erkennen der Interaktionspartner ist für alle diese Moleküle äußerst wichtig, um eine fehlerfreie Funktion zu gewährleisten. Als Alternative zur evolutiven Erzeugung von optimalen Bindern und Katalysatoren auf der Basis von Aminosäuren und Nukleotiden wurden von Wulff, Shea und Mosbach synthetische molekular geprägte Polymere (molecularly imprinted polymers, MIPs) konzipiert. Das Prinzip dieser künstlichen Erkennungselemente beruht auf der Tatsache, dass sich funktionelle Monomere spezifisch um eine Schablone (Templat) anordnen. Werden diese Monomere dann vernetzend polymerisiert, entsteht ein Polymer mit molekularen Kavitäten, in denen die Funktionalitäten komplementär zum Templat fixiert sind. Dadurch ist die selektive Bindung des Templats in diese Kavitäten möglich. Aufgrund ihrer hohen chemischen und thermischen Stabilität und ihrer geringen Kosten haben “bio-inspirierte” molekular geprägte Polymere das Potential, biologische Erkennungselemente in der Affinitätschromatographie sowie in Biosensoren und Biochips zu ersetzen. Trotz einiger publizierter Sensorkonfigurationen steht der große Durchbruch noch aus. Ein Hindernis für Routineanwendungen ist die Signalgenerierung bei Bindung des Analyten an das Polymer. Eine Möglichkeit für die markerfreie Detektion ist die Benutzung von Kalorimetern, die Bindungs- oder Reaktionswärmen direkt messen können. In der Enzymtechnologie wird der Enzym-Thermistor für diesen Zweck eingesetzt, da enzymatische Reaktionen eine Enthalpie in einer Größenordnung von 5 – 100 kJ/mol besitzen. In dieser Arbeit wird die Herstellung von katalytisch geprägten Polymeren nach dem Verfahren des Oberflächenprägens erstmalig beschrieben. Die Methode zur Immobilisierung des Templats auf der Oberfläche von porösem Kieselgel sowie die Polymerzusammensetzung wurden optimiert. Weiter wird die Evaluation der katalytischen Eigenschaften über einen optischen Test, sowie das erste Mal die Kombination eines kalorimetrischen Transduktors – des Thermistors – mit der Analyterkennung durch ein katalytisch aktives MIP gezeigt. Bei diesen Messungen konnte zum ersten Mal gleichzeitig die Bindung/Desorption, sowie die katalytische Umwandlung des Substrats durch konzentrationsabhängige Wärmesignale nachgewiesen werden. / Bio macromolecules are responsible in nature for many reactions in living organisms. This reaches from the structure of the extra cellular matrix and the cytoskeleton over the recognition of ligands by receptors up to the catalysis of the most diverse reactions in the cells themselves. These tasks are taken over to the largest part by proteins, and particularly specific recognizing of the interaction partners is extremely important for all these molecules, in order to ensure an error free function. As alternative to the evolutionary production of optimal binders and catalysts on the basis of amino acids and nucleotides, synthetic molecularly imprinted polymer (MIPs) were invented by Wulff, Shea and Moosbach. The principle of these artificial recognition elements is based on the fact that functional monomers specifically arrange themselves around a template. If these monomers are copolymerized with crosslinking monomers, a polymer with molecular cavities is created, in which the functionalities are fixed complementary to the template. Thus the selective binding of the template is possible into these cavities. Due to their high chemical and thermal stability and their small costs "bioinspired" molecularly imprinted polymers have the potential to replace biological recognition elements in affinity chromatography as well as in biosensors and biochips. Despite some published sensor configurations the large break-through is still pending. An obstacle for routine application of is the signal generation on connection of the analyte to the polymer. A possibility for marker-free detection is the use of calorimeters, which can measure heats of reaction or adsorption directly. In enzyme technology the enzyme thermistor is used for this purpose, as enzymatic reactions possess enthalpies in an order of 5 - 100 kJ/mol. In this work the production of catalytically imprinted polymers is described for the first time by the procedure of surface imprinting. The method for immobilization of the template on the surface of porous silicagel as well as the polymer composition were optimized. The evaluation of the catalytic characteristics is shown by an optical test, as well as the first time the combination of a calorimetric transducer - the thermistor - with the analyte recognition by a catalytically active MIP. With these measurements for the first time the binding/desorption, as well as the catalytic transformation of the substrate could be proven at the same time by concentration-dependent heat signals.

In the field of chemical and biological sensors, the increased need for better sensitivity, faster response and higher selectivity during an analysis process, requires the development of more and more efficient transducing sensing layers. In this context, and with the aim to detect small non-electroactive molecules, such as atrazine (ATZ), we designed, characterized and developed sensing layers constituted by functionalized Molecularly Imprinted Conducting Polymers (MICP) and we integrated them into electrochemical and gravimetrical sensors. Starting from acetonitrile pre-polymerization media containing ATZ as template molecules in the presence of thiophene-based functional monomers (FM, namely TMA, TAA, EDOT, TMeOH or Th), differently functionalized and structurally different polythiophene-based FM-MICP films were electrosynthesized onto gold substrates and used for ATZ detection. The sensing properties of FM-MICP layers were shown to result from the presence in their backbones of pre-shaped FM-functionalized imprinted cavities which keep the memory of the targets. Nevertheless, non-specific adsorption onto the surface of the sensing layers takes place systematically, which affects the selectivity of the recognition process. Thanks to surface characterization techniques, we highlighted the influence of the thickness and of the structural properties of the layers on the efficiency of the recognition process. Besides, this latter was shown to operate in the bulk of the polymer matrixes thanks to layers porosity. On another hand, electrochemical measurements correlated with semi-empirical calculations demonstrated the influence of the nature of FM on the strength of the ATZ-FM interaction in the pre-polymerization medium, and then on the number of ATZ molecular imprints and on the sensitivity towards ATZ of the FM-MICP layers. We showed that TAA-MICP, which presents a low limit of detection (10-9 mol L-1) and a large dynamic range (10-8 to 10-4 mol L-1), is the best sensing layer since it offers the best compromise between high level of specific detection of ATZ and low level of non-specific adsorption. Finally, TAA-MICP was used as sensitive layer in an original Electrochemical Surface Acoustic Wave sensor (ESAW) which enabled simultaneous coupled gravimetric and electrochemical measurements.

L'industrie cosmétique a un recours croissant aux espèces végétales comme sources de principesactifs naturels. Leur extraction nécessite des supports sélectifs tels que les polymères à empreintesmoléculaires (MIP). Les travaux de cette thèse reposent sur le développement de MIP pourl’extraction sélective de la glucosamine, de la fructosazine et de la 2,5-déoxyfructosazine.Dans une première partie, trois approches ont été développées pour extraire la glucosamine par desMIP : l’approche covalente, semi-covalente et non covalente. Pour chacune, les différents paramètresintervenant dans la synthèse des MIP ont été optimisés. Les meilleurs résultats ont été obtenus avecun MIP synthétisé selon une approche non covalente ionique reposant sur la complexation de laglucosamine par un acide sulfonique. Les performances du MIP se sont avérées supérieures à cellesde supports commerciaux et des extractions à partir de végétaux ont été réalisées. Le potentielindustrialisable du MIP a été validé lors de premiers tests à plus grande échelle.Dans une deuxième partie, l’extraction simultanée de la fructosazine et de la 2,5-déoxyfructosazine aété réalisée suite au développement d’un MIP synthétisé selon une approche covalente reposant surla formation d’esters boroniques. Une méthode de synthèse originale est exposée puisque lestemplates ont été formés in situ lors de la polymérisation. Le MIP obtenu s’est avéré sélectif dechaque composé et a permis de purifier et de séparer la fructosazine et la 2,5-déoxyfructosazine dematrices végétales et alimentaires.Tous ces travaux ont été réalisés dans une démarche éco-responsable s’appuyant sur l’emploi desolvants aqueux lors de la polymérisation et de l’extraction. / The cosmetic industry uses plants as sources of natural active ingredients. The extraction of theseactive ingredients requires selective extraction method such as molecularly imprinted polymers (MIP)technique. This thesis describes the development of MIP for the selective extraction of glucosamine,fructosazine and 2,5-déoxyfructosazine.In the first part, three approaches were developed to extract glucosamine by MIP technique: thecovalent approach, semi-covalent and noncovalent. For each approach, the various parametersinvolved in the synthesis of the MIP were optimized. The best results were obtained with a MIPsynthesized with a non-covalent ionic approach based on the complexation of glucosamine by asulfonic acid. The MIP exhibits higher performance than commercial media and extractions fromplants were performed. The potential for industrialization of the MIP was validated during initial testson a larger scale.In the second part, the simultaneous extraction of fructosazine and 2,5-déoxyfructosazine wasperformed following the development of a MIP synthesized using a covalent approach based on theformation of boronic esters. An original synthesis method is exposed since the templates were formedin situ during the polymerization. The MIP obtained showed good selectivity for each compound andallowed to separate and purify fructosazine and 2,5-déoxyfructosazine from plant and food matrices.All these works were performed according to an eco-friendly approach based on the use of aqueoussolvents as solvents for polymerization and extraction.

Ce mémoire présente une exploration des polymères à empreintes moléculaires (MIP) comme outils d’une libération contrôlée de bioactifs olfactifs ou pour le criblage/préselection de composés à activité antivirales ou anti-tumorales sur le site actif d’une enzyme. La première partie est une étude de la complexation de la vanilline sur des billes polymériques sphériques en vue d’une libération contrôlée (pH, salinité, …). Ces études portent sur les caractéristiques de l'absorption et la libération de la molécule d'intérêt dans le milieu aqueux sur les microsphères fonctionnalisées fourni par Merck ESTAPOR® Microsphères. Nous avons ensuite synthétisé divers MIP de vanilline au format monolithique. Plusieurs stratégies d’impression ont été étudiées: non covalente, covalente et semi-covalente. La composition du MIP préparé dans chaque approche a été optimisée pour obtenir les meilleures propriétés et performances. L'affinité, la sélectivité et la capacité du MIP ont été déterminées. Les MIPs ont été évalués par extraction en phase solide (SPE) d'analogues structuraux de la vanilline dans des échantillons naturels (extrait de vanille, vin). La deuxième partie de ce mémoire concerne l’évaluation de MIPs de l’adénosine 5’-monophosphate (AMP) Le polymère a été préparé par une approche non-covalente et son efficacité de recapture a été caractérisée par analyse frontale (FA). L’analyse frontale est une technique qui permet de discriminer des interactions spécifiques des non spécifiques et de comprendre les mécanismes de liaison dans des cavités spécifiques. / This thesis report presents the exploration of molecularly imprinted polymers (MIP) for the application in controlled release and targeting antivirus and anticancer drugs. The first part of this study describes the imprinting of vanillin as a monolith. Several strategies were studied: non-covalent, covalent and semi-covalent. The composition of the MIP prepared in each approach was optimized to obtain the best properties and performance. The affinity, selectivity and capacity of MIP were determined. MIPs were evaluated in solid-phase extraction (SPE) of structural analogues in natural samples (vanilla extract, wine). We also present the study of the exploration of spherical beads as potential tools for the controlled release of vanillin. These studies concern the characteristics of uptake and release of the molecule of interest in the aqueous medium on functionalised microspheres supplied by Merck ESTAPOR Microspheres®. The second part of this thesis is devoted to studies on the evaluation of MIP of adenosine 5'-monophosphate (AMP). The polymer was prepared in non-covalent approach and efficiency of binding was characterised using frontal analysis (FA). FA is a useful technique that allows discriminate specific and nonspecific interactions and to understand the binding mechanisms in specific cavities.