Monolithische Phasen werden seit ca. 20 Jahren entwickelt und sind in den letzten Jahren eine attraktive Alternative zu etablierten mit Partikeln gefüllten Säulen geworden. Sie werden in anorganische Phasen und organische Polymermonolithe unterteilt. Monolithe bestehen aus einem einzigen, durchgehenden Stück. Charakteristisch ist das sie durchziehende Porennetzwerk, durch das der Eluent mit geringerem hydraulischen Widerstand fließen kann und das somit schnellere Flussraten ermöglicht. Polymermonolithe werden vorwiegend für die Separation großer Biomoleküle aufgrund eines durch Konvektion beschleunigten Massentransfers eingesetzt. Zudem sind sie über einen breiten pH-Wert-Bereich stabil und können direkt (in situ) im gewünschten Format polymerisiert werden. In der vorliegenden Arbeit gelang die Herstellung neuartiger epoxidbasierter Phasen nach einem von Weller et al. entwickelten Konzept, die im Affinitätsexperiment angewendet wurden. Die Herstellung erfolgte durch Autopolymerisation polyfunktioneller Glycidylether. Für die Funktionalisierung wurden nicht polymerisierte Epoxide genutzt. Als Monomere dienten TEPIC, GE 100 sowie GE 500. Die Arbeiten konzentrierten sich vor allem auf die bei Raumtemperatur durchführbaren Synthesen mit dem höher funktionellen GE 500. Die Polymerisationsbedingungen wurden hinsichtlich Porogenmischung und -anteil optimiert. Eine mit 75 Vol.-% Porogen (Dioxan/ MTBE (2:3)) hergestellte und mit rProtein A funktionalisierte Kapillarsäule (66 %, 12 µm, 7m2/g) ergab im Affinitätsexperiment eine Kapazität von 0,44 mg/mL aus Kaninchenserum isolierbarem IgG. Durch Beimischung von 60 % BDE konnte der Epoxidgehalt vervierfacht und die Porengröße auf 400 nm bei 59 % Porosität reduziert werden. Die spezifische Oberfläche wurde verdreifacht und die Kapazität präparierter Disks auf 0,90 mg/mL etwa verdoppelt. Die in dieser Arbeit entwickelten Disks können zur Isolierung von IgG aus einer komplexen Probe, wie beispielsweise Blutserum, eingesetzt werden. / Monolithic supports have been developed since 20 years and have become an attractive alternative to well-established columns packed with particles over the past years. They are classified into inorganic media and organic polymer monoliths. Monoliths consist of a single, continuous piece with an integrated characteristic porous network through which the eluent can flow with lower hydraulic resistance and which consequently offers higher flow rates. Due to an accelerated mass transfer caused by convection polymer monoliths are mainly used for separation of large biomolecules. In addition, they are stable over a wide pH range and can be polymerized directly (in situ) in the desired format. In the present work the successful preparation of new epoxide-based supports according to a concept introduced by Weller et al. as well as their application in affinity chromatography are reported. Their preparation was carried out by self-polymerization of polyfunctional glycidyl ethers and for functionalization non-polymerized epoxide groups were used. As monomers TEPIC, GE 100 and GE 500 were utilized. The work has focused especially on the polymerization of the higher functional GE 500, which can be perfomed at room temperature and was optimized in terms of both composition and amount of porogen. The extraction of IgG from rabbit serum with a capillary column (66 %, 12 µm, 7m2/g) prepared by 75 vol.-% porogen (dioxane/ MTBE (2:3)) and functionalized with rprotein A resulted in a capacity of 0,44 mg/mL. By addition of 60 % BDE the epoxide content was quadrupled and the pore size reduced to 400 nm while maintaining consistently high porosity of 59 %. The specific surface area was tripled and the capacity of prepared disks approximately doubled to 0,90 mg/mL. The disks developed in this work can be applied for the isolation of IgG from complex samples such as serum.

Background: Separation science is heavily reliant on materials to fulfill ever more complicated demands raised by other areas of science, notably the rapidly expanding molecular biosciences and environmental monitoring. The key to successful separations lies in a combination of physical properties and surface chemistry of stationary phases used in liquid chromatographic separation, and this thesis address both aspects of novel separation materials. Methods: The thesis accounts for several approaches taken during the course of my graduate studies, and the main approaches have been i) to test a wild-grown variety of published methods for surface treatment of fused silica capillaries, to ascertain firm attachment of polymeric monoliths to the wall of microcolumns prepared in silica conduits; ii) developing a novel porogen scheme for organic monoliths including polymeric porogens and macromonomers; iii) evaluating a mesoporous styrenic monolith for characterization of telomers intended for use in surface modification schemes and; iv) to critically assess the validity of a common shortcut used for estimating the porosity of monoliths prepared in microconduits; and finally v) employing plasma chemistry for activating and subsequently modifying the surface of rigid, monodisperse particles prepared from divinylbenzene. Results: The efforts accounted for above have resulted in i) better knowledge of the etching and functionalization parameters that determine attachment of organic monoliths prepared by radical polymerization to the surface of silica; ii) polar methacrylic monoliths with a designed macroporosity that approaches the desired "connected rod" macropore morphology; iii) estab¬lishing the usefulness of monoliths prepared via nitroxide mediated polymerization in gradient polymer elution chromatography; iv) proving that scanning electron microscopy images are of limited value for assessing the macroporous properties of organic monoliths, and that pore measurements on externally polymerized monolith cocktails do not represent the porous properties of the same cocktail polymerized in narrow confinements; and v) showing that plasma bromination can be used as an activation step for rigid divinylbenzene particles to act as grafting handles for epoxy-containing telomers, that can be attached in a sufficiently dense layer and converted into carboxylate cation exchange layer that allows protein separations in fully aqueous eluents.

Segundo a Organização Mundial da Saúde (OMS), a esquizofrenia é reconhecida como um transtorno neuropsiquiátrico grave que afeta mais de 21 milhões de pessoas em todo o mundo. Para diminuir os sintomas associados a esta doença, a maioria dos pacientes esquizofrênicos fazem uso concomitantemente de antipsicóticos, antidepressivos, ansiolíticos e anticonvulsivantes. O desenvolvimento de métodos analíticos para a determinação desses fármacos em fluidos biológicos é importante para ajustar as doses administradas, minimizar os efeitos adversos e verificar a anuência do paciente à terapia. A química analítica moderna tem se direcionado para a simplificação, através da miniaturização dos sistemas analíticos. Neste contexto, pode-se destacar a técnica de microextração em sorvente empacotado (MEPS). O desenvolvimento de novas fases extratoras para MEPS, como, os monolitos de sílica híbrida permitem pré-concentração seletiva dos analitos. Neste projeto monolitos de sílica híbrida funcionalizados com grupos aminopropil ou cianopropil foram sintetizados pelo processo sol-gel. Estes monolitos apresentaram estrutura contínua, uniforme e porosa, como evidenciado pelas imagens de microscopia eletrônica de varredura (MEV). As análises de espectroscopia vibracional na região do infravermelho por transformada de Fourier (FTIR) dos monolitos de sílica híbrida exibiram picos facilmente identificáveis, característicos dos grupos cianopropil e aminopropil. Estes monolitos foram avaliados como fase estacionária para a MEPS para a determinação de fármacos em amostras de plasma por cromatografia líquida acoplada a espectrometria de massa em Tandem (LC-MS/MS). As variáveis MEPS (influência do pH na sorção dos analitos, número de ciclos aspirar/dispensar, etapa de limpeza do sorvente e solvente de eluição) foram otimizadas para aumentar a eficiência da extração. Em comparação com a fase de sílica híbrida aminopropil, a fase de sílica híbrida cianopropil apresentou maior capacidade de sorção para os fármacos em estudo. O método desenvolvido, MEPS/LC-MS/MS, empregando a fase extratora de sílica híbrida cianopropil, apresentou linearidade adequada com concentrações que variaram desde o limite inferior de quantificação (0,05-1,00 ng.mL-1) até o limite superior de quantificação (40-10.500 ng.mL-1) com coeficientes de determinação (R2) maiores do que 0,9955. Este método apresentou precisão com coeficientes de variação (CV) inferiores a 14% e exatidão com erro padrão relativo (EPR) de -12 a 14%. O método MEPS/LC-MS/MS foi aplicado com êxito para análise simultânea de cinco antipsicóticos (olanzapina, quetiapina, clozapina, haloperidol e clorpromazina), em combinação com sete antidepressivos (mirtazapina, paroxetina, citalopram, sertralina, imipramina, clomipramina e fluoxetina), dois anticonvulsivantes (carbamazepina e lamotrigina) e dois ansiolíticos (diazepam e clonazepam) em amostras de plasma de pacientes esquizofrênicos para fins de monitorização terapêutica. / According to World Health Organization (WHO), schizophrenia is recognizably a devastating neuropsychiatric disorder that affects more than 21 million people worldwide. To lessen the symptoms associated with the disease, most schizophrenic patients use other classes of drugs such as antidepressants, anxiolytics, and anticonvulsants concomitantly with antipsychotics. Developing analytical methods to quantify these drugs in biological fluids is important in therapeutic drug monitoring (TDM) to adjust doses, minimize adverse effects, and check patient adherence to the therapy. Regarding miniaturization and automation, microextraction by packed sorbent (MEPS) is a promising sample preparation technique. Sample preparation of biological matrixes is an important step in analytical processes: it removes endogenous components from the sample and pre-concentrates trace-level analytes. The development of new phases for MEPS such as the hybrid silica monoliths allows selective pre-concentration of the analytes. The present study reports on the synthesis of two hybrid silica monoliths functionalized with aminopropyl or cyanopropyl groups by the solgel process; evaluates these monoliths as selective stationary phase for MEPS to determine drugs in plasma samples by liquid chromatography-tandem mass spectrometry (LC-MS/MS); and discusses important factors (influence of pH on the sorption of analytes, number of draw/eject cycles, washing step, and elution solvent) related to the optimization of MEPS efficiency. The prepared hybrid silica monoliths consisted of a uniform, porous, and continuous silica monolithic network, as shown by scanning electron microscopy (MEV) images. The Fourier-transform infrared spectroscopy (FTIR) spectra of the hybrid silica monoliths displayed readily identifiable peaks, characteristic of the cyanopropyl and aminopropyl groups. Compared with the aminopropyl hybrid silica phase, the cyanopropyl hybrid silica phase exhibited higher binding capacity for most of the target drugs. The developed method, MEPS/LC-MS/MS, afforded adequate linearity at concentrations ranging from the lower limit of quantification (0.05 to 1.00 ng.mL-1) to the upper limit of quantification (40 to 10500 ng.mL-1); the coefficients of determination (R2) were higher than 0.9955. The precision of the method presented coefficients of variation (CV) lower than 14%; the relative standard error (RSE) of the accuracy ranged from -12 to 14%. The developed MEPS/LC-MS/MS method allowed for simultaneous analysis of five antipsychotics (olanzapine, quetiapine, clozapine, haloperidol, and chlorpromazine) in combination with seven antidepressants (mirtazapine, paroxetine, citalopram, sertraline, imipramine, clomipramine, fluoxetine), two anticonvulsants (carbamazepine and lamotrigine), and two anxiolytics (diazepam and clonazepam) in plasma samples from schizophrenic patients, which should be valuable for TDM purposes.

To date, there has been little research into the role of microbial community structure in the functioning of the soil ecosystem and on the links between microbial biomass size, microbial activity and key soil processes that drive nutrient availability. The maintenance of structural and functional diversity of the soil microbial community is essential to ensure the sustainability of agricultural production systems. Soils of the same type with similar fertility that had been under long-term organic and conventional crop management in Canterbury, New Zealand, were selected to investigate relationships between microbial community composition, function and potential environmental impacts. The effects of different fertilisation strategies on soil biology and nitrogen (N) dynamics were investigated under field (farm site comparison), semi-controlled (lysimeter study) and controlled (incubation experiments) conditions by determining soil microbial biomass carbon (C) and N, enzyme activities (dehydrogenase, arginine deaminase, fluorescein diacetate hydrolysis), microbial community structure (denaturing gradient gel electrophoresis following PCR amplification of 16S and 18S rDNA fragments using selected primer sets) and N dynamics (mineralisation and leaching). The farm site comparison revealed distinct differences between the soils in microbial community structure, microbial biomass C (conventional > organic) and arginine deaminase activity (organic > conventional). In the lysimeter study, the soils were subjected to the same crop rotation (barley (Hordeum vulgare L.), maize (Zea mays L.), rape (Brassica napus L. ssp. oleifera (Moench)) plus a lupin green manure (Lupinus angustifolius L.) and two fertiliser regimes (following common organic and conventional practice). Soil biological properties, microbial community structure and mineral N leaching losses were determined over 2½ years. Differences in mineral leaching losses were not significant between treatments (total organic management: 24.2 kg N per ha; conventional management: 28.6 kg N per ha). Crop rotation and plant type had a larger influence on the microbial biomass, activity and community structure than fertilisation. Initial differences between soils decreased over time for most biological soil properties, while they persisted for the enzyme activities (e.g. dehydrogenase activity: 4.0 and 2.9 µg per g and h for organic and conventional management history, respectively). A lack of consistent positive links between enzyme activities and microbial biomass size indicated that similarly sized and structured microbial communities can express varying rates of activity. In two successive incubation experiments, the soils were amended with different rates of a lupin green manure (4 or 8t dry matter per ha), and different forms of N at 100 kg per ha (urea and lupin) and incubated for 3 months. Samples were taken periodically, and in addition to soil biological properties and community structure, gross N mineralisation was determined. The form of N had a strong effect on microbial soil properties. Organic amendment resulted in a 2 to 5-fold increase in microbial biomass and enzyme activities, while microbial community structure was influenced by the addition or lack of C or N substrate. Correlation analyses suggested treatment-related differences in nutrient availability, microbial structural diversity (species richness or evenness) and physiological properties of the microbial community. The findings of this thesis showed that using green manures and crop rotations improved soil biology in both production systems, that no relationships existed between microbial structure, enzyme activities and N mineralisation, and that enzyme activities and microbial community structure are more closely associated with inherent soil and environmental factors, which makes them less useful as early indicators of changes in soil quality.

In der vorliegenden Arbeit wird die kationische Polymerisation neuer Furanmonomere beschrieben, die zu anorganisch-organischen Nanokompositen führt. Die kationische Polymerisation des Tetrafurfuryloxysilans steht dabei im Vordergrund. Ausgehend von den synthetisierten Kompositen wird die Herstellung von anorganischen Oxiden durch thermische Oxidation und von Kohlenstoffmaterialien durch thermische Behandlung in Schutzgasatmosphäre beschrieben. Die Charakterisierung der Komposite, Oxide und Kohlenstoffmaterialien erfolgt mittels Festkörper-NMR-Spektroskopie, Röntgenbeugung und Elektronenmikroskopie. Das Prinzip der neu entdeckten Zwillingspolymerisation wird vorgestellt und anhand verschiedener Beispiele auf seine weitere Anwendbarkeit zur Synthese anorganisch-organischer Kompositmaterialien überprüft.

Abstract The harmful emissions of chlorinated volatile organic compounds (CVOCs) originate only from man-made sources. CVOCs are used in a variety of applications from pharmaceuticals production to decaffeination of coffee. Currently, CVOC emissions are limited by strict legislation. For these reasons, efficient CVOC abatement technologies are required. Catalytic oxidation is very promising option for this purpose, since catalysts can be tailored to each case to maximize the efficiency and minimize the formation of unwanted products, such as dioxins or Cl2. The goal of this thesis was to study the role of the physico-chemical properties of catalysts in dichloromethane (DCM) oxidation. To reach the aim, several catalytic materials were prepared and characterized, and their performance was tested in total oxidation of DCM. The catalytic materials used were powders of four single metal oxides (γ-Al2O3, TiO2, CeO2, MgO), three mixed oxides (Al2O3-xSiO2) washcoated on a cordierite monolith and four active phases (Pt, Cu, V, Mn). At first, support properties were studied. It was found that the DCM conversion and HCl production are dependent on support acidity when the studied single oxides are considered. The best DCM conversions and HCl yields were observed with the support having the highest total acidity (γ-Al2O3). Further, the quality of the by-products formed was dependent on the type of the acid sites present on the support surface. Secondly, the impregnation of the active compound was observed to improve the selectivity of the material. From the tested active phases, Pt presented the best performance, but also V2O5 and CuO showed almost equal performances. Especially CuO supported on γ-Al2O3, that had less formation of by-products and is less toxic than V-containing oxides, seems to be a promising alternative to Pt. Concerning stability, no deactivation was observed after 55h of testing of Pt/Al2O3. Furthermore, in the used reaction conditions, the formation of CuCl2 is not thermodynamically favoured. Finally, the good characteristics of the powder form catalysts were successfully transferred to the monolith. The performance of the Pt/90Al2O3-10SiO2 catalyst in DCM oxidation was improved when the channel density was increased due to an increase in geometric surface area and mechanical integrity factor, and a decrease in open fraction area and thermal integrity factor. / Tiivistelmä Haitallisten kloorattujen orgaanisten yhdisteiden (CVOC) päästöt ovat ihmisten aiheuttamia. CVOC-yhdisteitä käytetään mm. liuottimina lääkeaineiden valmistuksessa ja kofeiinin poistossa. Nykyisin CVOC-päästöjä rajoitetaan tiukalla lainsäädännöllä. Näistä syistä tehokas CVOC-yhdisteiden käsittelymenetelmä on tarpeen. Katalyyttinen hapetus on hyvä vaihtoehto tähän tarkoitukseen, koska katalyytit voidaan räätälöidä niin, että puhdistuksen tehokkuus saadaan maksimoitua samalla kun ei-haluttujen tuotteiden, kuten dioksiinit ja kloorikaasu, muodostuminen voidaan minimoida. Tämän väitöskirjatyön tavoitteena oli selvittää katalyyttien fysikaalis-kemiallisten ominaisuuksien yhteyksiä dikloorimetaanin (DCM) hapetukseen. Tavoitteen saavuttamiseksi valmistettiin useita katalyyttejä, jotka karakterisoitiin ja testattiin DCM:n kokonaishapetuksessa. Työssä tutkittiin neljää jauhemaista metallioksidia (γ-Al2O3, TiO2, CeO2 ja MgO), kolmea metallioksidiseosta (Al2O3-xSiO2), jotka pinnoitettiin kordieriittimonoliitille, sekä neljää aktiivista ainetta: Pt, Cu, V and Mn. Aluksi työssä keskityttiin tukiaineiden ominaisuuksiin. Työn tulokset osoittivat, että DCM:n konversio ja HCl:n tuotanto ovat riippuvaisia tukiaineen happamuudesta. Paras tulos saavutettiin alumiinioksidilla, jolla oli korkein kokonaishappamuus. Lisäksi havaittiin, että sivutuotteiden laatu riippuu tukiaineen pinnalla olevien happopaikkojen tyypistä. Aktiivisen aineen impregnointi tukiaineeseen paransi materiaalin selektiivisyyttä. Tutkituista aineista Pt osoittautui parhaimmaksi, mutta myös V2O5 ja CuO olivat lähes yhtä hyviä. Erityisesti CuO-katalyytti, joka tuotti vähemmän sivutuotteita ja joka on materiaalina vähemmän haitallinen kuin V2O5, osoittautui lupaavaksi jalometallikatalyyttien korvaajaksi. Materiaalien stabiilisuuteen liittyen Pt/Al2O3-katalyytin toiminnassa ei havaittu muutoksia 55 tunnin testauksen jälkeen. Lisäksi CuCl2:n muodostuminen ei mallinnuksen mukaan ole termodynaamisesti todennäköistä tutkituissa reaktio-olosuhteissa. Jauhemaisen katalyytin hyvät ominaisuudet pystyttiin pinnoituksessa siirtämään monoliittirakenteiseen katalyyttiin. Pt/90Al2O3-10SiO2 -katalyytin aktiivisuus DCM:n hapetuksessa tehostui, kun monoliitin aukkoluku kasvoi aiheutuen suuremmasta geometrisestä pinta-alasta ja mekaanisesta eheystekijästä sekä pienemmästä avoimen pinnan osuudesta ja termisestä eheystekijästä.

Att designa mjukvaruapplikationer med en viss struktur kan ofta framhäva efterfrågade egenskaper. För att välja rätt arkitektur behövs ofta övervägningar, och ibland till och med kompromisser, göras om applikationens planerade karaktär. Det är ofta bra att i detta stadie ha en klar bild om vilka attribut en applikation önskas ha. Ett av de viktigare attributen på sikt är skalbarhet. Kunskapen om olika arkitekturers skalbarhet spelar en definitiv roll i designfasen, vilket avgör hur en applikation senare skalas. På senare år har mikrotjänst-arkitektur blivit ett populärt sätt att bygga mjukvara på där den höga skalbarheten sägs vara en bidragande faktor. Detta arbete har till syfte att undersöka skalbarheten hos mikrotjänst-arkitektur i förhållande till monolitisk arkitektur och visa hur detta kvalitetsattribut påverkas när en transformering från en monolit till en mikrotjänst-arkitektur görs. Arbetet har valt att utgå ifrån en existerande modul i en E-handelsplattform med öppen källkod. Modulen som transformerades till en mikrotjänst, skalades horisontellt för respektive arkitektur och applikations-version. Vid användandet av lämpliga verktyg, såsom Docker, visar resultatet att horisontell skalbarhet finns i högre grad hos mikrotjänst-arkitekturen och fortsätter därefter vara hög. Skalning av mikrotjänster kan göras med en högre precision av det som önskas förändras. Detta står i kontrast till den monolitiska strukturen, där skalning begränsas av prestandan av den miljö där mjukvaruapplikationen körs. Efter transformationen till en mikrotjänst-arkitektur ökade skalbarheten, då skalningsmetoden gjordes med mer finkornighet och isolering av den utvalda modulen. För att individuellt skala den monolitiska modulen horisontellt behövdes förändringen göras virtuellt med hjälp av bakgrundsprocesser. Denna lösning visar sig vara en indirekt skalning av hela den monolitiska strukturen. Utöver horisontell skalbarhet fokuserar utvärderingen av resultatet på kvalitativa attribut i form av simplicitet, autonomi och modularitet. / In designing software applications, a chosen structure can often accentuate desired properties. To choose the correct architecture, one must often do considerations and sometimes even compromises, about the intended characteristics of the application. In that stage it is often well motivated to have a clear picture about which attributes the application shall possess. Over time, one of the most important attributes is scalability. The knowledge about the scalability of different architectures could play a crucial part in the design phase, determining how an application is scaled in the future. In recent years Microservice Architecture has been a popular way of building software and its high scalability is said to be a contributing factor. This work has the purpose of examine the scalability of microservice architecture relative to the monolithic architecture and how this quality attribute is affected after a transformation is done from a monolith to a microservice system. This work is based on an existing module from an open source E-commerce platform. The module was first transformed into a working microservice, then both architectures was horizontally scaled. Using suitable tools such as Docker, the result of this work shows that horizontal scalability exists in a high degree within the microservice architecture and continues being high there after. Scaling of microservices can be done with higher precision of what are to be changed. This stands in relation to the monolithic approach where scaling is limited to the performance of the environment where the software application is running. The transformation to a microservice architecture resulted in an increase of scalability. The scaling method was more fined-grained and isolated to the selected module. In contrast, individual horizontal scaling of the monolithic module was required to be done virtually with background processes. This concluded in an indirect scaling of the whole structure of the monolith. Besides horizontal scalability, the evaluation is focused on the system quality attributes of simplicity, autonomy and modularity.

Segundo a Organização Mundial da Saúde (OMS), a esquizofrenia é reconhecida como um transtorno neuropsiquiátrico grave que afeta mais de 21 milhões de pessoas em todo o mundo. Para diminuir os sintomas associados a esta doença, a maioria dos pacientes esquizofrênicos fazem uso concomitantemente de antipsicóticos, antidepressivos, ansiolíticos e anticonvulsivantes. O desenvolvimento de métodos analíticos para a determinação desses fármacos em fluidos biológicos é importante para ajustar as doses administradas, minimizar os efeitos adversos e verificar a anuência do paciente à terapia. A química analítica moderna tem se direcionado para a simplificação, através da miniaturização dos sistemas analíticos. Neste contexto, pode-se destacar a técnica de microextração em sorvente empacotado (MEPS). O desenvolvimento de novas fases extratoras para MEPS, como, os monolitos de sílica híbrida permitem pré-concentração seletiva dos analitos. Neste projeto monolitos de sílica híbrida funcionalizados com grupos aminopropil ou cianopropil foram sintetizados pelo processo sol-gel. Estes monolitos apresentaram estrutura contínua, uniforme e porosa, como evidenciado pelas imagens de microscopia eletrônica de varredura (MEV). As análises de espectroscopia vibracional na região do infravermelho por transformada de Fourier (FTIR) dos monolitos de sílica híbrida exibiram picos facilmente identificáveis, característicos dos grupos cianopropil e aminopropil. Estes monolitos foram avaliados como fase estacionária para a MEPS para a determinação de fármacos em amostras de plasma por cromatografia líquida acoplada a espectrometria de massa em Tandem (LC-MS/MS). As variáveis MEPS (influência do pH na sorção dos analitos, número de ciclos aspirar/dispensar, etapa de limpeza do sorvente e solvente de eluição) foram otimizadas para aumentar a eficiência da extração. Em comparação com a fase de sílica híbrida aminopropil, a fase de sílica híbrida cianopropil apresentou maior capacidade de sorção para os fármacos em estudo. O método desenvolvido, MEPS/LC-MS/MS, empregando a fase extratora de sílica híbrida cianopropil, apresentou linearidade adequada com concentrações que variaram desde o limite inferior de quantificação (0,05-1,00 ng.mL-1) até o limite superior de quantificação (40-10.500 ng.mL-1) com coeficientes de determinação (R2) maiores do que 0,9955. Este método apresentou precisão com coeficientes de variação (CV) inferiores a 14% e exatidão com erro padrão relativo (EPR) de -12 a 14%. O método MEPS/LC-MS/MS foi aplicado com êxito para análise simultânea de cinco antipsicóticos (olanzapina, quetiapina, clozapina, haloperidol e clorpromazina), em combinação com sete antidepressivos (mirtazapina, paroxetina, citalopram, sertralina, imipramina, clomipramina e fluoxetina), dois anticonvulsivantes (carbamazepina e lamotrigina) e dois ansiolíticos (diazepam e clonazepam) em amostras de plasma de pacientes esquizofrênicos para fins de monitorização terapêutica. / According to World Health Organization (WHO), schizophrenia is recognizably a devastating neuropsychiatric disorder that affects more than 21 million people worldwide. To lessen the symptoms associated with the disease, most schizophrenic patients use other classes of drugs such as antidepressants, anxiolytics, and anticonvulsants concomitantly with antipsychotics. Developing analytical methods to quantify these drugs in biological fluids is important in therapeutic drug monitoring (TDM) to adjust doses, minimize adverse effects, and check patient adherence to the therapy. Regarding miniaturization and automation, microextraction by packed sorbent (MEPS) is a promising sample preparation technique. Sample preparation of biological matrixes is an important step in analytical processes: it removes endogenous components from the sample and pre-concentrates trace-level analytes. The development of new phases for MEPS such as the hybrid silica monoliths allows selective pre-concentration of the analytes. The present study reports on the synthesis of two hybrid silica monoliths functionalized with aminopropyl or cyanopropyl groups by the solgel process; evaluates these monoliths as selective stationary phase for MEPS to determine drugs in plasma samples by liquid chromatography-tandem mass spectrometry (LC-MS/MS); and discusses important factors (influence of pH on the sorption of analytes, number of draw/eject cycles, washing step, and elution solvent) related to the optimization of MEPS efficiency. The prepared hybrid silica monoliths consisted of a uniform, porous, and continuous silica monolithic network, as shown by scanning electron microscopy (MEV) images. The Fourier-transform infrared spectroscopy (FTIR) spectra of the hybrid silica monoliths displayed readily identifiable peaks, characteristic of the cyanopropyl and aminopropyl groups. Compared with the aminopropyl hybrid silica phase, the cyanopropyl hybrid silica phase exhibited higher binding capacity for most of the target drugs. The developed method, MEPS/LC-MS/MS, afforded adequate linearity at concentrations ranging from the lower limit of quantification (0.05 to 1.00 ng.mL-1) to the upper limit of quantification (40 to 10500 ng.mL-1); the coefficients of determination (R2) were higher than 0.9955. The precision of the method presented coefficients of variation (CV) lower than 14%; the relative standard error (RSE) of the accuracy ranged from -12 to 14%. The developed MEPS/LC-MS/MS method allowed for simultaneous analysis of five antipsychotics (olanzapine, quetiapine, clozapine, haloperidol, and chlorpromazine) in combination with seven antidepressants (mirtazapine, paroxetine, citalopram, sertraline, imipramine, clomipramine, fluoxetine), two anticonvulsants (carbamazepine and lamotrigine), and two anxiolytics (diazepam and clonazepam) in plasma samples from schizophrenic patients, which should be valuable for TDM purposes.

Une des évolutions dans le domaine de l'analyse chimique concerne la miniaturisation des systèmes d'analyse. Cette tendance s'accompagne du développement de nouvelles approches expérimentales basées, par exemple, sur l'intégration de plusieurs étapes analytiques, couplées en ligne en système miniaturisé. Cette intégration, en ligne, d'étapes mettant en jeu des mécanismes de séparation différents et généralement orthogonaux, implique cependant d'être capable de définir des zones (segments de colonne vides et/ou remplis de phase stationnaire) présentant des chimies de surface adaptées. L'approche choisie pour la préparation de ces colonnes " multimodales ", repose sur (1) la synthèse d'un monolithe de silice poreux "générique " dans des tubes capillaires de quelques dizaines de microns de diamètre interne et (2) la modification de surface localisée dans le capillaire permettant d'apporter des propriétés de surface complémentaires. Dans le cadre de cette thèse, deux procédés de fonctionnalisation innovants, initiés par photochimie, ont été développés pour la préparation des colonnes multimodales miniaturisées : la photopolymérisation, basée sur des réactions de polymérisation radicalaire, et la " photoclick chemistry ", basée sur un greffage radicalaire contrôlé (et non plus une polymérisation). Un état de l'art de leur utilisation en sciences séparatives a été dressé pour chacun des procédés, afin de guider le choix des stratégies de greffage. Après une optimisation des conditions de greffage, les résultats présentés dans ce manuscrit montrent que ces procédés de fonctionnalisation sont rapides (fonctionnalisation en quelques minutes), efficaces, polyvalents (transposables à de multiples greffons) et localisables. Leurs potentiels respectifs dans la préparation des colonnes multimodales ont ensuite été démontrés pour la préconcentration/séparation en ligne de plusieurs composés. L'approche par " click chemistry " qui permet un meilleur contrôle du greffage, a été étendue au greffage de biomolécules pour la préparation de supports d'immunoaffinité. Ainsi, une colonne multimodale composée d'une première zone remplie de monolithe photogreffée avec des aptamères et une deuxième zone vide a été préparée pour la préconcentration/séparation électrocinétique en ligne de l'Ochratoxine A / Miniaturization of analytical processes is a general trend in analytical chemistry. Such trend is driven by the development of new experimental approaches based, for example, on hyphenated analytical steps or techniques. The in-line coupling of different and generally orthogonal/complementary separation mechanisms at the microscale, is dependent on the capability to define functional segments (open column segments and/or filled with stationary phase). Preparation of such "multimodal" capillary columns is based on (1) the in-capillary synthesis of a "generic" porous silica monolith and (2) on its localized chemical surface modification to define specific functional segments. Herein, two innovative photo-functionalization processes have been investigated for the preparation of multimodal miniaturized columns. The former, called photopolymerization is based on acrylate free radical polymerization reactions while the latter, called photografting, implements the thiol-ene "photoclick chemistry" reaction. These photo-initiated processes, after optimization, prove to be rapid (within few minutes), versatile (adapted to the grafting of various monomers) and localizable. Photopolymerization of acrylate monomers on activated silica monolith (using ?-methacryloxypropyltrimethoxysilane) gives rise to highly retentive columns due to the polymeric nature of the layer obtained. Photografting of octadecanethiol on vinylized silica columns leads to monolayer-like coating. The preparation of dedicated multimodal columns using such approaches was then successfully applied to the in-line preconcentration / separation of neuropeptides and preconcentration / fractionation of various neutral and charged compounds. The "click chemistry" approach which allows a better control of the reaction, has been extended to the grafting of biomolecules for the preparation of immunoaffinity supports. Thus, a multimodal column composed a 1-cm length aptamer-functionalized monolith at the entrance of a CZE open capillary has been prepared and successfully applied to the in-line preconcentration/electrokinetic separation of Ochratoxin A in white wine and beer

La Réduction Catalytique Sélective (SCR) par l'ammoniac, ou l'urée, est un procédé connu de post-traitement permettant de réduire efficacement les oxydes d'azote émis par les motorisations Diesel, en azote et en eau. Les zéolithes échangées au cuivre sont parmi les meilleures formulations pour une application sur véhicules légers, puisque efficaces sur une large zone de température. Le but de cette thèse était d'étudier la stabilité hydrothermale de ce type de catalyseur. L'approche utilisée au cours de ce travail repose sur l'étude des différentes fonctionnalités d'un catalyseur commercial de NH3-SCR présent sous forme de monolithe, à l'état frais et pour différentes conditions de vieillissement, au Banc Gaz Synthétique couplée à une analyse physico-chimique précise de la phase active du catalyseur. Ceci nous a permis de comprendre les phénomènes de désactivation intervenant au cours d'un vieillissement hydrothermal et de mesurer leur impact sur l'activité et la sélectivité de ce type de catalyseur. Après traitement hydrothermal, on assiste à une désalumination plus ou moins importante de la zéolithe, pouvant conduire à l'effondrement de sa structure, ainsi que d'importantes modifications au niveau du cuivre dès les plus faibles températures de vieillissements. Les résultats ont montré l'importance de maintenir une teneur minimal de cuivre en position d'échange, afin de conserver une capacité de stockage en ammoniac suffisante, mais surtout pour garantir une bonne efficacité à basse température en SCR du NO. Le maintien de la structure de la zéolithe semble essentiel pour que le catalyseur conserve une bonne efficacité et sélectivité au cours du temps. / The Selective Catalytic Reduction (SCR) by ammonia, or urea, is a well-known after-treatment process used for converting efficiently the nitrogen oxides, emitted by Diesel engines, into nitrogen and water. Copper-exchanged zeolites are among the most efficient formulations for light-duty applications, since effective over a wide temperature-range. The aim of this thesis is to study the hydrothermal stability of this type of catalyst. The approach used is this work is based on the study of the catalytic properties of a fresh commercial monolith catalyst for NH3-SCR in fresh and after different ageing conditions, at synthetic gas test bench, coupled with a comprehensive physicochemical analysis of the catalyst active phase. This allowed us to understand the deactivation phenomena occurring during a hydrothermal ageing and the impact on the catalyst activity and selectivity. A hydrothermal treatment induces a dealumination of the zeolite, into a more or less significant extent, which can lead to its collapse, as well as important modifications of the copper sites, even at low ageing. The results showed the importance to maintain a minimal copper content into exchanged sites, in order to retain a sufficient ammonia storage capacity, and especially to provide a good efficiency for the SCR of NO at low temperature. The preservation of the zeolite structure seems to be essential in order to maintain the catalyst efficiency and selectivity over time.