Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biophysics and Physical Chemistry Candidate: Kateřina Brokešová Supervisor: Ing. Martin Drastík, Ph. D. Thesis title: The use of core-shell columns for fluconazole determination A novel HPLC method for determination of fluconazole in dissolution test samples was developed and partly validated. A matrix formed by lactic and glycolic acid copolymer branched by different compounds was used as a drug carrier. Fluconazol was incorporated as the model drug. The concentration profile of fluconazole was studied by developed HPLC method during the dissolution test. A modern core-shell column Ascentis Express RP-Amide, 10 cm × 3.0 mm; 2.7 μm was employed. A mixture of acetate buffer pH 5.0:methanol (80:20) served as the mobile phase. The flow rate was 0.70 ml/min and the detection wavelength was 260 nm. The temperature of analysis was 50 řC. The retention time of fluconazole was 3.3 min and the whole analysis took just 4 min. Keywords: fluconazole, core-shell column, HPLC, PLGA

Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biophysics and Physical Chemistry Candidate: Andrea Malovaná Supervisor: Ing. Martin Drastík, Ph. D. Diploma thesis: Simplification of terbinafine HPLC analysis of samples based on biodegradable polyesters A HPLC method for determination of terbinafine in samples consisting of copolymers of lactic and glycolic acid was optimized and validated. The development of the method was based on the finding of suitable chromatographic conditions for separation of terbinafine. The separation was performed on the Ascentis Express ES-CN, 15 cm × 4.6 mm; 2.7 μm core- shell column. The mixture of the citrate phosphate buffer pH 4 and acetonitrile in ratio 40:60 (v/v) was chosen as the mobile phase. The mobile phase flow rate was set to 1.4 ml/min and the temperature to 30 řC. The injection volume of samples containing terbinafine was 5 µl. The UV detection at 226 nm was employed. The retention time of terbinafine was 3.3 min. The whole analysis was completed within 4 min. The method was validated, following parameters were tested: column efficiency, factor of symmetry, LOD, LOQ, linearity, repeatability and robustness. Keywords: terbinafine, HPLC, core-shell column, PLGA

BSc. Martina Hloušková Supervisor: Ing. Martin Drastík, Ph.D. The aim of this diploma thesis was the optimization and validation of a HPLC method for miconazole determination in samples supplied by the Department of Pharmaceutical Technology. A gradual miconazole release dependent on the composition of the copolymere of glycolic and lactic acid was studied. HPLC analysis was performed using a modern core-shell Column Ascentis Express RP- Amide, 10 cm x 3.0 mm; 2.7 μm. Optimized analytical conditions were: mobile phase methanol:water 70:30, flow rate 0.8 ml/min, temperature 45 řC, injection 5 l and UV detection at 220 nm. Miconazole retention time was 5.65 min. The entire analysis was carried out in 7 minutes. When the optimal conditions of analysis were determined, the method could be validated. The following parameters were monitored during validation: linearity, selectivity, efficiency, LOD, LOQ, repeatability and tailing factor. All of the monitored parameters met the requirements of the Czech Pharmacopoeia.

Atualmente, as nanopartículas superparamagnéticas despertam enorme interesse científico devido sua grande variedade de aplicações em biomedicina, tanto na área de diagnóstico quanto no tratamento de enfermidades. Embora muitos materiais vem sendo estudados, os óxidos de ferro (magnetita e maghemita) apresentam maiores avanços nos estudos para aplicações em medicina. A preferência por óxidos de ferro se deve a baixa toxicidade destas partículas quando comparado as nanopartículas metálicas ou ligas. Entretanto, as nanopartículas destes óxidos possuem baixas magnetizações de saturação que diminuem ainda mais com as sucessivas etapas de recobrimento necessárias para conferir funcionalidade a estas partículas. Desse modo, há uma necessidade atual para o desenvolvimento de nanopartículas superparamagnéticas com elevada magnetização, baixa toxicidade e maior facilidade de funcionalização da sua superfície com biopolímeros e agentes funcionalizantes. Neste trabalho, nanopartículas superparamagnéticas de cobalto metálico e ferrita de cobalto foram sintetizadas e suas propriedades magnéticas foram comparadas com a magnetita. Nanopartículas de cobalto foram escolhidas, pois seu elevado comportamento ferromagnético é menor apenas que o ferro metálico, além do baixo custo de seus reagentes. As nanopartículas magnéticas foram sintetizadas pelos métodos de microemulsão e decomposição térmica (baseado no método poliol) e suas composições química, estrutural, tamanho e distribuição de tamanho foram devidamente determinadas. Além disso, as nanopartículas de cobalto metálico e ferrita de cobalto foram recobertas com ouro utilizando o método de crescimento mediado por semente. Os sistemas microemulsionados utilizados neste trabalho não foram eficientes nem na síntese de nanopartículas estáveis de cobalto metálico nem no seu esperado controle morfológico. Já o método de decomposição térmica resultou em um rigoroso controle de composição química, estrutural e morfológico para as diferentes nanopartículas sintetizadas. O recobrimento com ouro foi efetivo na proteção do núcleo magnético e adicionalmente conferiu estabilidade, baixa toxicidade e bifuncionalidade às nanopartículas magnéticas através do seu fenômeno de ressonância plasmônica de superfície o qual foi preservado na nanoestrutura core@shell. O comportamento superparamagnético das nanopartículas de cobalto metálico recobertas com ouro e sua elevada magnetização de saturação foram expressivamente intensificadas quando comparadas as nanopartículas de magnetita sem recobrimento. Portanto, as nanopartículas sintetizadas neste trabalho apresentam propriedades de superfície e magnéticas otimizadas demonstrando um bom potencial para aplicações em biomedicina como sensores bifuncionais óptico-magnético. / Superparamagnetic nanoparticles have been extensively studied because its wide range of biomedical applications in both diagnostic and therapy areas. Although different materials are currently investigated, superparamagnetic iron oxides nanoparticles (SPION), magnetite and maghemite, are the most extensively studied for applications in medicine. The lower toxicity profile of the SPION becomes the most attractive than metal or alloys nanoparticles. Nevertheless, iron oxides nanoparticles have low saturation magnetization, which further decreases due to successive coats to provide their functionality, leading the actual demand to develop superparamagnetic nanoparticles with high magnetization, low toxicity and easy surface functionalization with biocompatible agents. In this work, superparamagnetic nanoparticles of metallic cobalt and cobalt ferrite were synthesized and their magnetic properties were compared with the magnetite SPION. Cobalt nanoparticles were chosen because present high ferromagnetic behavior among chemical elements, second only to iron, besides their low cost. The magnetic nanoparticles were synthesized by both microemulsion and thermal decomposition (based on the polyol process) methods and their chemical composition, structure, size and size distribution were properly characterized. In addition, the ferrite and metallic cobalt nanoparticles were coated with gold by using the seed-mediated growth method. The used microemulsion systems were not efficient enough to synthesize stable metallic nanoparticles and to promote the expected morphological control even to ferrites. Instead, the thermal decomposition processes resulted in rigorous control of chemical compositional, structure and morphology in all different prepared samples. Au-coating process was effective to protect the magnetic nuclei also giving additional stability, low toxicity and a bifunctionality to the magnetic nanoparticle since their surface plasmon resonance phenomenon was preserved in the core@shell nanostructure. The superparamagnetic behavior of the Au-coated cobalt nanoparticle was preserved and their saturation magnetization was significantly increased compared with the naked magnetite SPION. In conclusion, the synthesized nanoparticles present enhanced magnetic and surface properties showing good potential to be used in biomedical application as bifunctional optical-magnetic sensor.

Submitted by Cássia Santos (cassia.bcufg@gmail.com) on 2017-07-10T11:42:20Z No. of bitstreams: 2 Dissertação - Raphael Lucas de Sousa e Silva - 2017.pdf: 3882688 bytes, checksum: 6b33233b331ed11ce579c9148910d0d4 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2017-07-10T11:46:08Z (GMT) No. of bitstreams: 2 Dissertação - Raphael Lucas de Sousa e Silva - 2017.pdf: 3882688 bytes, checksum: 6b33233b331ed11ce579c9148910d0d4 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-07-10T11:46:09Z (GMT). No. of bitstreams: 2 Dissertação - Raphael Lucas de Sousa e Silva - 2017.pdf: 3882688 bytes, checksum: 6b33233b331ed11ce579c9148910d0d4 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2017-03-03 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Among the various iron oxides studied, a magnetite (Fe3O4) stands out due to its superparamagnetic behavior and its high biocompatibility. Luminescent properties working cooperatively in magnetic nanoparticles with which the material converts certain types of energy into emission of electromagnetic radiation at the same time, which enables an interaction by magnetic fields. In the present work, an investigation and structural, elemental and morphological analysis of superparamagnetic / luminescent particles formed by coating of iron oxide by an SiO2 base network was carried out. The Fe3O4 oxide was synthesized by the coprecipitation method, and in the Eu3+ rare earth together with the 1,10-phenanthroline linker in the network, were responsible for energy transfer processes in systems that checked as optical materials for materials. The nanocomposites synthesized obeyed the formation of a core-shell system and was verified by micrographs obtained by Electron Transmission Electron Microscopy, evidencing a formation of nanoparticles of varied sizes. As the compositions of the phases of the materials were characterized by X-ray Diffraction, Radiated Spark Energy and Absorption Spectroscopy in the Infrared region, showing a crystal phase of the magnetic oxide and a formation of nanocomposite. The properties of interest (magnetism and photoluminescence) were evaluated and the respective results presented agreed with each other, with characteristic emissions of Eu3+ emitting a pink coloration, and an emission intensity for a concentration of this species in the material. The materials presented a superparamagnetic behavior, evidencing that the oxide coating did not change as Fe3O4 magnetic characteristics, and is in turn presented in a saturation magnetization of 55 emu / g with a magnetic domain of 8.9 nm. As nanoparticles have dimensions that indicate that a part has a magnetic domain. / Entre os diversos óxidos de ferro estudados, a magnetita (Fe3O4) destaca-se, devido ao seu comportamento superparamagnético e a sua alta biocompatibilidade. Propriedades luminescentes trabalhando de forma cooperativa em nanopartículas magnéticas faz com que o material converta certos tipos de energia em emissão de radiação eletromagnética ao mesmo tempo, que possibilita a interação por campos magnéticos. No presente trabalho, foi investigada a síntese e a caraceterização estrutural, elementar e morfológica de partículas superparamagnéticas/luminescentes formadas pelo revestimento do óxido de ferro por uma rede a base de SiO2. O óxido Fe3O4 foi sintetizado pelo método de coprecipitação, e o íon terra-rara Eu3+ juntamente com ligante 1,10-fenantrolina na rede, foram responsáveis por processos de transferência de energia inter e intrasistemas que conferiram as propriedades ópticas aos materiais. Os nanocompósitos sintetizados obedeceram a formação de um sistema core-shell sendo comprovado pelas micrografias obtidas por Microscopia Eletrônica de Transmissão, evidenciando a formação de nanopartículas de tamanhos variados. As composições das fases dos materiais foram caracterizadas por Difração de Raios X, Energia Dispersiva de Raios X e Espectroscopia de Absorção na região do Infravermelho, mostrando a fase cristalina do óxido magnético e a formação do nanocompósito. As propriedades de interesse (magnetismo e fotoluminescência) foram avaliadas e seus respectivos resultados apresentaram concordância entre si, com emissões características do íon Eu3+ emitindo uma coloração rosa, e a intensidade de emissão foi equivalente a concentração desta espécie no material. Os materiais apresentaram um comportamento superparamagnético, evidenciando que o recobrimento do óxido não alterou as características magnéticas do Fe3O4, e este por sua vez apresentou uma magnetização de saturação de 55 emu/g com domínio magnético de 8,9 nm. As nanopartículas apresentaram dimensões que indicaram que majoritariamente uma partícula possui apenas um domínio magnético.

Atualmente, as nanopartículas superparamagnéticas despertam enorme interesse científico devido sua grande variedade de aplicações em biomedicina, tanto na área de diagnóstico quanto no tratamento de enfermidades. Embora muitos materiais vem sendo estudados, os óxidos de ferro (magnetita e maghemita) apresentam maiores avanços nos estudos para aplicações em medicina. A preferência por óxidos de ferro se deve a baixa toxicidade destas partículas quando comparado as nanopartículas metálicas ou ligas. Entretanto, as nanopartículas destes óxidos possuem baixas magnetizações de saturação que diminuem ainda mais com as sucessivas etapas de recobrimento necessárias para conferir funcionalidade a estas partículas. Desse modo, há uma necessidade atual para o desenvolvimento de nanopartículas superparamagnéticas com elevada magnetização, baixa toxicidade e maior facilidade de funcionalização da sua superfície com biopolímeros e agentes funcionalizantes. Neste trabalho, nanopartículas superparamagnéticas de cobalto metálico e ferrita de cobalto foram sintetizadas e suas propriedades magnéticas foram comparadas com a magnetita. Nanopartículas de cobalto foram escolhidas, pois seu elevado comportamento ferromagnético é menor apenas que o ferro metálico, além do baixo custo de seus reagentes. As nanopartículas magnéticas foram sintetizadas pelos métodos de microemulsão e decomposição térmica (baseado no método poliol) e suas composições química, estrutural, tamanho e distribuição de tamanho foram devidamente determinadas. Além disso, as nanopartículas de cobalto metálico e ferrita de cobalto foram recobertas com ouro utilizando o método de crescimento mediado por semente. Os sistemas microemulsionados utilizados neste trabalho não foram eficientes nem na síntese de nanopartículas estáveis de cobalto metálico nem no seu esperado controle morfológico. Já o método de decomposição térmica resultou em um rigoroso controle de composição química, estrutural e morfológico para as diferentes nanopartículas sintetizadas. O recobrimento com ouro foi efetivo na proteção do núcleo magnético e adicionalmente conferiu estabilidade, baixa toxicidade e bifuncionalidade às nanopartículas magnéticas através do seu fenômeno de ressonância plasmônica de superfície o qual foi preservado na nanoestrutura core@shell. O comportamento superparamagnético das nanopartículas de cobalto metálico recobertas com ouro e sua elevada magnetização de saturação foram expressivamente intensificadas quando comparadas as nanopartículas de magnetita sem recobrimento. Portanto, as nanopartículas sintetizadas neste trabalho apresentam propriedades de superfície e magnéticas otimizadas demonstrando um bom potencial para aplicações em biomedicina como sensores bifuncionais óptico-magnético. / Superparamagnetic nanoparticles have been extensively studied because its wide range of biomedical applications in both diagnostic and therapy areas. Although different materials are currently investigated, superparamagnetic iron oxides nanoparticles (SPION), magnetite and maghemite, are the most extensively studied for applications in medicine. The lower toxicity profile of the SPION becomes the most attractive than metal or alloys nanoparticles. Nevertheless, iron oxides nanoparticles have low saturation magnetization, which further decreases due to successive coats to provide their functionality, leading the actual demand to develop superparamagnetic nanoparticles with high magnetization, low toxicity and easy surface functionalization with biocompatible agents. In this work, superparamagnetic nanoparticles of metallic cobalt and cobalt ferrite were synthesized and their magnetic properties were compared with the magnetite SPION. Cobalt nanoparticles were chosen because present high ferromagnetic behavior among chemical elements, second only to iron, besides their low cost. The magnetic nanoparticles were synthesized by both microemulsion and thermal decomposition (based on the polyol process) methods and their chemical composition, structure, size and size distribution were properly characterized. In addition, the ferrite and metallic cobalt nanoparticles were coated with gold by using the seed-mediated growth method. The used microemulsion systems were not efficient enough to synthesize stable metallic nanoparticles and to promote the expected morphological control even to ferrites. Instead, the thermal decomposition processes resulted in rigorous control of chemical compositional, structure and morphology in all different prepared samples. Au-coating process was effective to protect the magnetic nuclei also giving additional stability, low toxicity and a bifunctionality to the magnetic nanoparticle since their surface plasmon resonance phenomenon was preserved in the core@shell nanostructure. The superparamagnetic behavior of the Au-coated cobalt nanoparticle was preserved and their saturation magnetization was significantly increased compared with the naked magnetite SPION. In conclusion, the synthesized nanoparticles present enhanced magnetic and surface properties showing good potential to be used in biomedical application as bifunctional optical-magnetic sensor.

Different scenarios for assembling rod-like and spherical colloidal particles using binary mixtures of partially miscible liquids were investigated experimentally. Suitable rod-like colloids were developed first. The subsequent studies of colloids in binary liquids consisted, on one hand, of systems where particles were partially wetted by both phases and, on the other hand, of systems where particles were completely wetted by the minority phase. A simple method to prepare large quantities of micrometer-sized akagan eite-silica core-shell rods was developed. These were proven to be very versatile, with the possibility of modifying their properties on different levels. The aspect ratio is simply controlled by a gradual growth of the silica shells. From them, hollow silica rods and rods with an increased responsiveness to a magnetic field could be obtained in straightforward ways. Bijels were prepared by trapping rod-like particles on a percolating liquid-liquid interface. The familiar bicontinuous organization of liquid domains was observed after structural arrest. At a fixed volume per particle it is demonstrated that for rod-like particles the domain size decreases faster with increasing quantity of particles than in the case of spherical particles. Additionally, the packing of the rods at the interface was elucidated, revealing several characteristic features. In particle-stabilized droplet emulsions rapid evaporation of the continuous phase and eventual full mixing of the liquid phases can leave a cellular network of particles. The formation and eventual stability of these networks were investigated in detail with confocal microscopy. When colloids are completely wetted by the minority component of an asymmetric binary mixture there can be substantial temperature and composition regimes outside the binodal where shear-induced aggregation can take place. This happens as adsorbed layers present at the particle surfaces coalesce and bind particles through a liquid bridge. Depending on particle concentration, percolating networks can form of rods wetted by the minority phase after temperature quenching such a system just across the binodal.

Thesis advisor: Dunwei Wang / Metal nanocrystals with size and shape control have great potential in heterogeneous catalysis. Controllable encapsulation of well-defined metal nanoparticles into the novel porous materials results in new multifunctional nanomaterials. The core-shell nanostructure can enhance the selectivity, durability, or reactivity of the catalysts and even provide additional functionalities. Metal-organic frameworks (MOFs) are a class of novel crystalline nanoporous materials, with well-defined pore structures and distinctive chemical properties. Using MOFs as the encapsulating porous materials has drawn great interest recently due to their tunable structures and properties. However, it could be challenging to grow another porous material layer on metal surface due to the unfavorable interfacial energy. In this work we develop a new concept of colloidal synthesis to synthesize the metal@MOF core-shell nanostructures, in which a layer of self-assembled molecules directed the growth and alignment between two materials. Surfactant cetyltrimethylammonium bromide (CTAB) is designated to facilitate the overgrowth of MOF onto metal surface, and an alignment between the {100} planes of the metal and {110} planes of the MOF can be observed. By utilizing the same concept, a third layer of mesoporous silica could also be coated on the MOF shell with assistance of CTAB. And our method could be a general strategy to fabricate multiple-layer MOF materials. / Thesis (MS) — Boston College, 2015. / Submitted to: Boston College. Graduate School of Arts and Sciences. / Discipline: Chemistry.

In recent years, employing advanced oxidation processes (AOPs) as a means of wastewater remediation has emerged as a promising route towards maintaining a sustainable global water management program. The heterogeneous photocatalytic oxidation process has been of particular interest due to the prospective of utilizing solar radiation as the driving force behind the degradation of pollutants. Of the photocatalyst studied to date, TiO2 remains the most attractive material for environmental applications due to its affordability, stability, biocompatibility and high quantum yield. A key draw back however is roughly only 5% of solar radiation incident on earth can provide the energy required (3.0-3.2 eV) to generate the electron-hole pairs necessary for photo-oxidation. As a means to improve the process under solar irradiance, optical properties such as surface plasmon resonance of metallic nanoparticles and upconversion luminescence of rare earth ions have been exploited for improved light harvesting as well as the generation of more usable UV light from lower energy photons. In order to explore these phenomena and their role in the enhancement of this AOP, the photocatalytic degradation of organic dyes was studied under various conditions employing Degussa P25 TiO2 as the photocatalyst. Ag nanocubes, Ag-Pd core-shell nanoparticles and YAG:Yb+3,Er+3 served as the dopants for the various studies which resulted in enhanced degradation rates, insight into the applicability of utilizing Yb+3 as sensitizing ion under solar radiation and a novel core-shell nanoparticle synthesis.

Atom transfer radical polymerisation (ATRP) technique was used to synthesize poly(methacrylic acid-block-methyl methacrylate) (P(MAA₁₀₂-b-MMA₁₀)) copolymer in order to study the aggregation behavior in aqueous solution over the course of neutralization. A combination of static and dynamic light scattering (SLS, DLS) and potentiometric titration techniques were used to investigate the size and shape of the micelle at various degrees of neutralization. The hydrodynamic radius (Rh) determined from dynamic light scattering increases from ~26nm (for unneutralized) to ~42nm (for completely neutralized sample). Both potentiometric and laser light scattering studies indicate the formation of a core shell micelle. The weighted average molecular weights of the polymer and micelle are 1.18x10⁴ and 2.25 x 10⁵ g/mol respectively, which suggests that the aggregation number of the micelle is ~20. / Singapore-MIT Alliance (SMA)