High performance catalyst supports and packing materials are playing an increasing role in many reactions and separations. The dispersion in packed bed reactors and separation columns can be reduced by the development of new packing structures having open and connected pore geometries. The application of new materials in High Performance Liquid Chromatography (HPLC) with sub 5 micron particle sizes are growing. These small particles offer better performance and improved bed and column efficiencies. Recently developed, twinned Alumina Nanosheets (TAN) are 2D/3D nanomaterials that offer promising open geometries for use as column packings and catalysts supports. They have a small particle size (4 um in length, 1 um in width and 0.1 um in thickness) and excellent flow-through capabilities. TANs have recently been used to successfully produce high throughput dynamic membranes. However, their resistance to compaction is unknown and thought to be limited. A technique was developed to reinforce the TAN nanomaterial. Two binder materials were tested as reinforcing agents; SiO2 and AlH6O12P3. The binder-reinforced TANs were then packed into columns. Eleven columns having a 4 cm initial packing length were assembled. Tracer injection studies were performed to investigate the flow behavior and dispersion in these columns. SEM images were also taken to characterize the particles before and after compaction. The best results were obtained using a binding solution containing 7.5 (wt%) SiO2. The binder SiO2 offered a better resistance to compaction than the AlH6O12P3. The Peclet (Pe) number for the columns ranged from 22 to 648. When the content of SiO2 increased from 0 to 7.5 (wt%), the columns showed an increase in the Pe number. When SiO2 increased from 7.5 to 20 (wt%), the columns showed a decrease in the Pe number. However, AlH6O12P3 did not present any relation between the binder content and the Pe number. The results of this work demonstrate that reinforced TANs, are a new type of material that offers a packing with an open pore structure and improved channel connectivity. The new reinforced material offers considerable potential in many applications such as catalysis and separations over conventional materials. If they are used as packing materials in HPLC columns or packed bed reactors, they can contribute to a higher separation efficiency or an enhanced conversion rate or productivity, bringing more advantages and benefits than ordinary packing materials.

Objetivos: O objetivo deste estudo foi avaliar a influência do Diabetes Mellitus do tipo 2 na farmacocinética da glimepirida em ratos Wistar e descrever o perfil através de modelo farmacocinética populacional (popPK). Metodologia: Os experimentos com animais foram aprovados pelo CEUA/UFRGS (protocolo #27892). O diabetes foi induzido com administração intraperitoneal de 100 mg/kg de nicotinamida, 15 minutos antes da administração intravenosa de 65 mg/kg de STZ. Os animais com nível de glicemia > 250 mg/dL foram considerados diabéticos. A glimepirida foi administrada na dose de 5 mg/kg via i.v. nos animais sadios (n = 11) e diabéticos (n = 9) e quantificada por CLAE-UV. A ligação às proteínas plasmáticas foi determinada por método de ultracentrifugação (Centrifree®). A análise farmacocinética não compartimental (software Phoenix®) foi realizada, assim como a modelagem farmacocinética populacional (software Monolix ®). Resultados e Discussão: A metodologia analítica para quantificação da glimepirida em plasma foi desenvolvida e validada, seguindo os critérios do FDA, apresentou sensibilidade, exatidão e precisão. O modelo de indução da diabetes produziu glicemia > 250 mg/dL. A ligação às proteínas plasmáticas não foi afetada pela doença (LPPSaudáveis = 99,3 ± 0,09%, LPPDiabéticos = 99,13 ± 0,075%, p > 0,05). O modelo farmacocinético populacional estrutural de 2 compartimentos com eliminação de primeira-ordem com covariável categórica (diabetes), foi usado para descrever os perfis plasmáticos de concentração-tempo da glimepirida após administração intravenosa na dose de 5 mg/kg a ratos saudáveis e diabéticos. O CL e a ASC0-inf dos animais diabéticos foram estatisticamente diferentes dos animais saudáveis, CLpop Saudáveis= 0,066 L/h para CLpop diabéticos = 0,024 L/h e ASCpop saudáveis = 19,24 μg/mL.h para ASCpop diabéticos = 59,64 μg.h/mL, indicando que a eliminação foi alterada nos animais diabéticos induzidos STZ. Conclusões: A modela gempossibilitou identificação do parâmetro que atribuiu variabilidade entre os grupos. Desta forma, a variabilidade interindividual foi quantificada e incluída no modelo. O modelo popPK final, nos permitiu elucidar os fatores que afetam a farmacocinética da glimepirida e prever mudanças na exposição em uma população específica. / Objective: The aim of this study was to evaluate the influence of diabetes mellitus type 2 on the pharmacokinetics of glimepiride in rats and describe the profile in population pharmacokinetic model (popPK). Methods: The experiments with animals were approved by CEUA/UFRGS (protocol number). The diabetes was induced by intraperitoneal administration of NA (100 mg/kg) dissolved in saline 15 min before an intravenous administration of 65 mg/kg STZ in citrate buffer (pH 4.5) to overnight fasted rats. Animals with blood glucose level> 250 mg/dL were considered diabetic. After administered of glimepiride at a dose of 5 mg/kg i.v. bolus in healthy (n = 11) and diabetic animals (n = 9). Method HPLC-UV developed and validated quantified plasma concentrations. The plasma protein binding was determined by method ultracentrifugation (Centrifree®). Noncompartmental analysis of pharmacokinetic in Phoenix® software was performed, as well as the model pharmacokinetic population using Monolix®. Performed by Student's t-test for SigmaStat® software. Results and Discussion: The HPLC-UV method for quantification of glimepiride in plasma was developed and validated following requirements by FDA showing sensitivity, accuracy and precision. Induced diabetes model produced glucose> 250 mg / dL. The plasma protein binding was not affect by the disease (LPPSaudáveis = 99.3 ± 0.09%, LPPDiabéticos = 99.13 ± 0.075%, p> 0.05). The model pharmacokinetic population 2 compartments with eliminating first-order with categorical covariates diabetic was used to describe the plasma profile concentration-time glimepiride. The CL and AUC 0-inf of diabetic animals were significantly different. In healthy animals was Clpop Healthy = 0.066 L/h for diabetics Clpop = 0.024 L/h and healthy ASCpop = 19.24 g/mL.h ASCpop for diabetics = 59.64 g/mL.h, indicating that elimination was decreased in induced diabetic rats STZ. Conclusions: popPk enabled identification of the parameter assigned variability between the groups. Thus, the inter subject variability was measured and included in the model. The PBPK final model, allowed us to elucidate the factors that affect the pharmacokinetics of glimepiride and predict changes in exposure in a specific population.

Neste trabalho, foram desenvolvidos e validados métodos analítico e microbiológico, bem como estudo preliminar de estabilidade, cinética de degradação e citotoxicidade da Ceftarolina Fosamila em pó para solução injetável, um antibiótico da classe das cefalosporinas de quinta geração, indicado para pneumonias adquiridas na comunidade e infecções graves, de pele e tecidos moles. A validação do ensaio microbiológico pelo método de difusão em ágar cilindros em placa, delineamento 3x3, apresentou resultados satisfatórios, como especificidade, linearidade na faixa de 2,0 - 8,0 μg/mL, precisão (109,42 %), exatidão (102,3 %) e robustez. Soluções de Cefatarolina Fosamila do produto acabado expostas à radiação UVC (254 nm) e à degradação térmica a 60 °C foram utilizadas para avaliar a especificidade do bioensaio. A robustez foi avaliada através da alteração da concentração do meio inoculado (0,8 e 1,2 %). O desenvolvimento e validação de método por CLAE foi avaliado através da especificidade, linearidade, precisão, exatidão e robustez. No método cromatográfico foi utilizado cromatógrafo à liquido de alta eficiência SHIMADZU com coluna Agilent® C18, fase móvel (água com trietilamina 1,0% pH 5,0:acetonitrila 87:13 v/v). O método apresentou-se específico, linear, no intervalo de 5,0 - 60,0 μg/mL, preciso (110,0 %), exato (100,68 %) e robusto. Os métodos microbiológico e cromatográfico validados foram comparados estatisticamente e verificou-se não haver diferença significativa entre eles quando comparados através do teste “t” de Student. No estudo preliminar de estabilidade constatou-se ser estável em hidrólise ácida (0,1 M) e luz UVA no período avaliado, e instável frente à degradação térmica (40 e 60 °C), oxidativa com peróxido de hidrogênio, básica em NaOH (0,1 M e 0,01 M) e luz UVC. As cinéticas de degradação frente à luz UVC e degradação térmica 60 °C mostraram que as amostras possuem cinética de degradação de ordem zero e de segunda ordem, respectivamente. O ensaio de citotoxicidade demonstrou não haver diferença entre a condição normal e a amostra submetida à degradação forçada, sugerindo que os possíveis produtos de degradação formados não alteraram o resultado. / In this work, analytical and microbiological methods were developed and validated, as well as a preliminary study of the stability, degradation kinetics and cytotoxicity to Ceftaroline Fosamil powder for injectable solution, this is a fifth generation cephalosporin antibiotic indicated for community-acquired pneumonia and severe infections of the skin and soft tissues. The validation of the microbial assay by diffusion method in 3x3 cylinder agar delineated showed satisfactory results in specificity, linearity in the range of 2.0 - 8.0 μg / mL, precision (109.42 %), accuracy (102.3 %) and robustness. The development and validation of the method by HPLC was evaluated through specificity, linearity, precision, accuracy and robustness. In the chromatographic method was used high performance liquid chromatograph from SHIMADZU with Agilent® C18 column, mobile phase (water with triethylamine 1.0 % pH 5.0: acetonitrile 87:13 v/v). The method was linear, specific in the range of 5.0 - 60.0 μg/mL, accurate (110.0 %), exact (100.68 %) and robust. The validated microbiological and chromatographic methods were compared statistically and there was no significant difference between them when compared through Student's t-test. In the preliminary stability study, it was found stable in acid hydrolysis (0.1M) and UVA light in the period evaluated, and instable against thermal degradation (40 and 60 °C), oxidative with hydrogen peroxide, basic in NaOH (0.1 M and 0.0 1M) and UVC light. Samples exposed in UVC light an thermal degradation at 60°C showed degradation kinetics following zero order and second order, respectively. The cytotoxicity assay showed no difference between the normal condition and the sample submitted to forced degradation, suggesting that the possible degradation products formed did not change the result.

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Depuis plusieurs années, la recherche dans le domaine des bioénergies s'oriente de plus en plus vers l'utilisation des microorganismes (bactéries, levures, moisissures et microalgues) pour la production de biocarburants. Face au contexte actuel de développement durable, l'utilisation des microorganismes oléagineux photoautotrophes (cyanobactéries et microalgues) semble potentiellement plus intéressante d'un point de vue économique (énergie solaire) et environnemental (assimilation de CO2) malgré des productions plus faibles que celles des microorganismes oléagineux hétérotrophes (levures et moisissures). L'objectif général de ces travaux de thèse a donc été d'étudier l'accumulation de lipides neutres et d'hydrocarbures chez Botryococcus braunii race A (chlorophycée unicellulaire d'eau douce), sélectionnée pour son potentiel de production après une étude bibliographique et d'évaluer les mécanismes naturels d'exsudation de ces métabolites. Afin de fiabiliser la quantification et l'identification des composés lipidiques produits par la microalgue, des méthodes d'analyse qualitative et quantitative par HPLC ont été mises au point. Les résultats analytiques ont permis de montrer que les méthodes d'extraction et quantification décrites dans la littérature conduisaient à surévaluer la teneur en hydrocarbures synthétisés par la souche B. braunii Utex 572. Les études menées lors de ce travail de thèse ont également permis de déterminer l'impact des principaux facteurs abiotiques (milieu de culture, pH, CO2, intensité lumineuse, salinité) sur la production de biomasse, lipides neutres et hydrocarbures par B. braunii et d'en définir les conditions optimales. L'étude de la flore bactérienne naturellement présente dans les cultures de B. braunii en conditions non axéniques et de sa dynamique a montré que celle-ci n'avait pas ou peu d'impact sur la production de biomasse et de lipides neutres chez B. braunii dans les conditions de l'étude. L'application d'un stress oxydatif à une culture de B. braunii en phase stationnaire n'a pas engendré le phénomène de « milking » recherché mais a eu un impact positif sur l'accumulation de lipides intracellulaires et sur la synthèse de triglycérides. / Biofuels produced from vegetable oils or microbial cultures, are an alternative to fossil fuels that can reduce the human impact on the environment. For several years, research in the field of bioenergy is moving increasingly toward the use of microorganisms (bacteria, yeasts, fungi and microalgae) for the production of biofuels. In the current context of sustainable development, the use of photoautotrophic oleaginous microorganisms (cyanobacteria and microalgae) seems potentially more interesting economically (solar energy) and environmentally (CO2 assimilation) despite lower production rates than those of heterotrophic oleaginous microorganisms (yeasts and moulds). The overall objective of this thesis has been to study the accumulation of neutral lipids and hydrocarbons in the A-race Botryococcus braunii (unicellular green alga of freshwater), selected on its potential production after literature review, and to evaluate the mechanisms of natural exudation of these metabolites. To reliable quantification and identification of lipid compounds produced by microalgae, methods of qualitative and quantitative analysis by HPLC were developed. The analytical results have shown that the extraction and quantification methods described in the literature led to overestimate the amount of hydrocarbons synthesized by the strain B. braunii Utex 572. The studies in this thesis have also identified the impact of key abiotic factors (culture medium pH, CO2, light intensity, salinity) on biomass production, neutral lipids and hydrocarbons by B. braunii and to define the optimal conditions of production. The study of the bacterial flora naturally present in cultures of B. braunii under non axenic conditions and its dynamics showed that it had little or no impact on the production of biomass and neutral lipids in B. braunii in the conditions of the study. The application of oxidative stress to a culture of B. braunii at the stationary phase did not cause the sought phenomenon of "milking" but had a positive impact on the accumulation of intracellular lipid and triglyceride synthesis.

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Objetivos: O objetivo deste estudo foi avaliar a influência do Diabetes Mellitus do tipo 2 na farmacocinética da glimepirida em ratos Wistar e descrever o perfil através de modelo farmacocinética populacional (popPK). Metodologia: Os experimentos com animais foram aprovados pelo CEUA/UFRGS (protocolo #27892). O diabetes foi induzido com administração intraperitoneal de 100 mg/kg de nicotinamida, 15 minutos antes da administração intravenosa de 65 mg/kg de STZ. Os animais com nível de glicemia > 250 mg/dL foram considerados diabéticos. A glimepirida foi administrada na dose de 5 mg/kg via i.v. nos animais sadios (n = 11) e diabéticos (n = 9) e quantificada por CLAE-UV. A ligação às proteínas plasmáticas foi determinada por método de ultracentrifugação (Centrifree®). A análise farmacocinética não compartimental (software Phoenix®) foi realizada, assim como a modelagem farmacocinética populacional (software Monolix ®). Resultados e Discussão: A metodologia analítica para quantificação da glimepirida em plasma foi desenvolvida e validada, seguindo os critérios do FDA, apresentou sensibilidade, exatidão e precisão. O modelo de indução da diabetes produziu glicemia > 250 mg/dL. A ligação às proteínas plasmáticas não foi afetada pela doença (LPPSaudáveis = 99,3 ± 0,09%, LPPDiabéticos = 99,13 ± 0,075%, p > 0,05). O modelo farmacocinético populacional estrutural de 2 compartimentos com eliminação de primeira-ordem com covariável categórica (diabetes), foi usado para descrever os perfis plasmáticos de concentração-tempo da glimepirida após administração intravenosa na dose de 5 mg/kg a ratos saudáveis e diabéticos. O CL e a ASC0-inf dos animais diabéticos foram estatisticamente diferentes dos animais saudáveis, CLpop Saudáveis= 0,066 L/h para CLpop diabéticos = 0,024 L/h e ASCpop saudáveis = 19,24 μg/mL.h para ASCpop diabéticos = 59,64 μg.h/mL, indicando que a eliminação foi alterada nos animais diabéticos induzidos STZ. Conclusões: A modela gempossibilitou identificação do parâmetro que atribuiu variabilidade entre os grupos. Desta forma, a variabilidade interindividual foi quantificada e incluída no modelo. O modelo popPK final, nos permitiu elucidar os fatores que afetam a farmacocinética da glimepirida e prever mudanças na exposição em uma população específica. / Objective: The aim of this study was to evaluate the influence of diabetes mellitus type 2 on the pharmacokinetics of glimepiride in rats and describe the profile in population pharmacokinetic model (popPK). Methods: The experiments with animals were approved by CEUA/UFRGS (protocol number). The diabetes was induced by intraperitoneal administration of NA (100 mg/kg) dissolved in saline 15 min before an intravenous administration of 65 mg/kg STZ in citrate buffer (pH 4.5) to overnight fasted rats. Animals with blood glucose level> 250 mg/dL were considered diabetic. After administered of glimepiride at a dose of 5 mg/kg i.v. bolus in healthy (n = 11) and diabetic animals (n = 9). Method HPLC-UV developed and validated quantified plasma concentrations. The plasma protein binding was determined by method ultracentrifugation (Centrifree®). Noncompartmental analysis of pharmacokinetic in Phoenix® software was performed, as well as the model pharmacokinetic population using Monolix®. Performed by Student's t-test for SigmaStat® software. Results and Discussion: The HPLC-UV method for quantification of glimepiride in plasma was developed and validated following requirements by FDA showing sensitivity, accuracy and precision. Induced diabetes model produced glucose> 250 mg / dL. The plasma protein binding was not affect by the disease (LPPSaudáveis = 99.3 ± 0.09%, LPPDiabéticos = 99.13 ± 0.075%, p> 0.05). The model pharmacokinetic population 2 compartments with eliminating first-order with categorical covariates diabetic was used to describe the plasma profile concentration-time glimepiride. The CL and AUC 0-inf of diabetic animals were significantly different. In healthy animals was Clpop Healthy = 0.066 L/h for diabetics Clpop = 0.024 L/h and healthy ASCpop = 19.24 g/mL.h ASCpop for diabetics = 59.64 g/mL.h, indicating that elimination was decreased in induced diabetic rats STZ. Conclusions: popPk enabled identification of the parameter assigned variability between the groups. Thus, the inter subject variability was measured and included in the model. The PBPK final model, allowed us to elucidate the factors that affect the pharmacokinetics of glimepiride and predict changes in exposure in a specific population.

Insects represent very useful experimental model in various branches of biological research. The investigation is driven by economic importance of many insect species, and also by biological features of insects as model organisms such as short period of reproduction, easy breeding and manipulation and, in particular, the minimal regulatory requirements which are associated to the management of vertebrates. Here we report robust and efficient LC/MS/MS methodology for the determination of the physiologically important lipid molecular species in insects. The target metabolites represent polar glycerophos-phopholipids (GPL) and nonpolar lipids diacylglycerols (DG) and triacylglycerols (TG). Combination of the LC/MS data with the subsequent GC fatty acid analysis enables complete structural elucidation of particular lipid species including their fatty acid compositions. The developed methodology was applied to studies of the chill tolerance of the firebug Pyrhocorris appterus. Fields and laboratory experiments were conducted to separate the triggering effects of low temperature, desiccation and diapause progression on the physiological characteristics related to chill tolerance with emphasis on the restructuring of GPL composition. The same effect on the GPL composition was observed during acclimatization in the field and cold acclimation in laboratory. By contrast, the GPL changes related to desiccation and diapause progression were relatively small (Tomčala et al, 2006). In adults of Drosophila melanogaster it has been found that acclimation at 15, 20 and 25°C during preimaginal development affects thermal tolerance and composition of membrane GPLs. Low temperature acclimation was associated with increase in proportion of ethanolamine at the expense of choline in GPLS. Relatively small, but statistically significant changes in lipid molecular compositon were observed with decreasing acclimation temperature (Overgard et al, 2008). Hormonal treatment studies on insect model Locusta migratoria showed a heterogeneous distribution of individual DGs in haemolymph after the hormone application and revealed that mobilization of the DGs is molecular species-specific with the highest proportion of DG 16:0/18:1 and forming in summary about 20% of the total mobilized DG content. Additional analysis of fat body triacylglycerols revealed that the AKH mobilizes the DGs specifically with the preference of those possessing the unsaturated C18 fatty acids (FAs). The fat body FAs with more than 18 carbons did not participate on the mobilization (Tomcala et al, 2009). The LC/MS methodology was further applied to lipid composition studies of several samples with very diverse biological origin (fish, human blood etc.) and was proved to be universally applicable to the wide scope of biological samples.