Studies have been performed to evaluate the use of micellar electrokinetic chromatography (MEKC), one mode of capillary electrophoresis (CE), as an analytical technique in industrial pharmaceutical analysis. The potential for using chemometrics for the optimisation of MEKC methods has also been studied as well as the possibilities of coupling MEKC with mass spectrometry (MS). Two methods were developed, one for the determination of ibuprofen and codeine and another for pilocarpine, together with their degradation products and impurities in both cases. MEKC was found to be the most suitable mode of CE for the methods. Both methods were optimised by means of experimental design. Valuable information was gathered and optimum conditions were defined which resulted in fast systems with baseline-separated peaks. The ibuprofen-codeine method was validated according to the recommended validation procedures of the International Conference of Harmonisation. The validation was performed on a commercially available tablet formulation to verify the suitability of the method, i.e. for quantification of the two main compounds and to determine the degradation products and impurities in area% of each main peak. The following parameters were determined: selectivity, linearity, accuracy, precision, detection limit, quantitation limit, robustness and range. The results confirm that the method is highly suitable for its intended purpose, i.e. as a routine method for assay and impurity determination. The MEKC method for ibuprofen-codeine was coupled to a mass spectrometer in order to evaluate the potential of partial filling (PF)-MEKC-MS for identification of impurities in pharmaceutical substances and products. The so-called partial-filling technique was used to prevent the non-volatile micelles from entering the MS and was shown to fulfil its purpose of providing detection limits of about 10 pg. The study clearly shows that micellar electrokinetic chromatography is well-suited as an analytical technique in industrial pharmaceutical analysis.

Statistical experimental designs have been used for method development and optimisation of separation. Two reversed phase HPLC methods were optimised. Parameters such as the pH, the amount of tetrabutylammonium (TBA; co-ion) and the gradient slope (acetonitrile) were investigated and optimised for separation of erythromycin A and eight related compounds. In the second method, a statistical experimental design was used, where the amounts of acetonitrile and octane sulphonate (OSA; counter ion) and the buffer concentration were studied, and generation of an α-plot with chromatogram simulations optimised the separation of six analytes. The partial filling technique was used in capillary electrophoresis to introduce the chiral selector Cel7A. The effect of the pH, the ionic strength and the amount of acetonitrile on the separation and the peak shape of R- and S-propranolol were investigated. Microemulsion electrokinetic chromatography (MEEKC) is a technique similar to micellar electrokinetic chromatography (MEKC), except that the microemulsion has a core of tiny droplets of oil inside the micelles. A large number of factors can be varied when using this technique. A screening design using the amounts of sodium dodecyl sulphate (SDS), Brij 35, 1-butanol and 2-propanol, the buffer concentration and the temperature as factors revealed that the amounts of SDS and 2-propanol were the most important factors for migration time and selectivity manipulation of eight different compounds varying in charge and hydrophobicity. SDS and 2-propanol in the MEEKC method were further investigated in a three-level full factorial design analysing 29 different compounds sorted into five different groups. Different optimisation strategies were evaluated such as generating response surface plots of the selectivity/resolution of the most critical pair of peaks, employing chromatographic functions, simplex optimisation in MODDE and 3D resolution maps in DryLab™. Molecular descriptors were fitted in a PLS model to retention data from the three-level full factorial design of the MEEKC system. Two different test sets were used to study the predictive ability of the training set. It was concluded that 86 – 89% of the retention data could be predicted correctly for new molecules (80 – 120% of the experimental values) with different settings of SDS and 2-propanol. Statistical experimental designs and chemometrics are valuable tools for the development and optimisation of analytical methods. The same chemometric strategies can be employed for all types of separation techniques.

Les premières centaines de mètres du sous-sol sont au coeur d'enjeux sociétaux importants liés aux fluides auxquels la société doit encore affronter, tels que la détéction et la surveillance des ressources en eau et de la pollution, ainsi que le rôle des fluides dans les différents types d'évaluation des risques ou en géotechnique. Cette gamme de profondeur est marquée par une forte complexité et une forte hétérogénéité aussi bien structurale et lithologique qu'en termes de mélanges de phases fluides. Parmi les méthodes géophysique disponibles, le phénomène de couplage sismo-électrique (et électro-sismique) a le potentiel d'émerger comme une nouvelle technique d'imagerie haute résolution, naturellement sensible aux contrastes des fluides. Deux signaux sismo-électrique ont été prédits et parfois observés: les signaux co-sismique éléctrique à forte amplitude et les faibles perturbations électromagnétiques générées en profondeur lorque des ondes sismiques traversent une interface. Ce deuxième phénomène est le plus prometteur, en termes d'imagerie en raison de sa sensibilité aux contrastes originaux par rapport à la réflexion sismique. L'émergence d'une méthode sismo-électrique a cependant été ralentie en raison des difficultés dans l'enregistrement de ces faibles signaux sismo-électrique. Cette thèse, basée sur la combinaison d'approches expérimentales et numériques, vise à aborder des questions pratiques concernant l'acquisition et le traitement de données sismo-électrique et électro-sismique.Dans la première partie, afin d'augmenter le rapport signal-sur-bruit (S/N) des signaux sismo-électriques, nous étudions théoriquement et expérimentalement leur sensibilité à différents arrangements d'électrodes. Pour cela, nous avons développé une théorie des filtres conçue pour évaluer l'influence d'ensemble d'électrodes pour l'acquisition des signaux sismo-électriques. Cette théorie a été confrontée avec succès à des données expérimentales sismo-électriques acquises en utilisant diverses dispositions d'électrodes (c'est-à-dire l'espacement entre les électrodes, le nombre des électrodes et la vitesse sismique apparente) et à des simulations numériques des formes d'onde complète. En particulier, cette approche explique la difficulté d'enregistrement des ondes converties sismo-électromagnétique générées en profondeur converties à une interface, lorsque la configuration dipôle classique est utilisé comme récepteur. Nous encourageons ensuite l'utilisation d'une configuration de multi-électrodes dans des mesures sismo-électriques, puisque: 1) il réduit le bruit électrique omniprésent causé par l'homme, et 2) ses propriétés de filtrage amplifient les ondes électromagnétiques provenant des interfaces en profondeur par rapport aux événements co-sismiques dominants.Dans la deuxième partie, nous nous concentrons sur les phénomènes réciproques électro-osmotique, c'est-à-dire la génération de signaux sismiques par une source électrique dans des milieux saturés poroélastiques. Nous avons d'abord effectué des études numériques pour évaluer la nature et les propriétés des différentes arrivées sismiques générées par une source électrique dans un matériau homogène et hétérogène, à l'échelle du terrain. Ensuite, nous avons essayé d'acquérir des données de laboratoire électro-sismique dans un matériau homogène, données que nous avons de nouveau confrontées à des calculs numériques de forme d'onde complète. Les résultats montrent que les ondes électro-sismiques peuvent être observées et qu'elles sont générées localement autour de la source électrique. Nous montrons enfin que ces signaux électro-sismiques sont influencés par la conductivité des fluides. / The first few hundred meters of the subsurface is the seat of important issues related to fluids which society still has to face, such as the detection and monitoring of water ressources and pollution as well as the role of fluids in various types of hazard assessment or in geotechnics. This depth scale is highly complex and heterogeneous in terms of lithology and fluid content. Among the available geophysical methods, seismo-electric (or electro-seismic) coupling phenomena have the potential to emerge as a new high-resolution imaging technique, naturally sensitive to fluid contrasts. Two seismo-electric signals have been predicted and sometimes observed: co-seismic electric signals (high amplitude) and weak electromagnetic (EM) disturbances generated at depth when seismic waves cross an interface. This second phenomenon is the most promising in terms of imaging due to its sensitivity to original contrasts compared to seismic reflection. The emergence of the seismo-electric method was however slowed down due to difficulties in recording these weak seismo-electric signals. Based on the combination of experimental and numerical approaches, this PhD thesis aims at addressing practical questions concerning the acquisition and processing of seismo-electric and electro-seismic data.In the first part, in order to increase the signal-to-noise (S/N) ratio of seismo-electric signals, we theoretically and experimentally study their sensitivity to various electrode arrangements. For this, we developed a filter theory designed to assess the influence of complex electrode arrays for seismo-electric signals acquisition. This theory was successfully confronted to experimental seismo-electric data acquired using various electrode arrangements (namely the spacing between the electrodes, the number of the electrodes and the apparent seismic velocity) and to full waveform numerical simulations. This combined analysis shows that electrode configuration properties strongly influence seismo-electric amplitudes and waveforms. In particular, this approach explains the difficulty in recording depth-generated seismo-electromagnetic waves converted at an interface, when the conventional dipole configuration is used as receivers. We then promote the use of a 3-electrode configuration in seismo-electric measurements, since: 1) it reduces ubiquitous man-made electric noise, and 2) its filtering properties do amplify the EM waves originating from interfaces at depth with respect to dominant coseismic events.In the second part, we focus on the reciprocal electro-osmotic phenomena, i.e. the generation of seismic signals by an electric source in poroelastic saturated media. We first performed numerical investigations to assess the nature and properties of the different seismic arrivals generated by an electrical source in a homogeneous and an heterogeneous material, at the field scale. Then we tried to acquire electro-seismic laboratory data in a homogeneous material, data that we again confronted with numerical full waveform computations. The results show that electro-seismic waves can be observed and that they are locally generated around the electric source. We finally show that these electro-seismic signals are influenced by the fluid conductivity.

A combinação entre o delapril (DEL), um inibidor da enzima conversora de angiotensina e o manidipino (MAN), um antagonista dos canais de cálcio, produz um efeito anti-hipertensivo sinérgico, podendo ser considerado um ótimo tratamento para pacientes com hipertensão essencial leve e moderada. No presente trabalho foram desenvolvidos e validados métodos cromatográfico e eletroforético para avaliação simultânea de DEL e MAN em produto farmacêutico. As análises por cromatografia líquida em fase reversa (CL-FR) foram executadas utilizando coluna C8 (250 mm x 4,6 mm), mantida a 35 oC. A fase móvel foi constituída por acetonitrila e solução de trietilamina 0,3%, pH 3,0 (55:45; v/v), eluída na vazão de 1,2 mL/min com detecção a 220 nm. Paralelamente, desenvolveu-se método por eletroforese capilar, utilizando modo de separação por cromatografia eletrocinética micelar (MEKC) e ácido salicílico como padrão interno. Foi utilizado capilar de sílica fundida (comprimento efetivo de 72 cm) mantido a 35 °C, com solução eletrolítica composta de tampão borato 50 mM e dodecil sulfato de sódio (SDS) 5 mM, pH 9,0. Voltagem de 25 kV foi aplicada e a injeção foi de 50 mbar durante 5 s, com detecção a 208 nm. A especificidade e a capacidade dos métodos serem indicativos de estabilidade foram demonstradas através de estudos de degradação forçada dos fármacos e pela não interferência dos excipientes nas análises. Além disso, o desenho experimental Plackett-Burman foi utilizado para a avaliação da robustez, observando-se resultados adequados para ambos métodos. Os procedimentos foram validados de acordo com guias aceitos internacionalmente, observando-se resultados em uma faixa aceitável. Os métodos propostos foram aplicados com sucesso na determinação quantitativa simultânea de DEL e MAN em comprimidos, não havendo diferença significativa dos resultados (P>0,05), contribuindo, portanto, para aprimorar o controle da qualidade, assegurando a eficácia terapêutica. / The combination of delapril (DEL), an angiotensin converting enzyme inhibitor and manidipine (MAN), an antagonist of calcium channels, produces a synergic antihypertensive effect and may be regarded as an optimal antihypertensive drug treatment in mild to moderate essential hypertensive patients. The chromatographic and eletrophoretic methods for the simultaneous evaluation of DEL and MAN in pharmaceutical product were developed and validated in the present work. The reversed-phase liquid chromatography (RP-LC) method was carried out on a C8 column (250 mm x 4.6 mm i.d., 5 μm), maintained at 35 ºC. The mobile-phase consisted of acetonitrile and a solution of triethylamine 0.3% pH 3.0 (55:45; v/v), running at a flow rate of 1.2 mL/min, with detection at 220 nm. The capillary electrophoresis method was developed using the micellar electrokinetic chromatography (MEKC) as the separation mode, and salicylic acid as internal standard. The analysis were performed on a fused-silica capillary (effective length of 72 cm) maintained at 35 °C, with 50 mM of borate buffer and 5 mM of sodium dodecyl sulfate (SDS) at pH 9.0 as background electrolyte. The separation was achieved at 25 kV applied voltage and the injection was performed at 50 mbar for 5 s, with detection at 208 nm. The specificity and stability-indicating capability of the methods were demonstrated through forced degradation studies, which also show that there is no interference of the excipients in the analysis. Moreover, the Plackett- Burman experimental design was used for robustness evaluation, giving acceptable results for both methods. The procedures were validated according to Internationals guidelines, giving results within the acceptable range. Therefore, the proposed methods were successfully applied for the simultaneous quantitative analysis of DEL and MAN in the tablet dosage form, showing non-significant difference (P>0.05), contributing to improve the quality control and to assure the therapeutic efficacy.

The thesis deals with measuring the content of chosen phenolic substances in some specimen of the genera Chenopodium L. and Atriplex L. Two independent analytical methods were used to determine the content of phenolic substances, namely the Micellar Electrokinetic Capillary Chromatography (MECC) method and the High - Performance Liquid Chromatography (HPLC) method. Two cultured species of the genera Spinacia and Atriplex, three freely growing specimen of the genus Chenopodium and three freely growing species of the genus Atriplex were analysed. The analysis concerned the leaves and the inflorescence of these species.The total content of quercetin and rutin was determined by the MECC method. The highest total content of quercetin was found in the leaves of the Garden Orache (4240 mg/kg of dry matter), the lowest total content of quercetin was found in the inflorescence of the Atriplex prostrata DC. (19.6 mg/kg of dry matter). Rutin was only found in four samples, the rest of the samples contained rutin in quantities below the limit of quantification. The highest content of rutin was found in the leaves of the Lamb´s Quarters (868 mg/kg of dry matter).

A combinação entre o delapril (DEL), um inibidor da enzima conversora de angiotensina e o manidipino (MAN), um antagonista dos canais de cálcio, produz um efeito anti-hipertensivo sinérgico, podendo ser considerado um ótimo tratamento para pacientes com hipertensão essencial leve e moderada. No presente trabalho foram desenvolvidos e validados métodos cromatográfico e eletroforético para avaliação simultânea de DEL e MAN em produto farmacêutico. As análises por cromatografia líquida em fase reversa (CL-FR) foram executadas utilizando coluna C8 (250 mm x 4,6 mm), mantida a 35 oC. A fase móvel foi constituída por acetonitrila e solução de trietilamina 0,3%, pH 3,0 (55:45; v/v), eluída na vazão de 1,2 mL/min com detecção a 220 nm. Paralelamente, desenvolveu-se método por eletroforese capilar, utilizando modo de separação por cromatografia eletrocinética micelar (MEKC) e ácido salicílico como padrão interno. Foi utilizado capilar de sílica fundida (comprimento efetivo de 72 cm) mantido a 35 °C, com solução eletrolítica composta de tampão borato 50 mM e dodecil sulfato de sódio (SDS) 5 mM, pH 9,0. Voltagem de 25 kV foi aplicada e a injeção foi de 50 mbar durante 5 s, com detecção a 208 nm. A especificidade e a capacidade dos métodos serem indicativos de estabilidade foram demonstradas através de estudos de degradação forçada dos fármacos e pela não interferência dos excipientes nas análises. Além disso, o desenho experimental Plackett-Burman foi utilizado para a avaliação da robustez, observando-se resultados adequados para ambos métodos. Os procedimentos foram validados de acordo com guias aceitos internacionalmente, observando-se resultados em uma faixa aceitável. Os métodos propostos foram aplicados com sucesso na determinação quantitativa simultânea de DEL e MAN em comprimidos, não havendo diferença significativa dos resultados (P>0,05), contribuindo, portanto, para aprimorar o controle da qualidade, assegurando a eficácia terapêutica. / The combination of delapril (DEL), an angiotensin converting enzyme inhibitor and manidipine (MAN), an antagonist of calcium channels, produces a synergic antihypertensive effect and may be regarded as an optimal antihypertensive drug treatment in mild to moderate essential hypertensive patients. The chromatographic and eletrophoretic methods for the simultaneous evaluation of DEL and MAN in pharmaceutical product were developed and validated in the present work. The reversed-phase liquid chromatography (RP-LC) method was carried out on a C8 column (250 mm x 4.6 mm i.d., 5 μm), maintained at 35 ºC. The mobile-phase consisted of acetonitrile and a solution of triethylamine 0.3% pH 3.0 (55:45; v/v), running at a flow rate of 1.2 mL/min, with detection at 220 nm. The capillary electrophoresis method was developed using the micellar electrokinetic chromatography (MEKC) as the separation mode, and salicylic acid as internal standard. The analysis were performed on a fused-silica capillary (effective length of 72 cm) maintained at 35 °C, with 50 mM of borate buffer and 5 mM of sodium dodecyl sulfate (SDS) at pH 9.0 as background electrolyte. The separation was achieved at 25 kV applied voltage and the injection was performed at 50 mbar for 5 s, with detection at 208 nm. The specificity and stability-indicating capability of the methods were demonstrated through forced degradation studies, which also show that there is no interference of the excipients in the analysis. Moreover, the Plackett- Burman experimental design was used for robustness evaluation, giving acceptable results for both methods. The procedures were validated according to Internationals guidelines, giving results within the acceptable range. Therefore, the proposed methods were successfully applied for the simultaneous quantitative analysis of DEL and MAN in the tablet dosage form, showing non-significant difference (P>0.05), contributing to improve the quality control and to assure the therapeutic efficacy.

Terpenos são os principais constituintes dos óleos essenciais e vêm sendo explorados há mais de 3500 anos pela humanidade. Por conta das suas propriedades flavorizantes, são amplamente empregados na indústria de cosméticos e perfumaria. Apresentam ainda uma infinidade de funções biológicas, como promoção de polinização nas plantas, e proteção contra pragas e animais. Além dessas funções, muitos compostos possuem ainda atividade antimicrobiana, anti-inflamatória, antifúngica, entre outras. Tendo em vista a simplicidade estrutural dos terpenos e a alta hidrofobicidade que sugere fracas interações intermoleculares, é difícil de se imaginar como esses compostos conseguem desempenhar funções tão específicas e diversas. É de se esperar que quanto mais complexa a estrutura do composto, mais fácil seja seu reconhecimento pelo organismo. Isso mostra o grande poder de reconhecimento do meio biológico. Nesse trabalho, os parâmetros termodinâmicos de transferência da fase aquosa para a fase micelar de 10 terpenos (carvona, cânfora, cumeno, t-anetol, eugenol, limoneno, citronelal, linalol, terpineol e verbenona) e cumarina em dois sistemas, SDS 30 mmol.kg-1 + TBS 20 mmol.kg-1 e SDS 30 mmol.kg-1 + TBS 20 mmol.kg-1 + 10% v/v de etanol foram determinados buscando elucidar a incorporação micelar desses compostos. Micelas apresentam compartimentos com diferentes polaridades e podem servir como modelo para mimetizar as diferentes interações no meio biológico. Dessa forma, a utilização da cromatografia eletrocinética micelar (MEKC, do inglês Micellar Electrokinetic Chromatography) na determinação dos coeficientes de partição e dos parâmetros termodinâmicos de transferência entre as fases aquosa e micelar desses solutos pode contribuir para o entendimento da distribuição bem como auxiliar na compreensão das funções que os mesmos desempenham na natureza. A hipótese de que os parâmetros termodinâmicos podem elucidar detalhes da incorporação micelar foi ainda testada através da busca de relações lineares de energia de solvatação (LSER, do inglês Linear Solvation Energy Relashionships) com o intuito de evidenciar as principais características moleculares que contribuem para o processo detransferência. Os modelos LSER foram estudados através de regressão múltipla e análises multivariadas de PLS, SPLS, PLS-DA e SPLS-DA, com o objetivo de verificar as propriedades dos terpenos que explicam sua incorporação nas micelas. Outras análises estatísticas multivariadas, como análise de agrupamentos e PCA, foram utilizadas para estudar a variabilidade estrutural dos compostos selecionados, bem como, determinar se os descritores teóricos calculados conseguem descrever as características estruturais dos terpenos. O estudo da termodinâmica de transferência de solutos neutros da fase aquosa para a fase micelar demonstrou que mesmo pequenas diferenças estruturais das moléculas contêm informação sobre a distribuição dos compostos nos compartimentos micelares. Também podese inferir sobre o efeito do etanol nas partições e sobre a própria estrutura micelar. Os resultados para o limoneno mostraram a complexidade envolvida nas partições, levando a ideia de restrição de volume nas micelas modificadas por álcool. Resultados de LSER mostraram que a transferência da fase aquosa para a fase micelar desses compostos é governada principalmente pela interação hidrofóbica onde Vx (Volume de McGowan) foi selecionado como um dos descritores mais importantes para explicar lnP. A análise comparativa dos resultados obtidos pelos dois métodos (estudo dos parâmetros termodinâmicos e LSER) indicou similaridade de resultados. Isso demonstra a grande confiabilidade dos resultados e, então, que estudos similares usando outras soluções micelares e outras classes de compostos (hormônios, flavonoides, aminas, etc.) podem ser muito promissores. / Terpenes are the main constituents of essential oils and have been explored for more than 3,500 years. Because of their flavoring properties, terpenes are widely used in the cosmetics and perfumery industry. They also exert a multitude of ecological functions, such as the promotion of plant pollination and protection against pests and animals. In addition, many compounds have antimicrobial, antifungal, anti-inflammatory activities and others. Given the structural simplicity of terpenes and the high hydrophobicity that suggests weak intermolecular interactions, it is difficult to imagine how these compounds can perform such specific and diverse functions. It is expected that the more complex the structure of the compound, the easier it is its recognition by the organism, which does not seem to be true for this class showing the great power of recognition of the biological system. In this work, the thermodynamic parameters of aqueous and micellar phase transfer of ten terpenes (carvone, camphor, cumene, t-anethol, eugenol, limonene, citronellal, linalool, terpineol, and verbenone) and coumarin in two systems, 30 mmol.kg-1 of SDS + 20 mmol.kg-1 of TBS and 30 mmol.kg-1 of SDS, 20 mmol.kg-1 of TBS, and 10% v/v of ethanol were determined to elucidate the micellar distribution of these compounds. Micelles have compartments that possess different polarities and might be a model to mimic the different interactions that terpenes may have in the biological environment. Thus, the use of micellar electrokinetic chromatography (MEKC) in the determination of the partition coefficients and the thermodynamic parameters of transfer of the aqueous phase to the micellar phase of these solutes can contribute to the understanding of the distribution, as well as help in the understanding of the functions they perform in nature. The hypothesis that the thermodynamic parameters can elucidate details of the micellar incorporation was further analyzed through the search of Linear Solvation Energy Relashionships (LSER), in order to highlight the main molecular characteristics that contribute to the transfer process. The LSER models were studied through multiple regression and other multivariate analyzes, such as PLS, SPLS, PLS-DA and SPLS-DA, in order to verify the properties of terpenes that explain their incorporation into micelles.Other multivariate statistical analysis, such as cluster analysis and PCA were used to study the structural variability of the selected compounds, as well as to determine if the calculated theoretical descriptors can describe all the structural characteristics of the terpenes. The study of thermodynamics of transfer of neutral solutes from the aqueous phase to the micellar phase has shown that even small structural differences of the molecules contain information about the distribution of the compounds in the micellar compartments. It was also possible to infer about the effect of ethanol on the partitions and on the micellar structure. The results for limonene showed the complexity involved in the partitions, showing that occurs volume restriction in alcohol-modified micelles. Results from LSER showed that the transfer of these compounds is mainly governed by hydrophobic interactions where Vx (McGowan volume) was selected as one of the most important descriptors to explain partition. The comparative analysis of the results obtained by the two methods (thermodynamic parameters studies and LSER) indicated similarity of results. This demonstrates the great reliability of the methods, and that similar studies using other micellar solutions and other classes of compounds (hormones, flavonoids, amines, etc.) might be very promising.

Atualmente cerca de 38,6 milhões de pessoas estão infectadas pelo vírus da síndrome da imunodeficiência adquirida (AIDS) em todo mundo. O único modo de tratamento para esta doença é através da utilização de medicamentos responsáveis por atuarem em diferentes pontos do ciclo replicativo do vírus. No Brasil esta doença é tratada como de calamidade pública, sendo seu tratamento feito através do programa nacional DST/AIDS, o qual distribui gratuitamente os medicamentos necessários para o tratamento. Tendo em vista que vários desses medicamentos são formulados pela indústria local, esta dissertação tem como objetivo o desenvolvimento de métodos analíticos passiveis de aplicação na rotina farmacêutica para a qualificação de matérias primas, bem como o controle de qualidade dos produtos acabados. As determinações nas formulações de nevirapina e lamivudina foram realizadas por CZE, em eletrólitos ácidos com pH < 2,5. Para a lamivudina a determinação foi realizada em um eletrólito de 0,5 % de TEA, 20 mmol.L-1 de TRIS, pH = 2,5, ajustado com ácido fosfórico, com um tempo de análise inferior a 4 minutos. O método desenvolvido para a nevirapina foi conduzido em um eletrólito de 10 mmol.L-1 de fosfato de sódio (pH =2,5), com um tempo de análise de 3 minutos. Um outro método foi desenvolvido permitindo a determinação de efavirenz, estavudina e ritonavir por MEKC, utilizando-se um planejamento fatorial 23 com ponto central, um tempo inferior a 9 minutos em um eletrólito consituído de 20 mmol.L-1 de tetraborato de sódio, 20 mmol.L-1 de SDS e 30 % de acetonitrila. Os métodos desenvolvidos foram validados de acordo com os protocolos oficiais, mostrando que estes métodos apresentam características adequadas para a análise de formulações farmacêuticas. Outra abordagem foi feita utilizando o acoplamento da eletroforese capilar à espectrometria de massas, onde amostras de urina fortificadas foram analisadas. As análise foram conduzidas utilizando 400 mmol.L-1 de ácido fórmico e líquido auxiliar constituído de 0,5 % de ácido fórmico diluído com uma solução metanol:água (1:1), permitindo a identificação inequívoca dos fármacos. / There are approximately 38.6 million people infected by the immunodeficiency acquired virus (AIDS) over the world. The only way of treatment for this illness is administrating drugs that act in different points of the replicative cycle of the virus. In Brazil this illness is dealt as public calamity, being its treatment made through the national program DST/AIDS, which distributes free of charge necessary medicines for the treatment. Considering that many of these drugs are formulated by the local industries, this thesis has as objective the development of analytical methods to be applied in the pharmaceutical routine for the qualification of raw materials, as well as the quality control of the finished products. The analysis of drug formulations of nevirapine and lamivudine were carried by CZE, in acid electrolytes with pH < 2.5. For lamivudine the analysis was carried using an electrolyte composed of 0.5 % of TEA, 20 mmol.L-1 of TRIS, pH = 2.5, adjusted with phosphoric acid, with an analysis time less than 4 minutes. The method developed for the nevirapine, was lead in an electrolyte composed of 10 mmol.L-1 of phosphate buffer (pH = 2.5), with a time of analysis of 3 minutes. Another method was developed for efavirenz, estavudine and ritonavir by MEKC, using 23 a factorial design with central point, with an analysis time less then 9 minutes in an electrolyte of 20 mmol.L-1 of sodium tetraborate, 20 mmol.L-1 sodium dodecyl sulfate and 30 % acetonitrile. The developed methods were validated in accordance with official protocols, showing that these methods can be advantageously used in the analysis of pharmaceutical formulations. Another approach was to use the coupling of capillary electrophoresis with mass spectrometry, where fortified samples of urine had been analyzed. The analysis were performed using 400 mmol.L-1 formic acid and liquid sheath consisting of 0.5 % of formic acid diluted with a solution of (1:1) methanol:water, allowing the unequivocal identification detection of the drugs in the samples.

Submitted by Renata Lopes (renatasil82@gmail.com) on 2017-05-09T19:23:03Z No. of bitstreams: 1 carinadealmeidabastos.pdf: 1640331 bytes, checksum: 96c6f61dba624b337759b27baecc7a34 (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2017-05-17T14:34:40Z (GMT) No. of bitstreams: 1 carinadealmeidabastos.pdf: 1640331 bytes, checksum: 96c6f61dba624b337759b27baecc7a34 (MD5) / Made available in DSpace on 2017-05-17T14:34:40Z (GMT). No. of bitstreams: 1 carinadealmeidabastos.pdf: 1640331 bytes, checksum: 96c6f61dba624b337759b27baecc7a34 (MD5) Previous issue date: 2015-03-20 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Um método cromatográfico eletrocinético micelar para a determinação simultânea do mesilato de nelfinavir e das impurezas ácido 3-hidroxi-2-metilbenzóico e benzoato de (2R,3R)-4-((3S,4aS,8aS)-3-(terc-butilcarbamoil)octahidroisoquinolina-2(1H)-il)-3-hidroxi-1-(feniltio)butano-2-amônio, com tempo de análise de 25 minutos, foi proposto. O eletrólito composto por tampão tetraborato de sódio (pH 9,24; 25 mmol L−1), dodecil sulfato de sódio (9 mmol L−1) e metanol (10%, v/v) foi otimizado utilizando planejamento fatorial misto, com detecção direta em 200 nm. Após avaliação das figuras de mérito seletividade, linearidade, precisão, limite de detecção, limite de quantificação, exatidão e robustez (Teste de Youden), o método foi aplicado na análise do mesilato de nelfinavir e suas impurezas em uma formulação farmacêutica (comprimidos). O método otimizado pode ser útil na determinação desses analitos em processos de monitoramento de síntese, matérias-primas e formulações farmacêuticas, oferecendo como vantagens baixo consumo de solventes, pequena demanda de amostra e uso de colunas não específicas. / A methodology for the simultaneous determination of nelfinavir mesylate and the impurities 3-hydroxy-2-methylbenzoic acid and (2R,3R)-4-((3S,4aS,8aS)-3-(tert-butylcarbamoyl) octahydroisoquinolin-2(1H)-yl)-3-hydroxy-1-(phenylthio)butan-2-aminium benzoate by micellar electrokinetic chromatography, with an analysis time of 25 min, was proposed. An electrolyte composed of sodium tetraborate buffer (pH 9.24; 25 mmol L−1), sodium dodecyl sulphate (9 mmol L−1) and methanol (10%, v/v) was optimized using a mixed-level factorial design, with direct detection at 200 nm. After evaluating some figures of merit, such as selectivity, linearity, precision, limit of detection, limit of quantification, accuracy and robustness (Youden’s test), the method was successfully applied to the analysis of nelfinavir mesylate and its impurities in a pharmaceutical formulation (tablets). The optimized methodology is demonstrated to be useful in the determination of these analytes in a synthesis monitoring process, in raw materials and in pharmaceutical formulations, while offering low solvent consumption, requiring a small sample and using non-specific columns as advantages.

Le travail de thèse présenté dans ce manuscrit s’inscrit dans une dynamique d’intégration de matériaux non conventionnels en systèmes microfluidiques. Il vise à démontrer le potentiel du cPDMS, un matériau nanocomposite formé d’une matrice de polydiméthylsiloxane rendu conducteur par l’ajout de nanoparticules de carbone. Compatible avec les procédés technologiques habituels, le cPDMS peut être structuré dans une large gamme d’épaisseurs et de géométries mais présente surtout l’avantage de pouvoir être collé irréversiblement sur verre, PDMS et silicium. Son intégration est parfaitement étanche, rapide à mettre en oeuvre, et très économique. La première partie du manuscrit est consacrée à la caractérisation de ce matériau. Ses propriétés électriques et de surface, pouvant être critiques pour une utilisation en microfluidique, ont été particulièrement étudiées. Les champs électriques offrant de nombreuses possibilités pour réaliser des fonctions clés en microfluidique (détection, séparation, manipulation de fluides ou de particules), nous avons choisi d’évaluer l’intérêt d’électrodes de cPDMS dans deux types d’applications. Les aspects de détection ont d’abord été mis en évidence à l’aide de mesures électrochimiques. Cette méthode a permis à la fois de caractériser la surface du cPDMS tout en validant son utilisation potentielle pour des applications d’analyses électrochimiques. Dans la dernière partie du manuscrit, le matériau a été testé pour la manipulation de particules à travers l’observation de différents phénomènes électrocinétiques. Ceux-ci ont conduit à la mise au point de dispositifs microfluidiques (intégrant des lectrodes de cPDMS) dédiés à la lyse et à l’électrofusion de cellules. / The work presented in this thesis deals with the integration of non-conventional materials in microfluidic systems. It aims to demonstrate the potential of cPDMS, a conductive nanocomposite material made up of polydimethylsiloxane matrix mixed with carbon nanoparticles. Compatible with the usual technological processes such as soft lithography, cPDMS can be microstructured in a large range of thicknesses and geometries. Moreover, cPDMS can be quickly, irreversibly and perfectly sealed to glass, PDMS and silicon substrates, something that is not possible for conventional metallic electrodes. The first part of the manuscript is centered on the characterization of this material. Its electrical and surface properties that may turn out critical for microfluidic applications have been particularly studied. Electric fields present many opportunities to perform key functions in microfluidic (detection, separation, fluid or particles handling). We have chosen to assess the potential of cPDMS electrodes for two kinds of applications. Aspects of detection were first demonstrated using cyclic voltammetry measurements. This electrochemical method has enabled both to characterize the cPDMS surface while validating its potential as an electrochemical analysis tool. In the last part of this manuscript, cPDMS was tested for the electrokinetic manipulation of particles through thre study of different electrical fields with induced phenomena. This has led to the development of microfluidic devices (integrating cPDMS electrodes) designed for cell lysis and cells electrofusion.