Neste trabalho é desenvolvido um novo método para a análise de flúor em água, na região de concentrações de décimos de ppm, baseado na técnica PIGE (Particle-Induced Gamma-ray Emission). Mostra-se que a determinação de concentrações dessa ordem de grandeza pela técnica PIGE exige o uso de um método de pré-concentração. Algumas tentativas de pré-concentração do flúor por meio de sua adsorção em sólidos em pó (carvão ativado, sílica e óxido de zircônio, entre outros) são descritas. A não-uniformidade da distribuição de flúor observada nestes materiais, após a adsorção, constituiu um problema crítico para a análise PIGE, que inviabilizou a extração de informações quantitativas a respeito das concentrações. Assim, o método utilizado para pré-concentrar o flúor nas amostras de água foi a redução de volume por evaporação. As amostras pré-concentradas foram irradiadas por um feixe de prótons de 4 MeV, em ar. Para testar a validade da metodologia de análise desenvolvida, cinco amostras de água fuoretada da cidade de São Paulo foram coletadas para a realização de um exame intercomparativo com outros cinco laboratórios. A análise nestes laboratórios foi realizada por meio das técnicas do eletrodo íon-seletivo e de cromatografia iônica. Os resultados da intercomparação demonstram claramente que a metodologia proposta neste trabalho é adequada para a análise de água fluoretada. Não foram observados indícios de erros sistemáticos e a precisão final nos valores de concentração foi de 5 a 10%. Algumas perspectivas de tornar a análise mais precisa são apontadas e discutidas. / This work presents a new method for the analysis of fluorine in water, at the tenth of ppm concentration level, based on the PIGE (Particle-Induced Gamma-ray Emission) technique. It is shown that determination of concentrations of this order of magnitude by PIGE requires the use of a preconcentration method. Some attempts of preconcentration of fluorine by adsorption in powdered solids (activated carbon, silica and zirconium oxide, among others) are described. The non-uniformity of the fluorine distribution in these materials, after adsorption, was a critical problem for the PIGE analysis, hinding the extraction of quantitative information about fluorine concentrations. This, volume reduction by evaporation was used to preconcentrate fluorine in water. The preconcentrated samples were irradiated by a 4 MeV proton beam, in air. In order to test the validity of the analytical method developed in this work, five samples of fluoridated water of the city of São Paulo were collected for an intercomparative exam with ve other laboratories. Analysis in these laboratories was performed by the ion-selective electrode and ion cromatography techniques. The intercomparison results show with confidence that the methodology proposed in this work is adequate for the analysis of fluoridated water. No significant systematic errors were observed and the precision was at the 5-10% level. Possibilities for the improvement of precision are pointed and discussed.

O objetivo desta pesquisa foi desenvolver, validar e comparar métodos analíticos para separação e quantificação de citrato de colina e acetilmetionina em formulações injetáveis por cromatografia líquida de alta eficiência (CLAE) e eletroforese capilar (EC). O método por CLAE, com o qual foi possível quantificar e separar simultaneamente, o citrato de colina e a acetilmetionina, foi realizado empregando-se uma coluna cromatográfica C18, 250 x 4,6 mm, 5µm, Shim-Pack® VP-ODS e fase móvel constituída por tampão fosfato de potássio 25 mM pH 5,7: acetonitrila: metanol (88:10:2 v/v/v). O método por EC foi validado utilizando-se como eletrólito, tampão de tetraborato de sódio 20 mM a pH 9,2 e capilar de sílica fundida de 40,2 cm, sendo 30 cm efetivos, com 75 µm de diâmetro interno. O método por CLAE apresentou um coeficiente de correlação (r) de 0,9996 para o citrato de colina e 0,9998 para a acetilmetionina. No método por EC, o coeficiente de correlação (r) encontrado foi de 0,9996 para a acetilmetionina. Os métodos podem ser considerados eficientes e confiáveis para serem empregados em análises de rotina de controle de qualidade de soluções injetáveis contendo a associação dos dois fármacos selecionados para o estudo. / The objective of this research was to develop, to validate and to compare two analytical methods for separation and quantification of choline citrate and acetylmethionine in injectable formulations by high performance liquid chromatography (HPLC) and capillary electrophoresis (CE). The HPLC method which enables the separation and simultaneous quantitative determination of choline citrate and acetylmethionine was performed using a C18, 250 x 4.6 mm, 5µm-Shim-Pack®VP-ODS column and a mobile phase constituted of buffer 25 mM potassium phosphate, pH 5.7: acetonitrile: methanol (88:10:2 v/v/v). The CE method was validated using a solution of 20 mM sodium tetraborate buffer pH 9.2 as the electrolyte and a fused silica capillary of 40.2 cm, with 30 cm effective, and 75 mm of diameter. The HPLC method showed a correlation coefficient (r) of 0.9996 for choline citrate and 0.9998 for acetylmethionine. For CE method the correlation coefficient (r) was found to be 0.9996 for acetylmethionine. The methods showed to be efficient and reliable methods, and can be used in routine analysis for quality control of injectable solutions containing the association of the two compounds selected to be studied in this research.

O controle da fertilidade é obtido principalmente pela inibição da ovulação por meio da atividade combinada de dois componentes principais: estrogênio e progestina. Desta forma, novas formulações vêm sendo desenvolvidas com baixa dose de etinilestradiol associado a um novo agente progestógeno, a drospirenona, com o objetivo de garantir equilíbrio entre eficácia, segurança e controle de ciclo menstrual. Este trabalho tem como objetivo desenvolver, validar e comparar métodos analíticos para a identificação e quantificação de etinilestardiol e drospirenona presentes em uma mesma formulação por cromatografia em fase líquida de alta eficiência (CLAE) e a eletroforese capilar (EC). O método por CLAE, foi realizado empregando-se uma coluna LiChoCART® 100RP - C18, (125 x 4 mm) 5 µm, fase móvel constituída por MeCN/H2O 50:50 (v/v) e vazão de 1.0 mL min. Utilizou-se um detector UV 200 nm para drospirenona e um de fluorescência λex = 280 nm e λem = 310 nm, para o etinilestradiol. O método por EC foi validado utilizando-se como eletrólito, tampão de tetraborato de sódio 30 mM, dodecil sulfato de sódio 20 mM e acetonitrila 30% a pH 9,2, capilar de sílica fundida de 31,2 cm sendo 21 cm efetivos, com 75 µm de diâmetro interno. O método por CLAE apresentou um coeficiente de correlação (r2) de 0,9989 para o etinilestradiol e 0,999 para a drospirenona. No método por EC, o coeficiente de correlação (r2) encontrado foi de 0,9988 para o etinilestradiol e 0,998 para a drospirenona. Os métodos podem ser considerados eficientes e confiáveis para serem empregados em análise de rotina para controle de qualidade destes produtos farmacêuticos. / The fertility control is achieved mainly by inhibiting ovulation through the combined activity of two main components: estrogen and progestin. Thus, new formulations have been developed associating appropriately low dose of ethinylestradiol to a new drug, the drospirenone, in order to ensure the balance between efficacy, safety and cycle control. The objective of this research was to develop, to validate and to compare two analytical method for separation and quantification of ethinylestardiol and drospirenone using high performance liquid chromatography and capillary electrophoresis (CE). The HPLC method, was performed using a LiChoCART® 100RP - C18, (125 x 4 mm) 5 µm column, a mobile phase constituted of acetonitrile:water (50:50 v/v) and flow rate of 1.0 mL min. UV detection was made for drospirenone at 200 nm coupled with a fluorescence detector at λex= 280 nm and λem = 310 nm for ethinylestradiol. The CE method was validated using a solution of sodium tetraborate buffer 30 mM, sodium dodecil sulphate 20 mM and acetonitrile 30%, pH 9.2 and a fused silica capillary of 31,2 cm with 21 cm effective and 75 µm of diameter. The HPLC method showed a correlation coefficient (r2) of 0.9989 for ethinylestradiol and 0,999 for drospirenone. For CE method, the correlation coefficient was (r2) 0,9988 for ethinylestradiol and 0,9989 for drospirenone. The methods showed to be efficient and reliable, and can be used in routine analysis for quality control of these pharmaceutical preparations.

Há milhares de anos, as imagens visuais impactam significantemente o cotidiano do ser humano, caracterizando-se como um ótimo meio de comunicação e expressão. Com os avanços tecnológicos, evidenciam-se transformações significativas da linguagem visual, que se moldam aos novos contextos no qual se encontra. Os videogames, artefatos digitais amplamente difundidos na sociedade que permitem a imersão do usuário em ambientes lúdicos dotados de interatividade, são alvo de projetos estéticos cada vez mais sofisticados. Uma vez que utilizam linguagem predominantemente visual, tem-se como premissa que a imagem desempenha papel fundamental para que estes cumpram sua função adequadamente. Entretanto, as imagens em videogames nem sempre obedecem a um padrão de qualidade, carecendo de estudos e métodos que amparem seu desenvolvimento e compreensão. Com isso, o presente estudo tem como objetivo desenvolver um método para análise da imagem visual em videogames, considerando a ampla gama de funções que a mesma exerce em artefatos dessa natureza. Para isso, a fim de permitir o desenvolvimento do método e garantir sua replicabilidade, foram definidos determinados procedimentos metodológicos, que envolvem: a realização e avaliação de um primeiro modelo do método; desenvolvimento de um segundo modelo; coleta e análise de dados envolvendo sujeitos de pesquisa especialistas na área; e o desenvolvimento de um modelo final. Como resultados, pode-se perceber que a análise de imagens visuais em videogames pode ser realizada a partir de um método sistemático, todavia, foram apontadas inúmeras ressalvas e considerações a respeito de como o método pode tornar-se mais eficiente. / For thousands of years, the visual images significantly affect the daily life of the human being, characterized as a great means of communication and expression. With technological advances, are evident the significant changes in visual language, which are molded to the new contexts in which it is. Video games, digital artifacts widespread in society that allow the user's immersion in playful environments with interactivity. They are subject to increasingly sophisticated aesthetic designs. Once they predominantly use visual language, there is a premise that the image plays a key role for them to fulfill their function properly. However, images in video games does not always follow a standard of quality, lacking studies and methods that help its development and understanding. Thus, this study aims to develop a method for the analysis of visual image in video games, considering the wide range of functions that it carries on such artifacts. For this, in order to allow the development of the method and ensuring their replication, were defined certain methodological procedures that involve: implementation and evaluation of a first model of the method; development of a second model; collection and analysis of data involving research subjects experts in the field; and the development of the final model. As a result, it can be perceived that the analysis of visual images in videogames can be performed from a systematic method, however, were identified numerous considerations about how the method can become more efficient.

O controle da fertilidade é obtido principalmente pela inibição da ovulação por meio da atividade combinada de dois componentes principais: estrogênio e progestina. Desta forma, novas formulações vêm sendo desenvolvidas com baixa dose de etinilestradiol associado a um novo agente progestógeno, a drospirenona, com o objetivo de garantir equilíbrio entre eficácia, segurança e controle de ciclo menstrual. Este trabalho tem como objetivo desenvolver, validar e comparar métodos analíticos para a identificação e quantificação de etinilestardiol e drospirenona presentes em uma mesma formulação por cromatografia em fase líquida de alta eficiência (CLAE) e a eletroforese capilar (EC). O método por CLAE, foi realizado empregando-se uma coluna LiChoCART® 100RP - C18, (125 x 4 mm) 5 µm, fase móvel constituída por MeCN/H2O 50:50 (v/v) e vazão de 1.0 mL min. Utilizou-se um detector UV 200 nm para drospirenona e um de fluorescência λex = 280 nm e λem = 310 nm, para o etinilestradiol. O método por EC foi validado utilizando-se como eletrólito, tampão de tetraborato de sódio 30 mM, dodecil sulfato de sódio 20 mM e acetonitrila 30% a pH 9,2, capilar de sílica fundida de 31,2 cm sendo 21 cm efetivos, com 75 µm de diâmetro interno. O método por CLAE apresentou um coeficiente de correlação (r2) de 0,9989 para o etinilestradiol e 0,999 para a drospirenona. No método por EC, o coeficiente de correlação (r2) encontrado foi de 0,9988 para o etinilestradiol e 0,998 para a drospirenona. Os métodos podem ser considerados eficientes e confiáveis para serem empregados em análise de rotina para controle de qualidade destes produtos farmacêuticos. / The fertility control is achieved mainly by inhibiting ovulation through the combined activity of two main components: estrogen and progestin. Thus, new formulations have been developed associating appropriately low dose of ethinylestradiol to a new drug, the drospirenone, in order to ensure the balance between efficacy, safety and cycle control. The objective of this research was to develop, to validate and to compare two analytical method for separation and quantification of ethinylestardiol and drospirenone using high performance liquid chromatography and capillary electrophoresis (CE). The HPLC method, was performed using a LiChoCART® 100RP - C18, (125 x 4 mm) 5 µm column, a mobile phase constituted of acetonitrile:water (50:50 v/v) and flow rate of 1.0 mL min. UV detection was made for drospirenone at 200 nm coupled with a fluorescence detector at λex= 280 nm and λem = 310 nm for ethinylestradiol. The CE method was validated using a solution of sodium tetraborate buffer 30 mM, sodium dodecil sulphate 20 mM and acetonitrile 30%, pH 9.2 and a fused silica capillary of 31,2 cm with 21 cm effective and 75 µm of diameter. The HPLC method showed a correlation coefficient (r2) of 0.9989 for ethinylestradiol and 0,999 for drospirenone. For CE method, the correlation coefficient was (r2) 0,9988 for ethinylestradiol and 0,9989 for drospirenone. The methods showed to be efficient and reliable, and can be used in routine analysis for quality control of these pharmaceutical preparations.

O objetivo desta pesquisa foi desenvolver, validar e comparar métodos analíticos para separação e quantificação de citrato de colina e acetilmetionina em formulações injetáveis por cromatografia líquida de alta eficiência (CLAE) e eletroforese capilar (EC). O método por CLAE, com o qual foi possível quantificar e separar simultaneamente, o citrato de colina e a acetilmetionina, foi realizado empregando-se uma coluna cromatográfica C18, 250 x 4,6 mm, 5µm, Shim-Pack® VP-ODS e fase móvel constituída por tampão fosfato de potássio 25 mM pH 5,7: acetonitrila: metanol (88:10:2 v/v/v). O método por EC foi validado utilizando-se como eletrólito, tampão de tetraborato de sódio 20 mM a pH 9,2 e capilar de sílica fundida de 40,2 cm, sendo 30 cm efetivos, com 75 µm de diâmetro interno. O método por CLAE apresentou um coeficiente de correlação (r) de 0,9996 para o citrato de colina e 0,9998 para a acetilmetionina. No método por EC, o coeficiente de correlação (r) encontrado foi de 0,9996 para a acetilmetionina. Os métodos podem ser considerados eficientes e confiáveis para serem empregados em análises de rotina de controle de qualidade de soluções injetáveis contendo a associação dos dois fármacos selecionados para o estudo. / The objective of this research was to develop, to validate and to compare two analytical methods for separation and quantification of choline citrate and acetylmethionine in injectable formulations by high performance liquid chromatography (HPLC) and capillary electrophoresis (CE). The HPLC method which enables the separation and simultaneous quantitative determination of choline citrate and acetylmethionine was performed using a C18, 250 x 4.6 mm, 5µm-Shim-Pack®VP-ODS column and a mobile phase constituted of buffer 25 mM potassium phosphate, pH 5.7: acetonitrile: methanol (88:10:2 v/v/v). The CE method was validated using a solution of 20 mM sodium tetraborate buffer pH 9.2 as the electrolyte and a fused silica capillary of 40.2 cm, with 30 cm effective, and 75 mm of diameter. The HPLC method showed a correlation coefficient (r) of 0.9996 for choline citrate and 0.9998 for acetylmethionine. For CE method the correlation coefficient (r) was found to be 0.9996 for acetylmethionine. The methods showed to be efficient and reliable methods, and can be used in routine analysis for quality control of injectable solutions containing the association of the two compounds selected to be studied in this research.

Este trabalho apresenta estudo sobre o voriconazol, antifúngico de amplo espectro liberado para tratamento de infecções fúngicas invasivas no ano de 2002, focado no controle de qualidade e estudo de estabilidade. Foram realizados testes, ainda não citados na literatura, que visaram à caracterização do fármaco tais como: espectrofotometria na região do infravermelho, espectroscopia de ressonância magnética nuclear de hidrogênio e de carbono. Os seguintes métodos de análise quantitativa do fármaco em comprimidos foram desenvolvidos e validados: espectrofotometria na região do UV, cromatografia a líquido de alta eficiência e ensaio microbiológico - método de difusão em ágar. Todos os métodos foram avaliados frente aos parâmetros de linearidade, exatidão e precisão. A especificidade foi avaliada nos métodos de CLAE e UV, pela análise de placebo (CLAE e UV) e testes de degradação forçada (CLAE). A robustez foi avaliada no método de CLAE. Os resultados obtidos através destes métodos foram comparados estatisticamente por ANOVA, que indicou não haver diferenças estatisticamente significativas entre os mesmos; no entanto, os métodos por CLAE e ensaio microbiológico possibilitam a quantificação do voriconazol em amostras degradadas. Foram estudadas as estabilidades térmica, fotoquímica e química do voriconazol. Quimicamente, o fármaco é degradado intensamente em meio alcalino e em menor extensão em meios ácido, oxidante e neutro. Na temperatura testada (60 ºC), o voriconazol é estável em estado sólido e instável em solução metanólica (teor de 25% em 21 dias). O fármaco é instável às radiações UV-C, em maior grau, e UV-A (degradação menos intensa), tanto em solução quanto em estado sólido. Em ambas, a degradação é favorecida se o fármaco estiver em solução. Também foi estudada a estabilidade da formulação injetável, como produto reconstituído e em soluções para infusão. Verificou-se que o prazo de validade da solução reconstituída de 24 horas, proposto pelo laboratório produtor, pode ser estendido para oito dias. As soluções para infusão do voriconazol em cloreto de sódio 0,9% ou glicose 5%, preparadas em bolsas de PVC, devem ser administradas logo após sua preparação, se mantidas em temperatura ambiente. Mantidas sob refrigeração, são estáveis por 11 e nove dias, respectivamente. Isolou-se e identificou-se o produto de degradação majoritário observado durante os estudos de estabilidade, que corresponde a 1- (2,4-difluorfenil)-2-(1H-1,2,4-triazol-1-il)-1-etanona. Foi constatado que os produtos de degradação do voriconazol não apresentam atividade antifúngica; sendo assim, cuidados em relação à temperatura e luz devem ser tomados ao armazenar soluções do fármaco. / This study is focused on quality control and stability evaluation of voriconazole, a broad spectrum antifungal agent, approved for treatment of invasive fungal infections in 2002. The drug was characterized by tests as IV spectrophotometry, nuclear magnetic ressonance spectrometry of hydrogen and carbon, not refered until the moment. The following methods, applied to the assay of voriconazole in tablets were developed and validated: UV spectrophotometry, HPLC assay and microbiological assay, using the cylinder-plate method. All of them were evaluated in the following parameters: linearity, accuracy and precision. The specificity was evaluated in HPLC and UV assays, by analysis of excipients and/or degradated samples. The robustness was evaluated in the HPLC assay. The results obtained in the three methods were compared by ANOVA, which indicated that they are equivalent. However, only HPLC and microbiological assays could be used in the quantitation of voriconazole in degradated samples. The stability of voriconazole under temperature, radiation and different chemical mediums was evaluated. The drug is extensively degradated under alkaline medium, being degradated in less extension under acidic, neutral and oxidant mediums. Voriconazole is stable in the tested temperature (60 ºC) in solid state, but is unstable in solution (loss of 75%, in 21 days). The drug is unstable to UV-C and UV-A radiations, both in solution or in solid state, being the degradation on UV-C more intense. The degradation is higher if the drug is in solution. The stability of the injectable formulation was studied, as reconstituted product and infusion solutions in PVC bags. Voriconazole reconstituted product was stable for eight days, stored at 4-7 ºC. As infusion solution, in 0.9% sodium chloride or in 5% dextrose, stored at room temperature, the drug should be administered only just after preparation. Stored at 4-7 ºC, the infusion solutions were stable for 11 and 9 days, respectivelly. The main degradation product observed in the stability studies was isolated and identified. It corresponds to 1-(2,4-difluorophenyl)-2-(1H-1,2,4- triazol-1-yl)-1-ethanone. It was comproved that the voriconazole degradation xxx products don’t have antifungal activity. So, special care should be taken in relation to temperature and radiation, when solutions of voriconazole are stored.

Drug delivery technology is currently advancing faster than at any time in biotechnology history. The challenge of drug delivery is to achieve a controlled release of therapeutic agents over an extended period. Controlled release potentially offers significant advantages over conventional dosage forms, by eliminating both under- and overdosing while maintaining a desired range of drug concentrations. An existing drug regimen for induced calving produces a declining blood profile that does not mimic the naturally occurring, gradually increasing, cortisol blood level around parturition effectively, causing animal health issues. Likewise, while the existing progesterone controlled release systems for oestrous control successfully synchronise oestrus, it is however, associated with reduced fertility and as such other drugs have to be administered to improve the fertility during oestrus. Therefore, there is a need for a drug delivery system that is capable of delivering multiple drugs at various times and patterns. This research aimed to investigate, characterize, identify steroids with high absorption rates through vaginal mucosa and evaluate the potential of an electronic drug delivery system for the delivery of steroids for either the control of the bovine oestrous cycle or induced calving. In order to identify steroids with high absorption rates across the vaginal mucosa, an in vitro permeation method was developed to screen selected steroids for their ability to permeate artificial and biological membranes. The steroids were pre-formulated to enhance their solubility and permeation through these membranes. Analytical UV and HPLC assays to characterise the pure and formulated steroidal compounds were also developed and validated. An assessment of an intravaginal Electronically Modulated Intravaginal Device (EMID) for the control of the bovine oestrous cycle or induced calving was carried out. Five different release assessment methods were investigated and critically evaluated in order to identify the most appropriate release assessment method for the EMID. These were: 1) the Drug Dissolution Test, 2) a weight loss method, 3) the dispensed weight method, 4) the determination of piston travel distance method, and 5) the rod expulsion from the EMID method. The methods investigated were critically evaluated in terms of ease of use and automation, reproducibility and cost/time savings. Optimisation of various components and construction materials of the EMID were also investigated. Animal trials were carried out using the original EMID (manufactured from polypropylene polymer) and modified inserts (manufactured from high density polyethylene polymer) to determine their retention rate in the animals. Accelerated stability testing of progesterone in suspension, oestradiol-17 tablets, cloprostenol as a powder blend and the driving mechanism of the EMID were examined. The flux of the steroids was evaluated through poly-&-caprolactone and excised cow mucosa membranes using side-by-side permeation cells. Results indicated that progesterone followed by dexamethasone acetate and dexamethasone valerate showed higher permeability values through vaginal mucosa compared to dexamethasone or its other analogues. The weight loss method of the EMID proved to be an easy and appropriate method to measure the release rate from the EMID. A high density polyethylene polymer was identified as the most ideal body material for the insert compared to polypropylene body. Also double O-ring silicone, Elastollan WYO 1388-5 and solid silicone pistons were found to be amongst the best pistons tested and all performed well compared to other piston materials. There was a low retention rate with either the original EMID or the modified inserts. Further modification of retention wings of the EMID did not improve the retention rate, but a good blood profile response was obtained from cows treated with the complete EMID containing formulated progesterone. The formulations and driving mechanism were found to be stable under the tested conditions. Therefore, the EMID has potential for commercial application of induced calving or oestrous control per vaginum administration, reliant on improvement of its retention mechanism.

Polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) are two structurally related groups of chemicals, generally referred to as `dioxins´. These are of great concern due to their high toxicity and global spread. Other groups of compounds with similar chemical structure and toxicity mechanisms are the brominated analogues polybrominated dibenzo-p-dioxins (PBDDs) and polybrominated dibenzofurans (PBDFs), and the dioxin-like polychlorinated biphenyls (PCBs). Numerous studies have been undertaken to investigate sources and transport routes of dioxins. However, much remains to be done, including analytical, inventories of dioxin-like compounds, such as PBDD/Fs, and the development of more convenient analytical methods. The currently standard procedure for analyzing dioxins (and dioxin-like compounds) is to use Soxhlet extraction followed by multi-step clean-up and gas chromatography - high resolution mass spectrometry (GC- HRMS) for detection. Unfortunately, this method is very solvent, labor and time-consuming, making it very expensive. The main aim of the studies this thesis was to develop pressurized liquid extraction (PLE) with integrated clean up techniques for fast, convenient preparation of dioxin samples. PLE with integrated clean-up has previously been used for extracting dioxins from biological samples, but in these studies the possibility of extending its use to abiotic samples was explored. The results show that PLE with an integrated carbon trap is suitable for analyzing dioxins in various types of soil samples, sediment and flue gas samples. The results also showed that it has potential for analyzing dioxins in fly ash. The thesis focuses on developments of the methodology for dioxin analysis, but also includes results obtained from PBDDs and dioxin-like PCB analyses. In addition, the possibility of using various other kinds of detection techniques rather than GC-HRMS, such as enzyme-linked immunosorbent assays (ELISAs) or two-dimensional gas chromatography with micro electron capture detection (GCxGC-µECD) was explored. The results indicate that ELISA and GCxGC-µECD could serve as complementary detection systems in some cases. However, it is not yet possible to fully replace GC-HRMS. A further refinement of the PLE with in-cell clean-up technique is the modular approach developed in these studies. With this technique it is possible to include various steps for both clean-up and fractionation. For example, sulphuric acid impregnated silica could be combined with active carbon for the simultaneous removal of lipids (along with other interferences) and fractionation of PCBs and PCDD/Fs. It was shown that the method could provide data that agreed reasonably well with both reference values and values obtained using traditional methods. In general PLE proved to have high extraction efficiency and to yield very similar congener profiles to the reference method. In addition, it was shown that it allowed one-step extraction and clean-up of a salmon sample. Such single-step procedures are the ultimate goals for any extraction technique, and it would be highly desirable to develop one-step methods that could be extended to other types of samples. For the rest of the matrices tested (soil, sediment, mussel and crab tissue and flue gas) the method was successful, however a final polishing step is currently required, involving either dilution or clean-up using miniaturized multilayer silica columns, to obtain extracts that are pure enough for GC-HRMS analysis. Using the developed modular-PLE system substantial costs could be saved. It was estimated that the method could reduce the cost of preparing samples by up to 90%, which would greatly facilitate large-scale inventories.

Malaria is a parasitic disease claiming one million lives worldwide annually. Unfortunately, malaria-endemic countries in need of good quality medicines are also overwhelmed with counterfeit or substandard medicine. This results in treatment inefficacy, resistance towards treatment and death. Counterfeit or substandard quinine sulfate tablets are known to have infiltrated the market, however at this point in time, treatment efficacy of quinine sulfate has fortunately not yet been significantly impaired by resistance, but immediate action is required to prevent it from becoming obsolete. Validated analytical methods with justified specifications are effective in controlling the quality of medicines and to minimise the effect of poor quality medicines. Pharmacopoeia specifies analytical quality control procedures and accompanying specifications to standardise acceptable levels of product quality. Understandably, different monographs of different pharmacopoeias are developed by different independent laboratories and therefore their respective test procedures/specifications for the same FPP may differ from each other. Institutions such as the Pharmacopoeial Dicussion Group (PDG) aim to harmonise pharmacopoeia in order to synchronise final outcomes. This study evaluated the relevancy of differences in analytical procedures, results and specifications for quinine sulfate tablets set by the United States Pharmacopoeia (USP), British Pharmacopoeia (BP) and International Pharmacopoeia (Ph.Int.) in an aim to ensure that these different methods all provide with similar final outcomes and that they be effective in successfully evaluating the quality of quinine sulfate tablets. Four quinine sulfate tablet products were obtained from different manufacturers and were subjected to the tests of all three pharmacopoeia – BP, USP and Ph.Int. The results from identification, assay and related substance testing concluded that the outcomes were the same between the pharmacopoeia despite their differences in techniques/procedures/specifications. The assay, identification and related substances methods and specifications set by each respective monograph were deemed appropriate to evaluate the quality of quinine sulfate tablets. Even with differences in methodology, quantitative techniques and specifications, the USP and BP dissolution methods for quinine sulfate tablets shared the same final outcome at the first stage of dissolution, whereas none of the products achieved a compliant outcome using the Ph.Int. dissolution method. Possible reasons for the poor dissolution (when using the Ph.Int. method) were identified and investigated. Investigation into the solubility of quinine sulfate found the Ph.Int. dissolution method conditions to be too stringent, as the solubility of quinine sulfate in phosphate buffer pH 6.8 (dissolution medium specified by the Ph.Int.) was found to be much less than in acidic media (as proposed by the BP and USP dissolution methods). Several adapted dissolution methods (called developmental studies) were investigated to serve as potential alternatives for the Ph.Int. dissolution method. The developmental studies investigated an alternative dissolution medium, agitation rates (50 rpm, 75 rpm, 100 rpm) and medium volumes (500 ml, 750 ml, 900 ml and 1000 ml). Developmental study 6 was proposed as an alternative dissolution method. Developmental study 6 stipulates the use of the same medium as the original Ph.Int. method, as it was deemed the medium of choice for its discriminatory ability. To address the impaired solubility of quinine sulfate in phosphate buffer, the medium volume and agitation were increased (in reference to the original method) to 900 ml and 100 rpm respectively. The same analytical quantitation technique (UV-Vis spectroscopy) is proposed for Developmental study 6. The newly proposed method provided with final outcomes comparable to that of the USP and BP, however having more discriminatory power than the USP and BP. / MSc (Pharmaceutics), North-West University, Potchefstroom Campus, 2014