Ces travaux s’inscrivent dans une problématique actuelle de compréhension des effets, encore peu connus, des mélanges de pesticides dans le sol. Les formulations d’herbicides S-métolachlore (Dual Gold Safeneur®), mésotrione (Callisto®) et nicosulfuron (Milagro®) ont été appliquées en mélanges et à la dose agronomique recommandée. Les molécules formulées sont plus toxiques que les molécules actives seules (Microtox®) et aucun effet synergique ou antagoniste des mélanges n’a été observé. Cependant, ces mélanges impactent transitoirement et à court terme les communautés microbiennes du sol de Limagne, sans modifier les paramètres généraux de structure et d’abondance. Toutefois, des effets ont été mis en évidence sur des communautés sensibles telles que les phototrophes ou celles impliquées dans les processus de nitrification et de dénitrification. De plus, une comparaison des effets des mélanges sur les communautés phototrophes de deux sols souligne l’importante toxicité du Dual Gold Safeneur® et remet en cause son utilisation d’un point de vue écotoxicologique. / These experiments are part of the actual problematic of understanding the poorknown effects of pesticides mixtures in soil ecosystem. The herbicidal commercial formulations of S-metolachlor (Dual Gold Safeneur®), mesotrione (Callisto®) and nicosulfuron (Milagro®) were applied in mixtures at the recommended field rate. Formulated compounds are more toxic than active ingredients alone (Microtox®) and no synergistic or antagonistic effects were recorded. However, these mixtures impaired rapidly and transitory the Limagne soil microbial communities, without modifying the global parameters of structure and abundance. Effects were also recorded on sensitive communities, such as phototrophic, nitrifying and denitrifying communities. A comparison of mixtures effects on phototrophic communities from two soils underlined the high toxicity of Dual Gold Safeneur® and challenged its use from an ecotoxicological point of view..

Active pharmaceutical ingredients (APIs), which are incorporated in different formulations, i.e. creams, gels, foams, etc., are applied to the skin for a therapeutic effect. This therapeutic effect could either be required in the top layer of the skin (topical drug delivery) or deeper layers to reach the blood capillaries (transdermal drug delivery). Transdermal delivery avoids oral administration route limitations, such as first pass metabolism which is the rapid clearance of the drug in the gastrointestinal tract and degradation by enzymes. This delivery targets the drugs to skin sites, where there are significant advantages which include: improved patient compliance, a steady drug delivery state, less frequent dosing, adverse effects are minimal, it is less invasive and issues with the gastrointestinal absorption are avoided by eliminating the first pass metabolism (Perrie et al., 2012:392). This type of delivery is not free from limitations even though the skin can be employed for targeted drug delivery and is a readily available and large accessible surface area for adsorption of drugs. The most upper layer of the human skin, the stratum corneum, which is a watertight barrier, offers defence against hazardous exterior materials such as fungi, allergens, viruses and other molecules. This indicates the stratum corneum controls the drug penetration of most drugs to permeate the skin barrier (Lam & Gambari, 2014:27). Pravastatin is hydrophilic and is a 3-hydroxy-3-methyl-glutaryl coenzyme A (HMG-CoA) reductase inhibitor which inhibits cholesterol synthesis, a rate-limiting step, in the liver, thus decreasing the level of plasma low density lipoprotein cholesterol (LDL-C) (Heath et al., 1998:42). It can also slow the progression of atherosclerosis and can lower the incident of coronary events (Haria & McTavish, 1997:299). The first aim of the study is to deliver pravastatin transdermally into the blood circulation. Currently, pravastatin is only administered in oral dosages and can cause highly negative adverse effects such as myopathy and increased liver enzymes. This increase in liver enzymes causes hepatotoxicity and therefore would be ideal if pravastatin could be delivered transdermally, as the first pass metabolic effect would be nullified and adverse effects would decrease drastically (Gadi et al., 2013:648). Prof JW Wiechers‟ Delivery Gap Principle was designed in attempt to effectively enhance transdermal drug delivery. This Delivery Gap Principle was incorporated in the computer programme he developed known as “Formulating for Efficacy” (FFE™). The transdermal delivery of suggested APIs, which in this case is pravastatin, when incorporated into a formulation, may be optimised transdermally. The FFE™ computer programme suggests that the oil phase can be optimised, which in turn leads to better permeation through the skin to the target site (transdermal). The formula can be manipulated to reach desired polarity. The second aim of this in vitro study was to investigate the implementation of Wiechers‟ Delivery Gap Principle in a semi-solid dosage form, for the transdermal delivery of pravastatin sodium (2%). Six formulations, of which three were cream and three were emulgel formulations incorporated with pravastatin sodium (2%), were formulated. Each formulation had a different polarity, i.e. hydrophilic cream (HC) and emulgel (HE), lipophilic cream (LC) and emulgel (LE) and optimised cream (OC) and emulgel (OE). A high performance liquid chromatography (HPLC) method was developed and validated to analyse the concentration of pravastatin. Both the octanol-buffer distribution coefficient (log D) and the aqueous solubility of pravastatin were determined. For the API to permeate through the skin into the blood circulation, certain physicochemical properties are important and according to Naik et al (2000:321), there are specific ideal limits for the API in the formulations which include log D (1 to 3) and a aqueous solubility of >1 mg/ml. The aqueous solubility of 197.5 mg/ml in phosphate buffer solution (PBS) (pH 7.4) at a temperature of 32 °C indicated penetration was favourable (Naik et al., 2000:321), whilst the log D value of -0.703 indicated the API was unfavourable for skin penetration (Naik et al., 2000:321). Membrane release studies were conducted using a synthetic membrane to determine whether pravastatin was released from the six formulations each containing 2% pravastatin prior to diffusion studies with. The OE yielded the highest median flux value (7.175 μg/cm2.h), followed the by LE (6.401 μg/cm2.h), HE (6.355 μg/cm2.h), HC (5.061 μg/cm2.h), OC (4.297 μg/cm2.h) and lastly, LC (3.115 μg/cm2.h). By looking at the aforementioned data values, it was concluded that the emulgels performed better than the cream formulations when median flux values were compared. By using dermatomed excised female Caucasian skin, an execution of Franz cell diffusion studies were performed over a period of 12 h, followed by a tape-stripping experiment to determine which semi-solid formulation delivered pravastatin best on the skin and the results of the different polarity formulations were compared. The median amount per area which permeated through the skin after 12 h was as follows: the OE formulation (2.578 μg/cm2) depicted the highest median amount per area, followed by OC (1.449 μg/cm2), HC (0.434 μg/cm2), LE (0.121 μg/cm2), HE (0.055 μg/cm2) and lastly LC (0.000 μg/cm2). These results validate Wiechers` theory that when the oil phase is optimised, with regard to the same polarity as the skin, permeation will be enhanced (Wiechers, 2011). During both the membrane studies and the skin diffusion studies it was evident the emulgel formulations performed better and pravastatin permeated better than the cream formulations. When skin diffusion and membrane median data values were compared, it was evident in both the membrane release studies and the skin diffusion studies that OE yielded the highest median values and LC the lowest median values. It was clear that all six different formulations released pravastatin, but LC displayed no permeation into the systemic circulation (receptor phase). The data of the different polarity formulations which yielded the best results with regards to median concentrations within the stratum corneum-epidermis and epidermis-dermis, were identified and are: within the stratum corneum-epidermis, HE (1.448 μg/ml) yielded the highest median concentration pravastatin, followed by LE (1.301 μg/ml), LC (0.676 μg/ml), HC (0.505 μg/ml), OE (0.505 μg/ml) and lastly OC (0.400 μg/ml). As emulgels (hydrophilic) contain more water than creams (lipophilic), the penetration enhancement effect can be explained by hydration, since the water hydrated the skin leading the lipids to open and the stratum corneum to swell (Williams & Barry, 2004:606). Therefore more API could permeate into the skin. Within the epidermis-dermis the highest median concentration median was yielded by OE (0.849 μg/ml), followed by LC (0.572 μg/ml), HC (0.524 μg/ml), OC (0.355 μg/ml), HE (0.309 μg/ml) and lastly LE (0.138 μg/ml). Different polarity formulations permeating the viable epidermis could be a result of the solubility characteristics of the formulations. It contained both lipid properties (formulations contained oil content), leading to permeation through the stratum corneum and aqueous properties, which lead to diffusion into the underlying layers of the epidermis (Perrie et al., 2012:392). According to Perrie (2012:392), formulations that need to be delivered transdermally, must permeate through the lipophilic stratum corneum and thereafter the hydrophilic dermal layers to reach the blood circulation, which means formulations must consist of both lipophilic and aqueous solubility properties. When comparing the stratum corneum-epidermis (lipophilic) with the epidermis-dermis (more hydrophilic) and receptor phase (hydrophilic; systemic circulation), it is evident that all formulations had lipophilic and hydrophilic properties, as the API permeated through the stratum corneum and penetrated the deeper layers of the skin (viable epidermis) When all polarity formulations were compared, i.e. optimised, hydrophilic and lipophilic, it was observed that the optimised formulations depicted the highest median concentration values in the receptor phase (skin diffusion), but lowest median concentration in stratum corneum-epidermis, therefore the optimised formulation permeated best through the stratum corneum-epidermis. The reason for this could be that the optimised formulations had the same polarity as the skin (17, 8, 8), thus permeating through the skin to the receptor fluid more efficiently (Wiechers, 2011). It was observed that LC penetrated both stratum corneum-epidermis and epidermis-dermis, but did not permeate through the skin to the receptor fluid (systemic circulation), making it a good delivery vehicle for topical delivery. Overall when the emulgel and cream formulations are compared, according to their ability to deliver pravastatin transdermally, it is evident the pravastatin diffused more from the emulgel formulations than the cream formulations. This could be due to the fact that emulgels are more hydrophilic as they contain more water, resulting in the emulgels diffusing to the deeper layers of the skin (more hydrophilic viable epidermis) (Benson, 2005:28). All formulations contained not only aqueous (hydrophilic) but also lipid (lipophilic) solubility properties, therefore making it lipophilic enough to permeate the stratum corneum and hydrophilic enough to penetrate to deeper skin layers (viable epidermis) (Perrie et al., 2012:392). All formulations could still permeate the viable epidermis despite different polarities being used and all were appropriate candidates, although some were more suitable than others. The understanding from this study is that: * pravastatin could be delivered topically by all formulations, * the best formulation to reach the systemic formulation is the optimised emulgel, * the best formulation to deliver pravastatin topically is the hydrophilic emulgel. / MSc (Pharmaceutics), North-West University, Potchefstroom Campus, 2015

Active pharmaceutical ingredients (APIs), which are incorporated in different formulations, i.e. creams, gels, foams, etc., are applied to the skin for a therapeutic effect. This therapeutic effect could either be required in the top layer of the skin (topical drug delivery) or deeper layers to reach the blood capillaries (transdermal drug delivery). Transdermal delivery avoids oral administration route limitations, such as first pass metabolism which is the rapid clearance of the drug in the gastrointestinal tract and degradation by enzymes. This delivery targets the drugs to skin sites, where there are significant advantages which include: improved patient compliance, a steady drug delivery state, less frequent dosing, adverse effects are minimal, it is less invasive and issues with the gastrointestinal absorption are avoided by eliminating the first pass metabolism (Perrie et al., 2012:392). This type of delivery is not free from limitations even though the skin can be employed for targeted drug delivery and is a readily available and large accessible surface area for adsorption of drugs. The most upper layer of the human skin, the stratum corneum, which is a watertight barrier, offers defence against hazardous exterior materials such as fungi, allergens, viruses and other molecules. This indicates the stratum corneum controls the drug penetration of most drugs to permeate the skin barrier (Lam & Gambari, 2014:27). Pravastatin is hydrophilic and is a 3-hydroxy-3-methyl-glutaryl coenzyme A (HMG-CoA) reductase inhibitor which inhibits cholesterol synthesis, a rate-limiting step, in the liver, thus decreasing the level of plasma low density lipoprotein cholesterol (LDL-C) (Heath et al., 1998:42). It can also slow the progression of atherosclerosis and can lower the incident of coronary events (Haria & McTavish, 1997:299). The first aim of the study is to deliver pravastatin transdermally into the blood circulation. Currently, pravastatin is only administered in oral dosages and can cause highly negative adverse effects such as myopathy and increased liver enzymes. This increase in liver enzymes causes hepatotoxicity and therefore would be ideal if pravastatin could be delivered transdermally, as the first pass metabolic effect would be nullified and adverse effects would decrease drastically (Gadi et al., 2013:648). Prof JW Wiechers‟ Delivery Gap Principle was designed in attempt to effectively enhance transdermal drug delivery. This Delivery Gap Principle was incorporated in the computer programme he developed known as “Formulating for Efficacy” (FFE™). The transdermal delivery of suggested APIs, which in this case is pravastatin, when incorporated into a formulation, may be optimised transdermally. The FFE™ computer programme suggests that the oil phase can be optimised, which in turn leads to better permeation through the skin to the target site (transdermal). The formula can be manipulated to reach desired polarity. The second aim of this in vitro study was to investigate the implementation of Wiechers‟ Delivery Gap Principle in a semi-solid dosage form, for the transdermal delivery of pravastatin sodium (2%). Six formulations, of which three were cream and three were emulgel formulations incorporated with pravastatin sodium (2%), were formulated. Each formulation had a different polarity, i.e. hydrophilic cream (HC) and emulgel (HE), lipophilic cream (LC) and emulgel (LE) and optimised cream (OC) and emulgel (OE). A high performance liquid chromatography (HPLC) method was developed and validated to analyse the concentration of pravastatin. Both the octanol-buffer distribution coefficient (log D) and the aqueous solubility of pravastatin were determined. For the API to permeate through the skin into the blood circulation, certain physicochemical properties are important and according to Naik et al (2000:321), there are specific ideal limits for the API in the formulations which include log D (1 to 3) and a aqueous solubility of >1 mg/ml. The aqueous solubility of 197.5 mg/ml in phosphate buffer solution (PBS) (pH 7.4) at a temperature of 32 °C indicated penetration was favourable (Naik et al., 2000:321), whilst the log D value of -0.703 indicated the API was unfavourable for skin penetration (Naik et al., 2000:321). Membrane release studies were conducted using a synthetic membrane to determine whether pravastatin was released from the six formulations each containing 2% pravastatin prior to diffusion studies with. The OE yielded the highest median flux value (7.175 μg/cm2.h), followed the by LE (6.401 μg/cm2.h), HE (6.355 μg/cm2.h), HC (5.061 μg/cm2.h), OC (4.297 μg/cm2.h) and lastly, LC (3.115 μg/cm2.h). By looking at the aforementioned data values, it was concluded that the emulgels performed better than the cream formulations when median flux values were compared. By using dermatomed excised female Caucasian skin, an execution of Franz cell diffusion studies were performed over a period of 12 h, followed by a tape-stripping experiment to determine which semi-solid formulation delivered pravastatin best on the skin and the results of the different polarity formulations were compared. The median amount per area which permeated through the skin after 12 h was as follows: the OE formulation (2.578 μg/cm2) depicted the highest median amount per area, followed by OC (1.449 μg/cm2), HC (0.434 μg/cm2), LE (0.121 μg/cm2), HE (0.055 μg/cm2) and lastly LC (0.000 μg/cm2). These results validate Wiechers` theory that when the oil phase is optimised, with regard to the same polarity as the skin, permeation will be enhanced (Wiechers, 2011). During both the membrane studies and the skin diffusion studies it was evident the emulgel formulations performed better and pravastatin permeated better than the cream formulations. When skin diffusion and membrane median data values were compared, it was evident in both the membrane release studies and the skin diffusion studies that OE yielded the highest median values and LC the lowest median values. It was clear that all six different formulations released pravastatin, but LC displayed no permeation into the systemic circulation (receptor phase). The data of the different polarity formulations which yielded the best results with regards to median concentrations within the stratum corneum-epidermis and epidermis-dermis, were identified and are: within the stratum corneum-epidermis, HE (1.448 μg/ml) yielded the highest median concentration pravastatin, followed by LE (1.301 μg/ml), LC (0.676 μg/ml), HC (0.505 μg/ml), OE (0.505 μg/ml) and lastly OC (0.400 μg/ml). As emulgels (hydrophilic) contain more water than creams (lipophilic), the penetration enhancement effect can be explained by hydration, since the water hydrated the skin leading the lipids to open and the stratum corneum to swell (Williams & Barry, 2004:606). Therefore more API could permeate into the skin. Within the epidermis-dermis the highest median concentration median was yielded by OE (0.849 μg/ml), followed by LC (0.572 μg/ml), HC (0.524 μg/ml), OC (0.355 μg/ml), HE (0.309 μg/ml) and lastly LE (0.138 μg/ml). Different polarity formulations permeating the viable epidermis could be a result of the solubility characteristics of the formulations. It contained both lipid properties (formulations contained oil content), leading to permeation through the stratum corneum and aqueous properties, which lead to diffusion into the underlying layers of the epidermis (Perrie et al., 2012:392). According to Perrie (2012:392), formulations that need to be delivered transdermally, must permeate through the lipophilic stratum corneum and thereafter the hydrophilic dermal layers to reach the blood circulation, which means formulations must consist of both lipophilic and aqueous solubility properties. When comparing the stratum corneum-epidermis (lipophilic) with the epidermis-dermis (more hydrophilic) and receptor phase (hydrophilic; systemic circulation), it is evident that all formulations had lipophilic and hydrophilic properties, as the API permeated through the stratum corneum and penetrated the deeper layers of the skin (viable epidermis) When all polarity formulations were compared, i.e. optimised, hydrophilic and lipophilic, it was observed that the optimised formulations depicted the highest median concentration values in the receptor phase (skin diffusion), but lowest median concentration in stratum corneum-epidermis, therefore the optimised formulation permeated best through the stratum corneum-epidermis. The reason for this could be that the optimised formulations had the same polarity as the skin (17, 8, 8), thus permeating through the skin to the receptor fluid more efficiently (Wiechers, 2011). It was observed that LC penetrated both stratum corneum-epidermis and epidermis-dermis, but did not permeate through the skin to the receptor fluid (systemic circulation), making it a good delivery vehicle for topical delivery. Overall when the emulgel and cream formulations are compared, according to their ability to deliver pravastatin transdermally, it is evident the pravastatin diffused more from the emulgel formulations than the cream formulations. This could be due to the fact that emulgels are more hydrophilic as they contain more water, resulting in the emulgels diffusing to the deeper layers of the skin (more hydrophilic viable epidermis) (Benson, 2005:28). All formulations contained not only aqueous (hydrophilic) but also lipid (lipophilic) solubility properties, therefore making it lipophilic enough to permeate the stratum corneum and hydrophilic enough to penetrate to deeper skin layers (viable epidermis) (Perrie et al., 2012:392). All formulations could still permeate the viable epidermis despite different polarities being used and all were appropriate candidates, although some were more suitable than others. The understanding from this study is that: * pravastatin could be delivered topically by all formulations, * the best formulation to reach the systemic formulation is the optimised emulgel, * the best formulation to deliver pravastatin topically is the hydrophilic emulgel. / MSc (Pharmaceutics), North-West University, Potchefstroom Campus, 2015

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

Compostos de origem vegetal podem apresentar inúmeros efeitos benéficos à saúde, havendo, entretanto, uma necessidade de desenvolver formulações que permitam viabilizar seu uso farmacêutico, alimentício, nutracêutico ou cosmecêutico. Syzygium aromaticum, conhecido popularmente como cravo da Índia, é uma espécie vegetal aromática com marcada atividade antioxidante, analgésica e antimicrobiana. A baixa solubilidade e estabilidade química, assim como a volatilidade dos principais compostos associados às atividades biológicas da planta justificam o desenvolvimento de formulações que melhorem suas propriedades físico-químicas e características de liberação. Formulações lipídicas têm sido cada vez mais usadas para o aumento da solubilidade de compostos no trato gastrointestinal e para o aumento da biodisponibilidade. O principal objetivo deste projeto foi investigar a produção de formulações lipídicas sólidas contendo compostos bioativos de S. aromaticum e avaliar o efeito da composição da formulação e das variáveis operacionais nas propriedades físico-químicas das partículas, estabilidade e permeação intestinal in vitro. O processo de extração dos compostos a partir da matéria-prima vegetal também foi estudado. A formulação lipídica foi otimizada com respeito ao tipo e a proporção dos lipídeos, do emulsificante e dos carreadores de secagem. Os processos de emulsificação e secagem também foram criteriosamente estudados. Os resultados mostraram que a composição da formulação teve efeitos significativos nas propriedades físico-químicas do produto e no desempenho da secagem. A formulação lipídica otimizada mostrou ser mais estável que a formulação não lipídica em condições de armazenamento de alta umidade. Com relação à permeação intestinal in vitro, utilizando eugenol como marcador, não foram observadas diferenças significativas entre estas duas formulações. Este projeto permitiu obter informações relevantes sobre a secagem por spray drying de formulações lipídicas contendo extratos vegetais. Esta rota tecnológica representa uma estratégia interessante na obtenção de formulações lipídicas estáveis que promovam o aumento da biodisponibilidade oral de compostos bioativos. / Plant-derived compounds can provide important benefits to human health. However, these compounds should be properly formulated in order to facilitate their pharmaceutical, nutraceutical, food or cosmetic applications. Syzygium aromaticum commonly known as Indian clove, is an aromatic tree with antioxidant, antimicrobial and analgesic properties. The poor water solubility and the volatility of the compounds associated to the biological activities, justify the development of formulations that improve its physicochemical and release properties. Lipid based formulations have gained special attention for oral delivery due to the improvement of solubility in the intestinal tract and increase of bioavailability. The main objective of this project was to investigate the production of solid lipidic formulations containing bioactive compounds of S. aromaticum and to test the effect of the formulation composition and the process variables on the physhicochemical properties of the particles, stability and in vitro intestinal permeation. The extraction methods of the compounds from the plant were also studied. The lipid formulation was optimized with regard to the type and proportion of the solid lipid, the surfactant and the drying carrier. The emulsification and the drying processes were carefully evaluated. Results showed that the formulation composition had significant effects on the physicochemical properties of the product and on the drying performance. The optimized lipid formulation was more stable than the formulation without lipids in high humidity stress storage conditions. With regard to the in vitro intestinal permeation using eugenol as marker compound, not significant differences were observed between the samples. This project allowed to obtain relevant information about the spray drying process of lipid formulations containing plant extracts. This technique could be an interesting strategy to obtain stable lipid formulations than enhance the oral bioavailability of bioactive compounds

A confecção e avaliação do desempenho voltamétrico de eletrodos compósitos à base de resina epóxi comercial Araldite ® utilizada como fase aglutinante e pó de grafite como fase condutora, na confecção dos eletrodos compósitos foi realizada. A resina foi preparada seguindo instruções do fabricante e adicionada a uma quantidade de pó de grafite para obter eletrodos contendo 50, 60 e 70% de grafite (m/m). Tais compósitos foram caracterizados fisicamente quanto à reprodutibilidade da resposta voltamétrica, resistência ôhmica e área efetiva em função da composição grafite/Araldite ®, sendo 70% (grafite, m/m) a que ofereceu melhores resultados, em termos de repetibilidade de resposta e corrente de pico. O teor real de grafite foi determinado por termogravimetria. Os intervalos úteis de potenciais para uso dos eletrodos compósitos também foram avaliados em diferentes eletrólitos. Finalmente, os eletrodos compósitos foram avaliados na determinação de atenolol, uma droga antihipertensiva, e dopamina, um neurotransmissor central do grupo das catecolaminas. Em todos os casos, o eletrodo compósito foi mais sensível na determinação dos dois analitos, quando comparado ao eletrodo de carbono vítreo. A dopamina foi determinada por voltametria de pulso diferencial (DPV). O limite de detecção foi de 0,61 µmol L-1e a resposta linear foi observada no intervalo de 4,0 a 95 µmol L-1. Os resultados obtidos na determinação da dopamina em formulação farmacêutica concordaram com os resultados obtidos com o método comparativo cromatografia líquida de alta eficiência (HPLC), com 95% de confiança. O atenolol também foi determinado por DPV, com limite de detecção da ordem de 2,23 µmol L-1, com resposta linear entre 4,45 e 84,7 µmol L-1. A determinação de atenolol, em formulações farmacêuticas, apresentou resultados que concordam com o método comparativo HPLC, com 95% de confiança. O atenolol também foi determinado em amostras de águas naturais, com recuperações entre 92 e 115%. / The development and evaluation of the voltammetric behavior of composite electrodes prepared with graphite as a conducting phase and commercial epoxy resin Araldite ® as an insulating phase was performed. The resin was prepared according to instructions of the manufacturer and mixed with a suitable amount of graphite in order to prepare composites with 50, 60 and 70% of graphite (w/w). The composites were characterized in relation to their reproducibility of the cyclic voltammetric response, ohmic resistance, and repeatability of the effective area as a function of the Araldite ®/graphite ratio. The best repeatability of area as well as the higher peak currents was obtained with the 70% (graphite, w/w) composition using K3[Fe(CN)6] in KCl medium. The useful potential windows were also evaluated in different supporting electrolytes and pH ranges. Finally the composite electrodes were applied in the determination of the anti-hypertensive atenolol and the cathecolamine neurotransmitter dopamine. The composite electrodes were more sensitive to both analytes when compared to the glassy carbon. Dopamine was determined using differential pulse voltammetry (DPV) with linear dynamic range between 4.0 and 95 µmol L-1 and limit of detection 0.61 µmol L-1. The results agreed with those from high performance liquid chromatography (HPLC) within a 95% confidence interval. Atenolol was also determined by DPV with a 4.45 - 84.7 µmol L-1 linear range and 2.23 µmol L-1. The determination of atenolol in pharmaceutical formulations agreed with the HPLC method in a 95 % confidence interval. When applied to natural water samples spiked with the analyte, recoveries of 92 to 115% were found.

Os corticosteróides são substância largamente empregadas em dertatologia devido ao seu potente efeito anti-inflamatório na pela. Entretanto, associado a este efeito benéfico tem-se o risco da ocorrência de efeitos colaterais, decorrentes principalmente da absorção sistêmica dos corticosteróides pela via cutânea. A penetração transcutânea e retenção cutânea dos corticosteróides é influenciada pelo veículo no qual estes princípios ativos são incorporados. Assim sendo, os objetivos desta pesquisa foram investigar, in vitro, formulações que proporcionassem uma alta retenção cutânea dos corticosteróides na pele e também mínima penetração transcutânea. O estudo foi realizado com géis de Poloxamer 407 contendo diferentes concentrações dos promotores de absorção cutânea, uréia ou lecitina. Os parâmetros liberação, penetração transcutânea e retençâo cutânea dos acetatos de hidrocortisona e dexametasona, desonida e triamcinolona acetonida foram avaliados por metodologia in vitro, utilizando-se de célula de difusão e membranas. A quantificação dos corticosteróides nas diferentes fases do experimento foi realizada por cromatografia líquida de alta eficiência. Os esultados obtidos mostraram a influência do veículo nos parâAmetros avaliados. As formulações obedeceram cinética de 1ª. ordem para a liberação e penetração transcutânea. As preparações contendo lecitina promoveram uma maior retenção cutânea dos corticosteróides, sugerindo, assim, que géis de Poloxamer 407, associados com lecitina, formam veículos adequados para a incorporação de corticosteróides. / Corticoids are drugs often used in dermatology due to its anti-inflamatory effect in the skin. Meanwhile, together with this beneficial effect it can have side effects, due to the systemic absorption of the corticoids by cutaneous way. Transcutaneous penetration and cutaneous retention of the corticoids is influenciated by the vehicle in which this drogs are incorporated. In this way, the objetive of this research was to investigate, in vitro, formulations that would provide both, high cutaneous retention of the corticoids in the Skin and minimmn transcutaneous penetration. For this work was used Poloxamer 407 gels containing different concentrations of the absorptions enhancers, urea and lecithin. The release transcutaneous penetration and cutaneous retention of hydrocortisone and dexamethasone acetate, desonide and triamcinolone acetonide was evaluated by in vitro methodology using difiusion cells and membranes. The corticoids was analysed by HPLC. The results obtained showed the influence of the vehicle in the evaluated parameters. The corticoid relesse and transcutaneous penetration appeared to fit first order kinetic. The formulations containing lecithin promoved higher cutaneous retetion of the corticoids than the containing urea, sugesting, in this way, that Poloxamer 407 gels in the presence of lecithin are adequated preparations to the corticoids.

A exposição à radiação ultravioleta (RUV) pode levar a um desequilíbrio no balanço oxidante/antioxidante da pele causando prejuízos à sua integridade e levando a diversas alterações, entre as quais o envelhecimento precoce e o câncer de pele. Na tentativa de diminuir os efeitos biológicos mediados pelos radicais livres gerados pela RUV na pele, tem sido proposto a fotoquimioproteção com a utilização de antioxidantes tópicos. Dentre a gama de compostos disponíveis para serem empregados na fotoquimioproteção, a própolis, por sua pronunciada atividade antioxidante, entre suas inúmeras atividades biológicas, é uma matéria-prima com promissora ação tópica. Desta forma, extratos de própolis alcoólico e glicólico (EPA e EPG) foram caracterizados quanto à sua composição polifenólica e quanto à sua capacidade antioxidante frente a diversos radicais livres. Formulações adicionadas destes extratos foram desenvolvidas e submetidas a estudos de estabilidade física e funcional, estudos de liberação, permeação e retenção cutânea in vitro, bem como estudos preliminares de eficácia in vivo. Os resultados demonstraram que os extratos de própolis são capazes de seqüestrar eficientemente diversos radicais livres, principalmente radicais superóxido. Quando estes extratos foram adicionados em formulações de produtos para uso tópico, a atividade antioxidante foi mantida. Nos estudos prévios de estabilidade física foi observado que as formulações mais estáveis foram desenvolvidas com Hostacerin® SAF (menor conteúdo graxo) e Polawax® (maior conteúdo graxo). No entanto, somente a formulação desenvolvida com Polawax® apresentou estabilidade satisfatória por 1 ano quando armazenada à temperatura ambiente e a 40º.C 2º.C/70%UR 5%. Nos estudos de liberação, permeação e retenção cutânea foi observado a influência do conteúdo graxo na performance das formulações. A cinética de liberação das formulações tanto do ácido p-cumárico (utilizado como marcador), como dos compostos equivalentes ao extrato de própolis (EEP) demonstraram seguir o modelo de Higuchi. A formulação desenvolvida com Polawax® apresentou a melhor retenção cutânea, com retenção de 0,013 e 0,030 µL de EEP.cm-2 para as peles de camundongo e de porco, respectivamente. Em adição, esta formulação apresentou baixos níveis de permeação cutânea, sendo adequada para aplicação tópica fotoquimioprotetora. Nos estudos in vivo, esta formulação adicionada de EPA, foi capaz de diminuir o eritema, inibir o edema e aumentar a cicatrização de camundongos sem pêlo expostos à radiação UVB. Além disso, também foi observado a proteção da depleção da glutationa endógena (GSH). Os resultados preliminares de eficácia in vivo sugerem que formulações contendo o extrato de própolis apresentam boas perspectivas para serem utilizadas para prevenir e tratar os danos causados na pele pela radiação UV. / The ultraviolet radiation (UVR) exposition may lead to the skin oxidant/antioxidant imbalance injuring its integrity and leading to several disorders, such as ageing and skin cancer. In order to improve the biological effects caused by free radicals generated by UVR in skin, it has been suggested the photochemoprotection by using topical antioxidants. Among the available compounds to be employed in hotochemoprotection, propolis, due to its important antioxidant activity, among its innumerous biological activities, is a promising topical raw-material. Next, ethanolic and glycolic propolis extracts (EPE, GPE) were characterized in relation to their polyphenolic composition, and in relation to their antioxidant activity against several free-radicals. Formulations added with these extracts were developed and undergone to physical and functional stability studies, in vitro release and skin permeation and retention studies, as well as in vivo preliminary efficacy studies. The results showed that the propolis extracts are able to scavenge several free radicals efficiently, mainly superoxide radicals. When these extracts were added to formulations of topical products, their antioxidant activity were maintained. In the physical stability studies, it was observed that the most stable formulations were developed with Hostacerin® SAF (lower fat content) and Polawax® (higher fat content). However, only the formulation developed with Polawax® showed satisfactory stability for 1 year stored at room temperature and at 40º.C 2º.C/70%UR 5%UR. In the release, permeation and retention studies, it was observed the fat content influence in the formulation performance. The release profile of p-coumaric acid (used as marker compound) and the compounds equivalent to propolis extract (EPE) followed the Higuchi model. The formulation developed with Polawax® showed the best skin retention, retaining 0,013 and 0,030 L EPE.cm-2 in hairless mouse skin and in pig skin, respectively. In addition, this formulation presented low permeation, which is desired for photochemoprotective topical employment. In the in vivo studies, this formulation added with EPE was able to diminish erithema, inhibit oedema and increase cicatrisation in hairless mice exposed to UVB radiation. In addition, it was also observed the protection of the endogenous glutathione (GSH) depletion. The in vivo preliminary efficacy results suggest that formulations added with propolis extract present good perspectives to be employed to prevent and treat the injuries caused in skin by UV radiation.

A exposição à radiação ultravioleta (RUV) pode levar a um desequilíbrio no balanço oxidante/antioxidante da pele causando prejuízos à sua integridade e levando a diversas alterações, entre as quais o envelhecimento precoce e o câncer de pele. Na tentativa de diminuir os efeitos biológicos mediados pelos radicais livres gerados pela RUV na pele, tem sido proposto a fotoquimioproteção com a utilização de antioxidantes tópicos. Dentre a gama de compostos disponíveis para serem empregados na fotoquimioproteção, a própolis, por sua pronunciada atividade antioxidante, entre suas inúmeras atividades biológicas, é uma matéria-prima com promissora ação tópica. Desta forma, extratos de própolis alcoólico e glicólico (EPA e EPG) foram caracterizados quanto à sua composição polifenólica e quanto à sua capacidade antioxidante frente a diversos radicais livres. Formulações adicionadas destes extratos foram desenvolvidas e submetidas a estudos de estabilidade física e funcional, estudos de liberação, permeação e retenção cutânea in vitro, bem como estudos preliminares de eficácia in vivo. Os resultados demonstraram que os extratos de própolis são capazes de seqüestrar eficientemente diversos radicais livres, principalmente radicais superóxido. Quando estes extratos foram adicionados em formulações de produtos para uso tópico, a atividade antioxidante foi mantida. Nos estudos prévios de estabilidade física foi observado que as formulações mais estáveis foram desenvolvidas com Hostacerin® SAF (menor conteúdo graxo) e Polawax® (maior conteúdo graxo). No entanto, somente a formulação desenvolvida com Polawax® apresentou estabilidade satisfatória por 1 ano quando armazenada à temperatura ambiente e a 40º.C 2º.C/70%UR 5%. Nos estudos de liberação, permeação e retenção cutânea foi observado a influência do conteúdo graxo na performance das formulações. A cinética de liberação das formulações tanto do ácido p-cumárico (utilizado como marcador), como dos compostos equivalentes ao extrato de própolis (EEP) demonstraram seguir o modelo de Higuchi. A formulação desenvolvida com Polawax® apresentou a melhor retenção cutânea, com retenção de 0,013 e 0,030 µL de EEP.cm-2 para as peles de camundongo e de porco, respectivamente. Em adição, esta formulação apresentou baixos níveis de permeação cutânea, sendo adequada para aplicação tópica fotoquimioprotetora. Nos estudos in vivo, esta formulação adicionada de EPA, foi capaz de diminuir o eritema, inibir o edema e aumentar a cicatrização de camundongos sem pêlo expostos à radiação UVB. Além disso, também foi observado a proteção da depleção da glutationa endógena (GSH). Os resultados preliminares de eficácia in vivo sugerem que formulações contendo o extrato de própolis apresentam boas perspectivas para serem utilizadas para prevenir e tratar os danos causados na pele pela radiação UV. / The ultraviolet radiation (UVR) exposition may lead to the skin oxidant/antioxidant imbalance injuring its integrity and leading to several disorders, such as ageing and skin cancer. In order to improve the biological effects caused by free radicals generated by UVR in skin, it has been suggested the photochemoprotection by using topical antioxidants. Among the available compounds to be employed in hotochemoprotection, propolis, due to its important antioxidant activity, among its innumerous biological activities, is a promising topical raw-material. Next, ethanolic and glycolic propolis extracts (EPE, GPE) were characterized in relation to their polyphenolic composition, and in relation to their antioxidant activity against several free-radicals. Formulations added with these extracts were developed and undergone to physical and functional stability studies, in vitro release and skin permeation and retention studies, as well as in vivo preliminary efficacy studies. The results showed that the propolis extracts are able to scavenge several free radicals efficiently, mainly superoxide radicals. When these extracts were added to formulations of topical products, their antioxidant activity were maintained. In the physical stability studies, it was observed that the most stable formulations were developed with Hostacerin® SAF (lower fat content) and Polawax® (higher fat content). However, only the formulation developed with Polawax® showed satisfactory stability for 1 year stored at room temperature and at 40º.C 2º.C/70%UR 5%UR. In the release, permeation and retention studies, it was observed the fat content influence in the formulation performance. The release profile of p-coumaric acid (used as marker compound) and the compounds equivalent to propolis extract (EPE) followed the Higuchi model. The formulation developed with Polawax® showed the best skin retention, retaining 0,013 and 0,030 L EPE.cm-2 in hairless mouse skin and in pig skin, respectively. In addition, this formulation presented low permeation, which is desired for photochemoprotective topical employment. In the in vivo studies, this formulation added with EPE was able to diminish erithema, inhibit oedema and increase cicatrisation in hairless mice exposed to UVB radiation. In addition, it was also observed the protection of the endogenous glutathione (GSH) depletion. The in vivo preliminary efficacy results suggest that formulations added with propolis extract present good perspectives to be employed to prevent and treat the injuries caused in skin by UV radiation.

Os corticosteróides são substância largamente empregadas em dertatologia devido ao seu potente efeito anti-inflamatório na pela. Entretanto, associado a este efeito benéfico tem-se o risco da ocorrência de efeitos colaterais, decorrentes principalmente da absorção sistêmica dos corticosteróides pela via cutânea. A penetração transcutânea e retenção cutânea dos corticosteróides é influenciada pelo veículo no qual estes princípios ativos são incorporados. Assim sendo, os objetivos desta pesquisa foram investigar, in vitro, formulações que proporcionassem uma alta retenção cutânea dos corticosteróides na pele e também mínima penetração transcutânea. O estudo foi realizado com géis de Poloxamer 407 contendo diferentes concentrações dos promotores de absorção cutânea, uréia ou lecitina. Os parâmetros liberação, penetração transcutânea e retençâo cutânea dos acetatos de hidrocortisona e dexametasona, desonida e triamcinolona acetonida foram avaliados por metodologia in vitro, utilizando-se de célula de difusão e membranas. A quantificação dos corticosteróides nas diferentes fases do experimento foi realizada por cromatografia líquida de alta eficiência. Os esultados obtidos mostraram a influência do veículo nos parâAmetros avaliados. As formulações obedeceram cinética de 1ª. ordem para a liberação e penetração transcutânea. As preparações contendo lecitina promoveram uma maior retenção cutânea dos corticosteróides, sugerindo, assim, que géis de Poloxamer 407, associados com lecitina, formam veículos adequados para a incorporação de corticosteróides. / Corticoids are drugs often used in dermatology due to its anti-inflamatory effect in the skin. Meanwhile, together with this beneficial effect it can have side effects, due to the systemic absorption of the corticoids by cutaneous way. Transcutaneous penetration and cutaneous retention of the corticoids is influenciated by the vehicle in which this drogs are incorporated. In this way, the objetive of this research was to investigate, in vitro, formulations that would provide both, high cutaneous retention of the corticoids in the Skin and minimmn transcutaneous penetration. For this work was used Poloxamer 407 gels containing different concentrations of the absorptions enhancers, urea and lecithin. The release transcutaneous penetration and cutaneous retention of hydrocortisone and dexamethasone acetate, desonide and triamcinolone acetonide was evaluated by in vitro methodology using difiusion cells and membranes. The corticoids was analysed by HPLC. The results obtained showed the influence of the vehicle in the evaluated parameters. The corticoid relesse and transcutaneous penetration appeared to fit first order kinetic. The formulations containing lecithin promoved higher cutaneous retetion of the corticoids than the containing urea, sugesting, in this way, that Poloxamer 407 gels in the presence of lecithin are adequated preparations to the corticoids.