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

Este trabalho teve como objetivo propor novo método para estudar a permeabilidade de fármacos, utilizando epitélio intestinal de rã-touro (Rana catesbeiana) em método ex vivo, empregando células de Franz. Por utilizar epitélio intestinal, um material de descarte proveniente de um animal utilizado como alimento humano, pode ser considerado um método alternativo, pois não implica no sacrifício de animais. A quantidade de fármaco permeada foi determinada por método de eletroforese capilar com detecção ultravioleta e validado para os antivirais lamivudina, zidovudina e aciclovir, na presença de metoprolol e floridizina. O fármaco escolhido como modelo nos ensaios de permeabilidade foi a lamivudina. Para estabelecimento do protocolo experimental dos estudos de permeabilidade, foi proposta uma análise de variância three-way para verificar a influência na permeabilidade dos fármacos, das seguintes variáveis: secção intestinal, pH da solução de Ringer e temperatura. Foram determinados a quantidade total de fármaco permeado (Qt), o coeficiente de permeabilidade aparente (Papp) e a constante de absorção de primeira ordem (ka). A partir da análise do planejamento experimental, os efeitos das variáveis não foram significativos, exceto para a secção intestinal. Os resultados de coeficiente de permeabilidade aparente (Papp) obtidos foram de 0,09 x 10-4 cm/s para lamivudina e de 0,22 x 10-4 cm/s para o metoprolol. O valor de Papp obtido de para o metoprolol é próximo dos valores encontrados na literatura para outras técnicas. Para a lamivudina, entretanto, a diferença encontrada em comparação às células Caco-2 pode ser devida às diferentes técnicas empregadas. / This work aimed to propose a new method for studying drug permeability using frog intestinal epithelium (Rana catesbeiana) in ex vivo method, using Franz cells. By using intestinal epithelium, a disposal material from an animal used as human food, can be considered an alternative method, because it doesn\'t involve the sacrifice of animals. The amount of permeated drug was determined by capillary electrophoresis method with UV detection and validated for antiviral drugs lamivudine, zidovudine and acyclovir in the presence of metoprolol and floridizina. The drug chosen as a model in permeability studies was lamivudine. To establish the experimental protocol for the permeability studies, a three-way analysis of variance was proposed to check the influence of intestinal section, pH of Ringer\'s solution and the temperature on the permeability. Total amount of drug permeated (Qt), apparent permeability coefficient (Papp) and first-order constant absorption (ka) were determined. By the analysis of experimental design, the effects of the variables were not significant, except for intestinal section. The results of apparent permeability coefficient (Papp) obtained were 0.09 x 10-4 cm/s for lamivudine and 0.22 x 10-4 cm/s for metoprolol. The value of Papp obtained for metoprolol is quite close to the values found in literature for other methods. For lamivudine, however, the difference found in comparison to Caco-2 cells may be due to different techniques.

A genisteína é uma isoflavona da soja que vem sendo investigada pelo seu potencial antienvelhecimento, baseado nas suas atividades antioxidante, estrogênica e inibidora de proteínas tirosina-quinase. A associação da genisteína com β- ciclodextrina com a formação de um complexo elevou a hidrossolubilidade da isoflavona. A permeabilidade intrínseca da genisteína foi avaliada, bem como sua permeabilidade quando aplicada a partir de gel de hidróxi propilmetilcelulose (HPMC) a 3 %, pelo método de célula de Franz. As associações com β-ciclodextrina, em géis de HPMC 3 %, também foram avaliadas e foi observado um incremento da penetração em favor do complexo produzido em meio aquoso. Devido ao seu alto coeficiente de partição (log P) - 4,36 - a genisteína demonstrou a capacidade de formar reservatórios nas estruturas internas da pele favorecendo sua ação antienvelhecimento na pele. / Genistein is a soy isoflavone that has been investigated for its antiaging potencial, based on antioxidant, estrogenic and proteins tirosin-kinase inhibitor activities. The association of genistein with β-cyclodextrin resulted in a complex formation enhanced the isoflavone hidrosolubity. Genistein instrinsic permeability was evaluated, as well permebility from hydroxypropyl methylcelullose (HPMC) 3 % gel, wtih Franz cell method. The associations with β-cyclodextrin, in HPMC 3 % gels were also evaluated with the observation of na enhancing effect of cyclodextrin in the complex produced in aqueous media. Because its high partition coefficient (log P) - 4,36 - genistein was able to form reservoir in the internal skin layers, favorable to its antiagin action.

A genisteína é uma isoflavona da soja que vem sendo investigada pelo seu potencial antienvelhecimento, baseado nas suas atividades antioxidante, estrogênica e inibidora de proteínas tirosina-quinase. A associação da genisteína com β- ciclodextrina com a formação de um complexo elevou a hidrossolubilidade da isoflavona. A permeabilidade intrínseca da genisteína foi avaliada, bem como sua permeabilidade quando aplicada a partir de gel de hidróxi propilmetilcelulose (HPMC) a 3 %, pelo método de célula de Franz. As associações com β-ciclodextrina, em géis de HPMC 3 %, também foram avaliadas e foi observado um incremento da penetração em favor do complexo produzido em meio aquoso. Devido ao seu alto coeficiente de partição (log P) - 4,36 - a genisteína demonstrou a capacidade de formar reservatórios nas estruturas internas da pele favorecendo sua ação antienvelhecimento na pele. / Genistein is a soy isoflavone that has been investigated for its antiaging potencial, based on antioxidant, estrogenic and proteins tirosin-kinase inhibitor activities. The association of genistein with β-cyclodextrin resulted in a complex formation enhanced the isoflavone hidrosolubity. Genistein instrinsic permeability was evaluated, as well permebility from hydroxypropyl methylcelullose (HPMC) 3 % gel, wtih Franz cell method. The associations with β-cyclodextrin, in HPMC 3 % gels were also evaluated with the observation of na enhancing effect of cyclodextrin in the complex produced in aqueous media. Because its high partition coefficient (log P) - 4,36 - genistein was able to form reservoir in the internal skin layers, favorable to its antiagin action.

A genisteína é uma isoflavona da soja que vem sendo investigada pelo seu potencial antienvelhecimento, baseado nas suas atividades antioxidante, estrogênica e inibidora de proteínas tirosina-quinase. A associação da genisteína com β- ciclodextrina com a formação de um complexo elevou a hidrossolubilidade da isoflavona. A permeabilidade intrínseca da genisteína foi avaliada, bem como sua permeabilidade quando aplicada a partir de gel de hidróxi propilmetilcelulose (HPMC) a 3 %, pelo método de célula de Franz. As associações com β-ciclodextrina, em géis de HPMC 3 %, também foram avaliadas e foi observado um incremento da penetração em favor do complexo produzido em meio aquoso. Devido ao seu alto coeficiente de partição (log P) - 4,36 - a genisteína demonstrou a capacidade de formar reservatórios nas estruturas internas da pele favorecendo sua ação antienvelhecimento na pele. / Genistein is a soy isoflavone that has been investigated for its antiaging potencial, based on antioxidant, estrogenic and proteins tirosin-kinase inhibitor activities. The association of genistein with β-cyclodextrin resulted in a complex formation enhanced the isoflavone hidrosolubity. Genistein instrinsic permeability was evaluated, as well permebility from hydroxypropyl methylcelullose (HPMC) 3 % gel, wtih Franz cell method. The associations with β-cyclodextrin, in HPMC 3 % gels were also evaluated with the observation of na enhancing effect of cyclodextrin in the complex produced in aqueous media. Because its high partition coefficient (log P) - 4,36 - genistein was able to form reservoir in the internal skin layers, favorable to its antiagin action.

Este trabalho teve como objetivo propor novo método para estudar a permeabilidade de fármacos, utilizando epitélio intestinal de rã-touro (Rana catesbeiana) em método ex vivo, empregando células de Franz. Por utilizar epitélio intestinal, um material de descarte proveniente de um animal utilizado como alimento humano, pode ser considerado um método alternativo, pois não implica no sacrifício de animais. A quantidade de fármaco permeada foi determinada por método de eletroforese capilar com detecção ultravioleta e validado para os antivirais lamivudina, zidovudina e aciclovir, na presença de metoprolol e floridizina. O fármaco escolhido como modelo nos ensaios de permeabilidade foi a lamivudina. Para estabelecimento do protocolo experimental dos estudos de permeabilidade, foi proposta uma análise de variância three-way para verificar a influência na permeabilidade dos fármacos, das seguintes variáveis: secção intestinal, pH da solução de Ringer e temperatura. Foram determinados a quantidade total de fármaco permeado (Qt), o coeficiente de permeabilidade aparente (Papp) e a constante de absorção de primeira ordem (ka). A partir da análise do planejamento experimental, os efeitos das variáveis não foram significativos, exceto para a secção intestinal. Os resultados de coeficiente de permeabilidade aparente (Papp) obtidos foram de 0,09 x 10-4 cm/s para lamivudina e de 0,22 x 10-4 cm/s para o metoprolol. O valor de Papp obtido de para o metoprolol é próximo dos valores encontrados na literatura para outras técnicas. Para a lamivudina, entretanto, a diferença encontrada em comparação às células Caco-2 pode ser devida às diferentes técnicas empregadas. / This work aimed to propose a new method for studying drug permeability using frog intestinal epithelium (Rana catesbeiana) in ex vivo method, using Franz cells. By using intestinal epithelium, a disposal material from an animal used as human food, can be considered an alternative method, because it doesn\'t involve the sacrifice of animals. The amount of permeated drug was determined by capillary electrophoresis method with UV detection and validated for antiviral drugs lamivudine, zidovudine and acyclovir in the presence of metoprolol and floridizina. The drug chosen as a model in permeability studies was lamivudine. To establish the experimental protocol for the permeability studies, a three-way analysis of variance was proposed to check the influence of intestinal section, pH of Ringer\'s solution and the temperature on the permeability. Total amount of drug permeated (Qt), apparent permeability coefficient (Papp) and first-order constant absorption (ka) were determined. By the analysis of experimental design, the effects of the variables were not significant, except for intestinal section. The results of apparent permeability coefficient (Papp) obtained were 0.09 x 10-4 cm/s for lamivudine and 0.22 x 10-4 cm/s for metoprolol. The value of Papp obtained for metoprolol is quite close to the values found in literature for other methods. For lamivudine, however, the difference found in comparison to Caco-2 cells may be due to different techniques.

Bergström, A Utvärdering av hudvänliga lösningsmedel för ökad löslighet av NSAID i topikala formuleringar. Examensarbete i Farmakologi 15 poäng. Malmö Universitet: Fakulteten för Hälsa och Samhälle, Institutionen för Biomedicinsk Vetenskap, 2019.Introduktion Smärta har behandlats med topikala eller transdermala läkemedel i många år, men det är en utmaning att uppnå höga koncentrationer som kan transporteras över huden. För att kunna ersätta användandet av opioider vid behandling av måttlig till allvarlig smärta behövs mer effektiva formuleringar av transdermala läkemedel. Målet med denna studie var att undersöka möjligheten att öka mängden aktiv substans som transporteras över huden. För detta ändamål, bestämdes lösligheten hos diklofenak i olika lösningsmedel. Vidare bestämdes transporten av diklofenak över silikonmembran från dessa lösningsmedel med hjälp av Franz celler.Metod Lösligheten hos diklofenak studerades vid 32ºC i vatten, fosfatbuffrad saltlösning (PBS), saltlösning, polyetylenglykol (PEG) 400, polyetylenglykol (PEG) 1500 (60 vikt % i vatten), glycerol, propylenglykol, 1-propanol och 2-propanol. Diffusion hos diklofenak över silikonmembran från nämnda formuleringarna, inklusive Voltaren Gel som jämförelse, studerades med hjälp av Franzceller under sex timmar. För både löslighet- och diffusionsexperimenten bestämdes koncentrationen av diklofenak med en analysmetod baserad på UV-spektrofotometri.Resultat Lösligheten av diklofenak var högst i PEG 400 och propylenglykol med ungefär 40 vikt %. Den lägsta lösligheten uppmättes i 2-propanol med ca 1 %. I diffusionsstudierna kunde diklofenak enbart detekteras i receptorkammaren, dvs diklofenas transporterades över membranet, hos de formuleringar som innehöll enbart vatten eller Voltaren.Diskussion Resultatet av löslighetstesterna kunde förklaras genom att ta de kemiska och fysikaliska egenskaperna hos den olika lösningsmedlen som användes i beaktande samt genom att använda begrepp som utsaltningseffekt och vätedonatorer/acceptorer. Resultaten av diffusionsstudierna var ofullständiga men då enbart ett begränsat antal experiment kunde utföras inom ramen för detta arbete behöver resultaten vidare styrkas.Nyckelord: Diffusion, diklofenak, Franz cell, löslighet, transdermal / Bergström, A Evaluation of skin-friendly solvents for increased solubility of a model NSAID in topical formulations. Degree project in Pharmacology 15 credits. Malmö University: Faculty of Health and Society, Department of Biomedical Science, 2019.Introduction Pain has been treated with topical or transdermal drugs for many years, but the concentration of active substance that can be transported over the skin remains low. To be able to substitute the use of opioids in treatment of moderate to severe pain, more efficient formulations of transdermal drugs are needed. The aim of this study was to investigate the possibility to increase the concentration of diclofenac in topical formulations. For this reason, the solubility of diclofenac in different skin-friendly solvents was determined. In addition, the diffusion of diclofenac across silicone membranes from these solvents was investigated using Franz cells.Methods The solubility of diclofenac was studied at 32ºC in water, phophate buffered saline solution (PBS), saline solution, polyethylene glycol (PEG) 400, polyethylene glycol (PEG) 1500 (60 wt% in water), glycerol, propylene glycol, 1-propanol and 2-propanol. The diffusion of diclofenac over silicon membrane, used as a model for skin membranes, from six formulations, including Voltaren for comparison, was studied using Franz diffusion cells over six hours. Both for the solubility and diffusion experiments, the concentration of diclofenac was determined by an analytical method based on UV-spectroscopy.Results The solubility of diclofenac was highest in PEG 400 and propylene glycol with approximately 40 wt. %. The lowest solubility was seen in 2-propanol with approximately 1 %. In the diffusion study, only the formulation containing water as solvent and Voltaren gel resulted in detection of dicofenac in the receptor chamber, i.e. transported over the membrane in the time frame studied.Discussion The results of the solubility experiments can be rationalized based on the chemical and physical properties of the solvents and by considering concepts such as the salting-out efffect and appropriate number of hydrogen bond donors and acceptors. The results of the diffusion studies were inconclusive and since only a limited number of experiments could be performed within the scope of this work, the results need to be authenticated.Keywords: Diclofenac, diffusion, Franz cell, solubility, transdermal

In pharmaceutical sciences, topical delivery is a transport process of an active pharmaceutical ingredient (API) from a formulated dosage form to the target site of action. For most topical delivery systems, the skin surface, or the specific skin layers, such as the outermost layer of the stratum corneum, the lipids amid the corneocytes within the stratum corneum, the corneocytes themselves, the epidermis, dermis, Langerhans cells, Merckle cells or the appendageal structures can be the target delivery location. When an API is delivered to the skin, it has to firstly diffuse from the formulation in which it is applied, to the skin surface. From there the API may partition into the stratum corneum, permeate across the stratum corneum and partition into the viable epidermis, from where it may partition further into the dermis and permeate across the dermis into the bloodstream (Wiechers, 2008:1-3, 7). With respect to the barrier function of the skin, the intercellular spaces within the stratum corneum contain lipids and its main purpose is to operate as a barrier to water-loss and to provide an imperative diffusional barrier to the absorption of APIs. This resistance is comprised of a complex interaction of lipids that creates a hydrophilic and lipophilic penetration pathway. The fundamental aspect underlying the impermeability of the skin, therefore, is the lipophilic nature of the stratum corneum (Bouwstra et al., 2003:4; Franz & Lehman, 2000:25; Walker & Smith, 1996:296). A common approach for the promotion of poorly penetrating APIs in transdermal delivery is the incorporation of chemical penetration enhancers in delivery systems, in order to promote the partitioning of an API into the stratum corneum. These chemicals are also referred to as accelerants, promoters and absorption promoters. Penetration enhancers are added to topical formulations and usually also partition into the stratum corneum, where they temporarily and reversibly disrupt its fundamental diffusional barrier properties, hence facilitating the absorption of an API through the skin (Büyüktimkin et al., 1997:358-359; Sinha & Kaur, 2000:1131; Walker & Smith, 1996:296). The mechanisms for the enhancement of diffusion of the API should therefore increase the solubility and partitioning of the drug from the formulation into the skin. It should further increase the solubility of the API within the skin and promote its permeability and diffusion coefficient (Rajadhyaksha et al., 1997:489). Fatty acids are recognised to effectively enhance the penetration of transdermally delivered hydrophilic and lipophilic APIs. Many penetration enhancers contain saturated and unsaturated hydrocarbon chains, and a popular fatty acid that has been used in this regard is oleic acid (Williams & Barry, 2004:609-610). It is believed that fatty acids disrupt the lipid organisation of the intercellular lipids within the stratum corneum to cause fluidisation of these bilayers, making the stratum corneum more permeable to APIs. Excipients with polar (hydrophilic) head groups and long hydrophobic chains i.e. fatty acids, can penetrate into the intercellular lipids of the stratum corneum and disrupt these endogenous lipid components, thereby increasing diffusion of an API within the skin (Barry, 2006:9-10; Hadgraft & Finnin, 2006:367-368; Kanikkannan et al., 2006:18; Williams & Barry, 2004:610). Natural oils are widely used in topical formulations and were an obvious choice in this study. Oils are liquids at room temperature, whereas fats are in solid form. They are relatively easy to obtain from both plants and animals. The main constituents of fats and oils are triglycerides comprising of fatty acids and a glycerol. Oils control the evaporation of moisture from the skin, spread easily and evenly and are partly metabolised in the skin to release valuable fatty acids (Fang et al., 2004:170,173; Lautenschläger, 2004:46; Mitsui, 1997:121-122). The focus of this study was not formulation per se, but included the formulation of avocado-, grapeseed-, emu-, crocodile, olive and coconut oil into semisolid emulgel- and two foam formulations. This was done in order to investigate the penetration enhancing properties of their fatty acid content on flurbiprofen which was chosen as the marker API. The emulgels containing the natural oils were compared to the same emulgel formulation containing liquid paraffin, and a hydrogel without the inclusion of an oil. Six natural oils were analysed by gas chromatography (GC) in order to quantify their fatty acid compositions, whilst also providing qualitative information by indicating the retention times of the materials with an alkyl chain composition (Mitsui, 1997:260). Data obtained with the GC indicated that olive- (76%), avocado- (68%), emu- (46%) and crocodile oil (40%) presented with high levels of oleic acid, also known as a mono-unsaturated fatty acid (MUFA). Lower levels of oleic acid were observed within grapeseed- (27%) and coconut oil (8%). The only oil demonstrating high levels of the poly-unsaturated fatty acid (PUFA), linoleic acid, was grapeseed oil (61%), whereas the remainder of the oils showed levels below 24%. Contrary, coconut oil seemed to have been the only oil high in saturated fatty acids (SFAs) and consisted of a lauric acid content of 52% and medium levels of myristic acid (21%). Average levels of palmitic acid (SFA) were found in crocodile- (21%) and in emu oil (21%), both of animal origin, whereas avocado-, grapeseed-, olive- and coconut oils from plants presented with levels below 15%. Stearic acid was also present in levels below 10% in all of these oils, with the oils of animal origin portraying the highest values. A method was developed and validated to determine the concentration of the marker flurbiprofen after diffusion from the formulations into the skin, as well as concentrations of the marker that diffused through the skin, by means of high performance liquid chromatography (HPLC). Franz cell membrane diffusion studies were conducted prior to the skin diffusion studies in order to verify the actual release of the marker from the semisolid formulations. Skin diffusion experiments were performed using dermatomed excised, human skin to which the six emulgel formulations, containing the natural oils, were applied. A comparative study was performed utilising liquid paraffin and a hydrogel, in order to compare the diffusion of the marker, flurbiprofen, into and through the skin. The two oil emulgel formulations that had indicated the best flux values were subsequently formulated into foam preparations in order to compare the penetration enhancement properties on flurbiprofen of these two oils in a foam preparation, to those in the equivalent emulgels. The data generated for all ten the formulations were compared, and the formulations that yielded the best results with regards to median flux values and the flurbiprofen concentrations within the stratum corneum-epidermis and epidermis-dermis, were identified. Application of the liquid paraffin emulgel (21.29 μg/ml) depicted the highest average concentration of the diffused lipophilic flurbiprofen within the stratum corneum-epidermis, followed by the olive oil foam (21.47 μg/ml), olive oil emulgel (17.82 μg/ml) and grapeseed oil emulgel (17.78 μg/ml). Very similar concentrations for the marker were demonstrated by the hydrogel (16.73 μg/ml) and crocodile oil emulgel (14.89 μg/ml), whereas a lower concentration was shown for coconut oil emulgel (7.18 μg/ml). The remainder of the formulations yielded concentrations below 3%, i.e. the avocado oil emulgel (2.72 μg/ml), the coconut oil foam (1.57 μg/ml) and finally the emu oil emulgel (1.25 μg/ml). The penetration of the marker, flurbiprofen, being trapped within the skin seemed to have been enhanced more by the oleic acid (UFA) containing emulgels and foam, especially. This was followed by oils containing high linoleic acid values, which indicated that the more kinked shaped the fatty acids, the more difficult it became to insert themselves within the lipid structures of the stratum corneum, with a resulting accumulation of the marker (Fang et al., 2003:318-319). It therefore seemed that those oils that predominantly consisted of unsaturated fatty acids (UFAs) (grapeseed-, crocodile- and olive oils) seemed to have increased the concentration of the diffused marker more significantly than those oils containing an almost even combination of MUFAs and PUFAs (avocado oil), or those mainly consisting of SFAs (coconut oil). Average concentrations of the diffused flurbiprofen found in the epidermis-dermis region of the skin for all of the formulations demonstrated low concentrations, ranging between 0.97 - 5.39 μg/ml, with the exception of the emu oil emulgel that presented with a higher concentration of 16.15 μg/ml. The reason for the high accumulation of the marker might have been as a result of epidermal proliferation, with subsequent accumulation of the marker within the epidermis-dermis due to high oleic- and linoleic acid values, as well as small amounts of palmitoleic acid present within this oil (Katsuta et al., 2005:1011). The resistance of the epidermis-dermis region to the general permeation of flurbiprofen might have been caused by its lipophilic nature, resulting in a reduced solubility within the hydrophilic environment of this region (Hadgraft, 1999:5). Median results from the skin diffusion studies demonstrated that the hydrogel (23.79 μg/cm2.h) had the highest flux, followed by the olive oil- (17.99 μg/cm2.h), liquid paraffin- (15.70 μg/cm2.h), coconut oil- (13.16 μg/cm2.h), grapeseed oil- (11.85 μg/cm2.h), avocado oil- (8.31 μg/cm2.h), crocodile oil- (6.68 μg/cm2.h) and emu oil emulgels (4.41 μg/cm2.h). The fact that the hydrogel presented a higher flux value for the marker could have been as a result of its high water content that had caused hydration of the skin. Hydration opens up the dense lipid structures inside of the stratum corneum, due to swelling of the corneocytes, with a subsequent increase in the marker‘s flux (Benson, 2005:28; Ranade & Hollinger, 2004:213). The high flux value of flurbiprofen with the liquid paraffin emulgel might also have resulted from the fact that it occluded the skin, which increased the hydration of the stratum corneum, with a subsequent increase in the flux (Mitsui, 1997:124; Thomas & Finnin, 2004:699). Results from the skin diffusion studies could be explained by the fact that the fatty acids differ in their hydrocarbon chain by (1) the length of the chain, and (2) the position- and number of the double bonds (Babu et al., 2006:144). It is suggested that fatty acids with hydrocarbon (lipophilic) chains between C12 to C14 (also present within coconut oil) have an optimal balance of the partition coefficient and its affinity for the skin (Ogiso & Shintani, 1990:1067). It appears as though the branched UFAs, especially oleic acid, present in high quantities in olive oil, were more powerful enhancers of the diffusion of the marker, flurbiprofen (Chi et al., 1995:270). Foam formulations were manufactured with the olive- and coconut oil emulgels that had demonstrated the best median flux values of flurbiprofen from the natural oil emulgel formulations. These formulated foams, however, did not significantly increased flux values for flurbiprofen through the skin, but only achieved values of 5.56 μg/cm2.h for the olive oil foam and 4.36 μg/cm2.h for the coconut oil foam formulations. The low flux values could have been attributed to the nature of the formulation itself, which was filled with trapped air that could have resulted in the formulation not making optimal direct contact with the available skin surface. Throughout this study, it became evident that olive oil, predominantly consisting of oleic acid (UFA), was most effective in enhancing the flux of the lipophilic marker, flurbiprofen, through the skin, closely followed by coconut oil consisting of SFAs, with lauric- and myristic acid as its main constituents. Better enhancement effects were observed with those oils containing high amounts of oleic acid (MUFA), than oils consisting of almost equal amounts of both PUFAs and MUFAs (avocado-, emu- and crocodile oil), or oils mainly consisting of PUFAs (grapeseed oil) as its main components, but their effect was not more significant than the oil containing SFAs (coconut oil) as its key components. / Thesis (MSc (Pharmaceutics))--North-West University, Potchefstroom Campus, 2013.

In pharmaceutical sciences, topical delivery is a transport process of an active pharmaceutical ingredient (API) from a formulated dosage form to the target site of action. For most topical delivery systems, the skin surface, or the specific skin layers, such as the outermost layer of the stratum corneum, the lipids amid the corneocytes within the stratum corneum, the corneocytes themselves, the epidermis, dermis, Langerhans cells, Merckle cells or the appendageal structures can be the target delivery location. When an API is delivered to the skin, it has to firstly diffuse from the formulation in which it is applied, to the skin surface. From there the API may partition into the stratum corneum, permeate across the stratum corneum and partition into the viable epidermis, from where it may partition further into the dermis and permeate across the dermis into the bloodstream (Wiechers, 2008:1-3, 7). With respect to the barrier function of the skin, the intercellular spaces within the stratum corneum contain lipids and its main purpose is to operate as a barrier to water-loss and to provide an imperative diffusional barrier to the absorption of APIs. This resistance is comprised of a complex interaction of lipids that creates a hydrophilic and lipophilic penetration pathway. The fundamental aspect underlying the impermeability of the skin, therefore, is the lipophilic nature of the stratum corneum (Bouwstra et al., 2003:4; Franz & Lehman, 2000:25; Walker & Smith, 1996:296). A common approach for the promotion of poorly penetrating APIs in transdermal delivery is the incorporation of chemical penetration enhancers in delivery systems, in order to promote the partitioning of an API into the stratum corneum. These chemicals are also referred to as accelerants, promoters and absorption promoters. Penetration enhancers are added to topical formulations and usually also partition into the stratum corneum, where they temporarily and reversibly disrupt its fundamental diffusional barrier properties, hence facilitating the absorption of an API through the skin (Büyüktimkin et al., 1997:358-359; Sinha & Kaur, 2000:1131; Walker & Smith, 1996:296). The mechanisms for the enhancement of diffusion of the API should therefore increase the solubility and partitioning of the drug from the formulation into the skin. It should further increase the solubility of the API within the skin and promote its permeability and diffusion coefficient (Rajadhyaksha et al., 1997:489). Fatty acids are recognised to effectively enhance the penetration of transdermally delivered hydrophilic and lipophilic APIs. Many penetration enhancers contain saturated and unsaturated hydrocarbon chains, and a popular fatty acid that has been used in this regard is oleic acid (Williams & Barry, 2004:609-610). It is believed that fatty acids disrupt the lipid organisation of the intercellular lipids within the stratum corneum to cause fluidisation of these bilayers, making the stratum corneum more permeable to APIs. Excipients with polar (hydrophilic) head groups and long hydrophobic chains i.e. fatty acids, can penetrate into the intercellular lipids of the stratum corneum and disrupt these endogenous lipid components, thereby increasing diffusion of an API within the skin (Barry, 2006:9-10; Hadgraft & Finnin, 2006:367-368; Kanikkannan et al., 2006:18; Williams & Barry, 2004:610). Natural oils are widely used in topical formulations and were an obvious choice in this study. Oils are liquids at room temperature, whereas fats are in solid form. They are relatively easy to obtain from both plants and animals. The main constituents of fats and oils are triglycerides comprising of fatty acids and a glycerol. Oils control the evaporation of moisture from the skin, spread easily and evenly and are partly metabolised in the skin to release valuable fatty acids (Fang et al., 2004:170,173; Lautenschläger, 2004:46; Mitsui, 1997:121-122). The focus of this study was not formulation per se, but included the formulation of avocado-, grapeseed-, emu-, crocodile, olive and coconut oil into semisolid emulgel- and two foam formulations. This was done in order to investigate the penetration enhancing properties of their fatty acid content on flurbiprofen which was chosen as the marker API. The emulgels containing the natural oils were compared to the same emulgel formulation containing liquid paraffin, and a hydrogel without the inclusion of an oil. Six natural oils were analysed by gas chromatography (GC) in order to quantify their fatty acid compositions, whilst also providing qualitative information by indicating the retention times of the materials with an alkyl chain composition (Mitsui, 1997:260). Data obtained with the GC indicated that olive- (76%), avocado- (68%), emu- (46%) and crocodile oil (40%) presented with high levels of oleic acid, also known as a mono-unsaturated fatty acid (MUFA). Lower levels of oleic acid were observed within grapeseed- (27%) and coconut oil (8%). The only oil demonstrating high levels of the poly-unsaturated fatty acid (PUFA), linoleic acid, was grapeseed oil (61%), whereas the remainder of the oils showed levels below 24%. Contrary, coconut oil seemed to have been the only oil high in saturated fatty acids (SFAs) and consisted of a lauric acid content of 52% and medium levels of myristic acid (21%). Average levels of palmitic acid (SFA) were found in crocodile- (21%) and in emu oil (21%), both of animal origin, whereas avocado-, grapeseed-, olive- and coconut oils from plants presented with levels below 15%. Stearic acid was also present in levels below 10% in all of these oils, with the oils of animal origin portraying the highest values. A method was developed and validated to determine the concentration of the marker flurbiprofen after diffusion from the formulations into the skin, as well as concentrations of the marker that diffused through the skin, by means of high performance liquid chromatography (HPLC). Franz cell membrane diffusion studies were conducted prior to the skin diffusion studies in order to verify the actual release of the marker from the semisolid formulations. Skin diffusion experiments were performed using dermatomed excised, human skin to which the six emulgel formulations, containing the natural oils, were applied. A comparative study was performed utilising liquid paraffin and a hydrogel, in order to compare the diffusion of the marker, flurbiprofen, into and through the skin. The two oil emulgel formulations that had indicated the best flux values were subsequently formulated into foam preparations in order to compare the penetration enhancement properties on flurbiprofen of these two oils in a foam preparation, to those in the equivalent emulgels. The data generated for all ten the formulations were compared, and the formulations that yielded the best results with regards to median flux values and the flurbiprofen concentrations within the stratum corneum-epidermis and epidermis-dermis, were identified. Application of the liquid paraffin emulgel (21.29 μg/ml) depicted the highest average concentration of the diffused lipophilic flurbiprofen within the stratum corneum-epidermis, followed by the olive oil foam (21.47 μg/ml), olive oil emulgel (17.82 μg/ml) and grapeseed oil emulgel (17.78 μg/ml). Very similar concentrations for the marker were demonstrated by the hydrogel (16.73 μg/ml) and crocodile oil emulgel (14.89 μg/ml), whereas a lower concentration was shown for coconut oil emulgel (7.18 μg/ml). The remainder of the formulations yielded concentrations below 3%, i.e. the avocado oil emulgel (2.72 μg/ml), the coconut oil foam (1.57 μg/ml) and finally the emu oil emulgel (1.25 μg/ml). The penetration of the marker, flurbiprofen, being trapped within the skin seemed to have been enhanced more by the oleic acid (UFA) containing emulgels and foam, especially. This was followed by oils containing high linoleic acid values, which indicated that the more kinked shaped the fatty acids, the more difficult it became to insert themselves within the lipid structures of the stratum corneum, with a resulting accumulation of the marker (Fang et al., 2003:318-319). It therefore seemed that those oils that predominantly consisted of unsaturated fatty acids (UFAs) (grapeseed-, crocodile- and olive oils) seemed to have increased the concentration of the diffused marker more significantly than those oils containing an almost even combination of MUFAs and PUFAs (avocado oil), or those mainly consisting of SFAs (coconut oil). Average concentrations of the diffused flurbiprofen found in the epidermis-dermis region of the skin for all of the formulations demonstrated low concentrations, ranging between 0.97 - 5.39 μg/ml, with the exception of the emu oil emulgel that presented with a higher concentration of 16.15 μg/ml. The reason for the high accumulation of the marker might have been as a result of epidermal proliferation, with subsequent accumulation of the marker within the epidermis-dermis due to high oleic- and linoleic acid values, as well as small amounts of palmitoleic acid present within this oil (Katsuta et al., 2005:1011). The resistance of the epidermis-dermis region to the general permeation of flurbiprofen might have been caused by its lipophilic nature, resulting in a reduced solubility within the hydrophilic environment of this region (Hadgraft, 1999:5). Median results from the skin diffusion studies demonstrated that the hydrogel (23.79 μg/cm2.h) had the highest flux, followed by the olive oil- (17.99 μg/cm2.h), liquid paraffin- (15.70 μg/cm2.h), coconut oil- (13.16 μg/cm2.h), grapeseed oil- (11.85 μg/cm2.h), avocado oil- (8.31 μg/cm2.h), crocodile oil- (6.68 μg/cm2.h) and emu oil emulgels (4.41 μg/cm2.h). The fact that the hydrogel presented a higher flux value for the marker could have been as a result of its high water content that had caused hydration of the skin. Hydration opens up the dense lipid structures inside of the stratum corneum, due to swelling of the corneocytes, with a subsequent increase in the marker‘s flux (Benson, 2005:28; Ranade & Hollinger, 2004:213). The high flux value of flurbiprofen with the liquid paraffin emulgel might also have resulted from the fact that it occluded the skin, which increased the hydration of the stratum corneum, with a subsequent increase in the flux (Mitsui, 1997:124; Thomas & Finnin, 2004:699). Results from the skin diffusion studies could be explained by the fact that the fatty acids differ in their hydrocarbon chain by (1) the length of the chain, and (2) the position- and number of the double bonds (Babu et al., 2006:144). It is suggested that fatty acids with hydrocarbon (lipophilic) chains between C12 to C14 (also present within coconut oil) have an optimal balance of the partition coefficient and its affinity for the skin (Ogiso & Shintani, 1990:1067). It appears as though the branched UFAs, especially oleic acid, present in high quantities in olive oil, were more powerful enhancers of the diffusion of the marker, flurbiprofen (Chi et al., 1995:270). Foam formulations were manufactured with the olive- and coconut oil emulgels that had demonstrated the best median flux values of flurbiprofen from the natural oil emulgel formulations. These formulated foams, however, did not significantly increased flux values for flurbiprofen through the skin, but only achieved values of 5.56 μg/cm2.h for the olive oil foam and 4.36 μg/cm2.h for the coconut oil foam formulations. The low flux values could have been attributed to the nature of the formulation itself, which was filled with trapped air that could have resulted in the formulation not making optimal direct contact with the available skin surface. Throughout this study, it became evident that olive oil, predominantly consisting of oleic acid (UFA), was most effective in enhancing the flux of the lipophilic marker, flurbiprofen, through the skin, closely followed by coconut oil consisting of SFAs, with lauric- and myristic acid as its main constituents. Better enhancement effects were observed with those oils containing high amounts of oleic acid (MUFA), than oils consisting of almost equal amounts of both PUFAs and MUFAs (avocado-, emu- and crocodile oil), or oils mainly consisting of PUFAs (grapeseed oil) as its main components, but their effect was not more significant than the oil containing SFAs (coconut oil) as its key components. / Thesis (MSc (Pharmaceutics))--North-West University, Potchefstroom Campus, 2013.