The major objectives of this research were to produce a phenolic rich isolate from two locally grown Saskatchewan fruits, chokecherries and saskatoons, develop an encapsulation system for the phenolic isolate, and test this system for the delivery of the phenolic isolate in an animal (rat) model. Natural phenolic compounds present in plants such as fruits have antioxidant and free radical scavenging activities, which have been proposed to have health benefits. The extraction of these compounds from plants is commonly performed using methanol despite being toxic to both humans and animals. As such, ethanol was investigated for its ability to extract phenolics from plants as a food safe alternative to methanol. Phenolic extraction from chokecherries with ethanol:formic acid:water (EFW) resulted in higher concentrations (9.83 mg gallic acid equivalents (GAE)/g fresh weight) than with methanol:formic acid:water (MFW) (7.97 mg GAE/g fresh weight). Results from saskatoons showed similar phenolic levels of 4.26 and 4.21 mg GAE/g fresh weight with MFW and ethanol (EFW), respectively. These results showed that EFW was a suitable substitute for MFW in phenolic compound isolation from chokecherries and saskatoons, and could be used to produce extracts that were safe for use in foods and feeds. High performance liquid chromatography with photodiode array detection (HPLC-PDA) was used to determine the phenolic compound composition of the raw fruits and their phenolic rich isolates. Chlorogenic acid was identified in both chokecherry and saskatoon samples, and rutin was also shown to be present in saskatoons. These identifications were based on the relative retention time and ultra violet-visual spectra comparisons to standards. Solid phase extraction (SPE) using Amberlite XAD-16 was employed to produce phenolic isolates from chokecherries and saskatoons. HPLC-PDA results determined that there was a ~2.7x and ~1.6x increase in peak area for chokecherries and saskatoons, respectively when SPE was employed. The antioxidant activity of the extracts and isolates was determined using in vitro radical scavenging tests including 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2´-azinobis-3-ethylbenzthiazoline-sulphonic acid (ABTS). The EFW chokecherry extract and isolate had the highest overall free radical scavenging activity. Crude fruit extracts exhibited lower free radical scavenging values compared to the isolate samples in both of the assays performed. The fruit phenolic isolates were encapsulated in chitosan (CH) sodium tripolyphosphate (TPP) nanoparticles at a ratio of 4.0:1.0 (CH:TPP). HPLC-PDA was used to determine the entrapment efficiency of phenolic isolates to be 15.9 ± 2.7% and 23.0 ± 7.1% for chokecherries and saskatoons, respectively. Characteristics such as the size, surface potential and phenolic release were determined for the two fruit isolate containing nanoparticles. The size of the nanoparticles were 527.90 ± 74.57 nm and 443.03 ± 15.79 nm for chokecherries and saskatoons, respectively. Both of the nanoparticle systems had positive surface charges at 52.70 ± 2.93 mV and 54.43 ± 1.27 mV for chokecherries and saskatoons, respectively. The release properties of the CH:TPP nanoparticles containing fruit phenolics were examined in enzymatic simulated intestinal fluid and resulted in ~23% and ~28% release of chokecherry and saskatoon phenolics, respectively. Saskatoon phenolic isolates and isolates encapsulated in CH:TPP were gavage fed to rats (six animals in each of the two groups) at a dosage rate of 276.36 ± 9.74 mg/kg body weight. The saskatoon isolate contained 12.44 ± 0.44 mg/kg body weight anthocyanins (~3.30 mg anthocyanin per rat). These animals were sacrificed after 1 h and all stomach tissue samples in each of the treatment groups contained detectable levels of anthocyanins. In the small intestine tissues all six of the saskatoon isolate and three of the encapsulated isolate groups had detectable amounts of anthocyanins, while in the large intestine tissue, only one sample from the isolate group showed detectable amounts of anthocyanins. Although other tissues were tested (brain, heart, kidney and liver), anthocyanins were not detected. Therefore anthocyanins were detected in the gastrointestinal tract of both of the treatment groups. The research performed therefore illustrated that phenolic compounds can be extracted from fruit sources using EFW and can be successfully encapsulated in chitosan tripolyphosphate capsules allowing for targeted delivery in an animal model.

The major objectives of this research were to produce a phenolic rich isolate from two locally grown Saskatchewan fruits, chokecherries and saskatoons, develop an encapsulation system for the phenolic isolate, and test this system for the delivery of the phenolic isolate in an animal (rat) model. Natural phenolic compounds present in plants such as fruits have antioxidant and free radical scavenging activities, which have been proposed to have health benefits. The extraction of these compounds from plants is commonly performed using methanol despite being toxic to both humans and animals. As such, ethanol was investigated for its ability to extract phenolics from plants as a food safe alternative to methanol. Phenolic extraction from chokecherries with ethanol:formic acid:water (EFW) resulted in higher concentrations (9.83 mg gallic acid equivalents (GAE)/g fresh weight) than with methanol:formic acid:water (MFW) (7.97 mg GAE/g fresh weight). Results from saskatoons showed similar phenolic levels of 4.26 and 4.21 mg GAE/g fresh weight with MFW and ethanol (EFW), respectively. These results showed that EFW was a suitable substitute for MFW in phenolic compound isolation from chokecherries and saskatoons, and could be used to produce extracts that were safe for use in foods and feeds. High performance liquid chromatography with photodiode array detection (HPLC-PDA) was used to determine the phenolic compound composition of the raw fruits and their phenolic rich isolates. Chlorogenic acid was identified in both chokecherry and saskatoon samples, and rutin was also shown to be present in saskatoons. These identifications were based on the relative retention time and ultra violet-visual spectra comparisons to standards. Solid phase extraction (SPE) using Amberlite XAD-16 was employed to produce phenolic isolates from chokecherries and saskatoons. HPLC-PDA results determined that there was a ~2.7x and ~1.6x increase in peak area for chokecherries and saskatoons, respectively when SPE was employed. The antioxidant activity of the extracts and isolates was determined using in vitro radical scavenging tests including 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2´-azinobis-3-ethylbenzthiazoline-sulphonic acid (ABTS). The EFW chokecherry extract and isolate had the highest overall free radical scavenging activity. Crude fruit extracts exhibited lower free radical scavenging values compared to the isolate samples in both of the assays performed. The fruit phenolic isolates were encapsulated in chitosan (CH) sodium tripolyphosphate (TPP) nanoparticles at a ratio of 4.0:1.0 (CH:TPP). HPLC-PDA was used to determine the entrapment efficiency of phenolic isolates to be 15.9 ± 2.7% and 23.0 ± 7.1% for chokecherries and saskatoons, respectively. Characteristics such as the size, surface potential and phenolic release were determined for the two fruit isolate containing nanoparticles. The size of the nanoparticles were 527.90 ± 74.57 nm and 443.03 ± 15.79 nm for chokecherries and saskatoons, respectively. Both of the nanoparticle systems had positive surface charges at 52.70 ± 2.93 mV and 54.43 ± 1.27 mV for chokecherries and saskatoons, respectively. The release properties of the CH:TPP nanoparticles containing fruit phenolics were examined in enzymatic simulated intestinal fluid and resulted in ~23% and ~28% release of chokecherry and saskatoon phenolics, respectively. Saskatoon phenolic isolates and isolates encapsulated in CH:TPP were gavage fed to rats (six animals in each of the two groups) at a dosage rate of 276.36 ± 9.74 mg/kg body weight. The saskatoon isolate contained 12.44 ± 0.44 mg/kg body weight anthocyanins (~3.30 mg anthocyanin per rat). These animals were sacrificed after 1 h and all stomach tissue samples in each of the treatment groups contained detectable levels of anthocyanins. In the small intestine tissues all six of the saskatoon isolate and three of the encapsulated isolate groups had detectable amounts of anthocyanins, while in the large intestine tissue, only one sample from the isolate group showed detectable amounts of anthocyanins. Although other tissues were tested (brain, heart, kidney and liver), anthocyanins were not detected. Therefore anthocyanins were detected in the gastrointestinal tract of both of the treatment groups. The research performed therefore illustrated that phenolic compounds can be extracted from fruit sources using EFW and can be successfully encapsulated in chitosan tripolyphosphate capsules allowing for targeted delivery in an animal model.

A malária é um problema de saúde mundial. Esta doença é causada pela infecção dos eritrócitos com o protozoário do gênero Plasmodium. A atovaquona (ATQ) é um análogo estrutural da coenzima Q, usado como monoterapia ou em combinação com proguanil no tratamento da malária. A incorporação de fármacos de reduzida hidrossolubilidade, como a ATQ, em veículos tradicionais, é frequentemente limitada devido a razões de solubilidade. Neste contexto, nanoemulsões oferecem uma alternativa promissora devido a sua efetividade na solubilização de fármacos, aumentando a eficácia, e reduzindo os efeitos adversos. Assim, o objetivo deste trabalho foi desenvolver nanoemulsões catiônicas contendo ATQ para administração intravenosa. Nanoemulsões compostas de ATQ, triglicerídeos de cadeia média, lecitina de gema de ovo, 1,2-dioleoil-3-trimetil amônio propano (DOTAP), glicerol, polissorbato 80 e água foram obtidas pelo método de emulsificação espontânea. Este método levou à obtenção de nanoemulsões monodispersas com um diâmetro médio de aproximadamente 200 nm, como confirmado pela microscopia eletrônica de transmissão. O potencial  das nanoemulsões foi positivo (> 30 mV em pH 5). Um método isocrático de cromatografia líquida de fase reversa foi validado para quantificar ATQ. O método foi específico, linear, preciso e exato para a quantificação de ATQ. O teor de fármaco para todas as formulações foi próximo a 95%. A ATQ liberada a partir das nanoemulsões foi avaliada em dois meios diferentes (RPMI-1640 + 0,5% albumina ou PBS + 0,5% polissorbato 80) após a separação de ATQ livre através das membranas de ultrafiltração (poro de 0,1m). Independente do meio utilizado, ATQ foi progressivamente liberada a partir das formulações em função do fator de diluição e permaneceu constante em função do tempo. A maior liberação de ATQ foi observada para nanoemulsões contendo polissorbato 80 e no meio contendo este tensoativo não iônico em condições sink. O diâmetro das nanoemulsões permaneceu inalterado nas condições de liberação para nanoemsulões contendo polissorbato 80 enquanto uma inversão do - potencial de valores positivos para negativos foi detectado. Tais formulações mostraram uma inibição do crescimento do parasita in vitro frente a cepas de P. falciparum resistentes à ATQ. / Malaria is a global public health problem. This disease is caused by infection of red blood cells with protozoan parasites of the genus Plasmodium. Atovaquone (ATQ) is a structural analogue of coenzyme Q used as monotherapy or in combination with proguanil in malaria treatment. The incorporation of poorly-water soluble drugs such as ATQ in well-accepted vehicles is frequently limited due to solubility reasons. In this context, nanoemulsions offer an appealing alternative due to their effectiveness in drug solubilization, improved efficacy, and reduced side effects. The main purpose of this study was to develop cationic nanoemulsions containing ATQ for intravenous administration. Nanoemulsions composed of ATQ, medium chain triglycerides, egg lecithin, dioleoyl trimethylammonium propane (DOTAP), glycerol, polysorbate 80 and water were obtained by means of spontaneous emulsification. This procedure led to obtaining monodisperse nanoemulsions with mean droplet size of approximately 200 nm, as attested by transmission electron microscopy. - potential of nanoemulsions was positive (> 30mV at pH 5). An isocratic reversed – phase liquid chromatography method was validated for ATQ quantification. The method was specific, linear, precise, and accurate for ATQ quantification. The drug content in all formulations was close 95%. The ATQ release from nanoemulsions was evaluated in two different media (RPMI-1640 + 0.5% albumin or PBS + 0.5% polysorbate 80) after separation of free ATQ on ultrafiltration membranes (cut off 0.1μm). Whatever the media used, ATQ was progressively released from formulations upon the dilution ratio and remained constant over time. A higher release of ATQ was observed for nanoemulsions containing polysorbate 80 and in the media containing this non-ionic surfactant in sink conditions. The droplet size of nanoemulsions remained unchanged in the release conditions for nanoemulsions containing polysorbate 80 whereas an inversion of the -potential from positive to negative values was detected. Such formulations showed in vitro an inhibition of parasite growth against an ATQ-resistant P. falciparum strain.

A malária é um problema de saúde mundial. Esta doença é causada pela infecção dos eritrócitos com o protozoário do gênero Plasmodium. A atovaquona (ATQ) é um análogo estrutural da coenzima Q, usado como monoterapia ou em combinação com proguanil no tratamento da malária. A incorporação de fármacos de reduzida hidrossolubilidade, como a ATQ, em veículos tradicionais, é frequentemente limitada devido a razões de solubilidade. Neste contexto, nanoemulsões oferecem uma alternativa promissora devido a sua efetividade na solubilização de fármacos, aumentando a eficácia, e reduzindo os efeitos adversos. Assim, o objetivo deste trabalho foi desenvolver nanoemulsões catiônicas contendo ATQ para administração intravenosa. Nanoemulsões compostas de ATQ, triglicerídeos de cadeia média, lecitina de gema de ovo, 1,2-dioleoil-3-trimetil amônio propano (DOTAP), glicerol, polissorbato 80 e água foram obtidas pelo método de emulsificação espontânea. Este método levou à obtenção de nanoemulsões monodispersas com um diâmetro médio de aproximadamente 200 nm, como confirmado pela microscopia eletrônica de transmissão. O potencial  das nanoemulsões foi positivo (> 30 mV em pH 5). Um método isocrático de cromatografia líquida de fase reversa foi validado para quantificar ATQ. O método foi específico, linear, preciso e exato para a quantificação de ATQ. O teor de fármaco para todas as formulações foi próximo a 95%. A ATQ liberada a partir das nanoemulsões foi avaliada em dois meios diferentes (RPMI-1640 + 0,5% albumina ou PBS + 0,5% polissorbato 80) após a separação de ATQ livre através das membranas de ultrafiltração (poro de 0,1m). Independente do meio utilizado, ATQ foi progressivamente liberada a partir das formulações em função do fator de diluição e permaneceu constante em função do tempo. A maior liberação de ATQ foi observada para nanoemulsões contendo polissorbato 80 e no meio contendo este tensoativo não iônico em condições sink. O diâmetro das nanoemulsões permaneceu inalterado nas condições de liberação para nanoemsulões contendo polissorbato 80 enquanto uma inversão do - potencial de valores positivos para negativos foi detectado. Tais formulações mostraram uma inibição do crescimento do parasita in vitro frente a cepas de P. falciparum resistentes à ATQ. / Malaria is a global public health problem. This disease is caused by infection of red blood cells with protozoan parasites of the genus Plasmodium. Atovaquone (ATQ) is a structural analogue of coenzyme Q used as monotherapy or in combination with proguanil in malaria treatment. The incorporation of poorly-water soluble drugs such as ATQ in well-accepted vehicles is frequently limited due to solubility reasons. In this context, nanoemulsions offer an appealing alternative due to their effectiveness in drug solubilization, improved efficacy, and reduced side effects. The main purpose of this study was to develop cationic nanoemulsions containing ATQ for intravenous administration. Nanoemulsions composed of ATQ, medium chain triglycerides, egg lecithin, dioleoyl trimethylammonium propane (DOTAP), glycerol, polysorbate 80 and water were obtained by means of spontaneous emulsification. This procedure led to obtaining monodisperse nanoemulsions with mean droplet size of approximately 200 nm, as attested by transmission electron microscopy. - potential of nanoemulsions was positive (> 30mV at pH 5). An isocratic reversed – phase liquid chromatography method was validated for ATQ quantification. The method was specific, linear, precise, and accurate for ATQ quantification. The drug content in all formulations was close 95%. The ATQ release from nanoemulsions was evaluated in two different media (RPMI-1640 + 0.5% albumin or PBS + 0.5% polysorbate 80) after separation of free ATQ on ultrafiltration membranes (cut off 0.1μm). Whatever the media used, ATQ was progressively released from formulations upon the dilution ratio and remained constant over time. A higher release of ATQ was observed for nanoemulsions containing polysorbate 80 and in the media containing this non-ionic surfactant in sink conditions. The droplet size of nanoemulsions remained unchanged in the release conditions for nanoemulsions containing polysorbate 80 whereas an inversion of the -potential from positive to negative values was detected. Such formulations showed in vitro an inhibition of parasite growth against an ATQ-resistant P. falciparum strain.

A malária é um problema de saúde mundial. Esta doença é causada pela infecção dos eritrócitos com o protozoário do gênero Plasmodium. A atovaquona (ATQ) é um análogo estrutural da coenzima Q, usado como monoterapia ou em combinação com proguanil no tratamento da malária. A incorporação de fármacos de reduzida hidrossolubilidade, como a ATQ, em veículos tradicionais, é frequentemente limitada devido a razões de solubilidade. Neste contexto, nanoemulsões oferecem uma alternativa promissora devido a sua efetividade na solubilização de fármacos, aumentando a eficácia, e reduzindo os efeitos adversos. Assim, o objetivo deste trabalho foi desenvolver nanoemulsões catiônicas contendo ATQ para administração intravenosa. Nanoemulsões compostas de ATQ, triglicerídeos de cadeia média, lecitina de gema de ovo, 1,2-dioleoil-3-trimetil amônio propano (DOTAP), glicerol, polissorbato 80 e água foram obtidas pelo método de emulsificação espontânea. Este método levou à obtenção de nanoemulsões monodispersas com um diâmetro médio de aproximadamente 200 nm, como confirmado pela microscopia eletrônica de transmissão. O potencial  das nanoemulsões foi positivo (> 30 mV em pH 5). Um método isocrático de cromatografia líquida de fase reversa foi validado para quantificar ATQ. O método foi específico, linear, preciso e exato para a quantificação de ATQ. O teor de fármaco para todas as formulações foi próximo a 95%. A ATQ liberada a partir das nanoemulsões foi avaliada em dois meios diferentes (RPMI-1640 + 0,5% albumina ou PBS + 0,5% polissorbato 80) após a separação de ATQ livre através das membranas de ultrafiltração (poro de 0,1m). Independente do meio utilizado, ATQ foi progressivamente liberada a partir das formulações em função do fator de diluição e permaneceu constante em função do tempo. A maior liberação de ATQ foi observada para nanoemulsões contendo polissorbato 80 e no meio contendo este tensoativo não iônico em condições sink. O diâmetro das nanoemulsões permaneceu inalterado nas condições de liberação para nanoemsulões contendo polissorbato 80 enquanto uma inversão do - potencial de valores positivos para negativos foi detectado. Tais formulações mostraram uma inibição do crescimento do parasita in vitro frente a cepas de P. falciparum resistentes à ATQ. / Malaria is a global public health problem. This disease is caused by infection of red blood cells with protozoan parasites of the genus Plasmodium. Atovaquone (ATQ) is a structural analogue of coenzyme Q used as monotherapy or in combination with proguanil in malaria treatment. The incorporation of poorly-water soluble drugs such as ATQ in well-accepted vehicles is frequently limited due to solubility reasons. In this context, nanoemulsions offer an appealing alternative due to their effectiveness in drug solubilization, improved efficacy, and reduced side effects. The main purpose of this study was to develop cationic nanoemulsions containing ATQ for intravenous administration. Nanoemulsions composed of ATQ, medium chain triglycerides, egg lecithin, dioleoyl trimethylammonium propane (DOTAP), glycerol, polysorbate 80 and water were obtained by means of spontaneous emulsification. This procedure led to obtaining monodisperse nanoemulsions with mean droplet size of approximately 200 nm, as attested by transmission electron microscopy. - potential of nanoemulsions was positive (> 30mV at pH 5). An isocratic reversed – phase liquid chromatography method was validated for ATQ quantification. The method was specific, linear, precise, and accurate for ATQ quantification. The drug content in all formulations was close 95%. The ATQ release from nanoemulsions was evaluated in two different media (RPMI-1640 + 0.5% albumin or PBS + 0.5% polysorbate 80) after separation of free ATQ on ultrafiltration membranes (cut off 0.1μm). Whatever the media used, ATQ was progressively released from formulations upon the dilution ratio and remained constant over time. A higher release of ATQ was observed for nanoemulsions containing polysorbate 80 and in the media containing this non-ionic surfactant in sink conditions. The droplet size of nanoemulsions remained unchanged in the release conditions for nanoemulsions containing polysorbate 80 whereas an inversion of the -potential from positive to negative values was detected. Such formulations showed in vitro an inhibition of parasite growth against an ATQ-resistant P. falciparum strain.

No description available.

Siekiant efektyvaus propolio fenolinių junginių poveikio odoje, svarbu sumodeliuoti veiksmingai šiuos junginius atpalaiduojančias ir tiekiančias į odą puskietes sistemas su propolio produktais. Darbo tikslas – pagrįsti puskiečių formų su propolio produktais modeliavimo principus, optimizuoti eksperimentines puskietes formas ir suformuluoti kokybės modelį, remiantis biofarmaciniais tyrimais. Įvertinta atskirų fenolinių rūgščių (vanilo, kavos, p-kumaro, ferulo rūgšties) ir vanilino skvarba į žmogaus odą ex vivo iš vandeninės propolio ištraukos ir eksperimentinių propolio puskiečių formų: tepalo, vanduo-aliejus tipo kremo, hidrogelio, aliejus-vanduo tipo gelifikuoto kremo. Patvirtintas aktyvus puskietės sistemos vaidmuo, atpalaiduojant ir darant įtaką propolio veikliųjų junginių skvarbai į odą. Įvertintas tirtų fenolinių junginių pasiskirstymas odos sluoksniuose (epidermyje ir dermoje), kuris susietas su skirtingu jų lipofiliškumu. Santykinai lipofiliniai junginiai (ferulo ir p-kumaro rūgštis) kaupėsi epidermyje, o santykinai hidrofiliniai junginiai (vanilo rūgštis ir vanilinas) migravo į dermą. Kavos rūgštis, kurios tirtuose propolio produktuose yra mažesnis kiekis negu kitų fenolinių rūgščių, nustatyta tik epidermyje. Pritaikytas eksperimentų planavimas vaisto formų sudėties optimizavimui. Optimizavimo parametrai leido prognozuoti propolio veikliųjų junginių atpalaidavimą iš tirtų puskiečių sistemų. / In order to achieve a desired effect of propolis phenolic compounds in the skin, it is important to develop semisolid systems with propolis products that would efficiently release and deliver these compounds into the skin. The aim of the study is to justify the modeling principles of semisolid formulations with propolis products, to optimize experimental semisolid formulations, and to formulate a quality model based on biopharmaceutical studies. The penetration of individual phenolic acids (vanillic, caffeic, p-coumaric, and ferulic) and vanillin from aqueous propolis extract and experimental propolis semisolid formulations (ointment, water-in-oil cream, hydrogel, and oil-in-water gelified cream) into the human skin ex vivo have been evaluated. An active role of the semisolid system in releasing and influencing the penetration of propolis active compounds into the skin has been confirmed. The distribution of investigated phenolic compounds in the layers of skin (epidermis and dermis) was evaluated and linked to different lipophilicity. Relatively lipophilic compounds (ferulic and p-coumaric acids) accumulated in the epidermis and relatively hydrophilic compounds (vanillic acid and vanillin) migrated to the dermis. Caffeic acid, which was present in analyzed propolis products at lower quantities as compared with other phenolic acids, was determined only in the epidermis. The design of experiments was applied for the optimization of the composition of dosage forms. Optimization... [to full text]