Reverse micelles systems AOT/isooctane was used for the concentration chymotrypsin from crude aqueous extract of red perch (intestine). The effects of pH and AOT concentration in the forward extraction step and pH and KCl concentration in the backward extraction step on the enzyme activity, purification fold and recovery yield were studied. The optimum conditions for the forward extraction were AOT concentration 20 mM and pH of 7.0 and optimum conditions for backward extraction were KCl concentration 1.0 M and pH of 7.5 which gave a good recovery yield (102.24%) and a purification (32.24-fold). The addition of 15% v/v alcohol in backward extraction dramatically improved recovery yield by 4.5 times and purification by 2.5 times. The enzyme activity and recovery yield obtained using reverse micelles method under its optimal conditions were 2 fold higher than those obtained using the ammonium sulphate precipitation method, while purification fold were 3 fold higher. / Fish processing waste can be used to produce commercially valuable by-products, such as chymotrypsin which has application in various industries including the food industry, leather production industry and chemical industry. My project is to produce valuable by-products from the fish processing.

Made available in DSpace on 2014-06-11T19:28:03Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-12-20Bitstream added on 2014-06-13T18:57:11Z : No. of bitstreams: 1 silva_gbrf_me_arafcf.pdf: 1472073 bytes, checksum: 558b88d78004c177c0d910e0ef0cde8a (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / As microemulsões (ME) apresentam grande potencial como sistemas de administração de fármacos, pelas propriedades de solubilizar fármacos hidrofílicos em meio lipofílico ou fármacos lipofílicos em meio aquoso. O objetivo principal desse projeto foi preparar sistemas microemulsionados contendo misturas de Fosfatidilcolina de Soja (FS) / Óleo de Rícino Polioxil-40-Hidrogenado (ORPH) / Oleato de Sódio (OS), Colesterol (CO) e tampão aquoso Tris-HCl pH 7,2 e incorporar a esses sistemas metiljasmonato (MJ), que é inibidor da proliferação in vitro e supressor da tumorgenicidade in vivo. A caracterização físico-química desses sistemas com e sem MJ foi realizada através das técnicas de espalhamento dinâmico de luz, reologia, microscopia de luz polarizada e difração de raios-x. De acordo com o espalhamento dinâmico de luz foi observada diminuição do raio hidrodinâmico com o aumento da proporção da razão óleo/tensoativos (O/T), tanto na presença como na ausência de MJ. Foi observado aumento do raio hidrodinâmico com o aumento da porcentagem de sistema tensoativo na ausência do fármaco, enquanto na presença de metiljasmonato ocorreu diminuição do raio hidrodinâmico. Independente da variação da proporção de O/T e de sistema tensoativo, ocorreu diminuição do raio hidrodinâmico com a incorporação do fármaco. De acordo com as análises do comportamento reológico todas as amostras apresentaram comportamento de fluído pseudoplástico, porém nas microemulsões isentas de MJ a maioria apresentou tixotropia enquanto na presença do fármaco a maioria apresentou anti-tixotropia... / Microemulsions (ME) have great potential as drug delivery systems because of their ability to improve the dissolution of drugs in environments with opposite lipophilicity or hydrophilicity. The main objective of this project was to prepare microemulsions systems containing soya phosphatidylcholine (SPC) / polyoxyethyleneglycerol trihydroxystearate 40 (ORPH) / Sodium Oleate (SO) as surfactant, cholesterol (CHO) as oil phase and 0.01M Tris-HCl pH 7.2 as aqueous phase. Methyljasmonate (MJ), an inhibitor of in vitro proliferation and in vivo suppressor of tumorigenesis, was incorporated into these systems. The physico-chemical characterization of the systems, containing or not MJ, was performed by light scattering, rheological behavior, polarized light microscopy, and X-ray diffraction. According to dynamic light scattering, a decrease in the hydrodynamic radius was observed with increasing proportion of oil phase/surfactant (O/S) in the presence and absence of MJ. In the absence of the drug, the hydrodynamic radius increased with increasing ratio of surfactant, while in the presence of MJ the hydrodynamic... (Complete abstract click electronic access below)

Made available in DSpace on 2014-06-11T19:24:11Z (GMT). No. of bitstreams: 0 Previous issue date: 2006-08-03Bitstream added on 2014-06-13T20:48:29Z : No. of bitstreams: 1 franzini_cm_me_arafcf.pdf: 1232208 bytes, checksum: d972d0938f33ec34d5b67691a2669a0a (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O objetivo esse trabalho foi o desenvolvimento de sistemas microemulsionados ou subemulsionados estabilizados por fosfatidilcolina de soja (FS) associada à tensoativos hidrofílicos para veiculação de AmB. Foi avaliada a utilização de pluronic (PLU) como co-tensoativo e observou-se o predomínio de sistemas mais viscosos. O estudo do comportamento reológico das MEs, mostrou que todas as amostras apresentaram comportamento Newtoniano, mantendo a viscosidade constante independente da velocidade de deformação. Os estados de agregação da AmB foram avaliados através de seus espectros de absorção relativos às diferentes concentrações, inclusão em ciclodextrinas e incorporação no sistema microemulsionado selecionado. Foi verificada uma modificação relativa na proporção de monômeros do fármaco. A técnica de nefelometria foi realizada e indicou aumento no tamanho das gotículas da ME com o aumento da concentração da AmB. Verificou-se que a incorporação de AmB nos sistemas desenvolvidos é dependente da proporção relativa da fase interna oleosa. A liberação de AmB, in vitro, é mais prolongada à medida que aumenta a proporção de fase interna oleosa. Os sistemas microemulsionados e subemulsionados mostraram-se eficazes na veiculação de AmB apresentando-se como sistema reservatório facilitando a liberação prolongada. / The objective of this work was the development of subemulsified and microemulsified systems stabilized by soya phosphatidylcholine (FS) and/or FS associated to hydrophylic surfactants to AmB veiculation. It was evaluated the use of pluronic (PLU) as co-surfactant and it was observed the predominancy of the more viscose systems. The study of the rheological behavior of MEs verified that all the samples have presented Newtonian behavior, maintaining the viscosity constant independent of the deformation speed. The states of aggregation of the AmB have been evaluated through its relative absorption specters to the different concentrations, inclusion in cyclodextrins and incorporation in the selected microemulsion system. A relative modification in the proportion of monomers of the drug was verified. The Nefelometry technique was performed and showed increase in the droplets size of the ME with increase to the AmB concentration. It was verified that the AmB incorporation in the developed systems is dependent on the oil phase. The in vitro release of AmB is more prolonged as long as the internal oil phase increases. The microemulsified and subemulsified systems appear as more effective in the AmB veiculation presenting themselves as reservatory system, facilitating the prolonged release.

Orientador: Anselmo Gomes de Oliveira / Banca: Maria Vitória Lopes Badra Bentley / Banca: Newton Andréo Filho / Resumo: As microemulsões (ME) apresentam grande potencial como sistemas de administração de fármacos, pelas propriedades de solubilizar fármacos hidrofílicos em meio lipofílico ou fármacos lipofílicos em meio aquoso. O objetivo principal desse projeto foi preparar sistemas microemulsionados contendo misturas de Fosfatidilcolina de Soja (FS) / Óleo de Rícino Polioxil-40-Hidrogenado (ORPH) / Oleato de Sódio (OS), Colesterol (CO) e tampão aquoso Tris-HCl pH 7,2 e incorporar a esses sistemas metiljasmonato (MJ), que é inibidor da proliferação in vitro e supressor da tumorgenicidade in vivo. A caracterização físico-química desses sistemas com e sem MJ foi realizada através das técnicas de espalhamento dinâmico de luz, reologia, microscopia de luz polarizada e difração de raios-x. De acordo com o espalhamento dinâmico de luz foi observada diminuição do raio hidrodinâmico com o aumento da proporção da razão óleo/tensoativos (O/T), tanto na presença como na ausência de MJ. Foi observado aumento do raio hidrodinâmico com o aumento da porcentagem de sistema tensoativo na ausência do fármaco, enquanto na presença de metiljasmonato ocorreu diminuição do raio hidrodinâmico. Independente da variação da proporção de O/T e de sistema tensoativo, ocorreu diminuição do raio hidrodinâmico com a incorporação do fármaco. De acordo com as análises do comportamento reológico todas as amostras apresentaram comportamento de fluído pseudoplástico, porém nas microemulsões isentas de MJ a maioria apresentou tixotropia enquanto na presença do fármaco a maioria apresentou anti-tixotropia... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Microemulsions (ME) have great potential as drug delivery systems because of their ability to improve the dissolution of drugs in environments with opposite lipophilicity or hydrophilicity. The main objective of this project was to prepare microemulsions systems containing soya phosphatidylcholine (SPC) / polyoxyethyleneglycerol trihydroxystearate 40 (ORPH) / Sodium Oleate (SO) as surfactant, cholesterol (CHO) as oil phase and 0.01M Tris-HCl pH 7.2 as aqueous phase. Methyljasmonate (MJ), an inhibitor of in vitro proliferation and in vivo suppressor of tumorigenesis, was incorporated into these systems. The physico-chemical characterization of the systems, containing or not MJ, was performed by light scattering, rheological behavior, polarized light microscopy, and X-ray diffraction. According to dynamic light scattering, a decrease in the hydrodynamic radius was observed with increasing proportion of oil phase/surfactant (O/S) in the presence and absence of MJ. In the absence of the drug, the hydrodynamic radius increased with increasing ratio of surfactant, while in the presence of MJ the hydrodynamic... (Complete abstract click electronic access below) / Mestre

Orientador: Watson Loh / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Química / Made available in DSpace on 2018-08-18T09:05:41Z (GMT). No. of bitstreams: 1 Ferreira_MairaSilva_D.pdf: 4524526 bytes, checksum: 98c8f3d75444a9697c4d47cc2b82c519 (MD5) Previous issue date: 2011 / Resumo: Surfatantes à base de silicona possuem propriedades diferentes e especiais e são amplamente empregados na indústria de produtos farmacêuticos e cosméticos. Neste trabalho o emprego deste tipo de surfatante é utilizado para o estudo de incorporação de água em solventes que exibem comportamento diferenciado daquele apresentado por solventes hidrocarbonetos, devido à sua baixa energia coesiva, como o óleo de silicona. Siliconas fazem parte de uma classe de materiais industrialmente importante que apresentam uma excelente estabilidade química e inúmeras aplicações, tais como, cosméticos, agente de controle de espumas, antiaderente, entre outras. Embora importante, existem poucas pesquisas relatando o equilíbrio de fases e a caracterização de sistemas com estes constituintes. Dado este fato, iniciou-se a primeira etapa deste trabalho que é um estudo com o objetivo de caracterizar sistemas que contém óleos e surfatantes à base de silicona em água, investigando as estruturas dos agregados formados utilizando a técnica de espalhamento de raios-X a baixo ângulo (SAXS). Observações importantes como a presença de estados de agregação (microemulsões e mesofases) foram identificadas e caracterizadas a partir da construção do diagrama de fases dos sistemas. A identificação das fases formadas por estes constituintes, em especial as microemulsões bicontínuas e fase lamelar, impulsionaram o desenvolvimento da outra etapa deste trabalho, o estudo de nanoemulsões utilizando um método de baixa energia. Este método faz uso de propriedades físico químicas do sistema e utiliza a inversão espontânea da curvatura do surfatante para obtenção de gotas com tamanho diminuto. As nanoemulsões com surfatantes e óleos à base de silicona obtidas neste trabalho apresentaram raio hidrodinâmico (rh) entre 26 e 40 nm, baixo índice de polidispersidade (< 0,1), uma boa estabilidade cinética (semanas) e podem ser utilizadas em aplicações que não requerem sua formação por longos períodos como dispersões na área agroquímica. A presença do SDS nessa formulação aumentou o rh para 150 a 300 nm, mas melhorou consideravelmente a sua estabilidade cinética, estendendo-a a meses. O desenvolvimento de tais sistemas nos mostra uma nova classe de dispersões que podem competir ou superar sistemas semelhantes e assumir um grande papel no setor científico e industrial / Abstract: Silicone surfactants display unusual properties and are widely employed in industries, in which they are used as basic material in pharmaceutical and cosmetic products. In this work, these types of surfactants have been employed to study the water incorporation into solvents that exhibit behavior different from that presented by hydrocarbon solvents due to their low cohesive energy, such as silicone oil. Silicone oils are important industrial substances that present an excellent chemical stability and numerous applications, for instance in cosmetics, foam control and nonstick agents, among others. Despite their importance, there are few studies reporting phase equilibrium and characterization of these systems. Given this fact, the first step of this work is to characterize systems containing oils and silicone surfactants using small angle x-ray scattering (SAXS). Important aggregates (microemulsions and mesophases) were identified and characterized through their phase diagrams. Thus, identification of phases formed, in particular, by surfactant aggregation in bicontinuous microemulsions and lamellar phases has driven the next stage of this work that consists of preparing nanoemulsions using a low energy method. This method uses physical and chemical properties of the system and exploits the reversal of spontaneous curvature of surfactant structures to obtain droplets with diminutive size. The nanoemulsions formed by silicone surfactants and silicone oils in this work displayed hydrodynamic radii (rh) between 26 and 40 nm, low polydispersion index (< 0.1), a good kinetic stability (weeks) and may be used in applications allow for shorter shelf-life periods, such as in agrochemical area. The presence of SDS in its formulation increased the rh to around for 150-300 nm, but improved considerably its kinetic stability from weeks to months. The development of such systems reveals us a new class of dispersions that can compete or exceed similar systems and assume a significant role in the scientific and industrial sector / Doutorado / Físico-Química / Doutor em Ciências

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Orientador: Anselmo Gomes de Oliveira / Banca: Marco Vinícius Chaud / Banca: Renata Fonseca Vianna Lopez / Resumo: O objetivo esse trabalho foi o desenvolvimento de sistemas microemulsionados ou subemulsionados estabilizados por fosfatidilcolina de soja (FS) associada à tensoativos hidrofílicos para veiculação de AmB. Foi avaliada a utilização de pluronic (PLU) como co-tensoativo e observou-se o predomínio de sistemas mais viscosos. O estudo do comportamento reológico das MEs, mostrou que todas as amostras apresentaram comportamento Newtoniano, mantendo a viscosidade constante independente da velocidade de deformação. Os estados de agregação da AmB foram avaliados através de seus espectros de absorção relativos às diferentes concentrações, inclusão em ciclodextrinas e incorporação no sistema microemulsionado selecionado. Foi verificada uma modificação relativa na proporção de monômeros do fármaco. A técnica de nefelometria foi realizada e indicou aumento no tamanho das gotículas da ME com o aumento da concentração da AmB. Verificou-se que a incorporação de AmB nos sistemas desenvolvidos é dependente da proporção relativa da fase interna oleosa. A liberação de AmB, in vitro, é mais prolongada à medida que aumenta a proporção de fase interna oleosa. Os sistemas microemulsionados e subemulsionados mostraram-se eficazes na veiculação de AmB apresentando-se como sistema reservatório facilitando a liberação prolongada. / Abstract: The objective of this work was the development of subemulsified and microemulsified systems stabilized by soya phosphatidylcholine (FS) and/or FS associated to hydrophylic surfactants to AmB veiculation. It was evaluated the use of pluronic (PLU) as co-surfactant and it was observed the predominancy of the more viscose systems. The study of the rheological behavior of MEs verified that all the samples have presented Newtonian behavior, maintaining the viscosity constant independent of the deformation speed. The states of aggregation of the AmB have been evaluated through its relative absorption specters to the different concentrations, inclusion in cyclodextrins and incorporation in the selected microemulsion system. A relative modification in the proportion of monomers of the drug was verified. The Nefelometry technique was performed and showed increase in the droplets size of the ME with increase to the AmB concentration. It was verified that the AmB incorporation in the developed systems is dependent on the oil phase. The in vitro release of AmB is more prolonged as long as the internal oil phase increases. The microemulsified and subemulsified systems appear as more effective in the AmB veiculation presenting themselves as reservatory system, facilitating the prolonged release. / Mestre

Various colloidal delivery systems, including polymeric nanoparticles, metal colloids, liposomes, and microemulsions have been reported to enhance the delivery of therapeutic agents following intranasal administration. However, the mechanisms involved in the uptake of these nanomaterials, especially those in the ultrafine size ranges (diameter < 20 nm) through nasal mucosa and their subsequent biodistribution in the body are not well characterized. The objectives of this study address the knowledge gap regarding ultrafine nanoparticle transfer in the nasal mucosa by quantifying nanoparticle uptake and biodistibution patterns in the presence and absence of known inhibitors of endocytic processes. The uptake of ~ 10 nm fluorescent quantum dots (QDs) was investigated by measuring the concentration of QDs following exposure to bovine respiratory and olfactory mucosal explants. An inductively coupled optical emission spectroscopy method was developed to measure the amount of QDs within the tissues. The results demonstrated that carboxylate-modified QDs (COOH-QDs) show ~2.5 fold greater accumulation in the epithelial and submucosal regions of the olfactory tissues compared to the respiratory tissues. Endocytic inhibitory studies showed that in respiratory tissues clathrin-dependent, macropinocytosis and caveolae-dependent endocytosis process were all involved in the uptake of COOH-QDs. Whereas in olfactory tissues, clathrin-dependent endocytosis was the major endocytic pathway involved in uptake of COOH-QDs. Additional energy-independent pathways appeared to also be active in the transfer of COOH-QDs into the olfactory mucosa. Interestingly, PEGylated quantum dots (PEG-QDs) of similar size ~15 nm were not internalized into the bovine nasal tissues. In vivo fluorescence imaging was used to study the biodistribution of quantum dots following nasal instillation in mice. These studies showed that majority of COOH-QDs remain in the nasal tissues for relatively long periods of time (up to 24 h) whereas PEG-QDs showed no such accumulation. Biodistribution studies of gold nanoparticles (~15 nm) in mice using micro-CT showed that gold nanoparticles were transferred to the posterior turbinate region and a fraction of the administered dose distributed to regions in close proximity to the olfactory bulb. Both NIR imaging and micro-CT imaging were useful tools for visualization of in vivo nanoparticle distribution. A diazepam-containing microemulsion (dispersed phase ~40 nm) was formulated to investigate the uptake mechanisms utilized for fluid-phase colloidal dispersions in the nasal mucosa. The resulting diazepam-containing microemulsion showed enhanced transfer of the drug into the bovine nasal respiratory and olfactory tissues. It is unclear if endocytosis of the fluid-phase nanodispersions played a role in drug absorption from the microemulsions in a manner similar to the uptake of solid-phase nanoparticles, however, since there was significant loss of the epithelial cell layer following exposure to the microemulsion formulation which likely altered the barrier properties of the epithelium. These studies have increased the fundamental understanding of ultrafine nanoparticle uptake in the nasal tissues and the resulting nanoparticle biodistribution patterns. While ultrafine nanoparticles may have limited application in the development of efficient drug delivery systems, an understanding of the size-dependent and tissue-dependent processes responsible for the uptake of particulates into mucosal tissues will contribute to the rational development of nanoparticulate drug delivery strategies investigating the nasal and other routes of administration.

No description available.