The objective of this research is to further define the relationship between the charge state of insulin, and the self assembly properties of insulin and PEGylated insulin in solution. Polyethylene glycol (PEG) chains were covalently attached to insulin in order to evaluate their impact on insulin's systemic duration of action after pulmonary dosing. This thesis will focus on the assembly properties of the PEG-insulin and insulin, and also demonstrate how the charge state, which was modified by the covalent attachment of PEG, relates to different modes of behavior by anion and cation exchange chromatography. In addition, explain how modifying the assembly state extends to improving formulation properties of spray-dried insulin powders. This thesis is an investigation into the relationship of insulin's charge state controlled by pH and how the charge state affects the self assembly of insulin, especially when the zinc ion is removed. Ionic interaction is one of the major forces affecting insulin assembly. The theory that a change in the charge state of insulin could modulate the ionic interaction and reduce hexamer formation at alkaline conditions was investigated. Experiments were designed to measure the level of hexamer with light scattering, and the amount of hexamer was then correlated with the pH and zinc content of the solutions. The importance of the charge state of the monomer and its behavior extends to chromatography and purification modes as well. Specifically, the purification of various species of PEGylated insulin presents a challenge. By varying mobile phase pH which induces the charge to insulin, an ion exchange method demonstrated very high resolution and controllable interaction between the ion exchange media and the insulin derivatives. A highly accurate method for determining molecular weight and thus the average associated state of insulin in solution has been developed using the MALS (Multi-Angle Light Scattering). Insulin concentration, pH, and metal ion concentrations, were in pharmaceutically relevant ranges. The MALS method was developed to evaluate how the parameters above affect the self-assembly properties of insulin, and use the assembly properties to improve formulations of insulin or PEGylated insulin. To use the light scattering technique the dn/dc (change in refractive index with change in concentration) is required. During the method development, the dn/dc of insulin was measured at 690 nm, and a value of 0.185 mL/g based on theory was confirmed. A novel approach for preparing insulin powders with improved chemical stability, based on maintaining the dissociation of hexamers in solution during the spray drying process was developed. The mode presented here is to remove the zinc ions from solution, increase the pH from 6.6 to 7.8, and maintain a low concentration of insulin approximately 2 to 15 mg/mL. Each of these factors alone decreases the hexamer population in solution, but by combining all three factors, hexamers are driven to very low levels of equilibrium. The increased stability of the powders is predominately related to the decrease in covalent insulin dimer (CID). The data presented correlates a reduced hexamer population in the solution with lower levels of CID's in the dry powder compared to controls. The CID formation rate was reduced by 40% compared to a control.

M.Sc. (Chemistry) / Human immunodeficiency virus (HIV) is continuously rewriting medical history as one of the diseases affecting humankind. Current treatments available for HIV, namely antiretrovirals (ARVs), do not completely eradicate the virus from the body, leading to life time commitment. Many ARVs suffer from high toxicities and unpleasant side effects; as a result many patients do not adhere to the treatment. Nanoparticles (NPs) used as drug delivery systems (DDS) hold tremendous potential, since they can easily protect the drug from external environment and enter the human cells to deliver drugs. Therefore, the main objective of this work was to load two ARVs, namely lamivudine (LAM) and efavirenz (EFV), into a biodegradable, biocompatible poly(epsilon-caprolactone) (PCL) polymer based NPs. LAM is a hydrophilic drug suffering from low half life of 5 to 7 hours and many unpleasant side effects. EFV is a hydrophobic drug suffering from low aqueous solubility (4 μg/ml), which leads to a limited oral absorption and low bioavailability (40-45%).

<p>Dry Powder inhalers (DPIs) have been one of the most promising developments in pulmonary drug delivery systems. In general, DPIs are more effective than systemic administrations and convenient to use. However, delivering high-dose antibiotics through a DPI is still a challenge because high powder load may need a very large inhaler or increase the incidence of local adverse effects. Spray drying has been increasingly applied to produce DPI formulations for high-dose antibiotics; nevertheless, many spray-dried particles are amorphous and physically unstable during storage, particularly under the humid environment. </p> <p> </p> <p>My research focuses on addressing critical challenges in physical stability of DPIs for spray-dried high-dose antibiotics. The effects of moisture-induced crystallization on physical stability and aerosol performance of spray-dried amorphous Ciprofloxacin DPI formulations stored at different humidity conditions were studied. Our study not only provided a mechanistic understanding in the impact of crystallization on aerosol performance but also developed novel approaches for improving stability of spray-dried formulations used in DPI.</p> <p> </p> <p>Our work has shown that recrystallization of amorphous spray-dried Ciprofloxacin led to significant changes in aerosol performance of DPIs upon storage, which cause critical quality and safety concerns. These challenges have been solved through co-spray-drying Ciprofloxacin with either excipient such as leucine or synergistic antibiotic like Colistin. Co-spray-drying Ciprofloxacin with Colistin not only improved physical and aerosol stability but also enhanced antibacterial activity which is a great advantage for treating ‘difficult to cure’ respiratory infections caused by multidrug resistant bacteria.</p> <p> </p> <p>My research work is a sincere effort to maximize the utility and efficacy of high-dose DPI, an effective delivery tool for treating severe resistant bacterial respiratory infections.</p>

Nous avons mis au point par atomisation-séchage un système vecteur innovant de corticostéroïdes destiné à être administré par voie pulmonaire pour le traitement des maladies inflammatoires des voies respiratoires dont la plus fréquente est l’asthme. Cette forme pharmaceutique appelée "grosse particule poreuse" consiste en une matrice de deux excipients biocompatibles et biodégradables, la 1,2-dipalmitoyl-sn-glycero-3-phosphatidylcholine (DPPC) et l'acide hyaluronique (AH) encapsulant le palmitate de dexaméthasone (DXP). La première partie de ce travail a consisté à optimiser l’encapsulation d’un glucocorticoïde modèle, la DXP, une prodrogue lipophile de la dexaméthasone (DXM) au sein de microparticules poreuses par atomisation séchage. Les résultats des tests physicochimiques ont permis d’isoler la poudre à 5% de DXP dont les propriétés morphologiques et aérodynamiques répondent bien aux critères des grosses particules poreuses. La cinétique de libération in vitro a montré une libération relativement lente avec moins de 5% de DXP dans le surnageant après 21 jours. Nous avons ensuite mis au point une méthode d’extraction et de de dosage de la DXP et de son métabolite actif, la DXM dans le plasma et le liquide de lavage bronchoalveolaire (BALF). La dernière partie de notre travail a été consacrée à l’étude pharmacocinétique après administration pulmonaire de la formulation optimisée chez le rat sain. Les résultats in vivo montrent une libération relativement lente de la DXP dans le BALF tandis que la concentration en DXM diminue rapidement en 4 heures. En revanche les concentrations dans le plasma restent faibles. Ces résultats indiquent une bonne distribution pulmonaire de la DXP et la DXM avec une faible absorption sanguine de ces molécules ce qui est prometteur pour le traitement local de l’asthme avec la possibilité de réduire le nombre d’administrations et d’améliorer l’observance thérapeutique des patients. / We have developed by spray drying an innovative carrier system of corticosteroids to be administered by pulmonary route for the treatment of airway inflammatory diseases, among which the most common is asthma. This pharmaceutical form known as "large porous particles" consists of a matrix composed of two biocompatible and biodegradable excipients, 1,2-dipalmitoyl-sn-glycero-3-phosphatidylcholine (DPPC) and hyaluronic acid (HA) encapsulating dexamethasone palmitate (DXP). The first part of this work was to optimize the encapsulation of a model glucocorticoid, DXP, a lipophilic prodrug of dexamethasone (DXM) within porous microparticles by spray drying. The results of the physicochemical characterization allowed to isolate microparticles loaded with 5% DXP for which morphological and aerodynamic properties meet the criteria of large porous particles. In vitro release kinetics showed a relatively slow release with less than 5% of DXP in the supernatant after 21 days. Then we developed an extraction method and a HPLC method for the determination of DXP and its active metabolite, DXM in plasma and bronchoalveolar lavage fluid (BALF). In vivo results show a relatively slow release of DXP in BALF while the DXM concentration decreases rapidly in 4 hours. By contrast, the plasma concentrations remained low. These results indicate good pulmonary delivery of DXP and DXM with low blood absorption of these molecules which is promising for the local treatment of asthma with the possibility of reducing the number of administrations and improve patient compliance.

Ce travail porte sur l’étude de l’évaporation d’une goutte présentant une interface liquide/gaz complexe. Nous choisissons de nous focaliser sur deux manières différentes dont cette interface peut se complexifier, par interaction avec une phase solide :soit la phase solide constitue le support sur lequel repose la goutte, soit la phase solide se situe au sein même de la goutte. En particulier, nous nous intéressons à l’étude de l’évaporation de gouttes sessiles sur des surfaces géométriquement texturées ainsi qu’à l’étude de l’évaporation de gouttes en suspension contenant des composés non volatils.Tout d’abord, nous étudions les dynamiques d’évaporation de gouttes d’un liquide complètement mouillant et très volatil déposées sur des surfaces géométriquement texturées mais chimiquement homogènes. La texturation de la surface consiste en un réseau de piliers circulaires organisés selon un schéma carré, construits par photolithographie de façon à pouvoir contrôler la rugosité de la surface. La dynamique de la ligne de contact et la forme de la goutte sont caractérisées par interférométrie. Une étude paramétrique est réalisée en faisant varier les paramètres géométriques de la texturation de surface, typiquement le rayon et la hauteur des piliers (en gardant la distance inter-piliers fixe). L’étude met en évidence trois types de dynamiques en fonction de la valeur de ces para- mètres géométriques :i) un régime dominé par l’évaporation, avec une ligne de contact en récession pendant toute la durée de l’évaporation de la goutte ;ii) un régime dominé par l’étalement, avec un rayon initialement croissant pendant le début de l’évaporation ;iii) une situation intermédiaire avec des effets d’accrochage de la ligne de contact importants. Un modèle mathématique est développé, qui met en évidence l’influence de la géométrie de la texturation de surface sur la compétition entre les phénomènes d’étalement et d’évaporation de la goutte. Le modèle parvient à reproduire qualitativement les résultats expérimentaux, montrant que le choix entre les deux régimes extrêmes dépend de la valeur d’un paramètre adimensionnel, qui compare les temps caractéristiques d’évaporation et d’étalement au sein de la texturation de surface. Des lois d’échelles sont aussi mises en évidence, à la fois expérimentalement et théoriquement.Ensuite, nous nous intéressons à l’évaporation de gouttes au sein d’un séchoir par atomisation, dans le cas particulier de la microencapsulation de polyphénols. Cette situation appliquée repose sur l’évaporation de gouttes en suspension contenant deux composés non volatils dissous. Une étude expérimentale de microencapsulation de polyphénols par atomisation est réalisée à l’échelle du laboratoire. Nous observons l’influence des paramètres opératoires (proportion en masse du composé d’intérêt et de l’agent encapsulant dans la solution initiale, débit de liquide, vitesse et température du gaz de séchage) sur l’efficacité du procédé et sur les caractéristiques de la poudre obtenue (taille et morphologie des grains, contenu en eau, conservation de l’activité phénolique). Sur le plan théorique, l’évaporation de ces gouttes est caractérisée grâce à des modèles existants permettant de déterminer la température de la goutte et son temps d’évaporation, ainsi que la dynamique des composés non volatils au sein de la goutte au cours du temps. De plus, une méthode d’évaluation de l’efficacité de la chambre de séchage est proposée, qui repose sur la comparaison entre le temps d’évaporation des gouttes et leur temps de parcours au sein de la chambre. Ces différents modèles sont en accord avec les résultats expérimentaux obtenus et permettent notamment de donner des conseils pour la conception et le dimensionnement de séchoirs par atomisation. / Doctorat en Sciences de l'ingénieur et technologie / info:eu-repo/semantics/nonPublished

As frutas nativas brasileiras têm grande potencial agroindustrial e farmacêutico por serem excelentes fontes de compostos bioativos fenólicos. A cagaita (Eugenia dysenterica DC.), típica do Cerrado, se destaca por conter compostos fenólicos com alta capacidade de inibir as enzimas envolvidas no metabolismo de carboidratos in vitro. Esta ação também ocorre com o consumo de suco, que causa redução da hiperglicemia pós-prandial em seres humanos, tanto saudáveis como resistentes a insulina. No entanto, devido à alta perecibilidade dos frutos, o armazenamento, transporte e ampla distribuição destes torna-se fator limitante para sua incorporação na dieta. Dessa forma, a aplicação de processos que garantam a conservação dos componentes bioativos e facilitem o transporte e o armazenamento do material pode tornar o fruto mais atrativo para seu uso industrial. Este trabalho teve como objetivos a obtenção de extratos secos a partir do fruto da cagaita através da secagem por atomização, comparando a utilização de goma arábica e inulina como agentes carreadores e avaliando o efeito dos parâmetros de processo sobre as propriedades físico-químicas e estabilidade dos compostos fenólicos nos pós produzidos, assim como seu potencial funcional in vitro. A secagem do extrato bruto foi realizada variando-se a temperatura do ar de entrada (120, 140 e 160°C) e o tipo e a concentração dos carreadores (goma arábica - GA ou inulina - I; 10, 20 ou 30%). Os melhores rendimentos de processo foram os dos pós obtidos usando goma arábica como agente carregador (41% a 81%). Todas as amostras apresentaram menores valores de umidade e atividade de água quando comparadas com a amostra utilizada como controle (amostra liofilizada). O processo de atomização incrementou a solubilidade e reduziu a higroscopicidade das amostras. No caso das amostras obtidas usando GA como carreador, a temperatura de transição vítrea foi principalmente influenciada pela temperatura de ar de entrada. A concentração de carreador foi o parâmetro que mais influenciou a temperatura de transição vítrea das amostras com inulina. A morfologia das partículas foi influenciada por ambos os parâmetros, temperatura do ar de entrada e concentração do carreador. Os valores de retenção de fenólicos totais dos pós com goma arábica variaram de 88% a 99%, e para os pós obtidos com inulina, de 78% a 98%. A retenção das proantocianidinas variou de 72% a 98%, para os pós obtidos com goma arábica, e de 63% a 98%, para os pós obtidos com inulina. Altos coeficientes de correlação de Pearson, entre o conteúdo de fenólicos totais e a capacidade antioxidante das amostras contendo GA e inulina, foram obtidos de 0,90 a 0,99 (p < 0,05). A GA foi o material mais adequado para estabilizar os extratos e evitar a perda de compostos fenólicos ao longo do armazenamento. Comparadas com a acarbose (CI50 = 0,013 mg/mL), as amostras apresentaram uma baixa inibição da enzima &#945;-amilase, sendo que a concentração média inibitória (CI50) dos extratos secos variou entre 11 a 107 mg/mL e entre 10 a 100 mg/mL, para os pós obtidos usando GA e inulina, respetivamente. Todas as amostras mostraram inibição da enzima &#945;-glicosidase (6 mg/mL a 159 mg/mL e 13 mg/mL a 154 mg/mL, para os pós obtidos usando GA e inulina, respetivamente), mas a inibição observada foi média a baixa em comparação com a atividade do controle acarbose (CI50 = 0,11 mg/mL). No entanto, a atividade dos fenólicos dos pós comparada com a acarbose foi boa, 0,3 a 1,1 mg GAE/mL para &#945;-amilase e de 0,3 a 1,7 mg GAE/mL para &#945;-glicosidase. Os pós apresentaram atividade inibitória contra Staphylococcus aureus e Listerya monocytogenes, semelhante aos antibióticos utilizados como controle positivo. Os resultados obtidos sugerem que os produtos do processo de secagem por atomização utilizando goma arábica apresentam melhores propriedades físicas que os pós obtidos utilizando inulina, e que o uso de secagem por atomização para obtenção de extratos secos de cagaita pode se tornar uma alternativa viável para reduzir os custos de transporte e evitar a deterioração dos compostos bioativos que estão relacionados com algumas propriedades benéficas para a saúde. / Brazilian native fruits have great agroindustrial and pharmaceutical potential for being excellent sources of phenolic bioactive compounds. The cagaita (Eugenia dysenterica DC.), fruit typical of the Cerrado, stands out for containing phenolic compounds with high ability to inhibit enzymes involved in carbohydrate metabolism in vitro. This action also occurs with the consumption of juice, which causes reduction in postprandial hyperglycemia in humans, both healthy and insulin resistant. However, due to the high perishability of the fruit, storage, transportation and broad distribution becomes a limiting factor for its incorporation in the diet. Thus, the application of processes that ensure the preservation of bioactive components and facilitate the transport and storage of the material can make the fruit more attractive for industrial use. This study aimed to obtaining dry extracts from the cagaita fruit by spray drying, comparing the use of gum arabic and inulin as carrier agents and evaluating the effect of process parameters on the physicochemical properties and stability of phenolic compounds produced in the dry extracts, as well as their functional potential in vitro. The drying of the crude extract was performed by varying the inlet air temperature (120, 140 and 160 ° C) and the type and concentration of carrier agent (gum arabic - GA or inulin - I; 10, 20 or 30%) . The best process yields were obtained using gum Arabic as the carrier agent (41% to 81%). All samples showed lower humidity values and water activity when compared with the lyophilized sample used as control. The spray drying process increased solubility and reduced hygroscopicity of the samples. In the case of the samples obtained using GA as a carrier, the glass transition temperature was mainly influenced by the inlet air temperature. The carrier concentration was the parameter that most influenced the glass transition temperature of the samples dried with inulin. The morphology of the particles was influenced by both inlet air temperature and concentration of the carrier. The retention values of total phenolics in gum arabic powders ranged from 88 to 99%, and the powders obtained with inulin, from 78 to 98%. The retention of proanthocyanidins ranged from 72 to 98% for powders obtained with gum arabic, and 63 to 98% for powders obtained with inulin. High Pearson correlation coefficients between the total phenolic content and antioxidant capacity of the samples containing GA and inulin were obtained from 0.90 to 0.99 (p < 0.05). GA was the most suitable material to stabilize the extracts and prevent the loss of phenolic compounds during storage. Compared with acarbose (IC50 = 0.013 mg/mL), samples showed a low inhibition of &#945;-amylase, the half maximal inhibitory concentrations (IC50) of the dried extract ranged from 11 to 107 mg/mL and 10 to 100 mg/mL, for the powders obtained by using Ga and inulin, respectively. All samples showed inhibition of &#945;-glucosidase (6 mg/mL to 159 mg/mL and 13 mg/ml to 154 mg/mL, for the powders produced using inulin and GA, respectively), but the inhibition was lower compared to the activity of acarbose used as a control (IC50 = 0.11 mg/mL). However, the activity of the phenolic compounds of the powders, when compared with acarbose was good 0.3 to 1.1 mg GAE/mL for &#945;-amylase and 0.3 to 1.7 mg GAE/mL for &#945;-glucosidase. The powders showed inhibitory activity against Staphylococcus aureus and Listerya monocytogenes, similar to antibiotics used as positive control. The results suggest that the products of the spray drying process using gum arabic have better physical properties than powders produced using inulin, and the use of spray drying to obtain dry extracts of cagaita can become a viable alternative to reduce transportation costs and avoid the deterioration of bioactive compounds that are related to properties beneficial to health.

A busca de novas soluções em produtos e processos guia o desenvolvimento da indústria de alimentos. Os produtos desidratados apresentam vantagens em relação à facilidade de manuseio, custos de transporte e armazenagem e, em geral, maior durabilidade quando comparados às suas formas hidratadas. O iogurte é muito bem aceito pela sociedade brasileira e, se desidratado, pode vir a ser utilizado como ingrediente em diversos alimentos industrializados, propiciando o desenvolvimento de novos produtos. Este trabalho procura estabelecer condições ótimas para a secagem de iogurte natural por atomização (spray drying) em equipamento com disco rotativo em escala piloto. A caracterização do sistema e de suas respostas foi feita com auxílio de projetos fatoriais e metodologia de superfícies de respostas. Por meio de otimização multiobjetivo foi determinada a fronteira de Pareto de soluções ótimas para um modelo ponderado com funções compostas de atributos de qualidade do produto cor, temperatura do ar de saída da câmara de secagem, span e umidade; e de índices de desempenho do processo, com base na quantidade de material seco quantidade de produto obtido, material retido na parede da câmara de secagem e perdas no equipamento. Em virtude da natureza do iogurte houve depósito de produto na parede da câmara, o que motivou a pesquisa de aditivos para auxiliar a secagem. Foi estudada a influência de maltodextrinas DE 5, DE 10 e EMPCAP -, goma acácia, leite em pó desnatado e sílica pirogênica. A sílica pirogênica interferiu na formação de pontes sólidas entre as partículas, reduzindo a quantidade de material depositado na parede e aumentando a recuperação de produto. Para condições operacionais de velocidade de rotação do disco de 26000 rpm, temperatura do ar de entrada na câmara de secagem de 160 °C e depressão na câmara de secagem de 140 mmH2O, as condições ótimas de processo foram encontradas nas seguintes vazões de alimentação de iogurte natural batido adicionado de 3% (m/m) de sílica pirogênica: para o ótimo de qualidade produto, 10,3 kg/h; para o ótimo de desempenho de processo, 3,5 kg/h; e para a condição de pesos iguais entre produto e processo, 6,2 kg/h. A comparação dos resultados previstos pela otimização e os verificados experimentalmente mostrou que o modelo proposto foi adequado. / The search for new solutions in products and processes guide the development of the food industry. Dried products have advantages for ease of handling, transportation and storage costs, and in general, better durability when compared to their hydrated forms. Brazilian society accepts Yoghurt very well and, if dehydrated, could be used as an ingredient in many processed foods, promoting the development of new products. The aim of this paper is to establish optimal conditions for spray drying yoghurt on a wheel pilot plant atomizer. The system characteristics and outputs were established through factorial design and surface response methodology. Multi-objective optimization produced a Paretos optimal solution set for a weighted model built of composite functions. One composite function was consisted of product quality attributes: color, drying chamber output air temperature, span and moisture. The other composite function was made by drying process performance indexes: dry mass recovery, dry mass adhered on the wall and dry mass loss in equipment. The stickiness behavior of yoghurt led the search for drying aid additives. The influence of maltodextrins DE 5, DE 10 and EMCAP, acacia gum, skimmed milk powder and fumed silica was studied. Fumed silica has a negative influence in solid bridges formation between particles, thereby reducing the amount of dry mass adhered on the wall and increasing product recovery. For operating conditions of rotating speed of 26000 rpm, inlet air temperature of 160 °C and 140 mmH2O depression in drying chamber and a feed rate of beaten yoghurt with 3% (w/w) fumed silica concentration, optimal process conditions were found: product quality optimum, 10.3 kg/h; process performance optimum, 3.5 kg/h, and equal weights for product and process optimum, 6.2 kg/h. Experimental verification of the proposed optimization model suggests its adequacy.

O ovo é importante como alimento e como matéria-prima industrial. Tem alto conteúdo de colesterol, o qual, por sua vez, está sujeito à oxidação. Os óxidos formados interferem na morfologia e função da membrana celular, inibem a biossíntese do colesterol, e são aterogênicos, citotóxicos, mutagênicos e cancerígenos. O processamento e o armazenamento levam à oxidação do colesterol. O presente trabalho teve como objetivos (a) a caracterização e avaliação do ovo líquido integral (OLI) em relação ao armazenamento da matéria-prima; (b) a avaliação do ovo líquido integral (OLI), do ovo líquido integral pasteurizado (OLIP), do ovo integral em pó atomizado (OIPA) e do ovo integral em pó atomizado armazenado, em relação à estabilidade oxidativa do colesterol; e (c) a avaliação do efeito da adição de antioxidantes naturais durante o processamento do ovo integral em pó atomizado, em relação à estabilidade oxidativa do colesterol. O OLI produzido a partir de matéria-prima armazenada por 30 dias, a 4ºC, não foi afetado quanto à estabilidade oxidativa do colesterol. A pasteurização e a atomização, realizadas em condições de laboratório, não levaram ao comprometimento oxidativo do colesterol. Foi constatada oxidação significativa do colesterol no OIPA, obtido em condições de laboratório, quando armazenado por 90 dias, a 25ºC, na ausência de luz. Os antioxidantes naturais comerciais GUARDIAN® Rosemary Extract (RE) e o GUARDIANTM TOCO 70 (TOCO 70) não foram efetivos na prevenção da oxidação induzida do colesterol, quando adicionados (antes ou depois da atomização) ao ovo integral em pó atomizado obtido em condições de laboratório, armazenado por 21 dias, a 25°C, na ausência de luz. / Egg is important as food and as industrial raw material. It has high cholesterol, which, in turn, is subject to oxidation. Cholesterol oxides interfere with morphology and function of cell membrane, inhibit cholesterol biosynthesis, and are atherogenic, cytotoxic, mutagenic and carcinogenic. Processing and storage lead to oxidation of cholesterol. Present study aimed (a) characterization and evaluation of liquid whole egg (OLI) in relation to storage of raw material; (b) evaluation of liquid whole egg (OLI), liquid pasteurized whole egg (OLIP), atomized powder whole egg (OIPA) and stored atomized powder whole egg, relative to cholesterol oxidative stability, and (c) evaluation of natural antioxidants addition\'s effect in processing of atomized powder whole egg, relative to cholesterol oxidative stability. OLI produced from raw materials stored for 30 days, at 4°C, was not affected on cholesterol oxidative stability. Pasteurization and atomization, performed under laboratory conditions, did not lead to significant cholesterol oxidation. It has been found significant cholesterol oxidation in OIPA, obtained under laboratory conditions, when stored for 90 days at 25°C in absence of light. Commercial natural antioxidants GUARDIAN® Rosemary Extract (RE) and GUARDIANTMTOCO 70 (TOCO 70) were not effective in prevention of cholesterol induced oxidation when added (before or after spraying) to atomized powder whole egg obtained under laboratory conditions, stored for 21 days at 25°C in absence of light.

Submitted by FERNANDA DA SILVA VON PORSTER (fdsvporster@univates.br) on 2017-01-02T19:21:47Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) 2016CamilaEckert.pdf: 2023285 bytes, checksum: f674c42a62e9aa153c1c16a5e33f2284 (MD5) / Approved for entry into archive by Ana Paula Lisboa Monteiro (monteiro@univates.br) on 2017-01-06T17:28:58Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) 2016CamilaEckert.pdf: 2023285 bytes, checksum: f674c42a62e9aa153c1c16a5e33f2284 (MD5) / Made available in DSpace on 2017-01-06T17:28:58Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) 2016CamilaEckert.pdf: 2023285 bytes, checksum: f674c42a62e9aa153c1c16a5e33f2284 (MD5) Previous issue date: 2017-01 / Muitas bactérias láticas (BALs) são classificadas como probióticas, provendo benefícios à saúde quando ingeridas em quantidades adequadas. Para serem assim consideradas, um dos critérios é sua capacidade de tolerar as condições adversas do trato gastrintestinal humano (TGI). Entretanto, algumas cepas apresentam baixa resistência, sendo necessário proporcionar uma barreira física externa como proteção. Uma das metodologias mais propostas para esta finalidade é o encapsulamento. A técnica de spray drying é um dos métodos mais difundidos na indústria de alimentos para este fim. Ainda, outra forma da obtenção de encapsulados é a extrusão por tecnologia de vibração. Os subprodutos da indústria de laticínios são uma alternativa como material encapsulante na aplicação às condições adversas, em função de sua composição e por apresentarem potencial para a proteção dos microrganismos. Portanto, o objetivo deste trabalho foi selecionar BALs tolerantes às condições ambientais do TGI simulado, visando o encapsulamento por spray drying e extrusão por tecnologia de vibração, empregando soros lácteos como material de parede. Desta forma, bactérias láticas isoladas de leite e queijo, disponíveis comercialmente e de coleções de cultura foram estudadas quanto ao potencial de sobrevivência às condições ambientais do TGI simulado. Bactérias com elevados níveis de viabilidade (acima de 70% de sobrevivência) às condições ácidas (pHs 2,5 e 3) e de alta concentração de sais biliares (0,5% (m/v)) foram selecionadas para encapsulamento. No microencapsulamento por spray drying, soro de queijo, permeado de soro e retentado de soro foram utilizados como material encapsulante para proteção de L. plantarum ATCC 8014. Soro de queijo e retentado de soro provaram ser o melhores materiais encapsulantes para proteger L. plantarum da alta temperatura aplicada no processo. As microcápsulas foram armazenadas por 56 dias a 20 ºC, ou inseridas em leite por 42 dias a 4 ºC, mantendo, ainda assim, a alta viabilidade célular. Quando expostas ao TGI simulado, apresentaram viabilidade acima de 8 UFC.mL-1. As microcápsulas formadas não apresentaram diferenças na morfologia e diâmetro de 7,0 ± 1,0 μm. No encapsulamento por tecnologia de vibração, soro de queijo e permeado de soro foram combinados com alginato de sódio 1,5% e pectina 1,25%, para proteção de L. plantarum ATCC 8014 e isolados L. paracasei ML33 e L. pentosus ML82. Não houve perda na viabilidade dos microrganismos após este processo de encapsulamento. Quando armazenadas por 28 dias a 4 ºC, apresentaram uma redução de 1 ciclo log, indiferente do isolado ou material encapsulante aplicado. Ainda, forneceu proteção na exposição ao TGI simulado, sendo o soro de queijo com melhor efeito protetor. Os encapsulamentos por spray drying e extrusão por tecnologia de vibração, assim como os soros lácteos, materiais encapsulantes aplicados, mostraram-se eficientes na proteção de BALs. São uma alternativa de aplicação, visando o aumento da vida de prateleira e proteção às condições adversas, num produto de alto valor agregado. / Many lactic acid bacteria (BALs) are classified as probiotic, providing health benefits when ingested in suitable amounts. To be thus considered, one of the criteria is its ability to tolerate the adverse conditions of the human gastrointestinal tract (GIT). However, some strains have low resistance, being necessary to provide an external physical barrier as protection. One of the most proposed methodologies for this purpose is encapsulation. The spray drying technique is one of the most widespread methods in the food industry for this. Still another way of obtaining encapsulations is the extrusion by vibration technology. Dairy by-products are an alternative as encapsulating material in application to adverse conditions, depending on their composition and potential for protection of microorganisms. Therefore, the objective of this work was to select BALs tolerant to the environmental conditions of the simulated GIT, aiming the encapsulation by spray drying and extrusion by vibration technology, employing dairy whey as a wall material. In this way, commercially available milk and cheese isolated lactic bacteria and culture collections were studied for the potential survival of the simulated GIT environmental conditions. Bacteria with high levels of viability (above 70% survival) to acid conditions (pH 2.5 and 3) and high concentration of bile salts (0.5% (w / v)) were selected for encapsulation. In microencapsulation by spray drying, whey, permeate and retentate were used as encapsulating material to protect L. plantarum ATCC 8014. Whey and retentate proved to be the best encapsulating materials to protect L. plantarum from high temperature applied in the process. The microcapsules were stored for 56 days at 20 ° C, or inserted into milk for 42 days at 4 ° C, while still maintaining high cell viability. When exposed to the simulated GIT, they presented viability above 8 CFU.g-1. The microcapsules formed did not present differences in morphology and diameter of 7.0 ± 1.0 μm. In the encapsulation by vibration technology, whey and permeate were combined with 1.5% sodium alginate and 1.25% pectin, to protect L. plantarum ATCC 8014, L. paracasei ML33 and L. pentosus ML82 isolates. There was no loss in viability of the microorganisms after this encapsulation process. When stored for 28 days at 4 ° C, they showed a reduction of 1 log cycle, regardless of the isolate or encapsulating material applied. Furthermore, it provided protection in the simulated GIT exposure, being whey with the best protective effect. Spray drying and extrusion by vibration technology, as well as dairy whey, applied encapsulating materials, have proved to be effective in protecting BALs. They are an alternative of application, aiming the increase of the shelf life and protection to the adverse conditions, in a product of high added value.

Submitted by FERNANDA DA SILVA VON PORSTER (fdsvporster@univates.br) on 2017-06-29T18:22:03Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) 2016CamilaEckert.pdf: 2023285 bytes, checksum: f674c42a62e9aa153c1c16a5e33f2284 (MD5) / Approved for entry into archive by Ana Paula Lisboa Monteiro (monteiro@univates.br) on 2017-07-04T20:19:13Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) 2016CamilaEckert.pdf: 2023285 bytes, checksum: f674c42a62e9aa153c1c16a5e33f2284 (MD5) / Made available in DSpace on 2017-07-04T20:19:13Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) 2016CamilaEckert.pdf: 2023285 bytes, checksum: f674c42a62e9aa153c1c16a5e33f2284 (MD5) Previous issue date: 2017-06 / CAPES / Muitas bactérias láticas (BALs) são classificadas como probióticas, provendo benefícios à saúde quando ingeridas em quantidades adequadas. Para serem assim consideradas, um dos critérios é sua capacidade de tolerar as condições adversas do trato gastrintestinal humano (TGI). Entretanto, algumas cepas apresentam baixa resistência, sendo necessário proporcionar uma barreira física externa como proteção. Uma das metodologias mais propostas para esta finalidade é o encapsulamento. A técnica de spray drying é um dos métodos mais difundidos na indústria de alimentos para este fim. Ainda, outra forma da obtenção de encapsulados é a extrusão por tecnologia de vibração. Os subprodutos da indústria de laticínios são uma alternativa como material encapsulante na aplicação às condições adversas, em função de sua composição e por apresentarem potencial para a proteção dos microrganismos. Portanto, o objetivo deste trabalho foi selecionar BALs tolerantes às condições ambientais do TGI simulado, visando o encapsulamento por spray drying e extrusão por tecnologia de vibração, empregando soros lácteos como material de parede. Desta forma, bactérias láticas isoladas de leite e queijo, disponíveis comercialmente e de coleções de cultura foram estudadas quanto ao potencial de sobrevivência às condições ambientais do TGI simulado. Bactérias com elevados níveis de viabilidade (acima de 70% de sobrevivência) às condições ácidas (pHs 2,5 e 3) e de alta concentração de sais biliares (0,5% (m/v)) foram selecionadas para encapsulamento. No microencapsulamento por spray drying, soro de queijo, permeado de soro e retentado de soro foram utilizados como material encapsulante para proteção de L. plantarum ATCC 8014. Soro de queijo e retentado de soro provaram ser o melhores materiais encapsulantes para proteger L. plantarum da alta temperatura aplicada no processo. As microcápsulas foram armazenadas por 56 dias a 20 ºC, ou inseridas em leite por 42 dias a 4 ºC, mantendo, ainda assim, a alta viabilidade célular. Quando expostas ao TGI simulado, apresentaram viabilidade acima de 8 UFC.mL-1. As microcápsulas formadas não apresentaram diferenças na morfologia e diâmetro de 7,0 ± 1,0 μm. No encapsulamento por tecnologia de vibração, soro de queijo e permeado de soro foram combinados com alginato de sódio 1,5% e pectina 1,25%, para proteção de L. plantarum ATCC 8014 e isolados L. paracasei ML33 e L. pentosus ML82. Não houve perda na viabilidade dos microrganismos após este processo de encapsulamento. Quando armazenadas por 28 dias a 4 ºC, apresentaram uma redução de 1 ciclo log, indiferente do isolado ou material encapsulante aplicado. Ainda, forneceu proteção na exposição ao TGI simulado, sendo o soro de queijo com melhor efeito protetor. Os encapsulamentos por spray drying e extrusão por tecnologia de vibração, assim como os soros lácteos, materiais encapsulantes aplicados, mostraram-se eficientes na proteção de BALs. São uma alternativa de aplicação, visando o aumento da vida de prateleira e proteção às condições adversas, num produto de alto valor agregado. / Many lactic acid bacteria (BALs) are classified as probiotic, providing health benefits when ingested in suitable amounts. To be thus considered, one of the criteria is its ability to tolerate the adverse conditions of the human gastrointestinal tract (GIT). However, some strains have low resistance, being necessary to provide an external physical barrier as protection. One of the most proposed methodologies for this purpose is encapsulation. The spray drying technique is one of the most widespread methods in the food industry for this. Still another way of obtaining encapsulations is the extrusion by vibration technology. Dairy by-products are an alternative as encapsulating material in application to adverse conditions, depending on their composition and potential for protection of microorganisms. Therefore, the objective of this work was to select BALs tolerant to the environmental conditions of the simulated GIT, aiming the encapsulation by spray drying and extrusion by vibration technology, employing dairy whey as a wall material. In this way, commercially available milk and cheese isolated lactic bacteria and culture collections were studied for the potential survival of the simulated GIT environmental conditions. Bacteria with high levels of viability (above 70% survival) to acid conditions (pH 2.5 and 3) and high concentration of bile salts (0.5% (w / v)) were selected for encapsulation. In microencapsulation by spray drying, whey, permeate and retentate were used as encapsulating material to protect L. plantarum ATCC 8014. Whey and retentate proved to be the best encapsulating materials to protect L. plantarum from high temperature applied in the process. The microcapsules were stored for 56 days at 20 ° C, or inserted into milk for 42 days at 4 ° C, while still maintaining high cell viability. When exposed to the simulated GIT, they presented viability above 8 CFU.g-1. The microcapsules formed did not present differences in morphology and diameter of 7.0 ± 1.0 μm. In the encapsulation by vibration technology, whey and permeate were combined with 1.5% sodium alginate and 1.25% pectin, to protect L. plantarum ATCC 8014, L. paracasei ML33 and L. pentosus ML82 isolates. There was no loss in viability of the microorganisms after this encapsulation process. When stored for 28 days at 4 ° C, they showed a reduction of 1 log cycle, regardless of the isolate or encapsulating material applied. Furthermore, it provided protection in the simulated GIT exposure, being whey with the best protective effect. Spray drying and extrusion by vibration technology, as well as dairy whey, applied encapsulating materials, have proved to be effective in protecting BALs. They are an alternative of application, aiming the increase of the shelf life and protection to the adverse conditions, in a product of high added value.