Les liposomes sont des nanovecteurs polyvalents et prometteurs quant à leur utilisation dans plusieurs domaines. Il y a une décennie, un nouveau type de liposome constitué d’amphiphiles monoalkylés et de stérols est né fortuitement dans notre groupe. Ils sont nommés Stérosomes puisqu’ils contiennent une grande proportion de stérols, entre 50 et 70 mol %. Les objectifs de cette thèse sont de développer de nouvelles formulations de Stérosomes ayant des caractéristiques spécifiques et d’acquérir une compréhension plus profonde des règles physicochimiques qui dictent leur comportement de phase. Nous avons spécifiquement examiné le rôle de motifs moléculaires des stérols, de la charge interfaciale et de la capacité à former des liaisons H dans les interactions intermoléculaires menant à l’autoassemblage. Le comportement de phase a été caractérisé par calorimétrie différentielle à balayage (DSC), par spectroscopie infrarouge (IR) et par spectroscopie de résonance magnétique nucléaire du deutérium (²H NMR). Premièrement, nous avons établi certaines corrélations entre la structure des stérols, leur tendance à former des bicouches fluides en présence d'amphiphile monoalkylé et la perméabilité des grandes vésicules unilamellaires (LUV) formées. La nature des stérols module les propriétés de mélange avec de l’acide palmitique (PA). Les stérols portant une chaîne volumineuse en position C17 sont moins aptes à induire des bicouches fluides que ceux qui ont une chaîne plus simple, comme celle du cholestérol. Un grand ordre de la chaîne alkyle de PA est un effet commun à tous les stérols investigués. Il a été démontré que la perméabilité des LUV peut être contrôlée en utilisant des stérols différents. Cependant, ces stérols n’ont aucun impact significatif sur la sensibilité des Stérosomes au pH. Afin de créer des liposomes qui sont sensibles au pH et qui ont une charge positive à la surface, des Stérosomes composés de stéarylamine et de cholestérol (Chol) ont été conçus et caractérisés. Il a été conclu que l’état de protonation de l’amine, dans ce travail, ou du groupe carboxylique, dans un travail précédent, confère une sensibilité au pH et détermine la charge à la surface du liposome. Les premiers Stérosomes complètement neutres ont été fabriqués en utilisant un réseau de fortes liaisons H intermoléculaires. Le groupe sulfoxyde est capable de former de fortes liaisons H avec le cholestérol et les molécules d’eau. Une bicouche fluide métastable a été obtenue, à la température de la pièce, à partir d'un mélange équimolaire d’octadécyl méthyl sulfoxyde (OMSO) et de Chol. Ce comportement distinct a permis d’extruder le mélange pour former des LUV à la température de la pièce. Après 30 h, le temps de vie de la phase métastable, des Stérosomes stables et imperméables existaient toujours sous une forme solide. Un diagramme de température-composition a été proposé afin de résumer le comportement de phase des mélanges d’OMSO/Chol. Finalement, nous avons élaboré des Stérosomes furtifs en incorporant du polyéthylène glycol (PEG) avec une ancre de cholestérol (PEG-Chol) à l’interface de Stérosomes de PA/Chol. Jusqu’à 20 mol % de PEG-Chol peut être introduit sans perturber la structure de la bicouche. La présence du PEG-Chol n’a aucun impact significatif sur la perméabilité de la LUV. L'encapsulation active de la doxorubicine, un médicament contre le cancer, a été réalisée malgré la faible perméabilité de ces LUV et la présence du PEG à l’interface. L’inclusion de PEG a modifié considérablement les propriétés de l’interface et a diminué la libération induite par la variation de pH observée avec des LUV nues de PA/Chol. Cette formulation inédite est potentiellement utile pour l’administration intraveineuse de médicaments. / Liposomes are promising and versatile nanocarriers suitable for potential applications in many fields. A decade ago, a new type of liposomes formed from monoalkylated amphiphiles and sterols was born somehow fortuitously in our group. They are referred to as Sterosomes, because they contain a large proportion of sterols, between 50 and 70 mol %. The objectives of the present thesis are to develop novel Sterosome formulations with specific features, and to gain a deeper understanding of the physicochemical rules that dictate their phase behavior. We have specifically examined the role of the molecular features of sterols, of the interfacial charges and of the H-bond capacity in the intermolecular interactions leading to the self-assembly. The phase behavior was characterized by differential scanning calorimetry (DSC), infrared spectroscopy (IR), and nuclear magnetic resonance spectroscopy of deuterium (2H NMR). First, we have established some correlations between the structure of the sterols, the propensity to form fluid bilayers, and the permeability of the resulting large unilamellar vesicles (LUVs). The nature of the sterol modulates the properties of the mixture with palmitic acid (PA). Sterols bearing a bulky tail chain at C17 are less capable to induce fluid bilayers than those with a non-bulky tail chain, like that of cholesterol. A large ordering of the alkyl chain of PA is an effect exhibited by all of the investigated sterols. It is shown that the permeability of the LUVs can be controlled using different sterols. However, these sterols have no significant impact on the pH-sensitivity of Sterosomes. In order to create liposomes that are pH-sensitive and that have a positive surface charge, Sterosomes composed of stearylamine and cholesterol (Chol) were designed and characterized. It is concluded that the protonation/deprotonation state of the amine (in this work) and carboxylic acid (in previous work) groups confers the pH-sensitivity and determines the surface charge of the liposomes. The first completely neutral Sterosomes were crafted based on the creation of strong intermolecular hydrogen bond networks. The sulfoxide group was capable of forming strong hydrogen bonds with cholesterol and water molecules. In an equimolar octadecyl methyl sulfoxide (OMSO)/Chol mixture, a metastable fluid bilayer was obtained at room temperature. This distinct phase behavior allowed extruding the mixtures to form LUVs at room temperature. After 30 h, the life-time of the metastable phase, stable and impermeable Sterosomes still existed in the solid form. A temperature–composition diagram was proposed to summarize the phase behavior of OMSO/Chol mixtures. Finally, a further step was made to prepare “stealth” Sterosomes by incorporating polyethylene glycol (PEG) with a cholesterol anchor (PEG-Chol) at the interface of PA/Chol Sterosomes. Up to 20 mol % PEG-Chol can be introduced without disturbing the bilayer structure. The presence of PEG-Chol had no significant impact on the permeability of the resulting LUVs. Active-loading of an anti-cancer drug, doxorubicin, can be achieved despite the low permeability of these LUVs and the presence of the PEG at the interface. The inclusion of PEG modified considerably the interface properties and decreased significantly the pH-triggered release observed with naked PA/Chol LUVs. This novel formulation is potentially useful for the application of intravenous administration in the drug delivery field.

Phytosterols have been shown to be effective in reducing serum cholesterol levels in numerous human clinical studies and regular consumption is recommended as part of therapeutic lifestyle changes aimed at reducing low density lipoprotein (LDL-C) in the treatment of hyperlipidaemia, a risk factor for cardiovascular disease. Fat based spreads have been shown to be a very successful vehicle for delivery of plant sterols, readily accepted by consumers and efficacious in reducing cholesterol levels. Alfa One™ Rice Bran Oil (RBO) spread is a new product entering into the market place. It is derived from rice bran oil and contains high levels of unsaponifiable material rich in phytosterols, triterpene alcohols, ferulic acid esters ([gamma]-oryzanol) and vitamin E isomers. As such it may have the potential to lower serum cholesterol levels when consumed on a daily basis. In order to establish the effectiveness of Alfa One™ Rice Bran Oil (RBO) spread compared with Flora pro-activ® margarine, a well established brand of plant sterol margarine already proven to lower cholesterol, a randomised double blind cross-over human clinical trial over 12 weeks was conducted. The study was divided into two treatment arms. The first arm of the study was to determine whether Alfa One™ RBO spread (containing 1.5% plant sterols) could lower total and LDL cholesterol levels to a greater extent than standard Flora margarine (containing no plant sterols) or Flora Pro-activ® margarine (containing 8% plant sterols). The second study arm tested the proposition that daily consumption of Alfa One™ Rice Bran Oil (RBO) spread in conjunction with rice bran oil (containing 0.5% plant sterols) would lower total and LDL cholesterol to a greater extent than Alfa One™ RBO spread in isolation and more than Flora margarine in conjunction with sunflower oil. Eighty mildly hypercholesterolaemic individuals (total cholesterol [greater than or equal to] 5 mmol/L and [less than or equal to] 7.5 mmol/L) were recruited and randomised into two groups of forty. Participants were asked to continue with their normal dietary pattern but to replace any margarine/butter/fat consumption with the trial products. One group of 40 were then assigned to the first treatment arm of the study (margarine-only group) and were randomised to consume 20 g (4 teaspoons) Alfa One™ RBO spread daily for 4 weeks, or 20 g Flora margarine daily for 4 weeks, or 20 Flora pro-activ® daily for 4 weeks. Phytosterol levels delivered in these amounts were: RBO margarine: 118mg phytosterol and 14 mg [gamma]-oryzanol; Flora proactiv® 1600 mg phytosterol; Flora margarine 0mg phytosterol. The second group of 40 were allocated to the second arm of the trial (margarine and oil group) and consumed 20 g Alfa One™ RBO spread and 30 ml rice bran oil (RBO) daily for 4 weeks, or 20 g Flora margarine and 30 ml sunflower oil daily for 4 weeks, or 20 g Alfa One™ RBO spread daily for 4 weeks, changing treatment at the end of each 4-week period. Phytosterol amounts delivered in these amounts were: RBO margarine: 118 mg phytosterol and 14 mg [gamma] oryzanol; RBO 222mg mg phytosterol, 150 mg [gamma] oryzanol. Each participant consumed all three treatments in a random order over a 12 week period. At baseline and following each 4 week intervention period, measurements were made of weight and blood pressure. Venous blood samples were collected for analysis of total cholesterol, low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), total cholesterol: HDL-C, triglycerides and plasma phytosterols. Three-day diet records from each individual were also collected for analysis of normal dietary intake. Results showed that compared to a standard Flora margarine, Alfa One™ RBO spread significantly reduced total cholesterol by 2.2% (P=0.045), total cholesterol:HDL by 4.1% (P=0.005) and LDL-C by 3.5% (P=0.016), but was not as effective overall as Flora Pro-activ® which reduced total cholesterol by 4.4% (P=0.001), total cholesterol:HDL by 3.4% (P=0.014) and LDL-C by 5.6% (P=0.001). Consumption of Flora margarine alone produced no significant decrease from baseline figures in any of the cholesterol parameters measured. Surprisingly, in group two, the addition of rice bran oil to the Alfa One™ RBO spread produced no differences in cholesterol levels. The reason for this unexpected result is being explored further. These results confirm that Alfa One™ RBO spread is effective in lowering serum cholesterol levels when consumed as part of a normal diet. Studies have shown that a 1% reduction in LDL-C can equate to a 2% decrease in coronary heart disease (CHD) risk thus suggesting that the 3.5% reduction demonstrated by Alfa One™ RBO spread in this study could be effective in reducing CHD risk as much as 6% in a mildly hypercholesterolaemic population.

Phytosterols have been shown to be effective in reducing serum cholesterol levels in numerous human clinical studies and regular consumption is recommended as part of therapeutic lifestyle changes aimed at reducing low density lipoprotein (LDL-C) in the treatment of hyperlipidaemia, a risk factor for cardiovascular disease. Fat based spreads have been shown to be a very successful vehicle for delivery of plant sterols, readily accepted by consumers and efficacious in reducing cholesterol levels. Alfa One™ Rice Bran Oil (RBO) spread is a new product entering into the market place. It is derived from rice bran oil and contains high levels of unsaponifiable material rich in phytosterols, triterpene alcohols, ferulic acid esters ([gamma]-oryzanol) and vitamin E isomers. As such it may have the potential to lower serum cholesterol levels when consumed on a daily basis. In order to establish the effectiveness of Alfa One™ Rice Bran Oil (RBO) spread compared with Flora pro-activ® margarine, a well established brand of plant sterol margarine already proven to lower cholesterol, a randomised double blind cross-over human clinical trial over 12 weeks was conducted. The study was divided into two treatment arms. The first arm of the study was to determine whether Alfa One™ RBO spread (containing 1.5% plant sterols) could lower total and LDL cholesterol levels to a greater extent than standard Flora margarine (containing no plant sterols) or Flora Pro-activ® margarine (containing 8% plant sterols). The second study arm tested the proposition that daily consumption of Alfa One™ Rice Bran Oil (RBO) spread in conjunction with rice bran oil (containing 0.5% plant sterols) would lower total and LDL cholesterol to a greater extent than Alfa One™ RBO spread in isolation and more than Flora margarine in conjunction with sunflower oil. Eighty mildly hypercholesterolaemic individuals (total cholesterol [greater than or equal to] 5 mmol/L and [less than or equal to] 7.5 mmol/L) were recruited and randomised into two groups of forty. Participants were asked to continue with their normal dietary pattern but to replace any margarine/butter/fat consumption with the trial products. One group of 40 were then assigned to the first treatment arm of the study (margarine-only group) and were randomised to consume 20 g (4 teaspoons) Alfa One™ RBO spread daily for 4 weeks, or 20 g Flora margarine daily for 4 weeks, or 20 Flora pro-activ® daily for 4 weeks. Phytosterol levels delivered in these amounts were: RBO margarine: 118mg phytosterol and 14 mg [gamma]-oryzanol; Flora proactiv® 1600 mg phytosterol; Flora margarine 0mg phytosterol. The second group of 40 were allocated to the second arm of the trial (margarine and oil group) and consumed 20 g Alfa One™ RBO spread and 30 ml rice bran oil (RBO) daily for 4 weeks, or 20 g Flora margarine and 30 ml sunflower oil daily for 4 weeks, or 20 g Alfa One™ RBO spread daily for 4 weeks, changing treatment at the end of each 4-week period. Phytosterol amounts delivered in these amounts were: RBO margarine: 118 mg phytosterol and 14 mg [gamma] oryzanol; RBO 222mg mg phytosterol, 150 mg [gamma] oryzanol. Each participant consumed all three treatments in a random order over a 12 week period. At baseline and following each 4 week intervention period, measurements were made of weight and blood pressure. Venous blood samples were collected for analysis of total cholesterol, low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), total cholesterol: HDL-C, triglycerides and plasma phytosterols. Three-day diet records from each individual were also collected for analysis of normal dietary intake. Results showed that compared to a standard Flora margarine, Alfa One™ RBO spread significantly reduced total cholesterol by 2.2% (P=0.045), total cholesterol:HDL by 4.1% (P=0.005) and LDL-C by 3.5% (P=0.016), but was not as effective overall as Flora Pro-activ® which reduced total cholesterol by 4.4% (P=0.001), total cholesterol:HDL by 3.4% (P=0.014) and LDL-C by 5.6% (P=0.001). Consumption of Flora margarine alone produced no significant decrease from baseline figures in any of the cholesterol parameters measured. Surprisingly, in group two, the addition of rice bran oil to the Alfa One™ RBO spread produced no differences in cholesterol levels. The reason for this unexpected result is being explored further. These results confirm that Alfa One™ RBO spread is effective in lowering serum cholesterol levels when consumed as part of a normal diet. Studies have shown that a 1% reduction in LDL-C can equate to a 2% decrease in coronary heart disease (CHD) risk thus suggesting that the 3.5% reduction demonstrated by Alfa One™ RBO spread in this study could be effective in reducing CHD risk as much as 6% in a mildly hypercholesterolaemic population.

Phytosterols have been shown to be effective in reducing serum cholesterol levels in numerous human clinical studies and regular consumption is recommended as part of therapeutic lifestyle changes aimed at reducing low density lipoprotein (LDL-C) in the treatment of hyperlipidaemia, a risk factor for cardiovascular disease. Fat based spreads have been shown to be a very successful vehicle for delivery of plant sterols, readily accepted by consumers and efficacious in reducing cholesterol levels. Alfa One™ Rice Bran Oil (RBO) spread is a new product entering into the market place. It is derived from rice bran oil and contains high levels of unsaponifiable material rich in phytosterols, triterpene alcohols, ferulic acid esters ([gamma]-oryzanol) and vitamin E isomers. As such it may have the potential to lower serum cholesterol levels when consumed on a daily basis. In order to establish the effectiveness of Alfa One™ Rice Bran Oil (RBO) spread compared with Flora pro-activ® margarine, a well established brand of plant sterol margarine already proven to lower cholesterol, a randomised double blind cross-over human clinical trial over 12 weeks was conducted. The study was divided into two treatment arms. The first arm of the study was to determine whether Alfa One™ RBO spread (containing 1.5% plant sterols) could lower total and LDL cholesterol levels to a greater extent than standard Flora margarine (containing no plant sterols) or Flora Pro-activ® margarine (containing 8% plant sterols). The second study arm tested the proposition that daily consumption of Alfa One™ Rice Bran Oil (RBO) spread in conjunction with rice bran oil (containing 0.5% plant sterols) would lower total and LDL cholesterol to a greater extent than Alfa One™ RBO spread in isolation and more than Flora margarine in conjunction with sunflower oil. Eighty mildly hypercholesterolaemic individuals (total cholesterol [greater than or equal to] 5 mmol/L and [less than or equal to] 7.5 mmol/L) were recruited and randomised into two groups of forty. Participants were asked to continue with their normal dietary pattern but to replace any margarine/butter/fat consumption with the trial products. One group of 40 were then assigned to the first treatment arm of the study (margarine-only group) and were randomised to consume 20 g (4 teaspoons) Alfa One™ RBO spread daily for 4 weeks, or 20 g Flora margarine daily for 4 weeks, or 20 Flora pro-activ® daily for 4 weeks. Phytosterol levels delivered in these amounts were: RBO margarine: 118mg phytosterol and 14 mg [gamma]-oryzanol; Flora proactiv® 1600 mg phytosterol; Flora margarine 0mg phytosterol. The second group of 40 were allocated to the second arm of the trial (margarine and oil group) and consumed 20 g Alfa One™ RBO spread and 30 ml rice bran oil (RBO) daily for 4 weeks, or 20 g Flora margarine and 30 ml sunflower oil daily for 4 weeks, or 20 g Alfa One™ RBO spread daily for 4 weeks, changing treatment at the end of each 4-week period. Phytosterol amounts delivered in these amounts were: RBO margarine: 118 mg phytosterol and 14 mg [gamma] oryzanol; RBO 222mg mg phytosterol, 150 mg [gamma] oryzanol. Each participant consumed all three treatments in a random order over a 12 week period. At baseline and following each 4 week intervention period, measurements were made of weight and blood pressure. Venous blood samples were collected for analysis of total cholesterol, low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), total cholesterol: HDL-C, triglycerides and plasma phytosterols. Three-day diet records from each individual were also collected for analysis of normal dietary intake. Results showed that compared to a standard Flora margarine, Alfa One™ RBO spread significantly reduced total cholesterol by 2.2% (P=0.045), total cholesterol:HDL by 4.1% (P=0.005) and LDL-C by 3.5% (P=0.016), but was not as effective overall as Flora Pro-activ® which reduced total cholesterol by 4.4% (P=0.001), total cholesterol:HDL by 3.4% (P=0.014) and LDL-C by 5.6% (P=0.001). Consumption of Flora margarine alone produced no significant decrease from baseline figures in any of the cholesterol parameters measured. Surprisingly, in group two, the addition of rice bran oil to the Alfa One™ RBO spread produced no differences in cholesterol levels. The reason for this unexpected result is being explored further. These results confirm that Alfa One™ RBO spread is effective in lowering serum cholesterol levels when consumed as part of a normal diet. Studies have shown that a 1% reduction in LDL-C can equate to a 2% decrease in coronary heart disease (CHD) risk thus suggesting that the 3.5% reduction demonstrated by Alfa One™ RBO spread in this study could be effective in reducing CHD risk as much as 6% in a mildly hypercholesterolaemic population.

Phytosterols have been shown to be effective in reducing serum cholesterol levels in numerous human clinical studies and regular consumption is recommended as part of therapeutic lifestyle changes aimed at reducing low density lipoprotein (LDL-C) in the treatment of hyperlipidaemia, a risk factor for cardiovascular disease. Fat based spreads have been shown to be a very successful vehicle for delivery of plant sterols, readily accepted by consumers and efficacious in reducing cholesterol levels. Alfa One™ Rice Bran Oil (RBO) spread is a new product entering into the market place. It is derived from rice bran oil and contains high levels of unsaponifiable material rich in phytosterols, triterpene alcohols, ferulic acid esters ([gamma]-oryzanol) and vitamin E isomers. As such it may have the potential to lower serum cholesterol levels when consumed on a daily basis. In order to establish the effectiveness of Alfa One™ Rice Bran Oil (RBO) spread compared with Flora pro-activ® margarine, a well established brand of plant sterol margarine already proven to lower cholesterol, a randomised double blind cross-over human clinical trial over 12 weeks was conducted. The study was divided into two treatment arms. The first arm of the study was to determine whether Alfa One™ RBO spread (containing 1.5% plant sterols) could lower total and LDL cholesterol levels to a greater extent than standard Flora margarine (containing no plant sterols) or Flora Pro-activ® margarine (containing 8% plant sterols). The second study arm tested the proposition that daily consumption of Alfa One™ Rice Bran Oil (RBO) spread in conjunction with rice bran oil (containing 0.5% plant sterols) would lower total and LDL cholesterol to a greater extent than Alfa One™ RBO spread in isolation and more than Flora margarine in conjunction with sunflower oil. Eighty mildly hypercholesterolaemic individuals (total cholesterol [greater than or equal to] 5 mmol/L and [less than or equal to] 7.5 mmol/L) were recruited and randomised into two groups of forty. Participants were asked to continue with their normal dietary pattern but to replace any margarine/butter/fat consumption with the trial products. One group of 40 were then assigned to the first treatment arm of the study (margarine-only group) and were randomised to consume 20 g (4 teaspoons) Alfa One™ RBO spread daily for 4 weeks, or 20 g Flora margarine daily for 4 weeks, or 20 Flora pro-activ® daily for 4 weeks. Phytosterol levels delivered in these amounts were: RBO margarine: 118mg phytosterol and 14 mg [gamma]-oryzanol; Flora proactiv® 1600 mg phytosterol; Flora margarine 0mg phytosterol. The second group of 40 were allocated to the second arm of the trial (margarine and oil group) and consumed 20 g Alfa One™ RBO spread and 30 ml rice bran oil (RBO) daily for 4 weeks, or 20 g Flora margarine and 30 ml sunflower oil daily for 4 weeks, or 20 g Alfa One™ RBO spread daily for 4 weeks, changing treatment at the end of each 4-week period. Phytosterol amounts delivered in these amounts were: RBO margarine: 118 mg phytosterol and 14 mg [gamma] oryzanol; RBO 222mg mg phytosterol, 150 mg [gamma] oryzanol. Each participant consumed all three treatments in a random order over a 12 week period. At baseline and following each 4 week intervention period, measurements were made of weight and blood pressure. Venous blood samples were collected for analysis of total cholesterol, low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), total cholesterol: HDL-C, triglycerides and plasma phytosterols. Three-day diet records from each individual were also collected for analysis of normal dietary intake. Results showed that compared to a standard Flora margarine, Alfa One™ RBO spread significantly reduced total cholesterol by 2.2% (P=0.045), total cholesterol:HDL by 4.1% (P=0.005) and LDL-C by 3.5% (P=0.016), but was not as effective overall as Flora Pro-activ® which reduced total cholesterol by 4.4% (P=0.001), total cholesterol:HDL by 3.4% (P=0.014) and LDL-C by 5.6% (P=0.001). Consumption of Flora margarine alone produced no significant decrease from baseline figures in any of the cholesterol parameters measured. Surprisingly, in group two, the addition of rice bran oil to the Alfa One™ RBO spread produced no differences in cholesterol levels. The reason for this unexpected result is being explored further. These results confirm that Alfa One™ RBO spread is effective in lowering serum cholesterol levels when consumed as part of a normal diet. Studies have shown that a 1% reduction in LDL-C can equate to a 2% decrease in coronary heart disease (CHD) risk thus suggesting that the 3.5% reduction demonstrated by Alfa One™ RBO spread in this study could be effective in reducing CHD risk as much as 6% in a mildly hypercholesterolaemic population.

Submitted by Biblioteca de Pós-Graduação em Geoquímica BGQ (bgq@ndc.uff.br) on 2017-05-11T17:24:08Z No. of bitstreams: 1 Dissertação Completa.pdf: 4492376 bytes, checksum: 676d473676c9c53901a55c556909987f (MD5) / Made available in DSpace on 2017-05-11T17:24:08Z (GMT). No. of bitstreams: 1 Dissertação Completa.pdf: 4492376 bytes, checksum: 676d473676c9c53901a55c556909987f (MD5) / Conselho Nacional de Desenvolvimento Científico e Tecnológico / Universidade Federal Fluminense. Instituto de Química. Programa de Pós-Graduação em Geoquímica, Niterói, RJ / Alterações na cobertura original do solo, devido o desenvolvimento de atividades antrópicas, e o lançamento de esgoto doméstico diretamente em rios, são fatores que causam a redução na qualidade de água e sedimentos em ecossis temas aquáticos. A bacia do rio Jacuecanga (Angra dos Reis, R.J) vem sofrendo com crescentes alterações no uso do solo, principalmente, pelo crescimento populacional e o desenvolvimento de diferentes atividades industriais , o que pode acarretar a degradaçã o dos seus recursos hídricos . Esse estudo propõe a caracterização da matéria orgânica presente no canal principal d esse sistema fluvial , e de seus principais afluentes e ainda na sua região estuarina. Foram avaliados parâmetros físico - químicos da água e marcadores orgânicos moleculares, elementares e isotópicos nos sedimentos com o objetivo de compreender a influência das modificações da cobertura primária do solo sobre o aporte de material orgânico terrígeno para esses sistemas aquáticos. Foram realizada s cinco campanhas para a coleta de águas em 16 estações nos rios Jacuecanga, Caputera e Vermelho, contemplando as diferentes situações climatológicas do ano e um evento de chuva. Coletas dos sedimentos superficiais foram realizadas em 14 estações no rio e 15 na enseada. A estação (RJ2) localizada a jusante da Vila Caputera apresentou tendências a eutrofização devido ao desmatamento, exp osição do solo e ocupação humana desordenada caracterizada pelo maior aporte de carbono, nitrogênio, MPS e DBO . N a zona est uarina, o adensamento urbano da Vila de Jacuecanga foi relacionado aos altos percentuais de carbono e nitrogênio, principalmente durante eventos de chuva. As águas pluviais que drenam o pátio da Transpetro e são lançadas no rio Jacuecanga no final do trech o de água doce apresentou características alternadas entre as campanhas, com períodos de alta condutividade, e o período s com elevada concentração de MPS, sem , no entanto , interferir de forma relevante na qualidade das águas do rio Jacuecanga. Da mesma for ma a entrada lateral de águas oriundas da lagoa de estabilização e as águas do rio Vermelho com baixa concentração de oxigênio e elevado teor de material orgânico não demonstr aram alterações significativas n o rio Jacuecanga. N os sedimentos do rio Jacuecang a , o predomínio da fração arenosa seguida do cascalho demonstra a intensa hidrodinâmica e ausência de depósitos orgânicos ao longo da bacia. Na porção estuarina foi registrado elevação nos teores de matéria orgânica com a identificação de fontes de restos vegetais e de coprostanol a partir do maior adensamento urbano que ocorre em sua margem direita. Os sedimentos da enseada localizados próximo à foz do rio Jacuecanga e influenciados pela dispersão da pluma apresentaram maiores teores de matéria orgânica de qualidade terrígena (restos vegetais e efluentes domésticos). Os sedimentos marinhos, mais distantes da foz do rio, apresentaram menores teores de matéria orgânica com características fitoplânctonicas, sendo os sedimentos localizados na zona da água de mi stura com valores intermediários. Foi verificada contaminação fecal na estação mais próxima a foz do rio e influência fecal em duas estações pró ximas, porém situadas em frente ao canal de drenagem da Vila de Jacuecanga pertencente à bacia hidrográfica vizi nha / Changes in the original land use and cover due to human activities development and and launch of domestic sewage directly into rivers, are factors causing the reduction in water quality and sediments in aquatic ecosystems. The Jacuecanga river basin (Angra dos Reis, RJ) has been suffering with growing changes in land use, mainly by population growth and development of different industrial activities, which can lead to degradation of its water resources. This study proposes the characterization of organic ma tter present in the main channel of this river system, its tributaries and its estuary. We evaluated the physical - chemical and sediments molecular markers as molecular, elemental and isotopic compositions with the aim of understanding the influence of chan ges in the primary coverage of the ground on the input of terrigenous organic material for these aquatic systems. A total of five samples were obtained at 16 stations on the rivers Jacuecanga, Caputera and Vermelho rivers , contemplating the various climate conditions of the year and a rain event. Sampling of surface sediments were performed at 14 stations in the river and 15 at the bay. The station (RJ2) located downstream of Vila Caputera showed trends of eutrophication due to deforestation, soil exposure and disorderly urban occupation characterized by greater intake of carbon, nitrogen, BOD and SPM . In the estuarine zone, the density of the urban Vila Jacuecanga was related to high percentages of carbon and nitrogen, especially during rain events. Rainwat er draining Transpetro Plant are discharged into the river at the end of the stretch of Jacuecanga freshwater and showed features alternating between the dry season with high conductivity, and the rainy season with high concentrations of SPM , but did not interfere relevant ly in water quality of river Jacuecanga. Likewise, the side entrance of water coming from the stabilization pond and the waters of the Vermelho River with low oxygen and high content of organic material did not show significant changes in river. In the sediments of Jacuecanga river were observed the p redominance of the sandy fraction demonstrat ing the intense hydrodynamics and lack of organic deposits along the basin. In the estuarine portion was recorded high er organic matter content with the identification of sources of vegetal remains and coprostanol from the greater urban density that occurs in the right margin. The sediments of bay located near the mouth of the river Jacuecanga and influenced by the plume showed higher levels of terrig enous organic matter quality ( vegetal remains and effluents). Marine sediments, most distant from the river mouth, had lower levels of organic matter with phytoplankton characteristic . S ediments located in the zone of mixing water presented intermediate va lues. Fecal contamination was checked at the station nearest the river mouth and fecal influence in two nearby stations, but located outside the drainage channel of the Village of Jacuecanga belonging to neighboring watershed.

<p>Podizanje novih zasada i povećanje kapaciteta prerade<br />grožđa povećava količinu otpada sa kojim se suočava<br />industrija proizvodnje vina i rakije. Najbolji i najodgovorniji<br />način upravljanja otpadom je njegovo dalje iskori&scaron;ćenje<br />kao nusproizvoda. U ovoj disertaciji sagledavana je mogućnost iskori&scaron;ćenja<br />nusproizvoda iz različitih faza proizvodnje u vinarijama i<br />destilerijama za dobijanje semenki grožđa koje su<br />kori&scaron;ćene za proizvodnju hladno presovanog ulja. Hipoteza<br />se zasniva na činjenici da semenke grožđa raznih sorti iz<br />različitih faza proizvodnje vina (bela, roze ili crvena vina),<br />odnosno, rakije mogu imati sasvim različit hemijski sastav i<br />kvalitet koji svakako može da se reflektuje na kvalitet,<br />bioaktivne komponente i održivost hladno presovanog ulja.<br />Ovakav proizvod, obzirom da je dobijen hladnim<br />presovanjem, može biti veoma atraktivan za potro&scaron;ače,<br />zbog svojih specifičnih senzorskih i nutritivnih<br />karakteristika.<br />Za potrebe izrade disertacije prikupljeni su nusproizvodi<br />i pripremljeni su uzorci iz vinarija i destilerija fru&scaron;kogorskog<br />vinogorja. Dobijene su semenke i proizvedeno hladno<br />presovano ulje od semenki crnog grožđa sorte Merlot,<br />belog grožđa sorte Italijanski rizling i belog grožđa sorte<br />Sila, kao autohtone sorte vinove loze. Hladno presovana<br />ulja su proizvedena od semenki grožđa koje nisu pro&scaron;le<br />nikakav tretman, odnosno dobijene su nakon presovanja,<br />prilikom proizvodnje belih (Italijanski rizling i Sila) i roze<br />(Merlot) vina, zatim od semenki koje su pro&scaron;le proces<br />fermentacije prilikom proizvodnje crvenih vina (Merlot) i od<br />semenki koje su pro&scaron;le proces fermentacije i destilacije<br />prilikom proizvodnje rakije (Merlot, Italijanski rizling i Sila).<br />Pored navedenih semenki i ulja, pripremljen je i prosečan<br />proizvodni uzorak koji predstavlja uzorak dobijen od svih<br />prikupljenih semenki. Kao uporedni uzorci kori&scaron;ćeno je<br />nerafinisano i rafinisano ulje od semenki grožđa nabavljeno<br />na trži&scaron;tu. Za realizaciju postavljenog cilja, rad na izvođenju ove<br />disertacije obuhvatio je različite faze. U prvoj fazi<br />prikupljene su semenke grožđa i ispitivane su njihove<br />tehničko-tehnolo&scaron;ke karakteristika i kvalitet. Zatim su od<br />navedenih semenki proizvedena hladno presovana ulja.<br />U sledećim fazama, ovako dobijena uja, zajedno sa<br />uzorkom nerafinisanog i rafinisanog ulja od semenki<br />grožđa sa trži&scaron;ta ispitivana su sa aspekta senzorskog i<br />nutritivnog kvaliteta i praćene su razlike u održivosti ulja.<br />Izvr&scaron;ena je senzorska analiza, određivane su frakcije<br />pigmenata, karotenoidi i hlorofili, merena je transparencija,<br />a parametri boje ulja određivani su i instrumentalno.<br />Nutritivni kvalitet ulja sagledavan je na osnovu sadržaja i<br />sastava bioaktivnih komponenti, pre svega tokoferola i<br />tokotrienola, fenola i sterola. Budući da ova jedinjenja<br />ispoljavaju značajne antioksidativne aktivnosti izvr&scaron;eno je i<br />ispitivanje antiradikalske aktivnosti uzoraka. Odživost ulja<br />od semenki grožđa sagledana je na osnovu početnog<br />kvaliteta i oksidativnog stanja, kao i rezultata ubrzanih<br />testova, kao &scaron;to je Rancimat test, Schaal-oven test i<br />fluorescentni test.<br />Na osnovu dobijenih rezultata, može se konstatovati da<br />su svi dobijeni uzorci semenki imali dobru skladi&scaron;nu vlagu,<br />koja, obzirom na mali sadržaj ulja u semenkama, može da<br />osigura čuvanje semenki u dužem periodu. Sadržaj ulja u<br />semenkama, pored uticaja sorte vinove loze, zavisi i od<br />uticaja procesa kome su podvrgnute semenke pre<br />presovanja (fermentacija, destilacija).<br />Senzorska analiza ulja od semenki grožđa pokazala je<br />značajne razlike u karakteristikama koje su posledica ne<br />samo načina dobijanja ulja (hladno presovano ili rafinisano), sorte vinove loze, već i postupka dobijanja,<br />porekla i kvaliteta semenki grožđa. Uslovi kojima su<br />semenke grožđa izložene u toku alkoholne fermentacije i<br />posebno destilacije utiču na formiranje specifične arome<br />hladno presovanog ulja. I pored toga &scaron;to se pojavljuje<br />izuzetno &scaron;irok spektar različitih aroma, mirisa i ukusa u ulju<br />i &scaron;to postoje značajne razlike u aromi ulja u zavisnosti od<br />sorte grožđa, moguće je prepoznati da li je hladno<br />presovano ulje dobijeno od semenki grožđa bez<br />fermentacije, posle fermentacije ili posle destilacije.<br />Takođe, sa aspekta boje ulja može se kazati da rafinisano<br />ulje od semenki grožđa ima svetlo žutu boju sa<br />zelenkastom nijansom, dok su hladno presovana ulja<br />intenzivnijih boja i kreću se od žuto-zelenkaste, preko<br />zelenkasto žute i svetlo zelenkaste do tamno zelene. Veći<br />udeo zelene boje imaju hladno presovana ulja od semenki<br />grožđa dobijenih posle destilacije.<br />U radu je pokazano da sadržaj pigmenata (karotenoida i<br />hlorofila) u velikoj meri zavisi od porekla semenki.<br />Postupak fermentacije doprinosi povećanju sadržaja<br />pigmenata, dok postupak destilacije, zbog visokih<br />temperatura ima negativan efekat.<br />Transparencija uzoraka hladno presovanih ulja od<br />semenki grožđa proizvedenih za potrebe disertacije kretala<br />se od 32,8% do 53,8%.<br />Sadržaj nezasićenih masnih kiselina u svim uzorcima<br />ulja od semenki grožđa veći je od 90%, pri čemu<br />dominantnu masnu kiselinu čini linolna, omega-6, masna<br />kiselina.<br />Sadržaj tokotrienola je veći od sadržaja tokoferola u uljima od semenki grožđa, a dominantni tokoferol je alfatokoferol.<br />Njegov sadržaj je veći u uzorcima ulja dobijenim<br />iz semenki nakon fermentacije i destilacije.<br />Sadržaj fenolnih jedinjenja, u zavisnosti od sorte, u<br />proseku je ne&scaron;to vi&scaron;i u hladno presovanim uljima dobijenim<br />od crvene sorte grožđa, ali uočava se i značajan porast<br />(akumulacija) fenola u hladno presovanim uljima dobijenim<br />iz semenki grožđa nakon procesa fermentacije i destilacije.<br />Najveći pojedinačni sadržaju u ulju od semenki grožđa ima<br />ursolna kiselina, a pored nje značajniji sadržaj, mada<br />mnogo manji, pokazuju rezveratrol, kemferol i vanilinska<br />kiselina.<br />Najzastupljeniji steroli ulja semenki grožđa su &beta;-<br />sitosterol sa udelom od 62,59-69,74%, stigmasterol sa<br />udelom od 12,13-15,00% i kampesterol sa udelom od 6,59-<br />11,94% u ukupnim sterolima. Na sadržaj fitosterola u<br />uljima od semenki grožđa nemaju uticaja procesi<br />fermentacije i destilacije kojima su podvrgnute semenke<br />pre preosvanja.<br />U radu je dokazan negativan uticaj procesa fermentacije<br />i destilacije kojima su povrgnute semenke grožđa na<br />antiradikalski potencijal dobijenih ulja. Antiradikalski<br />potencijal ulja od semenki grožđa zavisi i od sorte vinove<br />loze, kao i od načina dobijanja ulja (hladno presovano ili<br />rafinisano).<br />Procesi fermentacije i destilacije utiču i na osnovni<br />hemijski kvalitet dobijenih hladno presovanih ulja na način<br />da dovode do povećanja kiselinskog i peroksidnog broja.<br />Hladno presovana ulja od semenki grožđa pokazuju<br />dobru oksidativnu stabilnost. Rancimat test je pokazao razlike u indukcionom periodu hladno presovanih ulja od<br />semenki grožđa kao posledicu razlike u sorti, kao i u<br />načinu dobijanja semenki. Proces fermentacije utiče na<br />povećanje, a proces destilacije na smanjenje indukcionog<br />perioda.<br />Na osnovu svih dobijenih rezultata istraživanja može se<br />konstatovati da su hladno presovana ulja od semenki<br />grožđa pokazala različit nutritivni kvalitet i oksidativnu<br />stabilnost, zbog specifičnih razlika koje su posledica razlika<br />u sorti i poreklu semenki, tj. zbog specifičnog efekta<br />fermentacije i destilacije na semenke od kojih je ulje<br />proizvedeno, čime je i potvrđena hipoteza.</p> / <p>Raising new plantations and increasing the grape<br />processing capacity increases the amount of waste faced<br />by the wine and brandy industry. The best and most<br />responsible way of waste management is its further<br />exploitation as by-products.<br />In this dissertation, the possibility of using by-products from different stages of production in wineries and distilleries for<br />obtaining grape seeds, which were used for the production<br />of cold pressed oil, was examined. The hypothesis is<br />based on the fact that grape seeds of various varieties<br />from different stages of wine production (white, rose or red<br />wine), that is, brandy can have a completely different<br />chemical composition and quality that can certainly be<br />reflected on the quality, bioactive components and stability<br />of cold pressed oil.<br />Such a product, as it is obtained by cold pressing, can<br />be very attractive to consumers, due to its specific sensory<br />and nutritional characteristics.<br />For the needs of the dissertation, the by-products were<br />collected and samples were prepared from the wineries<br />and distilleries of the Fru&scaron;ka Gora vineyard. Seeds were<br />obtained and cold pressed oil produced from Merlot grape<br />seed, as representatives of red grape varieties, Italian<br />Riesling, as representatives of white grape varieties and<br />Sila, as new white grape varieties. Cold pressed oils were<br />produced from grape seeds that did not undergo any<br />treatment, that is, they were obtained after pressing, during<br />the production of white (Italian Riesling and Sila) and rose<br />(Merlot) wines, then from the seeds that were fermented<br />during the production of red wines (Merlot ) and seeds that<br />have undergone fermentation and distillation process<br />during the production of brandy (Merlot, Italian Riesling and<br />Sila). In addition to the mentioned seeds and oils, an<br />average production sample was prepared, which is a<br />sample obtained from all collected seeds. Unrefined and<br />refined grape seed oil purchased on the market was used<br />as comparative samples. For realization of the set goal, the work on the<br />performance of this dissertation encompassed different<br />phases. In the first phase, grape seed were collected and<br />their technical and technological characteristics and quality<br />were examined. Then cold-pressed oils were produced<br />from these seeds.<br />In the following phases, the resulting ear, together with a<br />sample of unrefined and refined grape seed oil from the<br />market, were examined from the aspect of sensory and<br />nutritional quality and differences in oil stability were<br />observed. Sensory analysis was performed, pigmentate<br />fractions, carotenoids and chlorophylls were determined,<br />transparency was measured, and oil color parameters<br />were also determined instrumentally. The nutritive oil<br />quality was examined based on the content and<br />composition of bioactive components, primarily tocopherols<br />and tocotrienols, phenols and sterols. Since these<br />compounds exhibit significant antioxidant activities, the<br />antiradical activity of the samples was also tested. The<br />stability of grape seed oils was examined based on the<br />initial quality and the oxidative state, as well as the results<br />of accelerated tests, such as Rancimat test, Schaal-oven<br />and fluorescence test.<br />On the basis of the obtained results, it can be concluded<br />that all the obtained seed samples had good storage<br />moisture, which, given the small content of oil in the seeds,<br />can ensure the storage of seeds for a longer period. The<br />content of oil in the seed, in addition to the influence of the<br />grape variety, depends on the influence of the process to<br />which the seeds are subjected to pressing (fermentation,<br />distillation). Sensory analysis grape seed oils showed significant<br />differences in characteristics that are due not only to the<br />method of obtaining oil (cold pressed or refined), grape<br />varieties, but also the method of obtaining, the origin and<br />quality of grape seed. The conditions for grape seeds<br />exposed during fermentation and especially distillation<br />affect the formation of a specific cold pressed oil. Although<br />an extremely wide range of different flavors, odors and<br />flavors in oil appear, and there are significant differences in<br />the aroma of the oil, depending on the grape variety, it is<br />possible to recognize whether the cold-pressed oil is<br />derived from the grape seed without fermentation, after<br />fermentation or after distillation. Also, from the aspect of oil<br />color it can be said that refined grape seed oil has a light<br />yellow color with a greenish shade, while cold pressed<br />grape seed oils have more intense colors range from<br />yellowish-greenish, over greenish yellow and light greenish<br />to dark green. A higher proportion of green color has cold<br />pressed grape seed oils obtained after distillation.<br />The thesis has shown that the content of pigments<br />(carotenoids and chlorophyll) depends to a great extent on<br />the origin of the seed. The fermentation process<br />contributes to increasing the content of pigments, while the<br />distillation process has a negative effect due to high<br />temperatures.<br />Transparency of cold pressed grape seed oils produced<br />for the dissertation ranged from 32.8% to 53.8%.<br />The content of unsaturated fatty acids in all samples of<br />grape seed oil is greater than 90%, with the dominant fatty<br />acid being linoleic, omega-6, fatty acid. The content of tocotrienols is higher than the content of<br />tocopherols in grape seed oils, and the dominant<br />tocopherol is alpha-tocopherol. Its content is higher in oil<br />samples obtained from the seed after fermentation and<br />distillation.<br />The content of phenol compounds, depending on the<br />variety, is somewhat higher in cold pressed oils obtained<br />from red grape varieties, but there is also a significant<br />increase in accumulation of phenol in cold pressed oils<br />obtained from the grape seed after the fermentation and<br />distillation process. The largest individual oil content of<br />grape seeds has ursolic acid, and besides it, significant<br />content, although much smaller, shows rezveratrol,<br />kemferol and vanillinic acid.<br />The most prevalent sterols of grape seed oils are &beta;-<br />sitosterol with a share of 62.59-69.74%, stigmasterol with a<br />share of 12.13-15.00% and campesterol with a share of<br />6.59-11.94% in total sterols. Fermentation and distilation<br />processes, to which the seeds have been exposed, have<br />no effect on the content of phytosterols in grape seed oils.<br />The paper has shown the negative influence of the<br />fermentation and distillation process, to which the seeds<br />have been exposed, onto the antiradical potential of the<br />obtained oils. The antiradical potential of grape seed oils<br />depends on grape varietes, as well as on the way oil is<br />obtained (cold pressed or refined).<br />Processes of fermentation and distillation also affect the<br />basic chemical quality of obtained cold pressed oils in such<br />a way as to increase the acid and peroxide values.<br />Cold pressed grape seed oils show good oxidative stability. The Rancimat test showed differences in the<br />induction period of cold pressed grape seed oils as a result<br />of the variation in the variety, as well as in the method of<br />obtaining the seed. The fermentation process affects the<br />increase, and the process of distillation decreases the<br />induction period.<br />Based on all the obtained results, it can be concluded<br />that cold pressed grape seed oils showed different<br />nutritional quality and oxidative stability due to specific<br />differences resulting from differences in variety and seed<br />origin, i.e. due to the specific effect of fermentation and<br />distillation processes on the seeds from which the oil was<br />produced, thus confirming the hypothesis.</p>