Biodiesel is rapidly growing as a fuel of interest due to the various advantages it has over conventional diesel fuel. While the pros – non-toxic, biodegradable, low green house gas emissions seem advantageous, the major issue that plagues the use of biodiesel is its cold weather operability. Biodiesel can present challenges in cold-weather operation, because certain of its constituent compounds can form precipitates in the fuel. These precipitates can cause undesired effects like plugging of fuel filters and deposits. This issue has been attributed to the presence of impurities (mostly saturated monoglycerides, di-glycerides, soap etc) in biodiesel and has been discussed in the literature. There is a move by users and standards associations to implement more stringent norms and quality control to avoid problems in the widespread use of biodiesel. This study involves ways to reduce MG’s in biodiesel by mitigating to a greater extent the possibility of side reactions (formation of soap). The effect of selective transesterification of oil as a function of alcohol, temperature and catalyst concentration was also studied. Although saturated MG’s with high melting points are a greater source of deposits, it can be hypothesized that the polymorphic nature of unsaturated Monoglycerides could also be contributing to cold flow issues. It is hence vital to make sure the biodiesel is free from all forms of monoglycerides. It was also seen that there is very little specificity of selection of fatty acid types in the transesterification reaction and that the amount and type of MGs present in the biodiesel is reflected by the relative amount of fatty acids types present in the oil. In biodiesel derived from Canola oil, a preponderance of monoolein was found for all runs. The initial runs carried out as a two stage process using the membrane followed by batch reactor gave very low MG concentrations, well below ASTM standards.

Biodiesel is rapidly growing as a fuel of interest due to the various advantages it has over conventional diesel fuel. While the pros – non-toxic, biodegradable, low green house gas emissions seem advantageous, the major issue that plagues the use of biodiesel is its cold weather operability. Biodiesel can present challenges in cold-weather operation, because certain of its constituent compounds can form precipitates in the fuel. These precipitates can cause undesired effects like plugging of fuel filters and deposits. This issue has been attributed to the presence of impurities (mostly saturated monoglycerides, di-glycerides, soap etc) in biodiesel and has been discussed in the literature. There is a move by users and standards associations to implement more stringent norms and quality control to avoid problems in the widespread use of biodiesel. This study involves ways to reduce MG’s in biodiesel by mitigating to a greater extent the possibility of side reactions (formation of soap). The effect of selective transesterification of oil as a function of alcohol, temperature and catalyst concentration was also studied. Although saturated MG’s with high melting points are a greater source of deposits, it can be hypothesized that the polymorphic nature of unsaturated Monoglycerides could also be contributing to cold flow issues. It is hence vital to make sure the biodiesel is free from all forms of monoglycerides. It was also seen that there is very little specificity of selection of fatty acid types in the transesterification reaction and that the amount and type of MGs present in the biodiesel is reflected by the relative amount of fatty acids types present in the oil. In biodiesel derived from Canola oil, a preponderance of monoolein was found for all runs. The initial runs carried out as a two stage process using the membrane followed by batch reactor gave very low MG concentrations, well below ASTM standards.

Biodiesel is rapidly growing as a fuel of interest due to the various advantages it has over conventional diesel fuel. While the pros – non-toxic, biodegradable, low green house gas emissions seem advantageous, the major issue that plagues the use of biodiesel is its cold weather operability. Biodiesel can present challenges in cold-weather operation, because certain of its constituent compounds can form precipitates in the fuel. These precipitates can cause undesired effects like plugging of fuel filters and deposits. This issue has been attributed to the presence of impurities (mostly saturated monoglycerides, di-glycerides, soap etc) in biodiesel and has been discussed in the literature. There is a move by users and standards associations to implement more stringent norms and quality control to avoid problems in the widespread use of biodiesel. This study involves ways to reduce MG’s in biodiesel by mitigating to a greater extent the possibility of side reactions (formation of soap). The effect of selective transesterification of oil as a function of alcohol, temperature and catalyst concentration was also studied. Although saturated MG’s with high melting points are a greater source of deposits, it can be hypothesized that the polymorphic nature of unsaturated Monoglycerides could also be contributing to cold flow issues. It is hence vital to make sure the biodiesel is free from all forms of monoglycerides. It was also seen that there is very little specificity of selection of fatty acid types in the transesterification reaction and that the amount and type of MGs present in the biodiesel is reflected by the relative amount of fatty acids types present in the oil. In biodiesel derived from Canola oil, a preponderance of monoolein was found for all runs. The initial runs carried out as a two stage process using the membrane followed by batch reactor gave very low MG concentrations, well below ASTM standards.

Call number: LD2668 .T4 1985 C475 / Master of Science

O efeito de diferentes agentes antioxidantes naturais lipossolúveis (óleo de copaíba, manteiga de tucumã, manteiga de cacau, óleo de buriti e especiarias como orégano, tomilho branco, alecrim, sálvia sclarea, cravo talo e cananga) foi determinado na síntese enzimática de monoglicerídeos (MAG) a partir da glicerólise do óleo de babaçu, tomando como parâmetro de comparação os resultados alcançados na reação isenta de estratégia antioxidante (controle negativo) e sob atmosfera inerte (controle positivo). As reações foram conduzidas em reatores operando em regime descontínuo e contínuo usando a lipase de Burkholderia cepacia imobilizada em SiO2-PVA como catalisador. O melhor desempenho em reatores descontínuos foi obtido usando N2 no meio reacional (60 % MAG) seguido do óleo de buriti (57,6 % MAG) e manteiga de cacau (56,6 % MAG). Entretanto, a incorporação do óleo de buriti no meio reacional alterou o perfil de formação de monoglicerídeos, ocasionando elevada concentração de monoleina. Tomando por base esses resultados, selecionou-se a manteiga de cacau como agente antioxidante para a condução dos testes em fluxo contínuo. Nesse contexto, visando estabelecer maior homogeneidade do meio reacional e alcançar rendimentos elevados de MAGs, foram realizados testes em reator de leito fixo em fluxo contínuo, sob atmosfera inerte, variando-se a proporção entre óleo e glicerol no meio de alimentação. Os resultados obtidos nesta série de experimentos demonstraram forte influencia da razão molar dos materiais de partida na síntese de MAGs e o melhor desempenho foi obtido empregando a razão molar intermediária (1:9), que proporcionou a formação de 31,5 % de MAGs e produtividade de 52,2 mgMAGs/gamostra.h. Testes empregando manteiga de cacau como agente antioxidante foram efetuados e desempenhos similares daqueles obtidos em atmosfera inerte foram obtidos, confirmando o efeito benéfico da utilização da manteiga de cacau no meio de alimentação como agente antioxidante. Isto sugere que a manteiga de cacau inibiu ou reduziu a oxidação do óleo de babaçu durante a síntese de MAGs, indicando a possibilidade de substituição da atmosfera inerte (N2) na reação. Análises espectrofotométricas dos produtos de oxidação primários (hidroperóxidos e dienos conjugados) e secundários (aldeídos e cetonas) validaram a eficiência da manteiga de cacau como agente antioxidante da matéria-prima lipídica durante todo o processo contínuo. / The effects of several natural antioxidants (copaiba oil, buriti oil, cocoa butter, tucuman butter, oregano, white thyme, rosemary, salvia sclarea, carnation stem and cananga) were assessed in the enzymatic synthesis of monoglycerides (MAG) from the glycerolysis of babassu oil. The reactions were catalyzed by Burkholderia cepacia lipase immobilized on SiO2-PVA and the assays carried out in batch and continuous runs. Results were compared with those attained in the control reactions (without any strategy to avoid oxidation and under inert atmosphere), and the best approach was tested in a continuous packed bed reactor. The best performance was achieved using N2 in the reaction medium (60 % of MAG) followed by buriti oil (57.6 % of MAG) and cocoa butter (56.6 % of MAG), preventing the oxidation of babassu oil in batch reaction. However, the incorporation of buriti oil in the medium influenced the MAG profile, leading to the largest formation of monoolein, unlike other runs. Based on these results, cocoa butter was chosen as antioxidant agent to carry out the assays under continuous flow. Therefore, aiming to establish greater homogeneous reaction media and to attain high MAGs yields, assays were performed in packed bed reactor under continuous flow, in inert atmosphere, varying the proportional of babassu oil and glycerol in the feed media. The results obtained in this set of runs demonstrated a strong influence of molar ratio between starting materials in the synthesis of MAGs and the best performance was achieved using the intermediate molar ratio (1:9), which afforded the formation of 31.5% of MAGs and productivity of 52.2 mgMAGs/gsample.h. Tests using cocoa butter as antioxidant were made and similar results were attained, confirming the beneficial effect of using cocoa butter in the feed medium as an antioxidant agent. This suggests that the cocoa butter inhibited or reduced babassu oil oxidation during synthesis of MAGs, indicating the possibility of replacing the inert atmosphere (N2) in the reaction. Spectrophotometric analysis of the primary oxidation products (conjugated dienes and hydroperoxides) and secondary (aldehydes and ketones) validated the efficiency of cocoa butter as an antioxidant agent of lipid a continuous process. Thereby, among the nine tested antioxidant agents, cocoa butter was the most effective in both systems, since it not interfere in the MAG profile and also reduced the cost of the process.

O presente projeto teve como objetivo selecionar a melhor rota enzimática para produção de monoglicerídeos empregando lipase imobilizada em matriz obtida pela técnica sol-gel, visando total aproveitamento do glicerol gerado como sub-produto na obtenção de biodiesel. Para alcançar o objetivo proposto foram analisadas duas metodologias: (1) esterificação do glicerol com ácidos graxos e (2) glicerólise do óleo de babaçu; testando lipases de diferentes fontes como Candida antarctica B, Thermomyces lanuginosus, Candida rugosa, Pseudomonas fluorescens, Burkholderia cepacia, Peniccillium camembertii e pâncreas de porco, imobilizadas em polissiloxano álcool polivinílico (POSPVA). As preparações de lipase imobilizada disponíveis comercialmente como Lipozyme IM20, Novozym 435, Lipozyme RM IM e Lipozyme TL IM foram também utilizadas. As atividades experimentais foram, inicialmente, direcionadas para o estabelecimento das condições adequadas da esterificação, investigando a influência de diversos parâmetros, tais como: proporção molar dos reagentes, tamanho da cadeia carbônica e grau de insaturação dos ácidos graxos e fonte de lipase. Verificou-se que meios reacionais contendo excesso de glicerol favoreceram a síntese de ésteres glicéricos, entretanto, tanto a velocidade reacional como a conversão molar foram dependentes do tamanho da cadeia carbônica e grau de insaturação dos ácidos graxos testados. Os experimentos realizados permitiram também selecionar a lipase Candida antarctica B como fonte de lipase mais adequada, sendo alcançada uma elevada concentração de monolaurina em 12 h de reação (24,15%). Para obtenção de monoglicerídeo pela rota de glicerólise do óleo de babaçu, o melhor desempenho foi alcançado pela lipase Burkholderia cepacia, revelando uma formação de 10% (m/m) de monoglicerídeos em apenas 3h de reação. A análise comparativa do desempenho das duas rotas indicou que a esterificação foi mais eficiente que a glicerólise, sendo então selecionada para estudos de otimização. Um planejamento experimental completo 22 foi proposto para verificar a influência simultânea das variáveis x1 (temperatura entre 45-65ºC) e x2 (razão molar glicerol/ ácido láurico na faixa 3:1 a 5:1) na formação de monolaurina, empregando a lipase Candida antarctica B imobilizada em POS-PVA. De acordo com a análise estatística, a formação de monolaurina foi fortemente influenciada pela variável (x2) razão molar entre glicerol e ácido láurico, enquanto que a influência da temperatura (x1) foi pouco significativa. O modelo matemático proposto: y = 24,16 + 2,51 x1 + 4,43x2 - 2,74 x1x2, permitiu prever as condições que favorecem o alcance de elevados rendimentos de formação de monolaurina, sendo a reação maximizada (31,35%) para meios reacionais constituídos de glicerol e ácido láurico numa razão molar de 5:1 e temperatura de incubação de 45ºC. Essas condições foram também adequadas para síntese de monolaurina empregando outras fontes de lipase, como a lipase de Penicillium camembertii cujo desempenho foi altamente eficiente para obtenção de monoglicerídeos, alcançando valores mais competitivos de formação de monolaurina (51%), podendo, portanto, ser uma alternativa interessante para ser considerada no desenvolvimento de futuros trabalhos. / The objective of this project was to select the best enzymatic route to produce monoglycerides using lipase immobilized on matrix obtained by sol-gel technique, aiming to contribute for a better utilization of the glycerol generated as by-product in the biodiesel synthesis. For this purpose two methodologies were analyzed: (1) direct esterification of the glycerol with fatty acids and (2) glycerolysis of the babassu oil, testing lipases from different sources as Candida antarctica B, Thermomyces lanuginosus, Candida rugosa, Pseudomonas fluorescens, Burkholderia cepacia, Penicillium camemberti and porcine pancreas immobilized on polysiloxane-polyvinyl alcohol particles (POS-PVA). The commercial immobilized lipase preparations as Lipozyme IM20, Novozym 435, Lipozyme RM IM and Lipozyme TL IM were also used. The experimental activities were, initially, addressed to establish appropriate conditions for the monoglycerides synthesis by direct esterification such as: reactants molar ratio, carbon chain size and insaturation degree of the fatty acids and lipase source. Reaction performance was found to be dependent on the glycerol/lauric acid molar ratio, requesting higher amount of glycerol to reach high reaction conversion. It was also verified that the molar conversion were strongly dependent on both carbon chain size and insaturation degree of the tested fatty acids. Experiments also allowed to select Candida antarctica B as catalyst to carry esterification reactions and the highest monolaurin formation occurred at 12 h reaction (24.15%). For the synthesis of monoglycerides by the glycerolysis route using babassu oil as a starting material, the best performance was attained by the lipase Burkholderia cepacia, revealing a formation of 10% (m/m) of monoglyceryde in 3h reaction. The comparative performance of the two routes for the monoglycerides synthesis demonstrated that the direct esterification was more efficient than the glycerolysis, being selected for additional optimization studies. A full factorial design 22 was proposed to verify the simultaneous influence of the variables: x1 (temperature in the range from 45 to 65ºC) and x2 (glycerol/ lauric acid molar ratio in the range from 3:1 to 5:1) in the monolaurin formation, using as catalyst lipase Candida antarctica B immobilized on POS-PVA. In agreement with the statistical analysis, the monolaurin formation was strongly influenced by the variable (x2) glycerol/ lauric acid molar ratio. The proposed mathematical model: y = 24.16 + 2.51 x1 + 4.43x2 - 2.74 x1x2 allowed to predict conditions that favored to attain high monolaurin yield. The reaction was maximized (31.35%) for substrate containing glycerol/lauric acid molar ratio of 5:1 and at incubation temperature of 45ºC. Those conditions were also appropriated for the monolaurin synthesis using other lipase sources, as the lipase of Penicillium camembertii that performance was found to be highly efficient for monoglycerides reaching more competitive values for monolaurin formation (51%) and can be turn out, therefore, an interesting alternative to be considered in futures works.

O presente projeto teve como objetivo selecionar a melhor rota enzimática para produção de monoglicerídeos empregando lipase imobilizada em matriz obtida pela técnica sol-gel, visando total aproveitamento do glicerol gerado como sub-produto na obtenção de biodiesel. Para alcançar o objetivo proposto foram analisadas duas metodologias: (1) esterificação do glicerol com ácidos graxos e (2) glicerólise do óleo de babaçu; testando lipases de diferentes fontes como Candida antarctica B, Thermomyces lanuginosus, Candida rugosa, Pseudomonas fluorescens, Burkholderia cepacia, Peniccillium camembertii e pâncreas de porco, imobilizadas em polissiloxano álcool polivinílico (POSPVA). As preparações de lipase imobilizada disponíveis comercialmente como Lipozyme IM20, Novozym 435, Lipozyme RM IM e Lipozyme TL IM foram também utilizadas. As atividades experimentais foram, inicialmente, direcionadas para o estabelecimento das condições adequadas da esterificação, investigando a influência de diversos parâmetros, tais como: proporção molar dos reagentes, tamanho da cadeia carbônica e grau de insaturação dos ácidos graxos e fonte de lipase. Verificou-se que meios reacionais contendo excesso de glicerol favoreceram a síntese de ésteres glicéricos, entretanto, tanto a velocidade reacional como a conversão molar foram dependentes do tamanho da cadeia carbônica e grau de insaturação dos ácidos graxos testados. Os experimentos realizados permitiram também selecionar a lipase Candida antarctica B como fonte de lipase mais adequada, sendo alcançada uma elevada concentração de monolaurina em 12 h de reação (24,15%). Para obtenção de monoglicerídeo pela rota de glicerólise do óleo de babaçu, o melhor desempenho foi alcançado pela lipase Burkholderia cepacia, revelando uma formação de 10% (m/m) de monoglicerídeos em apenas 3h de reação. A análise comparativa do desempenho das duas rotas indicou que a esterificação foi mais eficiente que a glicerólise, sendo então selecionada para estudos de otimização. Um planejamento experimental completo 22 foi proposto para verificar a influência simultânea das variáveis x1 (temperatura entre 45-65ºC) e x2 (razão molar glicerol/ ácido láurico na faixa 3:1 a 5:1) na formação de monolaurina, empregando a lipase Candida antarctica B imobilizada em POS-PVA. De acordo com a análise estatística, a formação de monolaurina foi fortemente influenciada pela variável (x2) razão molar entre glicerol e ácido láurico, enquanto que a influência da temperatura (x1) foi pouco significativa. O modelo matemático proposto: y = 24,16 + 2,51 x1 + 4,43x2 - 2,74 x1x2, permitiu prever as condições que favorecem o alcance de elevados rendimentos de formação de monolaurina, sendo a reação maximizada (31,35%) para meios reacionais constituídos de glicerol e ácido láurico numa razão molar de 5:1 e temperatura de incubação de 45ºC. Essas condições foram também adequadas para síntese de monolaurina empregando outras fontes de lipase, como a lipase de Penicillium camembertii cujo desempenho foi altamente eficiente para obtenção de monoglicerídeos, alcançando valores mais competitivos de formação de monolaurina (51%), podendo, portanto, ser uma alternativa interessante para ser considerada no desenvolvimento de futuros trabalhos. / The objective of this project was to select the best enzymatic route to produce monoglycerides using lipase immobilized on matrix obtained by sol-gel technique, aiming to contribute for a better utilization of the glycerol generated as by-product in the biodiesel synthesis. For this purpose two methodologies were analyzed: (1) direct esterification of the glycerol with fatty acids and (2) glycerolysis of the babassu oil, testing lipases from different sources as Candida antarctica B, Thermomyces lanuginosus, Candida rugosa, Pseudomonas fluorescens, Burkholderia cepacia, Penicillium camemberti and porcine pancreas immobilized on polysiloxane-polyvinyl alcohol particles (POS-PVA). The commercial immobilized lipase preparations as Lipozyme IM20, Novozym 435, Lipozyme RM IM and Lipozyme TL IM were also used. The experimental activities were, initially, addressed to establish appropriate conditions for the monoglycerides synthesis by direct esterification such as: reactants molar ratio, carbon chain size and insaturation degree of the fatty acids and lipase source. Reaction performance was found to be dependent on the glycerol/lauric acid molar ratio, requesting higher amount of glycerol to reach high reaction conversion. It was also verified that the molar conversion were strongly dependent on both carbon chain size and insaturation degree of the tested fatty acids. Experiments also allowed to select Candida antarctica B as catalyst to carry esterification reactions and the highest monolaurin formation occurred at 12 h reaction (24.15%). For the synthesis of monoglycerides by the glycerolysis route using babassu oil as a starting material, the best performance was attained by the lipase Burkholderia cepacia, revealing a formation of 10% (m/m) of monoglyceryde in 3h reaction. The comparative performance of the two routes for the monoglycerides synthesis demonstrated that the direct esterification was more efficient than the glycerolysis, being selected for additional optimization studies. A full factorial design 22 was proposed to verify the simultaneous influence of the variables: x1 (temperature in the range from 45 to 65ºC) and x2 (glycerol/ lauric acid molar ratio in the range from 3:1 to 5:1) in the monolaurin formation, using as catalyst lipase Candida antarctica B immobilized on POS-PVA. In agreement with the statistical analysis, the monolaurin formation was strongly influenced by the variable (x2) glycerol/ lauric acid molar ratio. The proposed mathematical model: y = 24.16 + 2.51 x1 + 4.43x2 - 2.74 x1x2 allowed to predict conditions that favored to attain high monolaurin yield. The reaction was maximized (31.35%) for substrate containing glycerol/lauric acid molar ratio of 5:1 and at incubation temperature of 45ºC. Those conditions were also appropriated for the monolaurin synthesis using other lipase sources, as the lipase of Penicillium camembertii that performance was found to be highly efficient for monoglycerides reaching more competitive values for monolaurin formation (51%) and can be turn out, therefore, an interesting alternative to be considered in futures works.

O efeito de diferentes agentes antioxidantes naturais lipossolúveis (óleo de copaíba, manteiga de tucumã, manteiga de cacau, óleo de buriti e especiarias como orégano, tomilho branco, alecrim, sálvia sclarea, cravo talo e cananga) foi determinado na síntese enzimática de monoglicerídeos (MAG) a partir da glicerólise do óleo de babaçu, tomando como parâmetro de comparação os resultados alcançados na reação isenta de estratégia antioxidante (controle negativo) e sob atmosfera inerte (controle positivo). As reações foram conduzidas em reatores operando em regime descontínuo e contínuo usando a lipase de Burkholderia cepacia imobilizada em SiO2-PVA como catalisador. O melhor desempenho em reatores descontínuos foi obtido usando N2 no meio reacional (60 % MAG) seguido do óleo de buriti (57,6 % MAG) e manteiga de cacau (56,6 % MAG). Entretanto, a incorporação do óleo de buriti no meio reacional alterou o perfil de formação de monoglicerídeos, ocasionando elevada concentração de monoleina. Tomando por base esses resultados, selecionou-se a manteiga de cacau como agente antioxidante para a condução dos testes em fluxo contínuo. Nesse contexto, visando estabelecer maior homogeneidade do meio reacional e alcançar rendimentos elevados de MAGs, foram realizados testes em reator de leito fixo em fluxo contínuo, sob atmosfera inerte, variando-se a proporção entre óleo e glicerol no meio de alimentação. Os resultados obtidos nesta série de experimentos demonstraram forte influencia da razão molar dos materiais de partida na síntese de MAGs e o melhor desempenho foi obtido empregando a razão molar intermediária (1:9), que proporcionou a formação de 31,5 % de MAGs e produtividade de 52,2 mgMAGs/gamostra.h. Testes empregando manteiga de cacau como agente antioxidante foram efetuados e desempenhos similares daqueles obtidos em atmosfera inerte foram obtidos, confirmando o efeito benéfico da utilização da manteiga de cacau no meio de alimentação como agente antioxidante. Isto sugere que a manteiga de cacau inibiu ou reduziu a oxidação do óleo de babaçu durante a síntese de MAGs, indicando a possibilidade de substituição da atmosfera inerte (N2) na reação. Análises espectrofotométricas dos produtos de oxidação primários (hidroperóxidos e dienos conjugados) e secundários (aldeídos e cetonas) validaram a eficiência da manteiga de cacau como agente antioxidante da matéria-prima lipídica durante todo o processo contínuo. / The effects of several natural antioxidants (copaiba oil, buriti oil, cocoa butter, tucuman butter, oregano, white thyme, rosemary, salvia sclarea, carnation stem and cananga) were assessed in the enzymatic synthesis of monoglycerides (MAG) from the glycerolysis of babassu oil. The reactions were catalyzed by Burkholderia cepacia lipase immobilized on SiO2-PVA and the assays carried out in batch and continuous runs. Results were compared with those attained in the control reactions (without any strategy to avoid oxidation and under inert atmosphere), and the best approach was tested in a continuous packed bed reactor. The best performance was achieved using N2 in the reaction medium (60 % of MAG) followed by buriti oil (57.6 % of MAG) and cocoa butter (56.6 % of MAG), preventing the oxidation of babassu oil in batch reaction. However, the incorporation of buriti oil in the medium influenced the MAG profile, leading to the largest formation of monoolein, unlike other runs. Based on these results, cocoa butter was chosen as antioxidant agent to carry out the assays under continuous flow. Therefore, aiming to establish greater homogeneous reaction media and to attain high MAGs yields, assays were performed in packed bed reactor under continuous flow, in inert atmosphere, varying the proportional of babassu oil and glycerol in the feed media. The results obtained in this set of runs demonstrated a strong influence of molar ratio between starting materials in the synthesis of MAGs and the best performance was achieved using the intermediate molar ratio (1:9), which afforded the formation of 31.5% of MAGs and productivity of 52.2 mgMAGs/gsample.h. Tests using cocoa butter as antioxidant were made and similar results were attained, confirming the beneficial effect of using cocoa butter in the feed medium as an antioxidant agent. This suggests that the cocoa butter inhibited or reduced babassu oil oxidation during synthesis of MAGs, indicating the possibility of replacing the inert atmosphere (N2) in the reaction. Spectrophotometric analysis of the primary oxidation products (conjugated dienes and hydroperoxides) and secondary (aldehydes and ketones) validated the efficiency of cocoa butter as an antioxidant agent of lipid a continuous process. Thereby, among the nine tested antioxidant agents, cocoa butter was the most effective in both systems, since it not interfere in the MAG profile and also reduced the cost of the process.

Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / CoordenaÃÃo de AperfeÃoamento de Pessoal de NÃvel Superior / O objetivo desse trabalho foi investigar a produÃÃo de mono (MG) e diglicerÃdeos (DG) a partir da hidrÃlise e/ou glicerÃlise enzimÃtica de um triglicerÃdeo (TG), utilizando-se a tÃcnica de miniemulsÃo. Utilizou-se como substrato o triglicerÃdeo do Ãcido caprÃico, a tricaprilina. Dois tipos de lipases foram utilizadas neste estudo a fim de se estudar a regioseletividade da reaÃÃo de hidrÃlise em miniemulsÃo. A lipase Rhizopus arrhizus (RAL) com regiosseletividade especÃfica sn-1,3 e a lipase Pseudomonas cepacia (PS) devido a sua ausÃncia de regioseletividade. Uma vez que hidrÃlise e glicerÃlise ocorrem na interface Ãleo-Ãgua, diferenÃas na Ãrea interfacial pela variaÃÃo da quantidade de surfactante e a influÃncia da concentraÃÃo das lipases tambÃm foram estudadas. AlÃm disso, a glicerÃlise foi estudada pela adiÃÃo de glicerol na preparaÃÃo da miniemulsÃo com o objetivo de direcionar a formaÃÃo dos produtos para uma maior quantidade de monoglicerÃdeos. Os produtos das reaÃÃes foram caracterizados e quantificados pelas tÃcnicas de H1-RMN e HPLC. Por HPLC obteve-se as quantidades totais de cada componente (MG, DG, Ãcido graxo livre e glicerol) enquanto que por RMN, pode-se calcular as quantidades individuais de cada produto formado (1-MG, 2-MG, 1,2-DG e 1,3-DG). Observando-se as concentraÃÃes dos produtos formados na hidrÃlise catalisada pela lipase PS, pÃde-se concluir que essa lipase, conhecida por sua nÃo-especificidade, catalisou a reaÃÃo preferencialmente na posiÃÃo sn-2, tendo como principais produtos 1,3-DG e 1-MG, com concentraÃÃes mÃximas de 29% e 22%, respectivamente. Por outro lado, a hidrÃlise catalisada pela lipase RAL, teve como principais produtos 1,2-DG e 2- MG, com concentraÃÃes mÃximas de 24% e 18%, respectivamente. Os resultados corroboram com a preferÃncia dessa lipase pela posiÃÃo sn-1,3. Observou-se para a glicerÃlise em miniemulsÃo catalisada pela lipase RAL, que a adiÃÃo de glicerol mudou o perfil de formaÃÃo de 1-MG e 2-MG, alcanÃando um mÃximo de 10-12% e 32-35%, respectivamente, apÃs 4h, comparado com 8 e 22-25% durante hidrÃlise. JÃ para glicerÃlise catalisada pela lipase PS, os resultados mostraram que essa lipase, que apresentou preferÃncia pela posiÃÃo sn-2 na hidrÃlise, passou a ter comportamento similar ao da lipase RAL, sn-1,3 especÃfica. / The aim of this study was to investigate the production of mono (MG) and diglycerides (DG) from the enzymatic hydrolysis and/or glycerolysis of a triglyceride (TG), using the technique of miniemulsion. As substrate, the triglyceride of the caproic acid, the tricaprylin was used. Two types of lipases were used in this study in order to examine the regioselectivity of the hydrolysis reaction in miniemulsion. The Rhizopus arrhizus lipase (RAL) known as specific sn-1,3 and the Pseudomonas cepacia (PS) known as non-specific lipase. Since hydrolysis and glycerolysis occur in the oil-water interface, differences in the interfacial area by varying the amount of surfactant concentration and the influence of the lipase were also studied. Additionally, glycerolysis was studied by adding glycerol to prepare the miniemulsion in order to increase amount of monoglycerides as products. The products were characterized and quantified by the 1H-NMR and HPLC techniques. HPLC afforded the quantification of total amount of each component (MG, DG, free fatty acid and glycerol) whereas by NMR, it was possible to calculate the amounts of each individual isomers (1-MG, 2-MG, 1,2-DG e 1,3-DG). The concentrations of the isomers formed in the lipase PS catalyzed hydrolysis showed that this lipase, known for its non-specificity, catalyzed the reaction preferentially at the sn-2 position, having as main products 1,3-DG and 1-MG, with maximum concentrations of 29% and 22%, respectively. On the other hand, hydrolysis catalyzed by lipase RAL has as main products 1,2-DG and 2-MG with maximum concentrations of 35% and 25% respectively, at 4h. The results corroborate the preference of the lipase for the sn-1,3 position. The initial addition of glycerol to the reaction catalyzed with lipase RAL did not significantly affect the reaction profiles and the formation rates of the diglycerides. However, the introduction of glycerol changes the profile of the formation of 1- and 2-monocaprylin, reaching the concentration maximum of 10-12% and 32-35% after 4h compared to 8% and 22-25% during the hydrolysis reaction, respectively. Lipase PS, an unspecific enzyme (slight preference for the sn-2-position in hydrolysis in miniemulsion) showed in presence of glycerol a behavior similar to RAL, a sn-1,3 specific lipase.

Orientadores: Maria Regina Wolf Maciel, Cesar Benedito Batistella / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia Quimica / Made available in DSpace on 2018-08-05T23:41:15Z (GMT). No. of bitstreams: 1 Fregolente_LeonardoVasconcelos_M.pdf: 590348 bytes, checksum: 2ba3d20d50e05dcaaf811d7862b9cb67 (MD5) Previous issue date: 2006 / Resumo: Monoglicerídeos são emulsificantes amplamente utilizados em alimentos, fármacos e cosméticos. Industrialmente, são produzidos através da interesterificação de triglicerídeos com glicerol, a altas temperaturas (> 200°C), na presença de catalisadores inorgânicos. Também conhecida como glicerólise, esta reação produz uma mistura com aproximadamente 50% de monoglicerídeos. Este nível de concentração é adequado a muitos tipos de aplicações, no entanto, para fabricação de alguns produtos como argarinas, sorvetes e recheios, devem ser utilizados monoglicerídeos de alta concentração (teor mínimo de 90%), os quais são normalmente produzidos através do processo de destilação molecular. Assim, neste trabalho, foi utilizado um destilador molecular centrífugo com área de evaporação igual a 0,0046m2, para a obtenção de monoglicerídeos de alta concentração (até 96,3%). A metodologia de análise de superficie de resposta foi utilizada para a identificação das condições operacionais ótimas de enriquecimento de monoglicerídeos na corrente de destilado. Além disso, um planejamento experimental 23 foi empregado para estudarem-se os efeitos de alguns parâmetros reacionais sobre a concentração de monoglicerídeos na condição de equilíbrio químico da reação de glicerólise de óleo de soja refinado, catalisada por hidróxido de sódio. Então, utilizando-se as condições de destilação otimizadas através da metodologia de análise de superficie de resposta, os produtos reacionais foram destilados, aumentando-se assim a concentração de monoglicerídeos / Abstract: Monoglycerides are emulsifiers widely used in food, harmaceutical, and cosmetic fields. Current industrial processes for monoglycerides production consist on the interesterification of triglycerides with glycerol in the presence of inorganic catalysts at high temperatures (>200°C). This reaction is known as glycerolysis and produces a mixture of approx 50 % of monoglycerides. This leveI of concentration is suitable for many applications, although, some specific uses like margarine, icing and cream filling require distilled monoglycerides, which are purified monoglycerides (min. 90%) normally obtained by molecular distillation processo Therefore, in this work, a centrífugal molecular distillator of evaporation area of 0.0046m2 was used to obtain monoglycerides with up to 96.3% of purity. Response surface methodology was applied as a tool to identifY operating conditions that can lead to higher monoglycerides purity in the distillate stream. Furthermore, a 23 factorial design was employed to evaluate the effects of reaction parameters on the monoglycerides concentration after the interesterification reaction of refmed soybean oil with glycerol in the presence of sodium hydroxide as catalyst. The monoglycerides content in the reaction product was enhanced, using the optimized distillation conditions obtained by the response surface methodology / Mestrado / Desenvolvimento de Processos Químicos / Mestre em Engenharia Química