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Production and fractionation of antioxidant peptides from soy protein isolate using sequential membrane ultrafiltration and nanofiltrationRanamukhaarachchi, Sahan January 2012 (has links)
Antioxidants are molecules capable of stabilizing and preventing oxidation. Certain peptides, protein hydrolysates, have shown antioxidant capacities, which are obtained once liberated from the native protein structure. Soy protein isolates (SPI) were enzymatically hydrolyzed by pepsin and pancreatin mixtures. The soy protein hydrolysates (SPH) were fractionated with sequential ultrafiltration (UF) and nanofiltration (NF) membrane steps. Heat pre-treatment of SPI at 95 degrees celsius (C) for 5 min prior to enzymatic hydrolysis was investigated for its effect on peptide distribution and antioxidant capacity. SPH were subjected to UF with a 10 kDa molecular weight cut off (MWCO) polysulfone membrane. UF permeate fractions (lower molecular weight than 10 kDa) were fractionated by NF with a thin film composite membrane (2.5 kDa MWCO) at pH 4 and 8. Similar peptide content and antioxidant capacity (α=0.05) were obtained in control and pre-heated SPH when comparing the respective UF and NF permeate and retentate fractions produced. FCR antioxidant capacities of the SPH fractions were significantly lower than their ORAC antioxidant capacities, and the distribution among the UF and NF fractions was generally different. Most UF and NF fractions displayed higher antioxidant capacities when compared to the crude SPI hydrolysates, showing the importance of molecular weight on antioxidant capacity of peptides. The permeate fractions produced by NF at pH 8 displayed the highest antioxidant capacity, expressed in terms of Trolox equivalents (TE) per total solids (TS): 5562 μmol TE/g TS for control SPH, and 5187 μmol TE/g TS for pre-heated SPH. Due to the improvement in antioxidant capacity of peptides by NF at pH 8, the potential for NF as a viable industrial fractionation process was demonstrated.
Principal component analysis (PCA) of fluorescence excitation-emission matrix (EEM) data for UF and NF peptide fractions, followed by multi-linear regression analysis, was assessed for its potential to monitor and identify the contributions to ORAC and FCR, two in vitro antioxidant capacity assays, of SPH during membrane fractionation. Two statistically significant principal components (PCs) were obtained for UF and NF peptide fractions. Multi-linear regression models (MLRM) were developed to estimate their fluorescence and PCA-captured ORAC (ORAC-FPCA) and FCR (FCR-FPCA) antioxidant capacities. The ORAC-FPCA and FCR-FPCA antioxidant capacities for NF samples displayed strong, linear relationships at different pH conditions (R-squared>0.99). Such relationships are believed to reflect the individual and relative combined contributions of tryptophan and tyrosine residues present in the SPH fractions to ORAC and FCR antioxidant capacities. Therefore, the proposed method provides a tool for the assessment of fundamental parameters of antioxidant capacities captured by ORAC and FCR assays.
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Production and fractionation of antioxidant peptides from soy protein isolate using sequential membrane ultrafiltration and nanofiltrationRanamukhaarachchi, Sahan January 2012 (has links)
Antioxidants are molecules capable of stabilizing and preventing oxidation. Certain peptides, protein hydrolysates, have shown antioxidant capacities, which are obtained once liberated from the native protein structure. Soy protein isolates (SPI) were enzymatically hydrolyzed by pepsin and pancreatin mixtures. The soy protein hydrolysates (SPH) were fractionated with sequential ultrafiltration (UF) and nanofiltration (NF) membrane steps. Heat pre-treatment of SPI at 95 degrees celsius (C) for 5 min prior to enzymatic hydrolysis was investigated for its effect on peptide distribution and antioxidant capacity. SPH were subjected to UF with a 10 kDa molecular weight cut off (MWCO) polysulfone membrane. UF permeate fractions (lower molecular weight than 10 kDa) were fractionated by NF with a thin film composite membrane (2.5 kDa MWCO) at pH 4 and 8. Similar peptide content and antioxidant capacity (α=0.05) were obtained in control and pre-heated SPH when comparing the respective UF and NF permeate and retentate fractions produced. FCR antioxidant capacities of the SPH fractions were significantly lower than their ORAC antioxidant capacities, and the distribution among the UF and NF fractions was generally different. Most UF and NF fractions displayed higher antioxidant capacities when compared to the crude SPI hydrolysates, showing the importance of molecular weight on antioxidant capacity of peptides. The permeate fractions produced by NF at pH 8 displayed the highest antioxidant capacity, expressed in terms of Trolox equivalents (TE) per total solids (TS): 5562 μmol TE/g TS for control SPH, and 5187 μmol TE/g TS for pre-heated SPH. Due to the improvement in antioxidant capacity of peptides by NF at pH 8, the potential for NF as a viable industrial fractionation process was demonstrated.
Principal component analysis (PCA) of fluorescence excitation-emission matrix (EEM) data for UF and NF peptide fractions, followed by multi-linear regression analysis, was assessed for its potential to monitor and identify the contributions to ORAC and FCR, two in vitro antioxidant capacity assays, of SPH during membrane fractionation. Two statistically significant principal components (PCs) were obtained for UF and NF peptide fractions. Multi-linear regression models (MLRM) were developed to estimate their fluorescence and PCA-captured ORAC (ORAC-FPCA) and FCR (FCR-FPCA) antioxidant capacities. The ORAC-FPCA and FCR-FPCA antioxidant capacities for NF samples displayed strong, linear relationships at different pH conditions (R-squared>0.99). Such relationships are believed to reflect the individual and relative combined contributions of tryptophan and tyrosine residues present in the SPH fractions to ORAC and FCR antioxidant capacities. Therefore, the proposed method provides a tool for the assessment of fundamental parameters of antioxidant capacities captured by ORAC and FCR assays.
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Drinking Water Disinfection Byproduct Formation Assessment Using Natural Organic Matter Fractionation and Excitation Emission MatricesJohnstone, David Weston 01 September 2009 (has links)
No description available.
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Study of the aquatic dissolved organic matter from the Seine River catchment (France) by optical spectroscopy combined to asymmetrical flow field-flow fractionation / Étude de la matière organique dissoute aquatique dans le bassin versant de la Seine (France) par spectroscopie optique combinée au fractionnement par couplage flux/force avec flux asymétriqueNguyen, Phuong Thanh 06 November 2014 (has links)
Le but principal de cette thèse était d'étudier les caractéristiques de la matière organique dissoute (MOD) dans le bassin versant de la Seine. Ce travail a été réalisé dans le cadre du programme de recherche PIREN-Seine. Les travaux présentés ici visaient plus particulièrement à identifier les sources de MOD et à suivre son évolution dans les zones d’étude. L’analyse des propriétés optiques (UV-Visible, fluorescence) de la MOD, couplée aux traitements PARAFAC et ACP, a permis de discriminer différentes sources de MOD et de mettre en évidence des variations spatio-temporelles de ses propriétés. L’axe Seine, en aval de Paris, a notamment été caractérisé par l'activité biologique la plus forte. La MOD du bassin de l’Oise a montré des caractéristiques plus "humiques", tandis que le bassin de la Marne a été caractérisé par un troisième type spécifique de MOD. Il a d’autre part été mis en évidence la présence de MODs spécifiques dans chaque zone pour les échantillons prélevés en périodes d’étiage, alors qu’une distribution homogène des composants a été obtenue pour l’ensemble des échantillons prélevés en période de crue.Le rôle environnemental des colloïdes naturels étant étroitement lié à leur taille, il a d’autre part été développé une technique analytique/séparative originale pour l’étude de ce matériel complexe, un fractionnement par couplage flux/force avec flux asymétrique (AF4). Le fractionnement par AF4 des échantillons a confirmé la variabilité spatio-temporelle en composition et en taille de la MOD d'un site de prélèvement à un autre et a permis de distinguer différentes sources de MOD colloïdale confirmant les résultats de l’étude de ses propriétés optiques. / The main goal of this thesis was to investigate the characteristics of dissolvedorganic matter (DOM) within the Seine River catchment in the Northern part of France. ThisPhD thesis was performed within the framework of the PIREN-Seine research program. Theapplication of UV/visible absorbance and EEM fluorescence spectroscopy combined toPARAFAC and PCA analyses allowed us to identify different sources of DOM andhighlighted spatial and temporal variations of DOM properties. The Seine River wascharacterized by the strongest biological activity. DOM from the Oise basin seemed to havemore "humic" characteristics, while the Marne basin was characterized by a third specifictype of DOM. For samples collected during low-water periods, the distributions of the 7components determined by PARAFAC treatment varied between the studied sub-basins,highlighting different organic materials in each zone. A homogeneous distribution of thecomponents was obtained for the samples collected in period of flood.Then, a semi-quantitative asymmetrical flow field-flow fractionation (AF4) methodology wasdeveloped to fractionate DOM. The following optimized parameters were determined: across-flow rate of 2 ml min-1 during the focus step with a focusing time of 2 min and anexponential gradient of cross-flow from 3.5 to 0.2 ml min-1 during the elution step. Thefluorescence properties of various size-based fractions of DOM were evaluated by applyingthe optimized AF4 methodology to fractionate 13 samples, selected from the three sub-basins.The fluorescence properties of these fractions were analysed, allowing us to discriminatebetween the terrestrial or autochthonous origin of DOM.
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Caracterização do carbono associado a horizontes espódicos profundos de solos da floresta amazônica, visando sua estabilidade / Characterization of carbon associated with deep podzolic horizons of soils from Amazon Forest, aiming their stabilityPaula, Bruno Santos de 16 July 2015 (has links)
Este projeto nasceu inserido num contexto de crescentes pesquisas em direção à construção de um cabedal de conhecimento acerca das ciências ambientais que tratam e estudam a dinâmica dos reservatórios de carbono e suas contribuições para o Efeito Estufa. Extrapolações de novos mapas digitalizados de solos sugerem que os espodossolos hidromórficos da Amazônia podem estar com uma contagem de carbono subestimada em até 12,3 Pg de C e trabalhos anteriores mostram que o carbono imobilizado neste perfil pode sofrer mineralização devido ao corte na rede de drenagem ou rebaixamento do nível freático, podendo assim contribuir em grande parte para o aumento de emissão dos Gases do Efeito Estufa. Nesta circunstância, coube a este trabalho, caracterizar espectroscopicamente, a matéria orgânica estocada em profundidade em 9 perfis de solo da região de São Gabriel da Cachoeira - AM, na floresta amazônica. Foram analisadas 127 amostras em quantidade de carbono e por fluorescência induzida por laser (FIL). A partir destas análises preliminares, foi possível selecionar 12 amostras de 2 perfis para análise mais detalhada através da extração das substâncias húmicas. Foram realizadas análises para obtenção da textura destes perfis, análise dos ácidos húmicos como relação C/N, espectroscopia de infravermelho com transformada de Fourier (FTIR), RMN de 13C no estado sólido, fluorescências bidimensional e tridimensional com aplicação do método estatístico análise dos fatores paralelos (PARAFAC) e também análise dos ácidos fúlvicos como carbono orgânico total (TOC), espectroscopia na região do ultra-violeta e visível e fluorescência 3D associada ao PARAFAC. Em geral, as amostras se dividiram em 3 grupos diferentes, onde a matéria orgânica em superfície se mostrou lábil e recente, a matéria orgânica nos horizontes B húmicos se mostraram com características recalcitrantes e a matéria orgânica contida nos horizontes onde existia água livre (lençol freático), se mostraram com características diferentes, indicando não serem tão jovens quanto as em superfície, porém mais jovens do que a armazenada no Bh, devido a uma possível migração por movimentos laterais do lençol freático. A correlação entre os resultados mostrou que a textura do solo é importante para o acúmulo da matéria orgânica e conseqüentemente sua humificação. Os resultados de fluorescência corroboraram a interpretação dos dados de RMN, FTIR e textura, apontando que o Perfil 4 (mais próximo ao rio) por ter C2 maior nos horizontes intermediários, contém maior quantidade de exsudatos microbianos frescos. Para o Perfil 5, que é uma mancha de Espodossolo dentro de um Latossolo e encontra-se mais longe do rio, os fluoróforos mais representativos são oriundos de anéis aromáticos mais antigos e conseqüentemente mais humificados, devido às maiores intensidades de C1 e C3. Vale ressaltar que esta interpretação se alinha com os resultados de datação do 14C desta região. Foi notório que a humificação ocorre a partir do começo do horizonte Bh e que seu acúmulo pode ocorrer tanto nos horizontes intermediários (Bh) quanto mais os profundos (úmidos). Dessa forma, pode-se concluir que uma drenagem excessiva das bordas dos rios nesta região que acumula carbono em profundidade, poderá expor a MO guardada e estocada a longo tempo, contribuindo para o aumento da temperatura global e por conseguinte catalisando um desequilíbrio no clima da sua maior reguladora no mundo: a Floresta Amazônica. / This project was born inserted in a context of increasing research towards building knowledge about environmental science that treat and study the dynamics of carbon reservoirs and their contribution to the greenhouse. Extrapolation of new digital soil maps suggests that Amazonian hydromorphic Spodosols may be with a carbon accounting underestimated by up to 12.3 Pg of C and previous work show that the carbon fixed in this profile may suffer mineralization due to the cut in the drainage network or lowering of the water level and can contribute largely to the increased emission of greenhouse gas. In this circumstance, it was instructed to this work, the spectroscopic characterization, of the stored organic matter in depth at 9 soil profiles of São Gabriel da Cachoeira - AM, in the Amazon rainforest. 127 samples were analyzed for amount of carbon and laser-induced fluorescence (LIF). From these preliminary analyzes, it was possible to select 12 samples of 2 profiles for humic substances extraction and further detailed analysis. Analyzes were conducted to obtain the texture of these profiles, analysis of humic acids as C/N ratio, Fourier Transform Infrared Spectroscopy (FTIR), solid state 13C NMR, two and three dimensional fluorescence with applying of the Parallel factors analysis (PARAFAC) statistical method and also fulvic acid analysis as Total Organic Carbon (TOC), ultra-violet and visible region spectroscopy and 3D fluorescence associated with PARAFAC. In general, the samples were divided into 3 different groups, wherein the surface organic matter evinced labile and recent. The B humic horizons organic matter proved recalcitrant characteristics and organic matter contained in horizons which had free water (groundwater), showed different characteristic, indicating they are not so young as the surface, but younger than stored in Bh, because of possible migration by lateral movements of the water table. The correlation between the results showed that soil texture is important for the accumulation of organic matter and therefore its humification. The fluorescence results confirmed the interpretation of the NMR, FTIR and texture datas, indicating that the profile 4 (closest to the river), presenting major C2 in intermediate horizons, contains larger amount of fresh microbial exudates. To profile 5, which is a Spodosol spot within a Ferrasol and is thither from the river, the most representative fluorophores came from older aromatic rings and therefore is more humified, due to higher intensities C1 and C3. It is noteworthy that this interpretation is in accord with the results of 14C dating of samples from this region. It was clear that the humification occurs from the beginning of the Bh horizon and their accumulation can occur both in the intermediate horizons (Bh) and in deeper (wet). Thus, it can be concluded that excessive drainage of the edges of rivers in this region that collects carbon in depth, may expose the OM stored for long time, contributing to increase the overall temperature and therefore catalyzing an imbalance in the climate of the biggest regulatory of it in the world: the Amazon rainforest.
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Caracterização do carbono associado a horizontes espódicos profundos de solos da floresta amazônica, visando sua estabilidade / Characterization of carbon associated with deep podzolic horizons of soils from Amazon Forest, aiming their stabilityBruno Santos de Paula 16 July 2015 (has links)
Este projeto nasceu inserido num contexto de crescentes pesquisas em direção à construção de um cabedal de conhecimento acerca das ciências ambientais que tratam e estudam a dinâmica dos reservatórios de carbono e suas contribuições para o Efeito Estufa. Extrapolações de novos mapas digitalizados de solos sugerem que os espodossolos hidromórficos da Amazônia podem estar com uma contagem de carbono subestimada em até 12,3 Pg de C e trabalhos anteriores mostram que o carbono imobilizado neste perfil pode sofrer mineralização devido ao corte na rede de drenagem ou rebaixamento do nível freático, podendo assim contribuir em grande parte para o aumento de emissão dos Gases do Efeito Estufa. Nesta circunstância, coube a este trabalho, caracterizar espectroscopicamente, a matéria orgânica estocada em profundidade em 9 perfis de solo da região de São Gabriel da Cachoeira - AM, na floresta amazônica. Foram analisadas 127 amostras em quantidade de carbono e por fluorescência induzida por laser (FIL). A partir destas análises preliminares, foi possível selecionar 12 amostras de 2 perfis para análise mais detalhada através da extração das substâncias húmicas. Foram realizadas análises para obtenção da textura destes perfis, análise dos ácidos húmicos como relação C/N, espectroscopia de infravermelho com transformada de Fourier (FTIR), RMN de 13C no estado sólido, fluorescências bidimensional e tridimensional com aplicação do método estatístico análise dos fatores paralelos (PARAFAC) e também análise dos ácidos fúlvicos como carbono orgânico total (TOC), espectroscopia na região do ultra-violeta e visível e fluorescência 3D associada ao PARAFAC. Em geral, as amostras se dividiram em 3 grupos diferentes, onde a matéria orgânica em superfície se mostrou lábil e recente, a matéria orgânica nos horizontes B húmicos se mostraram com características recalcitrantes e a matéria orgânica contida nos horizontes onde existia água livre (lençol freático), se mostraram com características diferentes, indicando não serem tão jovens quanto as em superfície, porém mais jovens do que a armazenada no Bh, devido a uma possível migração por movimentos laterais do lençol freático. A correlação entre os resultados mostrou que a textura do solo é importante para o acúmulo da matéria orgânica e conseqüentemente sua humificação. Os resultados de fluorescência corroboraram a interpretação dos dados de RMN, FTIR e textura, apontando que o Perfil 4 (mais próximo ao rio) por ter C2 maior nos horizontes intermediários, contém maior quantidade de exsudatos microbianos frescos. Para o Perfil 5, que é uma mancha de Espodossolo dentro de um Latossolo e encontra-se mais longe do rio, os fluoróforos mais representativos são oriundos de anéis aromáticos mais antigos e conseqüentemente mais humificados, devido às maiores intensidades de C1 e C3. Vale ressaltar que esta interpretação se alinha com os resultados de datação do 14C desta região. Foi notório que a humificação ocorre a partir do começo do horizonte Bh e que seu acúmulo pode ocorrer tanto nos horizontes intermediários (Bh) quanto mais os profundos (úmidos). Dessa forma, pode-se concluir que uma drenagem excessiva das bordas dos rios nesta região que acumula carbono em profundidade, poderá expor a MO guardada e estocada a longo tempo, contribuindo para o aumento da temperatura global e por conseguinte catalisando um desequilíbrio no clima da sua maior reguladora no mundo: a Floresta Amazônica. / This project was born inserted in a context of increasing research towards building knowledge about environmental science that treat and study the dynamics of carbon reservoirs and their contribution to the greenhouse. Extrapolation of new digital soil maps suggests that Amazonian hydromorphic Spodosols may be with a carbon accounting underestimated by up to 12.3 Pg of C and previous work show that the carbon fixed in this profile may suffer mineralization due to the cut in the drainage network or lowering of the water level and can contribute largely to the increased emission of greenhouse gas. In this circumstance, it was instructed to this work, the spectroscopic characterization, of the stored organic matter in depth at 9 soil profiles of São Gabriel da Cachoeira - AM, in the Amazon rainforest. 127 samples were analyzed for amount of carbon and laser-induced fluorescence (LIF). From these preliminary analyzes, it was possible to select 12 samples of 2 profiles for humic substances extraction and further detailed analysis. Analyzes were conducted to obtain the texture of these profiles, analysis of humic acids as C/N ratio, Fourier Transform Infrared Spectroscopy (FTIR), solid state 13C NMR, two and three dimensional fluorescence with applying of the Parallel factors analysis (PARAFAC) statistical method and also fulvic acid analysis as Total Organic Carbon (TOC), ultra-violet and visible region spectroscopy and 3D fluorescence associated with PARAFAC. In general, the samples were divided into 3 different groups, wherein the surface organic matter evinced labile and recent. The B humic horizons organic matter proved recalcitrant characteristics and organic matter contained in horizons which had free water (groundwater), showed different characteristic, indicating they are not so young as the surface, but younger than stored in Bh, because of possible migration by lateral movements of the water table. The correlation between the results showed that soil texture is important for the accumulation of organic matter and therefore its humification. The fluorescence results confirmed the interpretation of the NMR, FTIR and texture datas, indicating that the profile 4 (closest to the river), presenting major C2 in intermediate horizons, contains larger amount of fresh microbial exudates. To profile 5, which is a Spodosol spot within a Ferrasol and is thither from the river, the most representative fluorophores came from older aromatic rings and therefore is more humified, due to higher intensities C1 and C3. It is noteworthy that this interpretation is in accord with the results of 14C dating of samples from this region. It was clear that the humification occurs from the beginning of the Bh horizon and their accumulation can occur both in the intermediate horizons (Bh) and in deeper (wet). Thus, it can be concluded that excessive drainage of the edges of rivers in this region that collects carbon in depth, may expose the OM stored for long time, contributing to increase the overall temperature and therefore catalyzing an imbalance in the climate of the biggest regulatory of it in the world: the Amazon rainforest.
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