京都大学 / 0048 / 新制・課程博士 / 博士(農学) / 甲第19025号 / 農博第2103号 / 新制||農||1030(附属図書館) / 学位論文||H27||N4907(農学部図書室) / 31976 / 京都大学大学院農学研究科応用生命科学専攻 / (主査)教授 加納 健司, 教授 植田 和光, 教授 植田 充美 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DFAM

No description available.

No description available.

L'idée de base est de permettre à deux microorganismes de partager le même environnement tout en les maintenant séparées à l'aide d'une membrane perméable les retenant sélectivement. La principale contrainte résulte du transfert des composées d'intérêts limité par l'écoulement dans et autour des fibres ainsi que dans module et par le colmatage. Le double bioréacteur a membrane étudié dans cette thèse, de par son fonctionnement, alterne les cycles de filtration et rétrofiltration (ou rétrolavage), limitant ainsi en partie le colmatage. Ce travail de thèse s'est donc attaché à approfondir la connaissance des mécanismes de limitation au transfert mis en jeu lors de la filtration de fluide biologique complexes et évolutifs en fonction des conditions opératoires et des caractéristiques géométriques du module de filtration à fibres creuses. Dans cet objectif, sur la base des choix de configuration de module membranaire proposés dans cette étude, et afin de tendre vers une optimisation rationnelle de l'utilisation de ce dispositif, l'étude s'appuya sur l'utilisation d'outils de mécanique des fluides numériques, complétée par une approche expérimentale menée dans des conditions modèles. Les simulations réalisées par cette approche ont ainsi mis en évidence de grandes variations des vitesses de filtration le long de la fibre et ceci en lien direct avec une augmentation de la perte de charge à l'extérieur des fibres due au confinement induisant une baisse des performances de filtration. De manière similaire, un modèle numérique de formation de dépôt nous a permis d'évaluer l'effet du confinement de fibres. Il entraine une augmentation de pression dans la partie fluide externe induisant une forte variation de pertes de charges entrainant une répartition du dépôt le long de la fibre beaucoup plus inhomogène. Le retour du numérique à l'expérimental réalisé s'est attaché à décrire l'influence des conditions de mise oeuvre sur les performances de filtration du pilote. L'analyse méthodique de l'influence du sens de filtration et de la compacité dans le cas de fluides modèles (suspension de différents microorganismes / solutions de protéines modèles) et dans le cas de fluides biologiques évolutifs (milieux de fermentation + micro organismes) fut réalisée. L'ensemble de ces résultats nous permettent de donner des recommandations aux futurs utilisateurs du double bioréacteur à membranes. / This work presents a specific bioreactor previously designed to study microbial interactions. In this process, the microbial species in two tanks are physically separated by a microfiltration membrane. In order to give to the microorganisms a molecular environment in each compartment similar to the one that would be obtained if the microbial cells were cultivated in the same reactor, two criteria have to be considered: (i) the flow rates between compartments have to be sufficient with respect to the microbial kinetics and (ii) all the molecular compounds of the medium that have an effect on the microorganism behaviour must pass through the membrane. The main constrain is due to transfer of component limited by the fluid flow in and around the fiber of the filtration module. This thesis has therefore committed to deepening the understanding of the mechanisms limiting the transfer involved during the filtration of biological fluid complex according to operating conditions and geometric characteristics of the hollow fiber module of filtration. For this purpose, based on the choice of membrane module configuration proposed in this study, and to strive for a rational optimization of the use of this device, the study relied on the use of CFD tools, supplemented by an experimental approach conducted under models conditions. The numerical simulations of fluid flow have shown a modification of the axial filtration velocity profile with packing density. Similarly, a numerical model of cake deposit was developed and show difference of cake growth along the fiber with packing density. Two experimental hollow fiber modules with two packing densities were tested with clean water and biological fluid, and showed good agreement with the numerical data. These results underline the variations of filtration velocity along the fiber that will allow some predictions on fouling deposit to be done.

Cette thèse présente une approche originale du procédé de distillation membranaire avec balayage gazeux pour la concentration des solutions thermosensibles (SGMD). Pour ce faire, un nouveau contacteur membranaire avec des fibres creuses métalliques a été conçu afin réaliser le procédé de distillation à basse température. La chaleur nécessaire au procédé est produite au niveau des fibres par effet Joule, plutôt qu'à partir de chaleur latente de la phase aqueuse. La génération localisée de la chaleur a comme conséquence une réduction du phénomène de polarisation de la température. Des fibres creuses en acier inoxydable ont été synthétisées avec les propriétés structurales appropriées et une bonne résistance mécanique. La surface des pores des fibres a été rendue hydrophobe par le dépôt d'une fine couche d'un élastomère. En outre, une nouvelle méthode « verte » a été développée pour fabriquer des fibres creuses en alumine et acier inoxydable. Cette méthode est basée sur la gélification ionique des bio-polymères et ne n'utilise pas des solvants nocifs. L'étude expérimentale détaillée du SGMD a permis de déterminer l'influence de différents paramètres opérationnels sur les performances du procédé. Il a été démontré que l'effet Joule permet d'améliorer le flux et l'efficacité de la séparation non seulement pour le SGMD mais aussi pour la pervaporation. / This thesis presents an original approach for the concentration of thermo-sensitive solutions: the Sweep Gas Membrane Distillation (SGMD) process. A new membrane contactor with metallic hollow fibers has been designed and allows the distillation process to be operational at low temperature. Heat is generated in the fibers by the Joule effect, rather than being supplied as latent heat in the liquid bulk. The localized generation of heat results in a reduction of temperature polarization phenomena. The stainless-steel hollow fiber membranes have been synthetized with appropriate structural properties and sufficient mechanical strength. The pore surface of the fibers has been made hydrophobic by the deposition of a thin layer of an elastomer. Moreover, a novel and green method is presented to fabricate alumina and stainless-steel hollow fibers. This method is based on ionic gelation of a biopolymer and completely avoids the use of harmful solvents. By a detailed experimental study of the SGMD the influence of different operational parameters on the process performance has been investigated. The improvements in the flux and the separation efficiency using Joule effect have been successfully demonstrated, even in the case of pervaporation.

Estudos de metabolismo in vitro possuem o intuito de caracterizar e quantificar possíveis metabólitos, elucidar as vias metabólicas e sugerir modelos a serem seguidos para a realização de estudos in vivo. Com o intuito de estudar o metabolismo in vitro não estereosseletivo da rosiglitazona (RSG) empregando fração microssomal de fígado de ratos,foi desenvolvida uma metodologia por cromatografia líquida de alta eficiência (HPLC) com detecção UV em 245 nm, para analisar a RSG e seus principais metabólitos, p-hidroxi rosiglitazona (?-OH-R) e N-desmetil rosiglitazona (N-Dm-R). Os analitos foram separados em fase reversa, utilizando uma coluna X-Terra MS C-18 (partículas de 3,5 ?m) e fase móvel composta por água:acetonitrila:ácido acético (85:15:0,5, v/v/v), na vazão de 1 mL min-1. Matrizes biológicas contém um grande excesso de proteínas, lipídeos e outros materiais endógenos que interferem na análise de fármacos e metabólitos, tornando necessário um procedimento adequado de preparação das amostras antes da análise cromatográfica. A microextração em fase líquida com membrana cilíndrica oca (HF-LPME) é uma técnica promissora para a preparação de amostras em estudos de metabolismo in vitro, pois, além de promover o clean-up, promove também o enriquecimento dos analitos na amostra. A HF-LPME foi aplicada pela primeira vez para a extração simultânea desse fármaco e seus metabólitos. O sistema de três fases foi escolhido como o mais apropriado, empregando uma solução de ácido clorídrico como fase aceptora e 1-octanol como solvente orgânico. A otimização dos demais parâmetros foi realizada através de planejamento fatorial fracionário. O método foi validado e foi linear no intervalo de 50-6000 ng mL-1, apresentando limites de quantificação de 50 ng mL-1 e recuperações acima de 47 % para a RSG e seus metabólitos (?-OH-R e N-Dm-R). O método validado foi empregado em um estudo de metabolismo in vitro com fração microssomal de fígado de ratos. Nesse estudo, foi possível estimar as constantes de Michaelis-Menten (Km) e a velocidade inicial máxima (Vmax). N-Dm-R e ?-OH-R apresentaram valores de Vmax de 87,30 ± 8,04 e 51,64 ± 12,25 ?mol min-1 mg proteína-1, respectivamente, enquanto que os valores de Km foram de 58,14 ± 11,85 e 77,84 ± 36,77 mmol L-1, respectivamente. Outros metabólitos foram observados nos cromatogramas e a identificação foi feita por espectrometria de massas: ?rto-hidroxi-rosiglitazona e N-desmetil-hidroxi-rosiglitazona. A RSG é comercializada como uma mistura racêmica, apesar de possuir sua atividade antidiabética relacionada essencialmente com o enantiômero (S). O centro quiral desse fármaco possui um grupo carbonila, por isso, o enantiômero (R) pode se converter no enantiômero (S) ou vice-versa, via tautomerismo cetoenólico. Dados da literatura indicavam que essa racemização poderia ser lenta o suficiente para possibilitar o estudo dos enantiômeros isoladamente. Entretanto, até o momento não há dados sobre a disposição cinética ii e metabolismo enantiosseletivos desse fármaco. Sendo assim, propôs-se o desenvolvimento de metodologias analíticas para estudar a racemização da RSG e seus metabólitos e avaliar a possibilidade de estudar seu metabolismo in vitro de forma estereosseletiva. O método foi desenvolvido empregando HPLC com detecção em 245 nm. A separação dos enantiômeros do fármaco e metabólitos, também quirais, foi obtida empregando uma coluna Chiralcel OJ-H e fase móvel constituída por metanol:etanol (90:10; v/v), na vazão de 0,3 mL min-1. O estudo de racemização mostrou que o fármaco e seus metabólitos são racemizados nas condições em que o estudo de metabolismo é conduzido. Finalmente, para estudar o metabolismo in vitro da pioglitazona (PGZ), foi desenvolvido um método para análise desse fármaco e de seus principais metabólitos, a hidroxi-pioglitazona (M-IV) e a ceto-pioglitazona (M-III) empregando a eletroforese capilar (CE). As análises foram realizadas em capilar de sílica de 50 ?m de diâmetro interno e com comprimento efetivo de 40 cm, utilizando tampão fosfato de sódio 50 mmol L-1, pH 2,5, detecção em 190 nm, tensão de 30 kV e temperatura do capilar de 35 °C. A HF-LPME também foi empregada para a preparação das amostras. O sistema de três fases foi escolhido, empregando solução de ácido clorídrico como fase aceptora e o 1-octanol como solvente orgânico. A otimização dos demais parâmetros foi realizada através de planejamento fatorial fracionário. O método validado foi linear no intervalo de 200 - 25000 ng mL-1 para PGZ e 200 - 2000 ng mL-1 para os metabólitos, apresentando limites de quantificação de 200 ng mL-1 e recuperações acima de 19 % para a RSG e seus metabólitos M-IV e M-III. O método validado foi empregado em um estudo de metabolismo in vitro contendo fração microssomal de fígado de ratos, mas nesse estudo, não foi possível observar a formação dos metabólitos. Entretanto, esse método pode ser usado em outros modelos de metabolismo in vitro (microssomas humanos), nos quais se observa a formação desses metabólitos em concentrações maiores. / In vitro metabolism studies have been used to characterize and to quantify possible metabolites, to elucidate metabolic pathways and to suggest models to perform in vivo studies. So the purpose of this study was to evaluate the in vitro rosiglitazone (RSG) metabolism employing microsomal fraction obtained from rat livers. A high- performance liquid chromatography (HPLC) method with UV detection at 245 nm was developed to analyze RSG and the main metabolites, p-hydroxy rosiglitazone (?-OH-R) e N-desmethyl rosiglitazone (N-Dm-R). The analytes were separated under reversed phase conditions, using a X-Terra MS C-18 column (3.5 ?m particle size) and a mobile phase consisting of water:acetonitrile:acetic acid (85:15:0.5, v/v/v), at a flow rate of 1 mL min-1. Biological matrices contain a large excess of proteins, lipids and other endogenous compounds that interfere in the analysis of drugs and metabolites. So, a suitable sample preparation technique is required. Hollow-fiber liquid phase microextraction (HF-LPME) is a promising technique for the preparation of biological samples. Besides the clean-up, analytes enrichment is also achieved. HF-LPME for the simultaneous analysis of RSG and its main metabolites is described for the first time. The three-phase extraction was performed using hydrochloride acid solution as acceptor phase and 1-octanol as organic solvent. The other parameters were optimized by fractional factorial design. The method was validated and it was linear over the concentration range of 50-6000 ng mL-1, with quantification limits of 50 ng mL-1 and recoveries above 47 %. The validated method was used to estimate Michaelis-Menten (Km) constant and maximum initial velocity (Vmax). N-Dm-R e ?-OH-R showed Vmax values of 87.30 ± 8.04 and 51.64 ± 12.25 ?mol min-1 mg protein-1, respectively, while the Kmvalues were 58.14 ± 11.85 e 77.84 ± 36.77 mmol L-1, respectively. Other possible metabolites were observed in the chromatograms and they were identified by mass spectrometry: ?rtho-hydroxy-rosiglitazone e N-desmethyl-hydroxy-rosiglitazone. RSG is marketed as a racemic mixture although the antidiabetic activity is related essentially to the (S)-enantiomer. The chiral center has a carbonyl group, therefore the (R)-enantiomer could be transformed to the (S)-enantiomer or vice-versa by keto-enolic tautomerism. The literature indicates that this racemization is slow enough to allow the evaluation of the properties of the isolated enantiomers. However, there is no information in the literature about enantioselective RSG kinetic disposition and metabolism. Considering this facts, an analytical procedure was developed to study the racemization of RSG and its metabolites under different conditions and to determine if the enantioselective metabolism would be performed. The method was developed by HPLC with detection at 245 nm. The chiral separation of RSG and metabolites was achieved on a Chiralcel OJ-H column, with the mobile phase consisting of methanol:ethanol (90:10,v/v). The results obtained showed that the racemization occurs under the conditions used in in vitro iv metabolism studies. Finally, to study the in vitro metabolism of pioglitazone (PGZ), another method was developed by capillary electrophoresis (CE) to determine this drug and its main metabolites, hydroxy-pioglitazone (M-IV) and keto-pioglitazone (M-III). The analyses were conducted using a fused silica capillary (50 ?m inner diameter and 40 cm effective length), sodium phosphate buffer 50 mmol L-1, pH 2.5, detection at 190 nm, voltage of 30 kV and capillary temperature of 35°C. HF-LPME was also used for sample preparation with hydrochloride acid solution as acceptor phase and 1-octanol as organic solvent. The other parameters were optimized by fractional factorial design. The method was validated showing to be linear in the concentration range of 200 - 25000 ng mL-1 for PGZ and 200 - 2000 ng mL-1 for the metabolites. Quantification limits were 200 ng mL-1 for all analytes and the recoveries were higher than 19%. The validated method was used to study the in vitro metabolism of PGZ by rat liver microsomal fraction, but it was not possible to observe the formation of the metabolites in this study. However this method could be used in other in vitro metabolism models (human microssomes), in which higher concentrations of these metabolites are observed. Keywords:

La capture du dioxyde de carbone (CO2) en postcombustion est une stratégie importante pour la limitation de l’effet de serre. Le procédé de référence est l’absorption du CO2 dans des solutions aqueuses aminées, suivie par une étape de stripage du solvant. La technologie mature associée à ce procédé est la colonne à garnissage. Toutefois, afin de rendre le procédé plus attractif, il convient de l’intensifier en réduisant le volume des équipements et le coût énergétique associé. Les contacteurs membranaires à fibres creuses (CMFC) constituent une alternative aux colonnes à garnissage. Les CMFC permettent de développer d’importantes aires spécifiques conduisant potentiellement à une intensification des transferts gaz-liquide. Ainsi, l’utilisation des CMFC réduirait la taille des installations, mais aussi diminuerait la consommation énergétique par la diminution de la quantité de vapeur de stripage. Cependant, l’utilisation de CMFC dans les étapes d’absorption et de stripage dans des conditions industrielles a été peu étudiée. Afin de combler cette lacune, des modèles à différents niveaux de complexité : monodimensionnel, bidimensionnel, isotherme et adiabatique ont été développés, comparés et validés. Ceci afin d’identifier le niveau de complexité approprié. Les résultats de simulation ont mis en évidence le potentiel d’intensification des CMFC dans l’étape d’absorption et aussi de stripage, se traduisant par une réduction en volume de 4 à 10 fois par rapport aux colonnes à garnissage. Néanmoins, les CMFC peuvent difficilement réduire le coût énergétique du procédé étant donné que l’étape de stripage fonctionne dans des conditions très proches de la limite thermodynamique / Post-combustion CO2 capture (PCC) is an important strategy in mitigating greenhouse effect. The reference process in PCC is the CO2 absorption into amine aqueous solutions, followed by the regeneration (or stripping) of the solvent. The robustness of packed columns makes it the standard technology for both absorption and stripping steps. However, the treatment of large quantities of flue gases requires itself equipment of a large size. Hollow fibre membrane contactors (HFMC) are considered as one of the most promising strategies for intensified CO2 absorption process, due to their significantly higher interfacial area than that of packed columns, allowing to reduce the equipment size. In addition, this would reduce the energy penalty of the process by reducing the required amount of stripping steam. However, despite the potential advantages of HFMC, very few investigations have studied implementing this technology for PCC within an industrial framework. To fill this lack, the performances of both absorption and stripping steps using HFMC under industrial conditions were estimated by modelling and simulation. To identify the optimal modelling strategy, transfer models with different levels of complexity were developed ranging from one-dimensional isothermal single-component to two-dimensional adiabatic multi-component. Simulation results of both absorption and stripping steps revealed that, compared to traditional packed columns, contactor volume reduction factors comprised between 4 and 10 might be achieved using HFMC. However, since the stripping operating conditions are very close to thermodynamic equilibrium, HFMC can hardly reduce the energy consumption of the process

Commercial pure terephthalic acid (PTA) manufacturing generates process streams mainly containing acetic acid (HAc) and water. A large financial incentive exists to replace the costly and energy intensive distillation column used to recycle HAc-water mixtures. This work focuses on the development of pervaporation technology to separate HAc-water mixtures using a hollow fiber-based membrane unit. Currently a 250 m outer diameter Matrimid® hollow fiber is used in industry for gas separation. Due to the difference between gas and liquid separations, the fiber performance associated with high flux in pervaporation is limited by a pressure change inside the bore along the axial direction of the fiber. A mathematical model was developed to describe the bore pressure change in pervaporation in this work, which demonstrated that spinning a large bore size fiber was a good solution to minimize the bore pressure change. Spinning technology has been adapted to obtain a large bore size defect-free Matrimid® hollow fiber. In addition to a large bore size, the asymmetric fiber exhibits an intrinsically defect-free selective layer supported on an open porous substrate. This eliminates the post-treatment with a caulking layer and has a special advantage for aggressive liquid separation. A proof of concept was provided by testing both small and large bore size defect-free fibers with a model 20% wt HAc feed in a pervaporation system at 101.5oC. The membrane selectivity (~ 25) and water flux (~ 4.5 kg/m2hr) were increased by about 150% with a diameter (O.D. ~ 500 m) twice as large as the regular fiber. Further, a decrease in the HAc flux was observed with the increased bore size due to the reduction in HAc-induced plasticization. Sub-Tg thermal annealing was used to stabilize the fiber by suppressing HAc-induced plasticization. This improves the polymer discrimination of shape and size for penetrants although no chemical reaction occurs with thermal annealing. The resulting membrane selectivity was increased from 10 to about 95 using a large bore size defect-free annealed fiber with acceptable water flux (~ 1.5 kg/m2hr) for 20% wt HAc concentration feed streams. These improvements make Matrimid® hollow fiber membranes very attractive for future scale-up and commercial development.

Mixed matrix hollow fiber membranes maintain the ease of processing polymers while enhancing the separation performance of the pure polymer due to inclusion of molecular sieve filler particles. This work shows the development process of high loading mixed matrix hollow fiber membranes for butane isomer separation, from material selection and engineering of polymer-sieve interfacial adhesion to mixed matrix hollow fiber spinning. The matching of gas transport properties in polymer and zeolite is critical for forming successful mixed matrix membranes. The nC4 permeability in glassy commercial polymers such as Ultem® and Matrimid® is too low (< 0.1 Barrer) for commercial application. A group of fluorinated (6FDA) polyimides, with high nC4 permeability and nC4/iC4 selectivity, are selected as the polymer matrix. No glassy polymers can possibly match the high permeable MFI to make mixed matrix membranes with selectivity enhancement for C4s separation. Zeolite 5A, which has a nC4 permeability (~3 Barrer) and nC4/iC4 selectivity (essentially ∞), matches well with the 6FDA polymers. A 24% nC4/iC4 selectivity enhancement was achieved in mixed matrix membranes containing 6FDA-DAM and 25 wt% treated 5A particles. A more promising mixed matrix membrane contains 6FDA-DAM-DABA matrix and 5A, because of a better match of gas transport properties in polymer and zeolite. Dual layer hollow fibers, with cellulose acetate core layer and sheath layers of 6FDA polyimides, were successfully fabricated. Successive engineering of the 6FDA sheath layer and the dense skin is needed for the challenging C4s separation, which is extremely sensitive to the integrity of the dense skin layer. The delamination-free, macrovoid-free dual layer hollow fiber membranes provide the solution for the expensive 6FDA polyimides spinning. Mixed matrix hollow fiber membranes are spun base on the platform of 6FDA/Cellulose acetate dual layer hollow fibers. Preliminary results suggest that high loading mixed matrix hollow fiber membranes for C4s is feasible. Following research is needed on the fiber spinning with well treated zeolite 5A nanoparticles. The key aspect of this research is elucidating the three-step (sol-gel-precipitation) mechanism of sol-gel-Grignard treatment, based on which further controlling of Mg(OH)2 whisker morphologies is possible. A Mg(OH)2 nucleation process promoted by acid species is proposed to explain the heterogeneous Mg(OH)2 growing process. Different acid species were tried: 1) HCl solution, 2) AlClx species generated by dealumination process and 3) AlCl3 supported on zeolite surfaces. Acids introduced through HCl solution and dealumination are effective on commercial 5A particles to generate Mg(OH)2 whiskers in the sol-gel-Grignard treatment. Supported AlCl3 is effective on both commercial and synthesized 5A particles (150 nm-1 µm) during the sol-gel-Grignard treatment, in terms of promoting heterogeneous Mg(OH)2 whiskers formation. But the byproduct of Al(OH)3 layer separates the Mg(OH)2 whiskers from zeolite surface, and leads to undesirable morphologies for polymer-zeolite interfacial adhesion. The elucidation of sol-gel-Grignard mechanism and importance of zeolite surface acidity on Mg(OH)2 formation, builds a solid foundation for future development towards ''universal'' method of growing Mg(OH)2 whiskers on zeolite surfaces.

Olefin/paraffin separation is a large potential market for membrane applications. Carbon molecular sieve membranes (CMS) are promising for this application due to the intrinsically high separation performance and the viability for practical scale-up. Intrinsically high separation performance of CMS membranes for olefin/paraffin separations was demonstrated. The translation of intrinsic CMS transport properties into the hollow fiber configuration is considered in detail. Substructure collapse of asymmetric hollow fibers was found during Matrimidﾮ CMS hollow fiber formation. To overcome the permeance loss due to the increased separation layer thickness, 6FDA-DAM and 6FDA/BPDA-DAM polyimides with higher rigidity were employed as alternative precursors, and significant improvement has been achieved. Besides the macroscopic morphology control of asymmetric hollow fibers, the micro-structure was tuned by optimizing pyrolysis temperature protocol and pyrolysis atmosphere. In addition, unexpected physical aging was observed in CMS membranes, which is analogous to the aging phenomenon in glassy polymers. For performance evaluation, multiple "proof-of-concept" tests validated the viability of CMS membranes under realistic conditions. The scope of this work was expanded from binary ethylene/ethane and propylene/propane separations for the debottlenecking purpose to mixed carbon number hydrocarbon processing. CMS membranes were found to be olefins-selective over corresponding paraffins; moreover, CMS membranes are able to effectively fractionate the complex cracked gas stream in a preferable way. Reconfiguration of the hydrocarbon processing in ethylene plants is possible based on the unique CMS membranes.