A glicerina é um composto de grande utilidade em muitas áreas de aplicação. Atualmente, a maior fonte de glicerina é como subproduto da produção do biodiesel. Em 2010 o Brasil foi o segundo maior produtor de biodiesel no mundo com 2,4 milhões de m3. Estima-se que 10% da produção de biodiesel é glicerina bruta e destes 10%, aproximadamente entre 30% e 60% correspondem à glicerol. As impurezas são formadas por água, sais orgânicos e inorgânicos, ésteres e álcoois, e traços de glicerídeos. Como as aplicações mais nobres da glicerina requerem uma glicerina isenta de impurezas, novas rotas de purificação da glicerina bruta vem sendo estudadas. Neste sentido, o Laboratório de Separações Térmicas e Mecânicas da EPUSP propôs uma nova rota de purificação em 2008, na qual a ultima etapa é a desidratação de uma solução glicerol-água por meio de destilação trifásica usando tolueno como arrastador. O objetivo do presente trabalho foi estudar o comportamento deste tipo de destilação em uma coluna modulada com três pratos perfurados. Nesta coluna avaliou-se o layout de pratos e as melhores condições de operação. Os resultados, obtidos com este estudo, constituem uma contribuição importante para o projeto básico de coluna de destilação trifásica. As séries experimentais foram planejadas sequencialmente empregando-se planejamento estatístico de experimentos (DOE). Como variáveis de processo foram estudadas a vazão de vapor do tolueno (kg/h), vazão de alimentação de glicerina (kg/h) e concentração de glicerol na alimentação (% em massa). Como variáveis geométricas foram estudadas a área livre do prato () e a altura de vertedouro (Hw). O desempenho da coluna foi avaliado mediante o incremento da concentração de glicerol (em relação à concentração de glicerol na alimentação) e a concentração de glicerol no fundo da coluna (estas duas variáveis de resposta são dependentes entre si). A influência das variáveis de processo e geométricas nas respostas estudadas permitiram a construção de modelos estatísticos, e o melhor deles foi comparado com os modelos de não equilíbrio (NEQ) e de equilíbrio (EQ) obtidos por simulação no programa ASPEN PLUS. O modelo de não equilíbrio está baseado nas equações de Maxwell-Stefan, que utiliza a abordagem de Eckert e Vanek (2001) e a correlação de Chen-Chuang (1993), para estimar os coeficientes binários de transporte de massa. O modelo de equilíbrio utiliza as equações MESH (Material balance, Equilibrium, mole fraction Summation, and Heat balance). Conclusivamente, pode-se afirmar que as variáveis operacionais exercem maior influência do que as variáveis geométricas na desidratação da glicerina. As melhores condições foram: a vazão de vapor de tolueno de 23,5 kg/h, vazão de alimentação de glicerina de 2,2 kg/h e concentração de glicerol na alimentação de 50 % glicerol em massa, usando o layout L5 com área livre de 0,04 e altura de vertedouro de 70 mm. Os valores preditos pelo modelo estatístico (obtido com dados experimentais) e pelo modelo de NEQ representaram o comportamento da desidratação da glicerina por destilação trifásica à pressão atmosférica usando tolueno como arrastador, na coluna de pratos perfurados estudada. O modelo de EQ superestimou os valores reais. / Glycerine is a material of outstanding utility with many areas of application. Currently, the largest source of glycerine is as a by-product of biodiesel production. In 2010, Brazil was the second largest biodiesel productor in the world with 2.4 million m3. It is estimated that 10% of biodiesel is raw glycerine, and of this 10%, approximately between 30% and 60% corresponds to glycerol. The impurities are formed by water, organic and inorganic salts, esters and alcohols, and traces of glycerides. Although many attempts have been made to use raw glycerine, it is still necessary to purify it for of the most applications; hence new routes of glycerine purification have been studied. In this way, the Laboratory of Thermal and Mechanical Separations of EPUSP have proposed a new route of purification in 2008, where the last step is the dehydration of glycerol-water solution by three-phase distillation using toluene as entrainer. The aim of the present work was to study the behavior of the three-phase distillation using a modulated column with three sieve trays. In this column, the tray layout and the best operating conditions were evaluated. The results achieved in this study are an important contribution to the basic design of three-phase distillation column. Experimental series were designed sequentially employing a statistical design of experiments (DOE). The process variables studied were the vapor flow rate of toluene (kg/h), feed flow rate of glycerine (kg/h) and the feed glycerol concentration (% wt.). The geometric variables studied were the fractional hole area () and the weir height (Hw). The column performance was evaluated by the increment of glycerol concentration (in relation to the feed glycerol concentration) and the bottom glycerol concentration (these two variables are mutually dependent responses). The influence of process and geometric variables allowed the empirical models building, in which the best model was compared with the non-equilibrium (NEQ) and equilibrium (EQ) models obtained by the simulator ASPEN PLUS. The non-equilibrium model is based on Stefan-Maxwell equations, which uses the approach of Eckert and Vanek\'s (2001) and the Chen-Chuang correlation (1993), to estimate the binary coefficients of mass transport. The equilibrium model is based on the MESH equations (Material balance, Equilibrium, mole fraction Summation, and Heat balance). Finally, it can be stated that the operating variables have more influence than the geometric variables in the glycerine dehydration. The best conditions were: vapor flow rate of toluene of 23.5 kg/h, feed flow rate of glycerine of 2.2 kg/h and feed glycerol concentration of 50% wt., using the fractional hole area of L5 of 0.04 and weir height of 70 mm. The predicted values obtained by the statistical model and by the non-equilibrium model represented the behavior of the glycerine dehydration by three-phase distillation at atmospheric pressure using toluene as entrainer in the sieve tray column studied. The equilibrium model (EQ) overpredicted the real values.

CoordenaÃÃo de AperfeÃoamento de Pessoal de NÃvel Superior / Aromatic hydrocarbons are chemical compounds of great interest in the petrochemical industry. These are commonly derived from reformed naphtha, which are in multicomponent mixtures with aliphatic hydrocarbons. The separation of these components is realized, generally, the liquid-liquid extraction process. In this process, the sulfolane is the most widely used solvent. However, this has some disadvantages such as high cost. For this reason, many studies are developed by the academic community in order to propose alternative solvents to replace the exclusive use of sulfolane by compounds that are important features like good selectivity, easy recoverability and low costs. Given the above, this study main aims to determine liquid-liquid equilibrium data (LLE) to study the ability of solvents [diethylene glycol (DEG), triethylene glycol (TEG), glycerol and 2-hydroxyethyl amine format (2-HEAF)] in extracting aromatic mixtures with aliphatic. The physical properties also of great importance in the understanding of the behavior and interactions that occur in liquid mixtures. Thus, in this study were also studied the density and viscosity of solvent extractors aromatic. The experimental methodology for the determination of LLE data was initially validated by reproducing the literature, and the results showed good agreement between these data. After the method has been validated, solubility curves and tie-lines were constructed for systems containing decane + toluene + DEG (+ TEG, + glycerol, + 2-HEAF, + sulfolane) at T = (303.15 and 333.15) K. The composition data of tie-lines, obtained experimentally, were used to calculate the selectivity parameter and the distribution coefficient. With this data, it can be concluded that all the solvents studied have a good capacity for toluene extraction from mixtures with decane. However, the glycerol has presented as the most promising solvent for aromatics extraction. Finally, the experimental data were correlated through the thermodynamic activity coefficient models, NRTL and UNIQUAC. The results were considered satisfactory, noting that the NRTL was the model that presented minor deviations in compositions. Thus, best represented the LLE data for the systems studied. / Os hidrocarbonetos aromÃticos sÃo compostos quÃmicos de grande interesse na indÃstria petroquÃmica. Estes sÃo comumente originados da nafta reformada, onde encontram-se em misturas multicomponentes com hidrocarbonetos alifÃticos. A separaÃÃo destes componentes à realizada, em sua grande maioria, pelo processo de extraÃÃo lÃquido-lÃquido. Nesse processo, o sulfolano à o solvente mais utilizado. No entanto, este apresenta algumas desvantagens, tais como alto custo. Por esse motivo, inÃmeros estudos sÃo realizados pela comunidade acadÃmica a fim de propor solventes alternativos que substituam o uso exclusivo do sulfolano por compostos que tambÃm apresentem caracterÃsticas importantes, como boa seletividade, boa recuperabilidade e baixos custos. Diante do exposto, o presente trabalho tem como principal objetivo determinar dados de equilÃbrio lÃquido-lÃquido (ELL) para estudar a capacidade dos solventes [dietilenoglicol (DEG), trietilenoglicol (TEG), glicerina e 2-hidroxietilamina formato (2-HEAF)] em extrair aromÃticos das misturas com alifÃticos. As propriedades fÃsicas tambÃm assumem extrema importÃncia no conhecimento do comportamento e das interaÃÃes que ocorrem nas misturas de lÃquidos. Deste modo, neste trabalho foram estudados a densidade e a viscosidade de solventes extratores de aromÃticos. A metodologia experimental para obtenÃÃo dos dados de ELL foi inicialmente validada reproduzindo dados da literatura, e os resultados mostraram uma boa concordÃncia entre estes dados. ApÃs a metodologia ter sido validada foram construÃdas curvas de solubilidade e tie-lines para os sistemas contendo decano + tolueno + DEG (+ TEG, + glicerina, + 2-HEAF, + sulfolano) a T = (303,15 e 333,15) K. Os dados de composiÃÃo das tie-lines, obtidos experimentalmente, foram utilizados para calcular o parÃmetro de seletividade e o coeficiente de distribuiÃÃo. De posse desses dados, pode-se concluir que todos os solventes estudados apresentam boa capacidade para extraÃÃo de tolueno a partir de misturas com decano. No entanto, a glicerina destacou-se como o solvente mais promissor para a extraÃÃo de aromÃticos apresentando os maiores valores para seletividade. Finalmente, os dados experimentais foram correlacionados atravÃs dos modelos termodinÃmicos de coeficiente de atividade, NRTL e UNIQUAC. Os resultados foram considerados satisfatÃrios, observando-se que o NRTL foi o modelo que apresentou os menores desvios na composiÃÃo. Dessa forma, representou melhor os dados de ELL para os sistemas estudados.

Submitted by Fabio Sobreira Campos da Costa (fabio.sobreira@ufpe.br) on 2016-08-12T12:30:57Z No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Tese de Doutorado Josemar Guerra.pdf: 4223686 bytes, checksum: 5ef7d536e636cc8daf52f5ee42c401f1 (MD5) / Made available in DSpace on 2016-08-12T12:30:58Z (GMT). No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Tese de Doutorado Josemar Guerra.pdf: 4223686 bytes, checksum: 5ef7d536e636cc8daf52f5ee42c401f1 (MD5) Previous issue date: 2015-10-15 / FINEP / ANP / PRH28 / Os derivados de petróleo são largamente empregados na geração de energia em todo o mundo. Por serem constituídos por diversos componentes, seus derivados podem ser empregados como matéria-prima em várias áreas industriais. No processamento do petróleo e no uso de seus derivados identificam-se inúmeras possibilidades de contaminação ao meio ambiente. Os resíduos líquidos gerados nas indústrias petroquímicas possuem diferentes composições químicas que inclui óleos e graxas, benzeno, tolueno e metais pesados, tais como níquel, cádmio, cobre e chumbo. Diante disso, as indústrias buscam, cada vez mais, ajustar os processos existentes através da adoção de procedimentos que visam diminuir a geração de resíduo e uma maior remoção de elementos tóxicos dos efluentes. Assim, este trabalho estudou a utilização de argilas esmectitas in natura e tratadas como adsorventes no processo de adsorção para remoção de poluentes orgânicos e de íons metálicos presentes em efluentes gerados nas indústrias de petroquímicas. A argila esmectita foi empregada no processo de adsorção dos compostos orgânicos benzeno e tolueno e no processo de troca catiônica para a remoção dos íons de metais pesado Cu+2 e Pb+2 como um processo de pós-tratamento para efluentes de refinaria. A partir do tratamento químico da argila esmectita policatiônica (argila chocolate) in natura com carbonato de sódio foi alcançada uma boa troca de íons de cálcio e magnésio por íons de sódio. Com o tratamento observou-se um aumento do número de íons trocáveis de 81,34 meq/100g para 89,39 meq/100g. Pode-se considerar que as argilas apresentam uma estrutura em que os poros são classificados como mesoporosos. Os experimentos de adsorção e de troca catiônica foram realizados em batelada empregando as argilas sódicas como adsorventes no estudo cinético e de equilíbrio. Os resultados obtidos a partir do modelo de equilíbrio de Langmuir-Freundlich indicaram que a argila sódica apresentou capacidade adsortiva perante o benzeno e o tolueno de 7,28mg/g e 7,39mg/g, respectivamente. Para os íons Cu+2 e Pb+2 foi aplicado o modelo de troca Catiônica o qual indicou uma capacidade de 0,99mg/g e 1,10mg/g para os íons Cu+2 e Pb+2, respectivamente. Além disso, o comportamento das análises cinética mostrou que as constantes cinéticas obtidas para o benzeno e tolueno apresentaram os valores de 0,0016 L/mg.min e 0,019 L/mg.min, respectivamente. Para os íons Cu+2 e Pb+2 em argila sódica as constantes cinéticas foram de 0,049 L/mg.min e 0,063 L/mg.min, respectivamente. / The oil derivatives are widely used in power generation throughout the world. For be constituted of several components, their derivatives can be used as raw materials in various industrial areas. In the oil processing and in the use of their derivatives are identified numerous possibilities for contamination to the environment. Liquid wastes generated in the petrochemical industries have different chemical compositions that include oils and greases, benzene, toluene, and heavy metals such as nickel, cadmium, copper and lead. Before that, the industries has been seeking, increasingly, adjust the existing processes by adopting procedures to reduce the generation of waste and to reach a greater removal of toxic elements effluent. This work has studied the use of smectite clays in natura and treated as adsorbents in the adsorption process for removing organic pollutants and metal ions present in wastewater generated in the petrochemical industries. The smectite clay was used in the adsorption of organic compounds benzene and toluene and the cation exchange process for the removal of heavy metal ions Cu+2 and Pb+2 as a post-treatment process for refinery effluents. From the chemical treatment of chocolate clay in natura with sodium carbonate has been achieved a good exchange of calcium and magnesium ions for sodium ions. With treatment there was an increase in the number of exchangeable ions of 81.34 meq/100g to 89.39 meq/100g. It can be considered that the clays have a structure in which the pores are classified as mesoporous. The adsorption and cation exchange experiments were conducted in batch mode using the sodium clays as adsorbents in the kinetic and equilibrium study. The results obtained from the Langmuir-Freundlich equilibrium model indicated that sodium clay adsorptive capacity presented to the benzene and toluene 7,28mg.g-1 and 7,39mg/g, respectively. For Cu+2 and Pb+2 ions was applied to the ion exchange model which indicated a capacity of 0,99mg.g-1 and 1,10mg.g-1 for Cu+2 and Pb+2 ions, respectively. Moreover, the behavior of the kinetic analysis showed that the kinetic constants obtained for benzene and toluene showed values of 0.0016 L.mg-1.min-1 and 0.019 L.mg-1.min-1 respectively. For Cu+2 and Pb+2 ions in sodium clay the kinetic constants were 0.049 L.mg-1.min-1 and 0.063 L.mg-1.min-1, respectively.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The volatile organic compounds benzene, toluene and xylenes (BTX) are among the most produced chemicals worldwide and can be found in different environmental compartments. Contamination of aquatic environments by these substances may result in to adverse toxic effects on organisms of different biological organizations. The present study aimed to evaluate the toxicity of BTX isolated and in binary mixtures using the acute toxicity test (96h) with the microcrustacean Mysidopsis juniae. To achieve that, organisms were exposed to BTX concentrations in order to identify the effects on survival, as well as to study the modes of action of the substances in mixture with the application of the theoretical models of Addition Concentration (AC) and Independent Action (AI). In the experiments with the isolated compounds it was possible to notice that xylene was the most toxic compound (LC50 16,10 ±2,4mg.L-1) followed by toluene (LC50 38 ± 5,3mg.L-1) and benzene (LC50 78,03 ±2,9mg.L-1) respectively. When in binary mixture deviations of the reference models for CA were observed in all experiments performed (dose ratio and antagonism). In the exposure of xylene and benzene the deviation that fit the results was the antagonism, where as the mixture of xylene and toluene was better explained by the deviation dose ratio where the toxicity was caused mainly by xylene. In the experiment combining toluene and benzene, the dose ratio pattern was observed as well, and the benzene was the main responsible for the toxicity. We concluded that BTX were toxic to mysids, both isolated and in binary mixtures, evidencing the importance of studies in this subject that can serve as support for evaluations and monitoring of the marine environments, considering the imminent risks of contamination of this environment and the wide scale of use of these compounds. / Os compostos orgânicos voláteis benzeno, tolueno e os xilenos (BTX) estão entre os produtos químicos mais produzidos mundialmente e podem ser encontrados em diferentes compartimentos ambientais. A contaminação de ambientes aquáticos por essas substâncias pode acarretar em efeitos tóxicos adversos em organismos de diferentes organizações biológicas. Diante disso, o presente estudo teve como objetivo avaliar a toxicidade dos BTX isolados e em misturas binárias por meio do teste de toxicidade aguda (96h) com o microcrustáceo Mysidopsis juniae. Para isso, os organismos foram expostos às concentrações de BTX a fim de identificar os efeitos na sobrevivência, assim como estudar os modos de ação das substâncias em mistura com a aplicação dos modelos teóricos de concentração de adição (CA) e ação independente (IA). Nos experimentos com os compostos isolados foi possível perceber que o xileno foi o mais tóxico (CL50 16,10±2,4mg.L-1) seguido por tolueno (CL50 38±5,3mg.L-1) e benzeno (CL50 78,03±2,9mg.L-1). Quando em mistura binária foram observados desvios dos modelos de referência para CA em todos os experimentos realizados (razão das doses e antagonismo). Na exposição do xileno e benzeno o desvio que se enquadrou aos resultados foi o antagonismo, já a mistura de xileno e tolueno foi melhor explicada pelo desvio razão das doses onde a toxicidade foi causada principalmente pelo xileno, no experimento com o tolueno e benzeno também foi observado um padrão de razão das doses, sendo que nesse caso o benzeno foi o maior responsável pela toxicidade. Logo, os hidrocarbonetos analisados foram tóxicos para o misidáceo, tanto isolados como em misturas binárias, evidenciando a importância de estudos nessa temática que possam servir de suporte para avaliações e monitoramento dos ambientes marinhos, visto os riscos iminentes de contaminação deste meio e a ampla escala de utilização desses compostos.

O tolueno e o xileno são substâncias presentes, ao lado de outras, em vários produtos, entre eles as tintas, tíneres e colas, provocando exposição combinada dos trabalhadores a diversas substâncias químicas. Assim, foi objetivo deste trabalho desenvolver um método simples, rápido e confiável de cromatografia em fase gasosa que permitisse análise simultânea do ácido hipúrico (AH) e ácido metil-hipúrico (AMH) em urina, bioindicadores de dose interna utilizados na monitorização da exposição ocupacional ao tolueno e xileno respectivamente. Após extração líquido-líquido e derivação com trimetilfenilamôneo, foi realizada análise cromatográfica em coluna capilar 100% polidimetilsiloxano, usando-se o ácido heptadecanóico como padrão interno. Foram avaliados 69 trabalhadores que manipulavam tintas em atividades de repintura de veículos e 31 trabalhadores não-expostos (controle). O método mostrou-se linear entre 0,1 (limite de quantificação) e 5,0 g/L para o AH e 0,2 (limite de quantificação) e 5 g/L para o AMH. A recuperação média foi de 81 % e a inexatidão de aproximadamente - 8,0% (bias) para ambos os analitos. Os coeficientes de variação médios para as concentrações de 0,5; 2,0 e 4,0 g/L foram, para a precisão intra-ensaio, de 3,5% para o AH e de 3,7% para o AMH, e para a interensaio, de 10,5% para o AH e de 10,7% para o AMH. Os analitos permaneceram estáveis em até 3 ciclos de congelamento/descongelamento das amostras de urina. O valor médio obtido nas amostras de trabalhadores expostos foi de 0,55 g/g de creatinina (mediana = 0,36 g/g de creatinina). Apesar de diferença estatisticamente significativa (teste t Student, p ≤ 0,05) ter sido observada entre os valores médios de AH nos trabalhadores expostos e nos do grupo controle, esta diferença não foi detectada quando analisou-se as medianas. O hábito de fumar e de ingerir bebidas alcoólicas também não mostrou alterar significativamente a excreção dos bioindicadores no grupo avaliado. / Toluene and the xylene, among others substances, are solvents used in several products, such as inks, thiners and glues, and are responsible for mixed exposure in workers. Thus, the aim of this work was to develop a simple, fast and reliable gas chromatographic method to allow the simultaneous detection and quantitation of hippuric acid (HA) and methylhippuric acid (MHA) in urine, which are internai dose bioindicators in the monitoring of occupational exposure to toluene and the xylene, respectively. After liquid-liquid extraction and derivation with trimethylphenylammonium, the chromatographic analysis was accomplished using capillary column 100% methylsiloxane, and heptadecanoic acid as internai standard. 69 workers exposed to inks in activities such as vehicles painting and 31 nonexposed workers were evaluated. The method showed linearity range between 0,1 (quantitation limit) and 5,0 g/L for HA and 0,2 (quantitation limit) and 5 g/L for MHA. The recovery was 81 % and the inaccuracy of approximately - 8,0% (bias) for both analytes. The mean variation . coefficients for the concentrations of 0,5; 2,0 and 4,0 g/L were, for the intraassay precision, of 3,5% for HA and of 3,7% for MHA, and for the inter assay, 10,5% for HA and 10,7% for MHA. The analytes were stable for 3 freezing cycles of the urine samples. The medium value obtained in the exposed workers\' samples was of 0,55 creatinina g/g (medium = 0,36 creatinina g/g). In spite of the observed statistical significant differences (test t Student, p ≤ 0,05) among the medium values of HA in the exposed workers and the control groups, this difference was not detected when it was analyzed the respective median values. Smoking and drinking alcoholic beverages habits did not significantly alter the excretion of these bioindicators in the studied group.

O presente trabalho avaliou a degradação anaeróbia de tolueno por Bactérias Redutoras de Sulfato (BRS), utilizando como inóculo lodo anaeróbio proveniente de reator UASB. Os ensaios foram realizados com reatores em batelada de 2300 mL contendo meio de cultura específico para o crescimento de BRS. No ensaio de enriquecimento, com lactato de sódio (2230 mg/L) e acetato de sódio (670 mg/L) na ausência de tolueno, ocorreu consumo total de sulfato (aproximadamente 1070 mg/L) em 192 horas e percentual de metano de 31% em 72 horas. A degradação do tolueno foi realizada em reatores sulfetogênicos e metanogênicos. Nos reatores sulfetogênicos alimentados com 2,1 mg/L e 7,7 mg/L houve degradação total do tolueno em 264 e 792 horas, respectivamente. Nesses reatores foi observado consumo total de sulfato e percentual de metano de aproximadamente 20,9%. Nos reatores metanogênicos a degradação do tolueno foi de, no máximo, 53%, e percentual de metano de 27,3% em 1032 horas. Nas condições sulfetogênicas, a degradação do tolueno foi mais rápida se comparada às condições metanogênicas. E em ambas as condições, a produção de metano foi relacionada aos bacilos hidrogenotróficos. As alterações na diversidade da população microbiana presente nos reatores, observadas em microscopia ótica, foram confirmadas nas análises por biologia molecular. / The present work evaluated the anaerobic degradation of toluene by sulfate reducing bacteria (SRB), using anaerobic sludge from reactor UASB as inoculum. The essays were accomplished in batch reactors of 2300 mL containing specific culture medium for the growth of SRB. In the enrichment essay with sodium lactate (2230 mg/L) and sodium acetate (670 mg/L) without toluene accured total consumption of sulfate (approximately 1070 mg/L) in 192 hours and percentile of methane of 31% in 72 hours. The toluene degradation it was accomplished in sulfetogenics and methanogenics reactors. In the sulfetogenics reactors fed with 2,1 mg/L and 7,7 mg/L there was total degradation of toluene in 264 and 792 hours, respectively. In those reactors it was observed total consumption of sulfate and percentile of methane of approximately 20.9%. In the methanogenic reactors the degradation of toluene was to the utmost of 53%, accompanied of production of 27.3% of methane in 1032 hours. In the sulfetogenics conditions, the degradation of tolueno was faster if compared to the methanogenic conditions. In both conditions, the methane production was related to the hidrogenotrophic bacilli. The alterations in the diversity of the microbial population present in the reactors, observed by optic microscopy, were confirmed in the analysis by molecular biology.

An exceptionally important stress response of Pseudomonas putida strains to toxic chemicals is the induction of efflux pumps that extrude solvents, as well as other toxicants, into the surrounding medium. However, the bacterial tolerance mechanisms are still not fully understood, thus in this thesis metabolomic approaches were used to detect and identify metabolites involved in P. putida DOT-T1E tolerance to abiotic stresses, in particular focussing on the role of efflux pumps. To elucidate any metabolome alterations several strains of P. putida, including the wild type DOT-T1E, and the efflux pump knockouts DOT-T1E-PS28 and DOT-T1E-18, were challenged with different levels of propranolol. Fourier-transform infrared (FT-IR) spectroscopy, which provided a rapid, high-throughput metabolic fingerprint of P. putida strains, was used to investigate any phenotypic changes resulting from exposure to propranolol. FT-IR data illustrated phenotypic changes associated with the presence of propranolol within the cell that could be assigned to the bacterial protein components. To complement this phenotypic fingerprinting approach metabolic profiling on the same samples was performed using gas chromatography mass spectrometry (GC-MS) to identify metabolites of interest during growth of bacteria following this toxic perturbation with propranolol. GC-MS revealed significant changes in ornithine levels which can be directly linked to bacterial tolerance mechanisms, and alterations in the levels of several other metabolites which were also modified in response to propranolol exposure. Moreover, the effect of the organic solvent toluene was also investigated using the same approach. Examination of FT-IR data indicated that protein and fatty acids were the most affected components of P. putida strains due to the presence of toluene within the cell. Moreover, application of GC-MS allowed for the identification and quantification of several metabolites which were differentially produced or consumed in the presence of toluene. To investigate the role of efflux pumps in P. putida DOT-T1E, several analytical techniques were employed including Raman spectroscopy, gas and liquid chromatography to identify and quantify the level of propranolol or toluene in P. putida cells. These analyses showed that propranolol and toluene accumulated in the mutant P. putida DOT-T1E-18 (lacking the TtgABC pump) at higher levels in comparison with the levels found in the wild-type DOT-T1E and the mutant DOT-T1E-PS28 (lacking the TtgGHI pump), indicating the key role of efflux pumps in solvent tolerance. Furthermore, the effect of Mg2+ and Ca2+ on the stabilisation of the toluene tolerance of P. putida DOT-T1E strains was examined in order to elucidate whether divalent cations interact with efflux pumps or other resistant mechanisms to improve solvent tolerance. FT-IR analysis suggested that the influence of divalent cations on the stabilisation of the toluene tolerance could be due to the contribution of metal ions towards other tolerance mechanisms such as lipopolysaccharide (LPS) instead of enhancing the activity of efflux pumps. In conclusion, this thesis presents evidence that phenotypic fingerprinting and metabolic profiling approaches in combination with chemometric methods can generate valuable information on phenotypic responses occurring within microbial cultures subjected to abiotic stress.

This diploma thesis has been carried out as a part of the project Utilization of long term (passive) sampling methods combined with in situ microcosms for assessment of (bio)degradation potential (PASSES). In the frame of the project groundwater remediation took place in the premises of Farmak a.s. in Olomouc using a pilot photooxidation unit and efficiency of the remediation was monitored through passive and active sampling methods. Pilot photooxidation unit is a technology based on the H2O2/UV-C photochemical oxidation of organic pollutants. In this work optimization tests of the pilot photooxidation unit were performed. The residence time of the groundwater in the photoreactors, required for its sufficient decontamination from pharmaceuticals and aromatic hydrocarbons, was 2.5 hours. 91% degradation of the pharmaceuticals and 80% degradation of aromatic hydrocarbons were reached during this interval. Although the removal efficiency of the pharmaceuticals by the photooxidation unit was high, the pilot photooxidation unit was not able to effectively remove the pharmaceuticals at the studied locality. By comparing the results of the pharmaceuticals from active and passive groundwater sampling during the remediation attempt, passive Polar Organic Chemical Integrative Sampler (POCIS) was found to be...

The integration of organic solvent nanofiltration in processes of fine chemistry involving homogeneous metal catalysts has a great potentiel because this eco-friendly process, efficient at molecular scale, does not desactivate the catalyst contrary to conventionnal distillation. The aim of this study is to integrate organic solvent nanofiltration in the process of 10-undecenitrile hydroformylation in toluene. A one-step nanofiltration does not permit to fulfill the goals of the separation, which are to extract the product and to recycle the catalytic system (Rh, biphephos). Simulations of membrane cascades based on experimental data of flux and retention according to the concentration (acquired in the first part of the thesis) highlighted that a four stages cascades with recycling is realistic and competitive for an industrial plant. / L'intégration de la nanofiltration organique dans les procédés de chimie fine impliquant des catalyseurs organométalliques solubles a un fort potentiel car ce procédé éco-efficace de séparation à l'échelle moléculaire ne désactive pas les catalyseurs contrairement à la distillation classiquement utilisée. L'objectif de cette thèse est d'intégrer la nanofiltration organique dans le procédé d'hydroformylation du 10-undecenitrile dans le toluène. Un procédé en une étape de nanofiltration ne permet pas de remplir les objectifs de séparation qui sont l'extraction du produit et le recyclage du système catalytique (Rh, biphephos). Des simulations de cascades de membranes basées sur les données expérimentales de flux et de rétentions variables en fonction de la concentration, acquises en première partie de thèse, ont permis d'identifier un design de cascade à 4 étages avec recyclages réaliste et compétitif pour une conception industrielle.

Photoelectron Photoion Coincidence (PEPICO) spectroscopy is a robust tool for elucidating complex unimolecular dissociation mechanisms and for determining thermochemical and kinetic data of gas-phase ion dissociations with high accuracy. In this work, the dissociative photoionization of two sets of isomeric systems were analyzed with PEPICO: 1) C7H7+ ions of toluene (Tol) and 1,3,5-cycloheptatriene (CHT), and 2) two butyl alcohol isomers, 1-butanol and isobutanol. Threshold dissociative photoionization data on these four molecules of interest were collected on the imaging PEPICO apparatus at the VUV beamline of the Swiss Light Source. Data analysis was aided by ab initio calculations and Rice-Ramsperger-Kassel-Marcus (RRKM) statistical rate theory was employed to model the complex dissociation pathways of each system. Finally, thermochemical, reaction mechanism, and dissociation kinetics data were extracted from the modeled data and are reported here. In the first project, the dissociation of energy-selected 1,3,5-cycloheptatriene (CHT) and toluene (Tol) cations was investigated by imaging photoelectron photoion coincidence spectroscopy. In the measured energy ranges of 10.30−11.75 eV for CHT and 11.45−12.55 eV for Tol, only the hydrogen atom loss channels open up, leading to C7H7+ from both molecular ions, which are both metastable at the H-loss threshold. Our quantum chemical calculations showed that these ions can interconvert below their dissociation thresholds. Therefore, we constructed a single statistical model to describe both systems simultaneously. We determined 0 K appearance energies (E0) for the tropylium and benzyl fragment ions from CHT to be 9.520 ± 0.060 eV and 9.738 ± 0.082 eV, and from Tol to be 10.978 ± 0.063 eV and 11.196 ± 0.080 eV, respectively. Using the experimentally determined benzyl ion appearance energy, its 0 K heat of formation was calculated to be 937.9 ± 7.7 kJ mol–1. On the basis of this value and the recently determined benzyl ionization energy, we point out discrepancies concerning the benzyl radical thermochemistry. For the second project, the fragmentation processes of two internal energy-selected C4H10O+• cations, 1-butanol and isobutanol, were investigated. For both isomers, the first dissociation channel leads to the formation of C4H8+• ions (m/z = 56) by a water loss. Using statistical energy distribution and rate models, which include isomerization of the molecular ions, the 0 K appearance energies (E0) were determined to be 10.347 ± 0.015 eV and 10.566 ± 0.050 eV, for 1-butanol and isobutanol, respectively. The second dissociation channel, the formation of CH3OH2+, quickly overtakes the water-loss channel in isobutanol, with an E0 of 10.612 ± 0.020 eV, but appears only as a minor channel in 1-butanol with an E0 of 10.738 ± 0.080 eV. The methanol-loss channel, forming propylene ion, opens up at E0 = 10.942 ± 0.040 eV and 10.723 ± 0.020 eV in 1-butanol and isobutanol, respectively. The next two fragmentation pathways correspond to a complementary pair of C3H7+ through the loss of CH2OH, and CH2OH+ through the loss of C3H7. From both isomers, C3H7+ is the isopropyl ion, which is readily formed in isobutanol via a simple bond cleavage at E0 = 10.970 ± 0.050 eV and its pair, CH2OH+, at E0 = 11.11 ± 0.20 eV. However, there is an internal hydrogen shift necessary in 1-butanol and, therefore, the complementary ions appear at the same E0 of 11.104 ± 0.030 eV, which most likely corresponds to their common transition state. Finally, C3H5+, a product of sequential dissociation from m/z = 56, appears above 11.6 eV as a minor channel for both isomers.