Liquid chromatography (LC) coupled to mass spectrometry (MS) offers highly selective and sensitive analysis of a wide variety of compounds. However, the use of hyphenated experimental set-ups implies that many parameters may have an effect on the studied response. Therefore, in order to determine optimized experimental conditions it is of vital importance to incorporate systematic procedures during method development. In this thesis, a generic stepwise optimization strategy is proposed that aims at high chromatographic quality, as well as high mass spectrometric response. The procedure comprises (i) screening experiments to identify the most important parameters, (ii) LC studies to ensure sufficient chromatographic separation, (iii) extended infusion experiments in order to maximize precursor signal(s), and in the case of tandem MS (iv) extended infusion experiments to determine optimal conditions for collision induced dissociation and when applicable also ion trap settings. Experimental design and response surface methodology is used throughout the procedure. Further, the general applicability of LC-MS is demonstrated in this thesis. Specifically, a novel quantitative column-switched LC-MS method for ferrichrome, ferrichrysin and ferricrocin determination is presented. Using the method it was shown how the siderophore content varies with depth in podzolic soil profiles in the north and south of Sweden. The parallel approach using LC coupled to both inductively coupled plasma (ICP) mass spectrometry, and electrospray ionization (ESI) tandem MS is also evaluated as a tool to identify unknown siderophores in a sample. Additionally, different trypsin digestion schemes used for LC-ESI-MS peptide mapping were compared. By multivariate data analysis, it was clearly shown that the procedures tested induce differences that are detectable using LC-ESI-MS. Finally, the glutathione S-transferase catalyzed bioactivation of the prodrug azathioprine was verified using LC-MS.

Η ανάγκη αντιμετώπισης του προβλήματος της ρύπανσης των επιφανειακών και υπόγειων υδάτων έχει οδηγήσει στην ανάπτυξη νέων και αποτελεσματικών μεθόδων για την απομάκρυνση έμμονων και μη βιοαποικοδομήσιμων ενώσεων από το νερό και τα υγρά απόβλητα. Η παρούσα διατριβή πραγματεύτηκε τη συστηματική μελέτη της διάσπασης ρύπων προτεραιότητας και αναδυόμενων ρύπων που ανήκουν σε διαφορετικές χημικές κατηγορίες (DEET, metribuzin, 2-ισοπρόπυλο-3-μεθόξυ πυραζίνη, η πενταχλωροφαινόλη, βενζοϊκό οξύ, Cr(VI) και φαινολικές ενώσεις) με τη μέθοδο της ετερογενούς φωτοκατάλυσης και τη χρήση εμπορικά διαθέσιμων δραστικών μορφών TiO2 και τροποποιημένων σωματιδίων ΤiΟ2 με αμέταλλα που παρουσιάζουν φωτοκαταλυτική δραστικότητα στο ορατό φάσμα της ακτινοβολίας. Για το σύνολο των ενώσεων που μελετήθηκαν, η έρευνα επικεντρώθηκε: i) στη μελέτη της κινητικής της αποδόμησης και της ολικής ανοργανοποίησης τους με τη μέθοδο της ετερογενούς φωτοκατάλυσης, ii) στη μελέτη της επίδρασης λειτουργικών παραμέτρων στην απόδοση της φωτοκαταλυτικής απομάκρυνσης των ρύπων και στην εύρεση των βέλτιστων συνθηκών με την εφαρμογή των δύο ευρέως χρησιμοποιούμενων χημειομετρικών μεθόδων, της μεθοδολογίας επιφάνειας απόκρισης (RSM) και των τεχνητών νευρωνικών δικτύων (ANNs), iii) στην ανίχνευση και ταυτοποίηση ενδιάμεσων προϊόντων διάσπασης των οργανικών ρύπων με φασματομετρικές τεχνικές όπως υγρή χρωματογραφία-φασματομετρία μαζών υψηλής διακριτικής ικανότητας και ακρίβειας μάζας με αναλυτή τροχιακής παγίδας (Orbitrap) και αέρια χρωματογραφία–φασματομετρία μάζας με την τεχνική ιοντισμού με πρόσκρουση ηλεκτρονίων (Εlectron Impact, EI) και φασματοσκοπία ηλεκτρονικού παραμαγνητικού συντονισμού, iv) στη συνεισφορά των δραστικών ειδών οξυγόνου, οπών και ηλεκτρονίων στη φωτοκαταλυτική αποικοδόμηση των ρύπων, v) στη μελέτη του ρόλου των φυσικών συστατικών (χουμικά και φουλβικά οξέα) διαφόρων υδατικών συστημάτων καθώς και στην εκτίμηση της επίδρασης του υδατικού μέσου στην κινητική της διεργασίας, vi) στην εκτίμηση της τοξικότητας πριν και κατά τη διάρκεια της φωτοκαταλυτικής επεξεργασίας. Με την εφαρμογή της ετερογενούς φωτοκατάλυσης επετεύχθη πλήρης απομάκρυνση όλων των ρύπων-μοντέλων που μελετήθηκαν και υψηλός βαθμός ανοργανοποίησης των διαλυμάτων τους. H φωτοκαταλυτική διάσπαση λαμβάνει χώρα σε ποσοστό μεγαλύτερο του 90% σε χρόνους που κυμαίνονται από 20-180 λεπτά ανάλογα με το μελετώμενο ρύπο. Ένας μεγάλος αριθμός κύριων προϊόντων διάσπασης και τουλάχιστον ένα ισομερές για τα περισσότερα από αυτά, ταυτοποιήθηκε κατά τη φωτοκαταλυτική διάσπαση της PCP, του DEET, του ΜΕΤ και της ΙPMP με τη χρήση προηγμένων τεχνικών φασματομετρίας μάζας. Με βάση τα προϊόντα που ταυτοποιήθηκαν, προτάθηκαν οι μηχανισμοί της φωτοκαταλυτικής διάσπασης των μελετώμενων ρύπων που περιλαμβάνουν κυρίως αντιδράσεις υδροξυλίωσης, οξείδωσης και απαλκυλίωσης. Η οξείδωση μέσω ριζών υδροξυλίου (HO•)βρέθηκε να αποτελεί το κύριο οξειδωτικό είδος σε όλα τα μελετώμενα συστήματα. Η δοκιμή τοξικότητας που πραγματοποιήθηκε, πριν και κατά τη διάρκεια της φωτοκαταλυτικής επεξεργασίας των οργανικών ρύπων έδειξε ότι η μέθοδος της ετερογενούς φωτοκατάλυσης οδηγεί σε πλήρη αποτοξικοποίηση των διαλυμάτων. Σύμφωνα με τα αποτελέσματα της φωτοκαταλυτικής οξείδωσης των οργανικών ρύπων-μοντέλων και της φωτοκαταλυτικής αναγωγής του Cr(VI) που μελετήθηκαν στην παρούσα διατριβή, η μέθοδος της ετερογενούς φωτοκατάλυσης μπορεί να αποτελέσει μια αποδοτική εναλλακτική πρόταση αντιρρύπανσης έναντι των κλασσικών μεθόδων. Η ουσιαστική λύση σε προβλήματα ρύπανσης αποφεύγοντας τη μεταφορά των ρύπων από τη μία φάση στην άλλη, η δυνατότητα χρήσης και αξιοποίησης ήπιων μορφών ενέργειας όπως η ηλιακή ακτινοβολία και η σύζευξη της μεθόδου με άλλες βιολογικές ή/και φυσικοχημικές μεθόδους επεξεργασίας αποδεικνύουν τις μεγάλες προοπτικές που παρουσιάζει. / In order to avoid deterioration of water resources, considerable efforts have been devoted to develop suitable purification methods that can easily remove recalcitrant (persistent) and non-biodegradable contaminants from water and wastewater. In the present thesis the photocatalytic removal of selected priority pollutants and emerging contaminants belonging in different chemical categories has been investigated in detail. DEET, metribuzin, 2 isopropyl-3-methoxy pyrazine, pentachlorophenol (PCP), benzoic acid (BA), Cr(VI) and phenolic compounds were selected as target compounds. The commercial form of TiO2 (Degussa P25) and NF-codoped TiO2, a material with improved photo efficiency and visible light response were used as photocatalysts. The main objectives of this thesis were: i) to evaluate the kinetics of selected pollutants disappearance and mineralization, ii) to investigate the effect of significant parameters on the total process efficiency as well as to optimize the photocatalytic procedure by means of chemometric optimization tools such as central composite design, response surface methodology and artificial neural networks, iii) to identify the transformation products formed during the photocatalytic treatment by using powerful analytical techniques such as high resolution accurate mass LC-MS, GC–MS and EPR spectroscopy, iv) to assess the role of the reactive species in the reaction mechanism using different scavengers, v) to assess the macroscopic effects of DOM (HA, FA) and water matrix on the photocatalytic degradation and vi) to evaluate the toxicity along the photocatalytic process. By the application of heterogeneous photocatalysis almost compete removal of the selected model contaminants and high percentages of mineralization were achieved. Photocatalytic removal (> 90%) was succeeded after 20-180 min of irradiation time, depending on the studied pollutant. Numerous different structures of transformation products (TPs), with at least one isomer for the majority of them, were identified with high resolution accurate mass liquid chromatography (HR-LC–MS) and gas chromatography mass spectrometry (GC–MS). Based on by-product identification using HR-LC-MS and GC–MS techniques possible degradation pathways were proposed. The pathways mainly include hydroxylation, oxidation and dealkylation reactions. Hydroxyl radicals (HO•) were determined to be the predominant reactive species during photocatalysis in all the studied systems. Toxicity assessment revealed the efficiency of the photocatalytic treatment to achieve almost complete detoxification of the solution. According to the results obtained for the photocatlytic oxidation of the studied organic pollutants-models and photocatalytic reduction of Cr(VI), heterogeneous photocatalyis was shown to be a great potential as a sustainable treatment technology. Ιts inherent destructive nature, not involving mass transfer, the potential use of solar radiation as well as the combination of heterogeneous photocatalysis with biological and/or physicochemical methods make this method particularly attractive for environmental decontamination and detoxification.

Computational tools such as finite element analysis and simulation are commonly used for system performance analysis and validation. It is often impractical to rely exclusively on the high-fidelity simulation model for design activities because of high computational costs. Mathematical models are typically constructed to approximate the simulation model to help with the design activities. Such models are referred to as “metamodel.” The process of constructing a metamodel is called “metamodeling.” Metamodeling, however, faces eminent challenges that arise from high-dimensionality of underlying problems, in addition to the high computational costs and unknown function properties (that is black-box functions) of analysis/simulation. The combination of these three challenges defines the so-called high-dimensional, computationally-expensive, and black-box (HEB) problems. Currently there is a lack of practical methods to deal with HEB problems. This dissertation, by means of surveying existing techniques, has found that the major deficiency of the current metamodeling approaches lies in the separation of the metamodeling from the properties of underlying functions. The survey has also identified two promising approaches - mapping and decomposition - for solving HEB problems. A new analytic methodology, radial basis function–high-dimensional model representation (RBF-HDMR), has been proposed to model the HEB problems. The RBF-HDMR decomposes the effects of variables or variable sets on system outputs. The RBF-HDMR, as compared with other metamodels, has three distinct advantages: 1) fundamentally reduces the number of calls to the expensive simulation in order to build a metamodel, thus breaks/alleviates exponentially-increasing computational difficulty; 2) reveals the functional form of the black-box function; and 3) discloses the intrinsic characteristics (for instance, linearity/nonlinearity) of the black-box function. The RBF-HDMR has been intensively tested with mathematical and practical problems chosen from the literature. This methodology has also successfully applied to the power transfer capability analysis of Manitoba-Ontario Electrical Interconnections with 50 variables. The test results demonstrate that the RBF-HDMR is a powerful tool to model large-scale simulation-based engineering problems. The RBF-HDMR model and its constructing approach, therefore, represent a breakthrough in modeling HEB problems and make it possible to optimize high-dimensional simulation-based design problems.

Computational tools such as finite element analysis and simulation are commonly used for system performance analysis and validation. It is often impractical to rely exclusively on the high-fidelity simulation model for design activities because of high computational costs. Mathematical models are typically constructed to approximate the simulation model to help with the design activities. Such models are referred to as “metamodel.” The process of constructing a metamodel is called “metamodeling.” Metamodeling, however, faces eminent challenges that arise from high-dimensionality of underlying problems, in addition to the high computational costs and unknown function properties (that is black-box functions) of analysis/simulation. The combination of these three challenges defines the so-called high-dimensional, computationally-expensive, and black-box (HEB) problems. Currently there is a lack of practical methods to deal with HEB problems. This dissertation, by means of surveying existing techniques, has found that the major deficiency of the current metamodeling approaches lies in the separation of the metamodeling from the properties of underlying functions. The survey has also identified two promising approaches - mapping and decomposition - for solving HEB problems. A new analytic methodology, radial basis function–high-dimensional model representation (RBF-HDMR), has been proposed to model the HEB problems. The RBF-HDMR decomposes the effects of variables or variable sets on system outputs. The RBF-HDMR, as compared with other metamodels, has three distinct advantages: 1) fundamentally reduces the number of calls to the expensive simulation in order to build a metamodel, thus breaks/alleviates exponentially-increasing computational difficulty; 2) reveals the functional form of the black-box function; and 3) discloses the intrinsic characteristics (for instance, linearity/nonlinearity) of the black-box function. The RBF-HDMR has been intensively tested with mathematical and practical problems chosen from the literature. This methodology has also successfully applied to the power transfer capability analysis of Manitoba-Ontario Electrical Interconnections with 50 variables. The test results demonstrate that the RBF-HDMR is a powerful tool to model large-scale simulation-based engineering problems. The RBF-HDMR model and its constructing approach, therefore, represent a breakthrough in modeling HEB problems and make it possible to optimize high-dimensional simulation-based design problems.

This thesis deals with the near-field of confluent jets, which can be of interest in many engineering applications such as design of a ventilation supply device. The physical effect of interaction between multiple closely spaced jets is studied using experimental and numerical methods. The primary aim of this study is to explore a better understanding of flow and turbulence behavior of multiple interacting jets. The main goal is to gain an insight into the confluence of jets occurring in the near-field of multiple interacting jets. The array of multiple interacting jets is studied when they are placed on a flat and a curved surface. To obtain the boundary conditions at the nozzle exits of the confluent jets on a curved surface, the results of numerical prediction of a cylindrical air supply device using two turbulence models (realizable 𝑘 − 𝜖 and Reynolds stress model) are validated with hot-wire anemometry (HWA) near different nozzles discharge in the array. A single round jet is then studied to find the appropriate turbulence models for the prediction of the three-dimensional flow field and to gain an understanding of the effect of the boundary conditions predicted at the nozzle inlet. In comparison with HWA measurements, the turbulence models with low Reynolds correction (𝑘 − 𝜖 and shear stress transport [SST] 𝑘 − 𝜔) give reasonable flow predictions for the single round jet with the prescribed inlet boundary conditions, while the transition models (𝑘 − 𝑘l − 𝜔𝜔 and transition SST 𝑘 − 𝜔) are unable to predict the flow in the turbulent region. The results of numerical prediction (low Reynolds SST 𝑘 − 𝜔 model) using the prescribed inlet boundary conditions agree well with the HWA measurement in the nearfield of confluent jets on a curved surface, except in the merging region. Instantaneous velocity measurements are performed by laser Doppler anemometry (LDA) and particle image velocimetry (PIV) in two different configurations, a single row of parallel coplanar jets and an inline array of jets on a flat surface. The results of LDA and PIV are compared, which exhibit good agreement except near the nozzle exits. The streamwise velocity profile of the jets in the initial region shows a saddle back shape with attenuated turbulence in the core region and two off-centered narrow peaks. When confluent jets issue from an array of closely spaced nozzles, they may converge, merge, and combine after a certain distance downstream of the nozzle edge. The deflection plays a salient role for the multiple interacting jets (except in the single row configuration), where all the jets are converged towards the center of the array. The jet position, such as central, side and corner jets, significantly influences the development features of the jets, such as velocity decay and lateral displacement. The flow field of confluent jets exhibits asymmetrical distributions of Reynolds stresses around the axis of the jets and highly anisotropic turbulence. The velocity decays slower in the combined regio  of confluent jets than a single jet. Using the response surface methodology, the correlations between characteristic points (merging and combined points) and the statistically significant terms of the three design factors (inlet velocity, spacing between the nozzles and diameter of the nozzles) are determined for the single row of coplanar parallel jets. The computational parametric study of the single row configuration shows that spacing has the greatest impact on the near-field characteristics.

About 8 tons of dry undigested solid waste is generated by the MixAlco process for every 40 tons of food residue waste fed into the process. This MixAlco process produces liquid fuels and the sludge generated can be further converted into synthesis gas using the process of pyrolysis. The hydrogen component of the product synthesis gas may be separated by pressure swing adsorption and used in the hydrogenation of ketones into fuels and chemicals. The synthesis gas may also be catalytically converted into liquid fuels via the Fischer-Tropsch synthesis process. The auger-type pyrolyzer was operated at a temperature between 630-770 degrees C and at feed rates in the range of 280-374 g/minute. The response surface statistical method was used to obtain the highest syngas composition of 43.9 +/- 3.36 v % H2/33.3 +/- 3.29 v % CO at 740 degrees C. The CH4 concentration was 20.3 +/- 2.99 v %. For every ton of sludge pyrolyzed, 5,990 g H2 (719.3 MJ), 65,000 g CO (660 MJ) and 21,170 g CH4 (1055.4 MJ) were projected to be produced at optimum condition. At all temperatures, the sum of the energies of the products was greater than the electrical energy needed to sustain the process, making it energy neutral. To generate internal H2 for the MixAlco process, a method was developed to efficiently separate H2 using pressure swing adsorption (PSA) from the synthesis gas, with activated carbon and molecular sieve 5A as adsorbents. The H2 can be used to hydrogenate ketones generated from the MixAlco process to more liquid fuels. Breakthrough curves, cycle mass balances and cycle bed productivities (CBP) were used to determine the maximum hydrogen CBP using different adsorbent amounts at a synthesis gas feed rate of 10 standard lpm and pressure of 118 atm. A 99.9 % H2 purity was obtained. After a maximum CBP of 66 % was obtained further increases in % recovery led to a decrease in CBP. The synthesis gas can also be catalytically converted into liquid fuels by the Fischer-Tropsch synthesis (FTS) process. A Co-SiO2/Mo-Pd-Pt-ZSM-5 catalyst with a metal-metal-acid functionality was synthesized with the aim of increasing the selectivity of JP-8 (C10-C17) fuel range. The specific surface areas of the two catalysts were characterized using the BET technique. The electron probe microanalyzer (with WDS and EDS capabilities) was then used to confirm the presence of the applied metals Co, Mo, Pd and Pt on the respective supports. In addition to the gasoline (C4-C12) also produced, the synthesis gas H2:CO ratio was also adjusted to 1.90 for optimum cobalt performance in an enhanced FTS process. At 10 atm (150 psig) and 250 degrees C, the conventional FTS catalyst Co-SiO2 produced fuels rich in hydrocarbons within the gasoline carbon number range. At the same conditions the Co-SiO2-Mo-Pd-Pt/HZSM-5 catalyst increased the selectivity of JP-8. When Co-SiO2/Mo-Pd-Pt-HZSM-5 was used at 13.6 atm (200 psig) and 250 degrees C, a further increase in the selectivity of JP-8 and to some extent diesel was observed. The relative amounts of olefins and n-paraffins decreased with the products distribution shifting more towards the production of isomers.

Conselho Nacional de Desenvolvimento Científico e Tecnológico / The generation of residual materials and its unavoidable disposal in the environment has been a reason of great concern, not only environmentally but also economically. The use of these renewable residual feedstocks, not only for the production of heat and energy, but also for the generation of technological products, has become a priority. In this study, it was investigated the physico-chemical and fermentative processes for the good use of the agro-industrial residue rice husks. The husks, resulting from the rice processing, has been also used, even incipiently, for the generation of heat to dry grains and electrical energy. As a result of this process, rice husk ash (RHA) is available, very rich in silica (sílica), representing a potential future source of industrial raw material. Micronized RHA, obtained from micro grinding process, was used to produce three types of silica xerogels, which were tested as additives for rice cultivation. The good results demonstrated the feasibility of the use of the xerogel silicas in substitution of agrochemical products. The influence of the process parameters (pH, time, concentration) was determined by means of the response surface methodology (RSM). The maximum production yields of sodium, potassium and calcium silica xerogels were 98%, 95% and 75%, respectively. The silica produced was characterized and analyzed by the following techniques: X-ray fluorescence and X-ray diffractometry (XRF, XRD), Fourier-transform infrared spectrometry (FTIR), scanning electronic microscopy (SEM), particle size distribution determined by laser diffraction, specific surface area by BET method (Brunauer, Emmett e Teller), and determination of metals by inductively coupled plasma emission optical pectrometry (ICP-OES). The production of xylose was studied by acid hydrolysis of rice husk, under pressure, in autoclave. Then, the xylose was converted into xylitol by fermentation. The process parameters, temperature and reaction time, were adjusted by RSM. Fermentations were performed using the yeast species Candida guilliermondii and Candida tropicalis, with maximum yields of 40% and 8%, respectively. The influence of various types of pretreatment of the sample was evaluated, demonstrating the potential yields of 66% and 64%, respectively, when the husk was treated with hydrogen peroxide and ammonium hydroxide solutions. When using physical pretreatment (ultrasound), yields up to 62% can be obtained. The influence of the variation of acid concentration and pH of the hydrolyzate on the yields was also evaluated. The contact time and amount of activated carbon added to the hydrolyzate exerts great influence on the process yield. The products obtained - xylose and xylitol - were identified and quantified by liquid chromatography coupled to mass spectrometer (LC-MS/MS) with electrospray ionization in positive mode. Results of the second order RSM applied to obtain silica xerogel as well as xylitol were evaluated by analysis of variance (ANOVA). The efficiency of the processes studied here shows that they may be interesting alternatives of use of biomass residue rice husk, with considerable economic potential and positive environmental impact. / A geração de materiais residuais e o seu descarte no ambiente têm sido motivo de grande preocupação, tanto ambiental quanto econômica. O aproveitamento destas matérias-primas residuais renováveis, não apenas para a produção de energia e calor, mas também para a geração de produtos tecnológicos, tornou-se uma prioridade. Neste trabalho, investigaram-se processos físico-químicos e fermentativos para o aproveitamento do resíduo agroindustrial casca de arroz (CA). A casca, resultante do beneficiamento do arroz, tem sido utilizada, ainda que incipientemente, na produção de calor para secagem de grãos e produção de energia elétrica. Em consequência deste processo, tem-se a formação de cinza da casca de arroz (CCA), muito rica em sílica (SiO2), apresentando grande potencial como futura matéria-prima industrial. A CCA micronizada, obtida a partir do processo de micromoagem, foi utilizada na obtenção de três tipos de sílicas xerogéis, que foram testadas como aditivos para o cultivo de arroz. Os bons resultados alcançados demonstram a viabilidade do aproveitamento da sílica xerogel em substituição a derivados agroquímicos. A influência dos parâmetros de processo (pH, tempo, concentração) foi determinada por meio de metodologia de superfície de resposta (RSM). Os rendimentos máximos de produção das sílicas xerogéis base sódio, potássio e cálcio foram de 98%, 95% e 75%, respectivamente. As sílicas produzidas foram caracterizadas e analisadas por meio de técnicas de fluorescência e difração de raios-X (XRF, XRD), espectrometria no infravermelho com transformada de Fourier (FTIR), microscopia eletrônica de varredura (SEM), distribuição do tamanho de partículas segundo a técnica de difratometria a laser, área de superfície específica pelo método BET (Brunauer, Emmett e Teller) e determinação de metais pela técnica de espectrometria de emissão óptica com plasma indutivamente acoplado (ICP-OES). A produção de xilose foi estudada por meio de hidrólise ácida à pressão da CA. A xilose, por sua vez, foi convertida em xilitol por meio de processo fermentativo. Os parâmetros de processo, temperatura e tempo reacional, foram ajustados por meio de RSM. As fermentações foram feitas com auxílio das leveduras das espécies Candida guilliermondii e Candida tropicalis, com rendimentos máximos de 40% e 8%, respectivamente. Avaliou-se a influência de diversos tipos de pré-tratamento da amostra, evidenciando-se rendimentos de 66% e 64%, respectivamente, quando a CA foi tratada com solução de peróxido de hidrogênio e com solução de hidróxido de amônio. Quando se utiliza pré-tratamento físico (ultra-som), rendimento de até 62% pode ser obtido. A influência da variação da concentração de ácido e do pH do hidrolisado sobre o rendimento também foi avaliada. Comprovou-se, ainda, que o tempo de contato e a quantidade de carvão ativado adicionado ao hidrolisado, exercem grande influência sobre o rendimento. Os produtos obtidos - xilose e xilitol - foram identificados e quantificados por cromatografia líquida acoplada a espectrômetro de massas (LC-MS2) com ionização eletrospray, no modo positivo. Os resultados do modelo de superfície de resposta de segunda ordem, aplicado tanto ao processo de obtenção de sílicas xerogéis como ao de produção de xilitol, foram avaliados por meio de análise de variância (ANOVA). A eficiência dos processos estudados demonstra que estes podem constituir interessante alternativa para o aproveitamento da biomassa residual CA, com apreciável potencial econômico e positiva repercussão ambiental.

Les procédés de fabrication de pièces mécaniques par enlèvement de matière (tournage, fraisage, perçage, ...) connaissent une utilisation massive dans l’industrie aéronautique et l’automobile. Les pièces obtenues par ces procédés doivent satisfaire à des propriétés géométriques, métallurgiques et à des caractéristiques de qualité. Pour répondre à ces exigences, plusieurs essais expérimentaux basés sur le choix des conditions de coupe sont souvent nécessaires avant d’aboutir à une pièce satisfaisante. Actuellement, ces méthodes empiriques basées sur l’expérience des fabricants et des utilisateurs des outils coupants sont souvent très longues et coûteuses, donnent une large plage de choix des paramètres en fonction de leurs besoins. Toutefois, le coût très élevé d’un essai limite fondamentalement le nombre d’expériences, avoir une pièce respectant les caractéristiques souhaitées avec un coût acceptable devient une tâche difficile. / Manufacturing processes of mechanical parts by removal of material (turning, milling, drilling ...) have extensive use in aeronautic and automobile industry. The components obtained using these methods must satisfy geometric properties, metallurgical and quality characteristics. To meet these requirements, several experimental tests based on the selection of cutting conditions are often necessary before manufacturing. Currently, these empirical methods based on the experience of manufacturers and users of cutting tools (charts, diagrams with experimental findings, ...) are often very lengthy and costly. However, the high cost of a trial limits the number of experiments, so to have a deserted component with an acceptable cost is a difficult task. The importance of cutting conditions monitored by limitations is related to the type of material to be machined, since it determines the behavior of the machining.

Made available in DSpace on 2016-06-02T19:02:44Z (GMT). No. of bitstreams: 1 6412.pdf: 14036212 bytes, checksum: dc6e90045bca1050bc9fd76f94f1f30a (MD5) Previous issue date: 2014-06-30 / Financiadora de Estudos e Projetos / The increase in the production of antimicrobial metabolites by Paenibacillus polymyxa RNC-D was appraised through the study of cultivation variables. Two process variables, namely the glucose and inoculum concentrations, were evaluated in different levels (5 to 40 g/l, and 2.5% to 5.0% v/v, respectively), and their effects on biomass formation, minimal inhibitory concentration (MIC) against Escherichia coli and surface tension reduction (STR) were studied. The fermentation process was firstly carried out using non-optimized parameters, where the dependent variables biomass, MIC and STR reached the values of 0.6 g/l, 1.000,0 μg/ml and 18.4 mN/m, respectively. The optimum glucose (16 g/l) and inoculum concentrations (5.0% v/v) were defined in order to maximize the biomass formation, with low value of MIC and large STR of extract. Under these conditions, a biomass of 2.76 g/l, MIC of 15.8 μg/ml, and STR of 14.58 mN/m were predicted by the model. Data attained by experiments using optimized settings showed the following values: biomass 2.05 g/l; MIC 31.2 μg/ml; STR 10.7 mN/m. Thus, the percentage of improvement for each target response was: biomass 241.6%; MIC 96.88%; STR 41.85%. It was found that high concentrations of glucose substrate, although reflected in an increase in bacterial biomass, inhibited the microbial secondary metabolism, resulting in a low production of biomolecules associated with high values of MICs. Thus, initial concentrations of glucose and inoculum are shown as variables of strong influence in the production of antimicrobial metabolites by P. polymyxa RNC-D. Through the methods of experimental factorial design and surfaceresponse followed by graphical optimization it was possible to determine the optimum operating condition to achieve both maximum biomass and RTS as well as and lowest possible values of CIM. The validity of the proposed model was verified and confirmed. This is the first study on the optimization of culture conditions for the production of antimicrobial metabolites by P. polymyxa RNC-D, and constitutes an important step in the development of strategies to modulate the production of antimicrobial molecules by this microorganism in elevated levels. Novel antimicrobial compounds were isolated from the fermentation broth of P. polymyxa RNC-D, here named total extract (TE). It was possible to verify the presence of lipopeptide and peptide active compounds through enzymatic assays made with ET. Total extract was subjected to a two-phase system, resulting in lipopeptide extract (LPE) and aqueous fraction (AF). According to the results of bioassays, LPE has broad-spectrum activity against Gram-positive bacteria, Gram-negative bacteria and fungi. The mass spectrometry analysis of PLA revealed the existence of a novel compound that was named polycerradin. The purification of a novel antimicrobial peptide (AMP) from the AF was carried out by using chromatography. The compound was active against Gram-negative bacteria. Nterminal analysis determined the amino acid sequence, as well as MS / MS analysis confirmed the primary structure of this new compound. This research reports firstly the production of PAM PpRNCD that has an unusual amino acid in its constitution. It is an unprecedented fact considering the bacterial specie P. polymyxa. In terms of molecule size, PAM PpRNCD can be considered one of the smallest active natural peptide reported to date. It was also possible to isolate from FA the depsipeptides IL-F04a (m/z 883), LI-F04b (m/z 897), LI-F03a (m/z 947) and LI-F03b (m/z 961) previously described in the literature. The photoluminescence study of the LPE, TE, AF in both at room temperature (RT) and low temperature (T = 8K) was performed. In addition, this technique was applied to evaluate the action of the ELP on Staphylococcus aureus ATCC 29213, Enterococcus faecalis ATCC 29212, Escherichia coli ATCC 29212, Shigella sonnei ATCC 1578 and Candida albicans ATCC 10231 in two different situations: (a) immediately after mixing LPE with the bacterial and fungus cell suspension, and (b) after thirty minutes. The photoluminescence emission was collected by a triple spectrometer (three diffraction gratings) T64000 model from Jobin Yvon, equipped with an optical microscope. For the detection of the radiation emitted by the sample we used a CCD camera (charge coupled device) cooled by liquid nitrogen. The slits of the spectrometer were adjusted to produce a spectral resolution of the order of 10-4 nm. The excitation source used was the line of 457 nm (violet) from an argon laser. The behaviors here observed indicate a strong potential for applications in biosensors as well as molecular markers. / Através do estudo de variáveis do cultivo pretendeu-se aumentar a produção de metabólitos antimicrobianos por Paenibacillus polymyxa RNC-D. Duas variáveis do processo - glicose e concentração de inóculo - foram avaliadas em diferentes níveis e seus efeitos na formação de biomassa, concentração inibitória mínima (CIM) contra Escherichia coli e redução na tensão superficial (RTS) foram estudados. Utilizando parâmetros não-otimizados as variáveis dependentes biomassa, CIM e RTS atingiram valores de 0,6 g/l, 1.000,0 μg/ml e 18,4 mN/m, respectivamente. As concentrações ótimas de glicose (16 g/l) e inóculo (5,0% v/v) foram definidas no sentido de maximizar a formação de biomassa e RTS do extrato, bem como diminuir o valor de CIM do extrato. Experimentalmente 2,05 g/l de biomassa; 31,2 μg/ml de CIM e 10,7 mN/m de RTS foram obtidos sob condições otimizadas. Foi constatado que altas concentrações do substrato glicose, embora refletissem em aumento de biomassa bacteriana, inibiram o metabolismo secundário microbiano, resultando em baixa produção de biomoléculas associada a altos valores de CIM. Através dos métodos de design fatorial experimental e superfície-resposta seguidos por otimização gráfica foi possível determinar a condição operacional ótima das concentrações iniciais de glicose e inóculo, as quais se demonstraram como variáveis de grande influência na produção de metabólitos antimicrobianos por P. polymyxa RNC-D. O extrato total (ET), proveniente do caldo de fermentação de P. polymyxa RNC-D, foi utilizado para pesquisa e isolamento de novos compostos antimicrobianos. Através de ensaios enzimáticos feitos com ET foi possível verificar a natureza lipopeptídica e peptídica dos compostos antimicrobianos. O ET foi submetido a um sistema de duas fases, separandose então em extrato lipopeptídico (ELP) e fração aquosa (FA). Resultados de bioensaios revelaram que o ELP apresenta amplo espectro de atividade contra bactérias Grampositivas, Gram-negativas e fungo. A análise por espectrometria de massas de ELP revelou a presença de um composto peptídico inédito o qual foi denominado polycerradin. A partir da fração aquosa (FA) foi possível a purificação de um novo peptídeo antimicrobiano (PAM) através de etapas cromatográficas. A bioatividade do composto foi avaliada e confirmada frente às bactérias Gram-negativas. A determinação da sequência de aminoácidos foi realizada por análise do N-terminal, e a confirmação da estrutura primária deste novo composto foi feita por MS/MS. O presente estudo relata pela primeira vez a produção do PAM PpRNCD que possui um aminoácido não usual em sua constituição, relato primeiramente aqui descrito considerando-se a espécie bacteriana P. polymyxa. Em termos de tamanho de molécula, pode-se considerar que o PAM PpRNCD é um dos menores peptídeos naturais ativos relatados até o momento. Utilizando-se a FA também foi possível o isolamento dos depsipeptídeos LI-F04a (m/z 883), LI-F04b (m/z 897), LI-F03a (m/z 947) e LI-F03b (m/z 961) previamente descritos na literatura. O estudo da fotoluminescência do ELP, do ET e da FA foi realizado tanto em temperatura ambiente (RT) quanto em baixa temperatura (T=8K). Também se estudou, através desta técnica, a ação do ELP sobre as bactérias Staphylococcus aureus ATCC 29213, Enterococcus faecalis ATCC 29212, Escherichia coli ATCC 29212, Shigella sonnei ATCC 1578 e fungo Candida albicans ATCC 10231 em duas situações: (a) imediatamente após a mistura do ELP com a suspensão celular bacteriana, e (b) trinta minutos após a mistura. Detectou-se emissão fotoluminescente por ELP, ET e FA, e sinais de Raman a λ 699 nm (FA a baixa temperatura). Decorridos 30 min da mistura do ELP com as suspensões celulares microbianas houve alteração na emissão fotoluminescente, sendo que alguns sinais foram suprimidos (λ 470, 480 e 700 nm para S. sonnei, por exemplo). Isto evidencia a potencial aplicação destas frações (ELP, ET e FA) para a fabricação de sensores, detectores e marcadores moleculares.

Desde meados do século XX, métodos analíticos, numéricos e experimentais foram utilizados para quantificar a eficiência térmica na soldagem a arco, isto é, a razão entre a energia do arco elétrico e a energia que é entregue ao material durante a soldagem. Conhecendo-se a eficiência térmica, as dimensões da chapa e as propriedades térmicas do metal de base, torna-se possível prever as taxas e tempos de resfriamento a partir de modelo analíticos. Consequentemente, permitindo a previsão das transformações de fase, garantindo que os valores das propriedades da junta soldada se enquadrem dentro de um intervalo desejado. Os modelos de previsão de taxa de resfriamento derivados a partir dos modelos analíticos de Rosenthal são validos apenas no regime quase estacionário. Todavia, estes modelos costumam ser utilizados com pouco critério, mesmo em situações em que este regime não é alcançado. Portanto, para o desenvolvimento de modelos mais precisos e acurados para previsões das taxas de resfriamento, torna-se necessário entender como os termos do balanço de energia se desenvolvem durante a soldagem, até atingir o regime quase estacionário. Este trabalho tem como objetivo desenvolver um entendimento mais abrangente dos fatores que impactam nos valores de taxas de resfriamento em juntas soldadas. Realizaram-se ensaios de calorimetria com nitrogênio líquido para diferentes tempos de soldagem, como também, foram coletados ciclos térmicos em diferentes posições ao longo do cordão, buscando entender como as variações no balanço de energia, ao longo da soldagem, impactam no material. A partir da Metodologia de Superfície de Resposta, foram obtidos os modelos empíricos da energia entregue ao material e da eficiência térmica do arco. Um novo entendimento acerca do balanço de energia na soldagem foi alcançado, servindo de base para o desenvolvimento e validação de um modelo preditivo de taxa de resfriamento, válido para diferentes condições de fluxo de energia no material. / Since the early 20th century, numerical and experimental methods have been used to quantify the thermal arc efficiency in welding, i.e., the ratio between the electric arc energy and the energy delivered to the material during welding. Knowing the thermal arc efficiency, the sample dimensions and the thermal properties of the base metal, it is possible to predict the cooling rates by analytical models. Consequently, allowing the prediction of phase transformations in the material, ensuring that values of welded joint properties are within a desired range. Cooling rate prediction models derived from Rosenthal\'s analytical models are valid only in quasi-stationary state. However, these models are used with little criterion, even in situations where this state is not achieved. Therefore, to development of more accurate and precise models for the prediction of cooling rate it become necessary to understand the evolution of the energy balance during the welding until the quasi-stationary state was reached. This work aims to develop a more comprehensive understanding of the factors that affect the cooling rate in welded joints. Calorimetry tests with liquid nitrogen were carried out for different welding times, and thermal cycles were collected at different positions along the weld bead, to understand how the variations in the energy balance during the welding affect the material. From the Response Surface Methodology, the empirical models of delivered energy and the thermal arc efficiency were obtained. A new understanding about the energy balance in the welding was found, that was used for the development and validation of a predictive model of cooling rate, valid for different condition of energy flow in the material.