A successful supercritical fluid extraction method includes removal of the analyte from the matrix into the bulk fluid as well as trapping or concentration of the analyte prior to analysis. In the first phase of this research, the trapping capacities of three solid-phase traps (glass beads, 50/50 (w/w) glass beads/octadecylsilica), 50/50 (w/w) Porapak Q®/glass beads) were determined as a function of trap composition for a mixture of components varying in polarity and volatility. The Porapak Q®/glass beads mixture was found to be the most successful solidphase investigated exhibiting the highest trapping capacity. The use of the Porapak Q®/glass beads as a solid-phase trap was investigated in later extraction studies in this dissertation. The extraction of highly polar, multifunctional analytes may not be completely successful with modified carbon dioxide, therefore, a secondary modifier (i.e. additive) may be added directly to the extraction fluid in hopes of improving the recoveries. In the second phase of this research, the effect of secondary modifiers in the subcritical fluid extraction of lovastatin from in-house prepared tablet powder mixtures and MEVACOR® tablets was investigated. The effect of in-line methanol-modifier percentage, additive type (acidic, basic, neutral) to the in-line methanol, and additive concentration on the extraction efficiency were examined. The extraction recoveries of lovastatin from MEVACOR® tablets were shown to be highly dependent on methanol concentration and additive type. Isopropylamine was shown to be the most successful additive investigated. An optimized and reproducible extraction method was developed. The extraction of ionic compounds with carbon dioxide may be difficult due to the high polarity of the compounds. In the third phase of this research, the addition of ion-pairing additives to the matrix in hopes of forming an ion-pair complex of reduced analyte polarity was investigated. Therefore, a screening study consisting of a fractional-factorial design was performed in order to identify the factors which contribute most to the recovery of an anionic species, triphenylphosphinetrisulfonate (TPPTS), from a spiked-sand surface employing supercritical fluid extraction with carbon dioxide. The experimental parameters investigated were: type of ion-pairing additive (i.e. tetralkylammonium hydrogen sulfates) and its concentration, carbon dioxide density, extraction temperature, static extraction time, CO₂ mass used, liquid CO₂ flow rate, and the volume of methanol spiked into the matrix prior to extraction. Of the eight factors investigated, four factors were identified as significantly affecting the recovery of the anionic species. They were: 1) ion-pairing reagent added to the spiked sand surface and its concentration; 2) static extraction time; and 3) volume of methanol present in the extraction vessel. The experimental parameters and settings identified as influential by the statistical approach were later shown in concert to yield 100% recovery of TPPTS from the spiked-sand. In the fourth phase, the extraction of a cationic species, pseudoephedrine hydrochloride, from spiked-sand and Suphedrine tablets, with pure and methanol-modified CO₂ was examined. Once the extraction was shown to feasible, several strategies were compared in terms of their effectiveness in enhancing the analyte's extractability. The first strategy involved the addition of ion-pairing additives. Several sodium salts of alkylsulfonic acids varying in lipophilicity and concentration were investigated. The addition of 1-heptanesulfonic acid, sodium salt, in methanol, in a 5:1 mole ratio of reagent to analyte was shown to be the most useful in recovering the drug from the spiked-sand. The second strategy considered the influence of acids and bases and other modifier compositions such as a methanol/water mixture with or without 1-heptanesulfonic acid, sodium salt, on the pseudoephedrine recovery. The recoveries obtained from the drug spiked-sand were shown to comparable in the presence of a methanol/water solution, a tetrabutylammonium hydroxide in methanol solution, and a methanol solution with 1-heptanesulfonic acid, sodium salt. Next the extraction of pseudoephedrine hydrochloride from Suphedrine tablets was performed with pure and modified CO₂. Similar to the sand-spike studies, the effect of the addition of the ion-pairing reagent and other in-cell modifiers were examined. Once again, the recoveries obtained when the matrix was in the presence of a methanol/water mixture and a methanol solution containing 1-heptanesulfonic acid, sodium salt were similar. Finally, the identity of the extracted analyte was determined via IR analyses, and it was shown that pseudoephedrine hydrochloride was indeed extractable from the tablets with in-line modified CO₂ in the absence of any in-cell modifier. In the last phase of this research, a supercritical fluid chromatographic separation with evaporative light scattering detection was developed for the separation of five phospholipids varying in polarity and ionic characteristics. Several parameters were investigated and shown to be influential in the separation. They were: 1) stationary phase composition, 2) addition of an acidic additive and its concentration, 3) mobile phase ramp rate, and 4) column outlet pressure. / Ph. D.

No description available.

This thesis aims at a deeper understanding of Supercritical Fluid Chromatography (SFC). Although preparative SFC has started to replace Liquid Chromatography (LC) in the pharmaceutical industry - because of its advantages in speed and its less environmental impact - fundamental understanding is still lacking. Therefore there is no rigid framework to characterize adsorption or to understand the impact of changes in operational conditions.   In Paper I we demonstrated, after careful system verification, that most methods applied to determine adsorption isotherms in LC could not be applied directly in SFC. This was mainly due to operational differences and to the fact that the fluid is compressible which means that everything considered constant in LC varies in SFC.   In Paper II we showed that the most accurate methods for adsorption isotherm determination in LC, the so called plateau methods, do not work properly for SFC. Instead, methods based on overloaded profiles should be preferred.   In Paper III a Design of Experiments approach was successfully used to quantitatively describe the retention behavior of several solutes and the productivity of a two component separation system. This approach can be used to optimize SFC separations or to provide information about the separation system.   In Paper IV severe peak distortion effects, suspected to arise from injection solvent and mobile phase fluid mismatches, were carefully investigated using experiments and simulations. By this approach it was possible to examine the underlying reasons for the distortions, which is vital for method development.   Finally, in Paper V, the acquired knowledge from Paper I-IV was used to perform reliable scale-up in an industrial setting for the first time. This was done by carefully matching the conditions inside the analytical and preparative column with each other. The results could therefore provide the industry with key knowledge for further implementation of SFC. / This thesis aims at a deeper understanding of Supercritical Fluid Chromatography (SFC). Although preparative SFC has started to replace Liquid Chromatography (LC) in the pharmaceutical industry - because of its advantages in speed and its less environmental impact - fundamental understanding is still lacking. Therefore there is no rigid framework to characterize adsorption or to understand the impact of changes in operational conditions.   In Paper I-II it was demonstrated why most methods applied to determine adsorption isotherms in LC could not be applied directly for SFC. Methods based on extracting data from overloaded profiles should be preferred.   In Paper III a Design of Experiments approach was successfully used to quantitatively describe the behavior of several solutes in a separation system. This approach can be used to optimize SFC separations or to provide information about the separation system.   In Paper IV severe peak distortion effects often observed in SFC were carefully investigated and explained using experiments and simulations.   Finally, in Paper V, the prerequisites for performing reliable and predictable scale-up of SFC were investigated by small and large scale experiments. / <p>Paper 4 ("Evaluation of scale-up from analytical to preparative...") ingick som manuskript med samma titel i avhandlingen. Nu publicerad. </p>

No description available.

A novel chromatographic method was devised that makes use of the superb group separation power of normal phase supercritical fluid chromatography (SFC) combined with a fast second separation by a resistively heated gas chromatograph (GC). The SFC was operated isothermally with stopped flow to provide the time required for the GC analysis. The GC analysis had a typical cycle time of 1 minute. During this time the GC column was independently heated at a rate of 450°C/min to 250°C and actively cooled down again to -50°C before the next GC injection takes place. This was achieved with an in-house designed, resistively heated, temperature programmable gas chromatograph. Various temperature measurement circuits were also evaluated. An interface was developed that allows transfer between the SFC and the GC in such a way that the entire eluent from the first separation is analysed by the second separator. Chromatographic resolution was not lost during the transfer process from the first to the second separation stages. The interface also allows for the exchange of the carrier gas used in the second gas chromatographic separation to provide for the maximum separation speed. In the first separation, a silica gel packed column and the novel application of a silica gel porous layer open tubular capillary column was used for SFC group separation. The SFCxGCftp was applied to petrochemical samples and essential oils and the results were compared to that obtained with a commercially available GCxGC system. / Thesis (PhD)--University of Pretoria, 2003. / Chemistry / unrestricted

På fartyget M/S Calmare Nyckel finns en NOHAB-dieselmotor installerad ombord som används förundervisning på sjöfartshögskolan i Kalmar. Projektet har undersökt om det är möjligt att görauppstart av motorn säkrare för både handledare och elever. Arbetet har tagit fram ett nytt elskåp och styrsystem enligt DNV-Standard för att försäkra sig om att motorn inte går att starta utan att alla kringsystem som behövs är igång. I framtiden finns det även möjlighet att utveckla nya systemet om det skulle vara önskvärt. Arbetet har även konstaterat att det saknas ordentliga handlingar för elritningar och kabeluppmärkning ombord som bör åtgärdas i framtiden. / On the ship M/S Calmare Nyckel there is a NOHAB-Diesel Engine installed that are used foreducational uses on Kalmar Maritime Academy in Kalmar. This project has investigated if it’s possibleto make the startup of the engine safe both for teachers and students.The project has brought a new control cabinet and a new control system according to DNV-Standardto ensure that the engine doesn’t start without any auxiliary system that’s needed is running.In the future, there’s also possible to develop the system if that would be desirable.The project has also found that the lack of proper documents for electrical drawings and cablemarkings onboard should be addressed in the future.

The objective of this research was to develop new sample preparation procedures for the isolation of sulfonamides, as well as, to determine the applicability of employing on-line nitrogen selective and mass spectrometric detection methods. The first phase of this research investigated the effect of temperature and pressure on the supercritical fluid extraction (SFE) of sulfonamides from a spiked sand matrix. Temperature effects were either positive or negative with respect to extraction rate and total recovery, depending on the pressure and extraction fluid employed. The second portion of this research compared trifluoromethane (CHF3) and carbon dioxide (CO2) as fluids for the extraction of sulfonamides from spiked non-fat dry milk, beef liver, and egg yolk were found to be more selective using CHF3 than CO2. The polar trifluoromethane improved the extraction efficiency of the polar sulfonamides from the biological matrices and also reduced the amount of co-extractives. The next phase of this research considered the effect of organic modifier and CO2 in the SFE of sulfonamides from chicken liver, beef liver and egg yolk. Methanol, ethanol, acetone, acetonitrile were compared to determine optimum conditions. A SFE method employing 20% acetonitrile modified CO2 yielded quantitative recovery of sulfonamides from chicken liver, but 20% acetone modified CO2 was required to obtain quantitative recovery from beef liver. Either 20% acetone or 20% acetonitrile yielded quantitative recovery from egg yolk. The last phase of this research focused on the evaluation of selective detection methods for sulfonamide analysis. Chemiluminescence nitrogen detection (CLND) parameters were optimized for use with packed column supercritical fluid chromatography (SFC) yielding a minimum detectable quantity (MDQ) of 5 ng of sulfamethazine, on column. Improvements in the detector design decreased the MDQ to 0.5 ng, while, decreasing the column diameter further reduced the MDQ to 125 pg. The second part of this phase evaluated PLC/Atmospheric pressure chemical ionization (APCI) mass spectrometry for the detection of sulfonamides. Sensitivity in selective ion mode was found to be as low as 50 pg on column for sulfamethazine. Supercritical fluid extracts of sulfonamides spiked at 100μg/kg in chicken liver were found to be readily detected by this method. / Ph. D.

PolyHistidine repeats and DYRK1A: from the localization to the functionEl principal objectiu d'aquesta tesi ha estat el d'esbrinar noves funcions de la proteína quinasa DYRK1A en el nucli cel.lular. Donat que el domini de repetició d'histidines de DYRK1A dirigeix la proteína al compartiment d'speckles nuclears, aquesta propietat ha estat utilitzada per adreçar aquesta pregunta. Els resultats obtinguts en aquesta tesi han permès proposar els homopolímers d'histidina com una nova i general senyal de localització a speckles nuclears. Proteïnes amb segments de polihistidines, la majoria d'elles factors de transcripció, mostren un comportament intranuclear dinàmic, compatible amb un model en el quèl diferents dominis d'interacció competeixen entre ells pel reclutament de la proteína a diferents subcompartiments nuclears. El mecanisme molecular que media l'acumulació a speckles de les proteïnes amb polihistines s'ha estudiat utilitzant DYRK1A com a model. Els resultats obtinguts exclouen la unió a l'RNA com a mecanisme de reclutament i concloure que, aquest, ocorre mitjançant la interacció amb proteïnes residents. S'han identificat dues noves proteïnes interactores per a DYRK1A, l'RNA polimerasa II i el factor de transcripció Brn-3b. La fosforilació de DYRK1A sobre el domini C-terminal o CTD de l'RNA polimerasa II suggereix una funció directa de la quinasa en el procés de transcripció o del seu acoblament al processament d'RNAs missatgers. La fosforilació de DYRK1A sobre el domini d'activació de Brn-3b sembla regular positivament l'activitat transcripcional d'aquest factor. Aquests resultats indiquen una funció activa de DYRK1A en la regulació de la transcripció gènica, tant directament sobre la maquinària transcripcional com indirectament, modulant l'activitat de factors de transcripció. PolyHistidine repeats and DYRK1A: from the localization to the functionThe main objective of this thesis work has been to identify new roles for the protein kinase DYRK1A in the cell nucleus. Given that a histidine repeat in DYRK1A targets the protein to the nuclear speckle compartment, this property has been used as a tool to approach the question. The results obtained in this thesis work have allowed proposing homopolymeric histidine runs as a novel and general nuclear speckle-directing signal. Proteins with polyHistidine segments, mostly transcription factors, present a dynamic intranuclear behaviour compatible with a model in which distinct interacting domains compete for recruiting elements within the nucleus. The molecular mechanisms that mediate speckle accumulation have been studied in DYRK1A as a model system. The results allow excluding RNA binding as the recruiting mechanism and concluding that targeting is mediated by interaction with speckle-resident proteins. Two novel DYRK1A interactors have been identified during the study, the RNA polymerase II and the transcription factor Brn-3b. DYRK1A phosphorylation of the C-terminal domain or CTD of the RNA polymerase II suggests a direct role of DYRK1A on transcription or coupling of transcription with RNA processing. DYRK1A phosphorylation of Brn-3b within its activation domain seems to positively regulate Brn-3b transcriptional activity. These results confirm an active role for DYRK1A in gene transcription regulation both direct on the transcriptional machinery and indirect by modulating the activity of transcription factors.

Made available in DSpace on 2016-12-12T17:38:31Z (GMT). No. of bitstreams: 1 SUZANA RIBAS DE ALMEIDA.pdf: 2655112 bytes, checksum: 346dca905796b24136deb70b7263fb5b (MD5) Previous issue date: 2012-02-15 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Nowadays the competition between companies is increasing. In a production system, efficiency, speed and flexibility are important means for cost reductions. The complexity of automation systems has been increasing, which makes necessary the implementation of more efficient control programs, free of errors and easy to maintain. However, in most practical applications the solution of these problems is dependent on designer's experience and doesn t follow a formal methodology, which results in programs with errors and difficulties to understand and maintain, since only the designer understands the adopted solutions. The Supervisory Control Theory (SCT) is a method extensively researched in academic circles for the synthesis of control logic of automated systems. It allows control problems to be solved in a formal way, ensuring a minimally restrictive and no blocking solution that meets the control specifications. However, this theory is not widely used in industrial environments yet, once the methods for implementation of supervisors obtained by TCS are still deficient. Thus, the intention of this Dissertation consists of showing a methodology for implementation of supervisory control in Programmable Logic Controllers (PLCs). In this work, the synthesis of supervisors is made in accordance to local modular approach, which brings the benefits of a formal methodology and allows exploring the modularity of the plant and the control specifications. The implementation is based on an architecture structured in three levels: supervisor, interface and system to be controlled. For the implementation in CLP three different programming languages are applied: the high-level languages Sequential Function Charts (SFC) and Structured Text and the low-level Ladder Diagram language in some parts. The development of the methodology was based on two works: a doctoral thesis by Vieira (2007) and master thesis by Cruz (2011) and brought together the advantages presented in each one of these. To compare the proposed methodology and the two mentioned methods, several tests were made on a manufacturing cell available at PUCPR and simulations on a PLC and computer as well. The results show that the use of the methodology proposed in this work makes possible to implement PLC programs that are able to treat all non-controllable events occurred in the system to be controlled and treat one controllable event by each control cell in the same scan cycle. Furthermore, the use of high-level languages results in a program of easier interpretation and maintenance, also enabling reutilization of codes. In this work some problems related to CLPs implementations are also showed and proposals for solving them are presented, as properties as well that, if accepted, ensure that the problem does not occur. / Hoje em dia a competitividade entre as empresas está cada vez maior. Em um sistema de produção, a eficácia, a velocidade e a flexibilidade são importantes, pois significam redução de custos. Os sistemas de automação atuais estão cada vez mais complexos, justificando a necessidade de programas de controle mais eficientes, sem erros e de fácil manutenção. Entretanto, na maioria das aplicações práticas a solução destes problemas é feita com base na experiência do projetista, não seguindo uma metodologia formal, o que resulta em programas com erros e de difícil entendimento e manutenção, uma vez que só o projetista entende perfeitamente as soluções adotadas. A Teoria de Controle Supervisório (TCS) é um método para a síntese da lógica de controle de sistemas automatizados. Ela permite que problemas de controle sejam solucionados de modo formal, garantindo uma solução minimamente restritiva e não bloqueante e que atenda às especificações de controle. Entretanto, esta teoria ainda não é amplamente adotada em ambientes industriais, sendo a carência de métodos de implementação dos supervisores obtidos via TCS um elemento que contribui para que isso aconteça. Assim, esta dissertação se propõe a apresentar uma metodologia de implementação de controle supervisório em Controladores Lógicos Programáveis (CLPs). Neste trabalho, a síntese de supervisores é feita de acordo com a abordagem modular local, que traz os benefícios de uma metodologia formal e ainda permite explorar a modularidade da planta e das especificações de controle. A implementação é baseada numa arquitetura estruturada em três níveis: supervisor, interface e sistema a ser controlado. Para a implementação em CLP são empregadas três linguagens distintas de programação, as linguagens de alto nível Sequential Function Charts (SFC) e Structured Text, bem como a linguagem Ladder Diagram em algumas partes. O desenvolvimento da metodologia foi baseado em dois trabalhos: a tese de doutorado de Vieira (2007) e a dissertação de mestrado de Cruz (2011) e permitiu reunir as vantagens apresentadas em cada um destes. Para comparação entre a metodologia proposta e as duas metodologias citadas, foram feitos diversos testes em uma célula de manufatura existente na PUCPR, bem como simulações em CLP e em computador. Os resultados mostram que a metodologia proposta neste trabalho permite a implementação de programas em CLP que, num mesmo ciclo de varredura, são capazes de tratar todos os eventos não controláveis ocorridos na planta e ainda um evento controlável por célula de controle. Além disso, o uso de linguagens de alto nível resulta em um programa de mais fácil interpretação e manutenção, possibilitando ainda o reaproveitamento de códigos. Nesta dissertação também são abordados alguns problemas de implementação da estrutura de controle supervisório em CLPs, apresentando-se propostas para solucioná-los, bem como propriedades que, se atendidas, garantem que o problema não ocorra.

No description available.