The aim of this diploma thesis is detailed characterization of new diol-based column, i.e. Torus DIOL designed for supercritical fluid chromatography, in hydrophilic interaction liquid chromatography. This stationary phase contains diol ligands bonded on BEH ("bridged ethylene hybrid") particles. The BEH sorbent is more stable at high pH, pressure and temperature than conventional silica sorbents. Five fluorinated pesticides were successfully separated on Torus DIOL column. Retention mechanism and effect of different parameters were tested by analysis of small polar compounds (nucleobases, uridine and its derivatives, acidic, neutral and basic compounds). Both partitioning of analytes between an aqueous layer deposited on the surface of the stationary phase and the mobile phase and adsorption of analytes on the stationary phase participate in the retention mechanism. Effect of the mobile phase aqueous part pH was investigated with the following buffering solutions: formic acid (pH = 2.1), 10mM ammonium acetate (pH = 4.7 and pH = 9.5). Obtained results showed that aqueous part pH significantly affects retention of acidic compounds according to their pKA values. Retention of all tested analytes on Torus DIOL column slightly increased with increasing ionic strength of mobile phase. The retention...

In the production of removable facial prosthetics, Computer-Aided Design and Manufacture (CAD/CAM) is being increasingly explored. This PhD thesis investigates the application of CAD/CAM in the design and production of components that retain the prostheses to the anatomy. Conventional methods of hand-crafting the retention elements are well established but little research has considered producing these elements using CAD/CAM. A fully digital prosthetic workflow has not yet been developed, and the efficacy of using CAD/CAM for retention mechanism design and fabrication remains unclear. This study firstly focuses on defining the requirements for designing extra-oral prosthesis retention mechanisms, by mapping the various stages of conventional practice and obtaining the opinions of practicing clinicians. Secondly, the qualitative findings are applied to develop a fully CAD/CAM process using existing technologies. Scanning, reverse engineering, design and fabrication technologies are trialled and samples of bar-clip mechanisms are produced. The final stage focuses on developing objective methods to evaluate aspects of bar-clip design previously limited to subjective evaluation, and to make an initial comparison of conventional and CAD/CAM bar-clip mechanisms. This focuses on measuring surface and dimensional quality, accuracy of fit and clip retention forces. This study provides an increased knowledge-base of current prosthetic practice; CAD/CAM prosthesis production and evaluation methods; and insight into the attitudes of clinicians towards the integration & implementation of CAD/CAM. The thesis demonstrates that CAD/CAM can be used to design, produce, and integrate bar-clip retention mechanisms in all aspects of the prosthesis production workflow. Digital measurement methods allow an objective evaluation of the important aspects of bar-clip mechanism design, identifying a number of inaccuracies/design flaws that current evaluation techniques fail to identify. The study concludes that the overall CAD/CAM workflow is not yet appropriate for clinical practice but there is potential in the newly developed processes and this drives future work.

Hydrophilic interaction chromatography is well known as a powerful technique separation of polar and ionizable compound nowadays. However the retention mechanism of the technique is still under debate. Understanding retention mechanism would facilitate the method development using the technique and its future improvement. This was inspiring and became the goal of this thesis. This work involves the characterization of the water enriched layer regarding to water and buffer salt accumulation. Twelve HILIC stationary phase with a diverse surface chemistry regarding to function groups and modification type were studied. Effect of water and salt on regarding to the retention mechanism was investigated by correlating the adsorption data to the retention of selected solutes This also involved the characterization of interactions involve in the separation of 21 HILIC columns. Interactions was probe by retention ratio of pair solutes which are characteristic for each specific interaction. The data was evaluate using principle component analysis – a multivariable data analysis method. The model was comprehensive and its outcomes were confirmed by the studies on adsorptions of water and salts.

<p>Electrospray ionization (ESI) mass spectrometry (MS) in combination with liquid chromatography (LC) is an excellent tool for the identification of drug metabolites. Utilizing this hyphenated technique in combination with proper sample pretreatment, the metabolic pathways of the analgesic drug ketobemidone were investigated in human urine and rat microdialysate from blood and brain. Two novel phase I metabolites (ketobemidone N-oxide and meta-hydroxymethoxyketobemidone) and three novel phase II metabolites (glucuronic acid conjugates of ketobemidone, norketobemidone and hydroxymethoxyketobemidone) were identified in human urine. Further, norketobemidone and ketobemidone N-oxide were identified in rat microdialysate from brain after regional distribution of ketobemidone in striatum. This indicates that the brain itself has the possibility to metabolize ketobemidone. </p><p>Synthetic ketobemidone metabolites were used for comparison of retention times and tandem MS spectra with the possible metabolites recovered from the biological samples. The conjugated metabolites were identified by accurate mass measurements and tandem MS spectra of the aglycones. The accuracy of the estimated masses was better than 2.1 ppm for two out of three conjugates in presence of internal standard.</p><p>On-line micro-SPE was successfully used for trapping and desalting of the microdialysates. The small SPE pre-column made it possible to inject approximately 100 times more sample on the analytical column compared to injection without pre-column. Selective trapping was demonstrated for the polar catechol amine metabolite, dihydroxyketobemidone, which forms covalent complexes with phenylboronic acid (PBA). A fluorinated silica type stationary phase was the only column out of several tested that was able to separate ketobemidone and all relevant phase I metabolites. </p><p>Liquid chromatography and mass spectrometry are independently valuable tools in the field of analytical pharmaceutical chemistry. The present study showed that the combination of LC-MS, with its excellent selectivity and sensitivity, offers an outstanding tool in the qualitative analysis of drugs and metabolites in biological fluids. </p>

Electrospray ionization (ESI) mass spectrometry (MS) in combination with liquid chromatography (LC) is an excellent tool for the identification of drug metabolites. Utilizing this hyphenated technique in combination with proper sample pretreatment, the metabolic pathways of the analgesic drug ketobemidone were investigated in human urine and rat microdialysate from blood and brain. Two novel phase I metabolites (ketobemidone N-oxide and meta-hydroxymethoxyketobemidone) and three novel phase II metabolites (glucuronic acid conjugates of ketobemidone, norketobemidone and hydroxymethoxyketobemidone) were identified in human urine. Further, norketobemidone and ketobemidone N-oxide were identified in rat microdialysate from brain after regional distribution of ketobemidone in striatum. This indicates that the brain itself has the possibility to metabolize ketobemidone. Synthetic ketobemidone metabolites were used for comparison of retention times and tandem MS spectra with the possible metabolites recovered from the biological samples. The conjugated metabolites were identified by accurate mass measurements and tandem MS spectra of the aglycones. The accuracy of the estimated masses was better than 2.1 ppm for two out of three conjugates in presence of internal standard. On-line micro-SPE was successfully used for trapping and desalting of the microdialysates. The small SPE pre-column made it possible to inject approximately 100 times more sample on the analytical column compared to injection without pre-column. Selective trapping was demonstrated for the polar catechol amine metabolite, dihydroxyketobemidone, which forms covalent complexes with phenylboronic acid (PBA). A fluorinated silica type stationary phase was the only column out of several tested that was able to separate ketobemidone and all relevant phase I metabolites. Liquid chromatography and mass spectrometry are independently valuable tools in the field of analytical pharmaceutical chemistry. The present study showed that the combination of LC-MS, with its excellent selectivity and sensitivity, offers an outstanding tool in the qualitative analysis of drugs and metabolites in biological fluids.

Made available in DSpace on 2014-12-17T14:09:18Z (GMT). No. of bitstreams: 1 JulianaAA_TESE.pdf: 2312979 bytes, checksum: 31c1a19cbf37d537febd576ec54e5eb2 (MD5) Previous issue date: 2013-09-13 / Discrepancies between classical model predictions and experimental data for deep bed filtration have been reported by various authors. In order to understand these discrepancies, an analytic continuum model for deep bed filtration is proposed. In this model, a filter coefficient is attributed to each distinct retention mechanism (straining, diffusion, gravity interception, etc.). It was shown that these coefficients generally cannot be merged into an effective filter coefficient, as considered in the classical model. Furthermore, the derived analytic solutions for the proposed model were applied for fitting experimental data, and a very good agreement between experimental data and proposed model predictions were obtained. Comparison of the obtained results with empirical correlations allowed identifying the dominant retention mechanisms. In addition, it was shown that the larger the ratio of particle to pore sizes, the more intensive the straining mechanism and the larger the discrepancies between experimental data and classical model predictions. The classical model and proposed model were compared via statistical analysis. The obtained p values allow concluding that the proposed model should be preferred especially when straining plays an important role. In addition, deep bed filtration with finite retention capacity was studied. This work also involves the study of filtration of particles through porous media with a finite capacity of filtration. It was observed, in this case, that is necessary to consider changes in the boundary conditions through time evolution. It was obtained a solution for such a model using different functions of filtration coefficients. Besides that, it was shown how to build a solution for any filtration coefficient. It was seen that, even considering the same filtration coefficient, the classic model and the one here propposed, show different predictions for the concentration of particles retained in the porous media and for the suspended particles at the exit of the media / Discrep?ncias encontradas entre dados experimentais e previs?es feitas a partir do modelo cl?ssico foram relatadas por v?rios autores. Para entender essas discrep?ncias, um modelo anal?tico cont?nuo para a filtra??o profunda ? proposto. Neste modelo, cada mecanismo de reten??o est? associado a um coeficiente de filtra??o diferente. Foi mostrado que os coeficientes de filtra??o n?o podem ser somados e considerados em um ?nico coeficiente de filtra??o global como se apenas um mecanismo de reten??o atuasse no sistema, o que ? feito no modelo cl?ssico. Al?m disso, foram obtidas solu??es expl?citas para o sistema de equa??es que representam o modelo proposto. Tais solu??es foram usadas para ajustar os dados experimentais, e um bom ajuste foi obtido. Comparando os resultados obtidos com rela??es emp?ricas dispon?veis na literatura foi poss?vel identificar o mecanismo de reten??o mais atuante na filtra??o. Foi mostrado ainda, que quanto maior o tamanho das part?culas injetadas, mais atuante ? o mecanismo de exclus?o pelo tamanho e maiores as discrep?ncias entre o modelo proposto e o modelo cl?ssico. Os modelos cl?ssico e proposto foram comparados atrav?s de uma an?lise estat?stica. Tal an?lise mostrou que os ajustes feitos com o modelo proposto s?o significativamente melhores que os ajustes feitos com o modelo cl?ssico, principalmente quando o mecanismo de exclus?o pelo tamanho ? o mais atuante na filtra??o. Neste trabalho foi, tamb?m, estudado a filtra??o de part?culas em meio poroso com capacidade finita de filtra??o. Observou-se, neste caso, que ? necess?rio considerar mudan?as nas condi??es de fronteira com a evolu??o do tempo. Foi obtida a solu??o para tal modelo para diferentes fun??es de coeficiente de filtra??o. Al?m disso, foi mostrado como construir a solu??o para um coeficiente de filtra??o qualquer. Observou-se que, ainda que se considere o mesmo coeficiente de filtra??o, o modelo cl?ssico e o modelo proposto apresentam previs?es distintas para as concentra??es de part?culas retidas no meio e de part?culas em suspens?o na sa?da do meio

High Performance Liquid Chromatography (HPLC) is a widely used tech-nique both for detecting and purifying substances in academy and in the industry. In order to facilitate the use of, and knowledge in HPLC, character-ization of stationary phases is of utmost importance. Tailor made characteri-zation methods and workflows are steadily increasing the speed and accura-cy in which new separation systems and methods are developed. In the field fundamental separation science and of preparative chromatography there is always the need for faster and more accurate methods of adsorption isotherm determination. Some of that demand are met with the steadily increase of computational power, but the practical aspects on models and methods must also be further developed. These nonlinear characterization methods will not only give models capable of describing the adsorption isotherm but also actual values of local adsorption energies and monolayer saturation capacity of an individual interaction sites etc.The studies presented in this thesis use modern alkali stable stationary phas-es as a model phase, which will give an insight in hybrid materials and their separation mechanism. This thesis will include an update and expansion in using the Elution by Characteristic Points (ECP) method for determination of adsorption isotherms. The precision is even further increased due to the ability to use slope data as well as an increase in usability by assigning a set of guidance rules to be applied when determine adsorption isotherms having inflection points. This thesis will further provide the reader with information about stationary phase characterization and the power of using existing tech-niques; combine them with each other, and also what the expansion of meth-ods can revile in terms of precision and increased usability. A more holistic view of what benefits that comes with combining a non-linear characteriza-tion of a stationary phase with more common linear characterization meth-ods are presented.