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Development of Instrumentation and Methodology in Proteomic and Environmental Analysis
Hezinová, Věra ; Bobáľová, Janette (oponent) ; Lubal,, Přemysl (oponent) ; Čáslavský, Josef (oponent) ; Klepárník, Karel (vedoucí práce)
This work is focused on both targeted and comprehensive approaches of proteomic studies. The targeted proteomics brings information about protein presence and localization in a cell or tissue using luminescence labels based on quantum dots, while the comprehensive approach deals with the identification of changes in proteomes of two or more specimens of the same organism that have grown in various conditions. Since the proteomics requires high sensitive separation and identification techniques, various methods of sensitivity improvement in capillary electrophoresis - mass spectrometry were verified in this work. The use of a liquid junction interface as a hyphenation between these two techniques ensures a higher sensitivity of analyses. This was verified also by the analysis of ethanol and cocaine metabolites in human urine. The developed instrumentation and methodology are utilizable for the estimation of the influence of important environmental factors on living systems at the cellular as well as molecular levels.
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A novel nanospray liquid junction interface for versatile CE-MS
Křenková, Jana ; Haselberg, R. ; Somsen, G. W. ; Foret, František
In this work we present a new polyimide-based liquid junction nanospray interface, combining the advantages of sheath flow and sheathless designs. The liquid junction interface was prepared as a hybrid microfabricated plastic liquid junction sprayer allowing to perform analysis in standard (30-100 micro m ID) separation capillaries. The interface incorporates a self-aligning port for coupling of the separation capillary as well as ports for automated flushing of both the liquid junction and the separation capillary. The interface permits performing CE both with and without electroosmotic flow. The use of large bore capillaries allows injection and preconcentration of larger sample volumes, favoring detection sensitivity. Different compositions of the spray liquid and BGE can be used when needed.
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Development of Instrumentation and Methodology in Proteomic and Environmental Analysis
Hezinová, Věra ; Bobáľová, Janette (oponent) ; Lubal,, Přemysl (oponent) ; Čáslavský, Josef (oponent) ; Klepárník, Karel (vedoucí práce)
This work is focused on both targeted and comprehensive approaches of proteomic studies. The targeted proteomics brings information about protein presence and localization in a cell or tissue using luminescence labels based on quantum dots, while the comprehensive approach deals with the identification of changes in proteomes of two or more specimens of the same organism that have grown in various conditions. Since the proteomics requires high sensitive separation and identification techniques, various methods of sensitivity improvement in capillary electrophoresis - mass spectrometry were verified in this work. The use of a liquid junction interface as a hyphenation between these two techniques ensures a higher sensitivity of analyses. This was verified also by the analysis of ethanol and cocaine metabolites in human urine. The developed instrumentation and methodology are utilizable for the estimation of the influence of important environmental factors on living systems at the cellular as well as molecular levels.
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