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Multidimensional Liquid Phase Separations
Šesták, Jozef ; Čáslavský, Josef (referee) ; Pulkrabová, Jana (referee) ; Česla, Petr (referee) ; Kahle, Vladislav (advisor)
This dissertation is dedicated to the topic of multidimensional liquid phase separations. This separation techniques are developed for analysis of complex samples containing thermally labile, low volatile or high molecular weight components that can´t be analysed by two-dimensional (2D) gas chromatography. Concepts of peak capacity and orthogonality are explained and various methods of their determination are stated in theoretical part of dissertation. High performance column liquid chromatography (HPLC) and high performance capillary electrophoresis (HPCE) are suggested as the most suitable methods for automated multidimensional liquid phase separations on-line coupled to mass spectrometry. Configuration of simplified miniaturized liquid chromatograph is described in experimental part of this thesis. Original concept of the system has been extended by simple mobile phase gradient generation technique. Correct function was demonstrated on repeatable separation of alkylphenones, peptides, nitroaromatics, and nitroesters. This system has been utilized as a base for a couple of simple two-dimensional separation platforms for HILIC-MALDI-MS analysis of glycans, for separation of peptides based on off-line coupling of isoelectric focusing and capillary liquid chromatography, and finally for on-line IEC×RPLC, RPLC×RPLC, and HILIC×RPLC two-dimensional liquid chromatography. Correct operation of submitted platforms has been proved.
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Testing and optimization of pressure modulator conditions for GCxGC analyses
Ston, Martin ; Čabala, Radomír (advisor) ; Jelínek, Ivan (referee)
Pulse (fluid) pressure modulator was constructed for comprehensive gas chromatography. Optimal conditions of its connection to the gas chromatograph with a flame ionization detector were found for the analysis of selected mixture of volatile solvents. Pressure interdependences in the system and relations between dimensions of modulator capillaries and columns have been evaluated with respect to the length of the modulation period and pulse duration. Two non-polar columns (15 m × 0.25 mm DB-5MS, 10 m × 0.15 mm CP-Sil 5 CB) of the same stationary phase were tested to compare the separation efficiency in the first dimension and the DB-5MS column was selected. The Supelcowax 10 columns of the lengths 1, 2 and 3 m were compared in the second dimension. It was found that using of 3 m length column lead to the extension of analysis time, however the increase in separation efficiency in the second dimension was very evident. The experimental measurements have shown that the selected parameters for the evaluation of the separation efficiency (the peak capacity and total resolution product) were inappropriate. Gasoline, kerosene and diesel fuel were analyzed as examples of real samples. Amount of added ethanol as a biofuel additive was determined in the gasoline sample. Pulse pressure modulator was modified...
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Testing and optimization of pressure modulator conditions for GCxGC analyses
Ston, Martin ; Čabala, Radomír (advisor) ; Jelínek, Ivan (referee)
Pulse (fluid) pressure modulator was constructed for comprehensive gas chromatography. Optimal conditions of its connection to the gas chromatograph with a flame ionization detector were found for the analysis of selected mixture of volatile solvents. Pressure interdependences in the system and relations between dimensions of modulator capillaries and columns have been evaluated with respect to the length of the modulation period and pulse duration. Two non-polar columns (15 m × 0.25 mm DB-5MS, 10 m × 0.15 mm CP-Sil 5 CB) of the same stationary phase were tested to compare the separation efficiency in the first dimension and the DB-5MS column was selected. The Supelcowax 10 columns of the lengths 1, 2 and 3 m were compared in the second dimension. It was found that using of 3 m length column lead to the extension of analysis time, however the increase in separation efficiency in the second dimension was very evident. The experimental measurements have shown that the selected parameters for the evaluation of the separation efficiency (the peak capacity and total resolution product) were inappropriate. Gasoline, kerosene and diesel fuel were analyzed as examples of real samples. Amount of added ethanol as a biofuel additive was determined in the gasoline sample. Pulse pressure modulator was modified...
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Multidimensional Liquid Phase Separations
Šesták, Jozef ; Čáslavský, Josef (referee) ; Pulkrabová, Jana (referee) ; Česla, Petr (referee) ; Kahle, Vladislav (advisor)
This dissertation is dedicated to the topic of multidimensional liquid phase separations. This separation techniques are developed for analysis of complex samples containing thermally labile, low volatile or high molecular weight components that can´t be analysed by two-dimensional (2D) gas chromatography. Concepts of peak capacity and orthogonality are explained and various methods of their determination are stated in theoretical part of dissertation. High performance column liquid chromatography (HPLC) and high performance capillary electrophoresis (HPCE) are suggested as the most suitable methods for automated multidimensional liquid phase separations on-line coupled to mass spectrometry. Configuration of simplified miniaturized liquid chromatograph is described in experimental part of this thesis. Original concept of the system has been extended by simple mobile phase gradient generation technique. Correct function was demonstrated on repeatable separation of alkylphenones, peptides, nitroaromatics, and nitroesters. This system has been utilized as a base for a couple of simple two-dimensional separation platforms for HILIC-MALDI-MS analysis of glycans, for separation of peptides based on off-line coupling of isoelectric focusing and capillary liquid chromatography, and finally for on-line IEC×RPLC, RPLC×RPLC, and HILIC×RPLC two-dimensional liquid chromatography. Correct operation of submitted platforms has been proved.
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