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Polymethacrylate monolithic columns in capillary liquid chromatography
Grafnetter, Jan ; Coufal, Pavel (advisor) ; Pacáková, Věra (referee) ; Ventura, Karel (referee) ; Sýkora, David (referee)
3 Conclusions This Thesis gives chromatographic and structural characteristics of the butyl methacrylate monolithic columns prepared for the capillary liquid chromatography. Results of this Thesis clearly demonstrate the possibility of preparation of capillary monolithic columns with a good repeatability of the principal chromatographic parameters. A completely new technique for preparation of the monolithic columns for use in the on-column detection mode was introduced. Different photometric detection modes were applied to monolithic capillary columns in CLC. The advantages and disadvantages of the on-column detection mode in comparison to the additional detection capillary were studied and described. It was shown that the best separation and the most sensitive quantification in the short analysis time could be reached by using the monolithic columns with detection window placed directry on the separation capillary, just after the stationary phase or within the monolith. The attempts leading to preparation of chiral monolithic stationary phases, which would allow enantioseparation of selected chiral analýes' were not totally successful. No time was left to find the suitable chromatographic conditions for enantioseparations. \ I
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Analysis, separation and physico-chemical characterization of peptide hormones by capillary electrophoresis
Šolínová, Veronika ; Jelínek, Ivan (advisor) ; Feltl, Ladislav (referee) ; Coufal, Pavel (referee) ; Samcová, Eva (referee)
I 11 CoNcr,usroNs The work bas demonstrated that capinary zone elechophoresis and micelrar electokinetic cbromatography are high-perfomrance and high-sensitive methods for qualitative and quantitative anďysis and for physico-chemicď characÚerization ofpeptide hormones. using the strategy for the rational selection of experimental conditions for cE analysis and separations ofpeptides, ionogenic peptides were separated as cations in the classical and isoelectric buffers in the acidic region, pH 2- 2'5, and as anions in amphoteric buffer in weakry arkaline conditions, pH = 8'1. Non-ionogenic peptides were analyzed by miceilar electrokinetic chromatography with micelrar phase consisting of anionic detergent sodium dodecylsulphate in alkaline conditions, pH 8.8. The experimental conditions were selected with the aim to avoid the adsorption of peptides on the capillary wall, which was particularly reached with suppression of dissociation of si'anol gÍoups on the imer capillary wall in the acidic pH region and with electrostatic repursion negatively charged peptides from negatively charged dissociation silanol groups on the inner capillary wall in the alkaline pH region. The pH values of background electrolýes were chosen in the region, where the biggest differences in calculated specific charges of analyzed...
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Post-column derivatization in HPLC for analysis of amino acids
Vaňkátová, Petra ; Coufal, Pavel (advisor) ; Sobotníková, Jana (referee)
Proteinogenic amino acids are the basic structural building units of proteins. Their analysis is important in many fields, especially in medicine. This thesis deals with one of the methods of derivatization of amino acids in order to increase their absorption in the UV - the post-column derivatization using copper (II) oxide. It is quick, robust and easy-to-use method. Following the nowadays trend in the analysis of free amino acids, this thesis is focused on the HILIC mode separation environment.
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Development of capillary electrophoretic method for determination of phosphorylated components of bacterial cell membranes
Červinková, Jana ; Čabala, Radomír (advisor) ; Coufal, Pavel (referee)
Title: Development of capillary electrophoretic method for determination of phosphorylated components of bacterial cell membranes Author: Jana Červinková Department: Department of analytical chemistry Supervisor: RNDr. Radomír Čabala, Dr. Abstract: The bachelor work reviews the present state of knowledge of the analyses and determination of phosphorylated compounds present in the bacterial cell membranes. This group of compounds is represented mainly by phosphatidylglycerol, cardiolipine, and phosphatidylethanolamine and minority phosphatidylserine, phosphatidic acid and lysylphospatidylglycerol. Together with fatty acids these substances are the main components of cell membranes and they could be, to a certain degree, applied to either identification of bacteria or the study of bacterial behaviour at different conditions. Partially or fully non-aqueous capillary electrophoresis analytical method with spectrophotometric detection would be developed for their determination. Optimum experimental conditions for the separation of standards of selected compounds should be found within the framework of the bachelor thesis. Keywords: phosphorylated compounds, bacterial cytoplasmic membrane, capillary electrophoresis, thin layer chromatography, Bacillus subtilis.
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System peaks in elektromigration systems with complexing agents
Dvořák, Martin ; Gaš, Bohuslav (advisor) ; Coufal, Pavel (referee)
Capillary zone electrophoresis (CZE) is a widely used analytical method. CZE is described theoretically very well and there are many simulation programs, which enable one to predict results of electrophoretic separations, and alternatively to study phenomena taking place during the electrophoretic separation in detail. The CZE method is not only an analytical method, but is often used for determination of many physical parameters of compounds, such as stability constants or complex mobilities. Among methods most often used for determination of complexation parameters belongs the affinity capillary electrophoresis (ACE). Its alternative is the vacancy affinity capillary electrophoresis (VACE). Whereas by the ACE method the stability constant is determined from the dependence of the analyte effiective electrophoretic mobility on the background electrolyte (BGE) composition, in the case of the VACE system peaks are used for this purpose. In this work the legitimacy of using system peaks in the VACE method for determination of stability constants was investigated. Several approaches dealing with the concentrating of complexing agent in the peak area were compared, both for the ACE and the VACE method. Two different kinds of electrophoretic systems were studied. In the first one, neutral cyclodextrin was used as...
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