This work is based on the occurrence of drug residues in the environment. This study is focused on the development and optimization methods for determination of selected drugs in the surface water, aquatic sediment and sewage sludge from waste water treatment plant. From the group of drugs were chosen antibiotics. Antibiotics presented in the environment can cause adverse effects including toxic effects, immunity disorders and indirect bioalteration effects. Sulfonamide antibiotics, which are used in the treatment of urinary and respiratory tract infections as well as in the treatment of other infectious diseases, were chosen as a target compounds. Three optimized analytical methods for determination of sulfonamide antibiotics were developed. For the optimization of extraction were tested: solid phase extraction, pressurized solvent extraction, microwave extraction and ultrasonic extraction. For the final analysis was used liquid chromatography with two detectors - diode array detector and mass spectrometer. These optimized methods were applied for the analysis of real samples. The surface water and sediment samples were collected from two Moravian rivers (the Svratka river and the Svitava river). Samples of sewage sludge were collected from waste water treatment plant Brno-Modřice. Fish samples from the Svratka river were also collected. It was observed that all the selected sulfonamide antibiotics are present in real sediment samples (ug.kg-1). Simultaneously presence of some target analytes in real surface water (ug.l-1) and in sewage sludge (ug.kg-1) samples has been confirmed. In samples of surface water from the Svitava river and in fish samples sulfonamide antibiotics were not detected or their concentrations were below the limit of detection.

In the presented PhD. thesis, problems covering occurrence of polybrominated diphenyl ethers (PBDE) and perfluorinated compounds in the environment were solved. The study was focused on identification and further verification of optimal methods in order to determine PBDEs and perfluorinated compounds sampled from the aquatic ecosystem. Polybrominated diphenyl ethers belong to persistent compounds classified as main pollutants; within the environmental constituents, they have been observed particularly in the last decade. For these studies, there were selected surface water matrices and sediments taken in various localities within the river Svratka basin. The analyses were to demonstrate whether they accumulate and remain in these specific matrices for longer time. There we assessed the congeners of polybrominated diphenyl ethers as follows: BDE-28, 47, 66, 85, 99, 100, 153, 154 and 183. In order to isolate them from matrices, various extraction techniques were used, i.e., ultrasonic extraction, microwave extraction, and pressure solvent extraction. Gas chromatography method with electron capture detection (GC/ECD) was selected for determination. The accomplished studies also assessed basic chemical, physical and environmental characteristics of diphenyl ethers in the environmental constituents. The attention was also paid to perfluorooctanoic compounds (PFOA), (PFOS) and (FOSA); physico-chemical properties of these organic pollutants were characterised and their toxicological and environmental aspects were evaluated. Usability of extraction techniques (ultrasonic extraction, pressure solvent extraction, solid phase extraction) applied to PFCs isolation from sediments samples was assessed. Identification and quantification of these analytes were performed using a high performance liquid chromatography/mass spectrometry method (HPLC/MS). After that, the optimized methods applied to real samples. Surface water and sediments were sampled within the river Svratka basin, in particular, from 19 sampling localities. PBDE occurrence was confirmed in sediments samples from the Svratka river (g.kg-1), PBDE were not detected in the surface water samples, their concentrations were below the detection limit, resp.; perfluorinated compounds were not detected in sediments and water samples at all.

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.

9 Abstract In this Ph.D. thesis possibilities of using our proposed potential programs for a multiple-pulse amperometry and a fast scan differential pulse voltammetry in combination with flow systems are presented. The development of new sensitive amperometric and voltammetric methods for the determination of oxidisable biologically active organic compounds is another aim of this work. In the first part of the work, the flow injection system and multiple-pulse amperometric detection were employed to develop and optimize a simple, low-cost, and rapid method for the simultaneous determination of natural and synthetic antioxidants. This technique involves the application of an appropriate potential waveform consisting of a suitable sequence of pulses on a single working electrode, thus allowing distinguish the analytes in a mixture with no need of separation. Conditions for the determination of antioxidants and modelling of the potential program were tested and studied, respectively. Second part of the work describes and characterizes the application of the fast scan differential pulse voltammetry (FSDPV) in combination with the flow systems. FSDPV is the electroanalytical technique that use high scan rate to record voltammograms within several milliseconds and ensures high temporal resolution. This technique...

Presented Ph.D. Thesis is focused on the use of the boron-doped diamond (BDD) electrodes for voltammetric and amperometric determination of selected nitrophenols: 2-nitrophenol (2NP), 4-nitrophenol (4NP), and 2,4-dinitrophenol (2,4DNP). These compounds are listed as "priority pollutants" by United States Environmental Protection Agency (US EPA) due to their negative impact on living organisms and are mainly used in agriculture as plant growth stimulators. BDD electrodes are used for determination of wide range of electrochemically both reducible and oxidisable organic compounds and have become a popular electrode material thanks to its commercial availability and excellent mechanical and electrochemical properties. A differential pulse voltammetric method was developed for the determination of 2NP, 4NP and 2,4DNP at a BDD film electrode using electrochemical reduction and of 4NP and 2,4DNP using electrochemical oxidation. The method was successfully applied for the direct determination of these compounds in drinking and river water in the concentration range from 4×10-7 to 2×10-5 mol.L-1 . To improve the limit of quantification, a preconcentration by solid phase extraction from 100 mL (drinking and river water) and 1000 mL (drinking water) of water samples was used with limit of determination...

In the first part of the doctoral thesis, the capillary electrophoresis was used to test the potential chiral separation properties of monosubstituted cyclodextrin derivatives, namely PEMEDA- and PEMPDA-β-cyclodextrins for the group of selected analytes. Both selectors exhibited excellent enantioseparation properties for N-boc-D,L-tryptophan, where the enantiomers were completely separated even at 0.5 mmol·l-1 concentration of the cyclodextrin derivative in the background electrolyte. However, the differences between the enantiodiscrimination properties of individual derivatives were minimal. The second test group consisted of two cyclodextrin derivatives, namely 2-O- and 3-O- cinnamyl-α-cyclodextrins. These derivatives are able to form supramolecular polymers in aqueous solutions that disintegrate at elevated temperature. The formation of these polymers was tested by NMR and DLS experiments. None of the tested cyclodextrin derivatives showed enantiodiscrimination properties towards a group of selected analytes. In the frame of antipredatory study, HPLC-MS/MS method working in HILIC mode was used for separation of ten pterin derivatives and riboflavin, which can be present as pigments in insects, reptiles or amphibians as a part of their warning coloration. The developed methodology was applied for...

(AJ) Food contact materials pose a risk to human health due to a variety of contaminants which they can release into food. These compounds, migrants, include aromatic hydrocarbons, dialkyl phthalates, bisphenols, printing ink photoinitiators, and perfluoroalkyl compounds. The determination of all these substances can be expensive and time-consuming since universally applicable analytical approaches are not available nowadays. This work attempted to develop methods for the simultaneous determination of migrants from all of the mentioned groups in paper-based food contact materials and fatty food. A total of 68 contaminants were studied in paper products using liquid chromatography and gas chromatography coupled with tandem mass spectrometry detection. All analytes were isolated simultaneously using the modified "QuEChERS" method. This method demonstrated acceptable recovery and repeatability for most analytes in the validation study; LOQs ranged from 1.3 to 220 µg/kg. Analysis of 132 real paper products confirmed the occurrence of almost all studied analytes, which were often present in complex mixtures and at concentrations up to 628 mg/kg. A total of 41 potential contaminants were monitored in fatty foods by liquid chromatography with tandem mass detection. Contamination of the mobile phase by...

(AJ) Food contact materials pose a risk to human health due to a variety of contaminants which they can release into food. These compounds, migrants, include aromatic hydrocarbons, dialkyl phthalates, bisphenols, printing ink photoinitiators, and perfluoroalkyl compounds. The determination of all these substances can be expensive and time-consuming since universally applicable analytical approaches are not available nowadays. This work attempted to develop methods for the simultaneous determination of migrants from all of the mentioned groups in paper-based food contact materials and fatty food. A total of 68 contaminants were studied in paper products using liquid chromatography and gas chromatography coupled with tandem mass spectrometry detection. All analytes were isolated simultaneously using the modified "QuEChERS" method. This method demonstrated acceptable recovery and repeatability for most analytes in the validation study; LOQs ranged from 1.3 to 220 µg/kg. Analysis of 132 real paper products confirmed the occurrence of almost all studied analytes, which were often present in complex mixtures and at concentrations up to 628 mg/kg. A total of 41 potential contaminants were monitored in fatty foods by liquid chromatography with tandem mass detection. Contamination of the mobile phase by...

9 Abstract In this Ph.D. thesis possibilities of using our proposed potential programs for a multiple-pulse amperometry and a fast scan differential pulse voltammetry in combination with flow systems are presented. The development of new sensitive amperometric and voltammetric methods for the determination of oxidisable biologically active organic compounds is another aim of this work. In the first part of the work, the flow injection system and multiple-pulse amperometric detection were employed to develop and optimize a simple, low-cost, and rapid method for the simultaneous determination of natural and synthetic antioxidants. This technique involves the application of an appropriate potential waveform consisting of a suitable sequence of pulses on a single working electrode, thus allowing distinguish the analytes in a mixture with no need of separation. Conditions for the determination of antioxidants and modelling of the potential program were tested and studied, respectively. Second part of the work describes and characterizes the application of the fast scan differential pulse voltammetry (FSDPV) in combination with the flow systems. FSDPV is the electroanalytical technique that use high scan rate to record voltammograms within several milliseconds and ensures high temporal resolution. This technique...

Despite significant progress in the development of anti-cancer drugs, there is still a need for novel therapeutic strategies that would improve the outcome of cancer patients. Using proteomic technologies and cell lines with different phenotype of p53 tumour suppressor, we monitored cancer cell response to anti-cancer treatment with focus on the development of drug resistance. The different levels of metabolic proteins were identified in our study which may help to explain different anti-cancer activity of drugs with only a subtle difference in structure. More importantly, proteins associated with the development of drug resistance were identified and such expression changes have become a focus of interest. Our findings demonstrate a higher protein level of serine hydroxymethyltransferase, serpin B5 and calretinin in cancer cells resistant to Aurora kinase inhibitors. Such proteins promote the tumour growth with no apparent impact of p53 phenotype whilst voltage-dependent anion-selective channel protein 2 contributes to the development of resistance only in cells with functional p53 which is accompanied by the decreased level of elongation factor 2. On the other hand, cancer cells with loss of p53 appear to amplify alternative mechanisms such as protection against oxidative stress. The results...