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Molecularly imprinted polymers as a tool for the isolation of key biologically active molecules
Vodová, Milada ; Jaroslava, Bezděková (referee) ; Nejdl, Lukáš (advisor)
This bachelor thesis is focused on preparation and optimization of molecularly imprinted polymers that are selective for the selected nucleobase(uracil). Molecularly imprinted polymers, which wereused for selective uracil isolation, have been prepared non-covalent imprinting technique. As a function monomer was used in this case dopamine. The detection ofisolated uracil was made by capillary electrophoresis with absorption detection (–260 nm). Conditions for preparation of molecularly imprinted polymers,their binding properties, adsorption kinetics, and selectivity wereinvestigatedin detail. Mentioned polymer materials were as well used for uracil selective isolation and detection from complexsamples. Tomato products were used as complex samples. Uracil in those samples was usedasanindicator ofbacterial contamination.
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Study of metallothionein conjugates by electromigration methods
Johaníková, Klára ; Zemánková, Kristýna (referee) ; Vaculovičová, Markéta (advisor)
The bachelor thesis deals with the study of conjugates of metallothionein (MT) with fluorescent nanoparticles using electromigration methods. Particular attention is paid to the preparation and characterization of quantum dots (QDs) and their bioconjugates with the metallothionein protein, whose separation conditions (concentration and suitable electrolyte pH) have been carefully optimized. For the preparation of QDs, the action of UV radiation for one type of quantum dots and the thermal action of high temperature for the second type of species were used. Conjugation of QDs to the protein was performed via a carbodiimide-zero-length crosslinker (EDC / sulfo-NHS), which activates carboxyl groups and allows bioconjugation of the ligand by covalent bonding. The resulting bioconjugates were studied by capillary electrophoresis with absorption detection ( 214).
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Sucralose - a new permitted sweetener in Czech Republic
Chlebcová, Lenka ; Vránová, Dana (referee) ; Vespalcová, Milena (advisor)
This bachelor thesis deals with a topic of an artifical sweetner sucralose. The atention is given to process of production, using, its properties like sweetness, palatability and influence over human health, toxicity. In the food processing industry it is used by virtue of its high sweetness and low energy value. Determination methods of sucralose are capillary electrophoresis, liquid chromatography.
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Optimization of a method for the determination of inorganic ions using capillary electrophoresis
Špačková, Simona ; Bezděková, Jaroslava (referee) ; Vaculovičová, Markéta (advisor)
Inorganic ions are one of the most important group chemicals. Their importance is crucial for all living organisms on the planet and monitoring of their content is essential for this purpose in clinical or environmental analysis but also in the food industry, agriculture or many other industries. Many analytical methods for ion detection have been designed and used, depending on the area of their use. Recently, numerous methods have been adapted to areas where not primarily intended. One such methods may be, for example, laser ablation with inductively coupled plasma mass spectrometry, which has recently begun to affect the analysis of biological samples. As a complementary method, able to detect also non-metallic ions is capillary electrophoresis. In this master thesis, the possibility of applying capillary electrophoresis with indirect photometric detection for the separation of inorganic ions in cell lysates of HeLa cells by capillary electrophoresis was verified. Main attention was paid to chloride ions. The ability to separate inorganic compounds in cellular material using salicylic acid as an absorbing anion in the base electrolyte was tested.
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Application of polymer membranes in microextraction techniques for analysis of biological samples
Ryšavá, Lenka ; Diviš, Pavel (referee) ; Dvořák, Miloš (advisor)
The thesis presents experimental study on application of various polypropylene membranes (with different thicknesses, porosity and pore size) for direct coupling of membrane microextractions with capillary electrophoresis. No comprehensive study, which describes effect of these membrane parameters on extraction recoveries, was published in the past. Previous scientific works prefer application of 100 µm polypropylene membranes for their easy handling and satisfactory extraction efficiencies. Experimental part includes examination of selected polypropylene membranes as supported liquid membranes in-line coupled to capillary electrophoresis for analysis of basic drugs from complex samples. Membranes with three thicknesses (25, 100 and 170 µm) were tested. The highest extraction recoveries were achieved for the 25µm thick polypropylene membrane. Various pH conditions of donor and acceptor operational solutions were examined for extractions from real complex matrices (urine, plasma). The optimal extraction conditions were 10 mM NaOH as donor phase and 10 mM HCl as acceptor phase. 25µm membranes offer higher extraction recoveries, reduced consumption of organic solvents for membrane impregnation, similar mechanical stability and similar clean-up ability compared to thicker polypropylene membranes.
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Application of microextraction techniques for analysis of biological samples
Lučaj, Martin ; Vespalcová, Milena (referee) ; Dvořák, Miloš (advisor)
This bachelor thesis deals with the analysis of basic drugs in biological samples by in line coupling of supported liquid membrane (SLM) to capillary electrophoresis (CE). The tITP method was included as an on-line preconcentration step to achieve better detection limits with ammonium acetate as the tITP stacker and acetic acid as the CE background electrolyte. The practical part of the thesis focuses on the optimization of the tITP and the SLM extraction conditions in the analysis of basic drugs in complex biological samples (human urine). The best analytical results were achieved with hydrodynamic injection of ammonium acetate for 15 s and basic drugs for 15 s. The donor was alkalised with 10 mM sodium hydroxide and acceptor was acidified with 10 mM hydrochloric acid. From the results, the in-line coupling of SLM extraction to CE and stacking by tITP is an efficient method allowing direct analysis of complex samples with improved sensitivity of the analytical method.
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On-chip and capillary electrophoresis for cancer research
Hájková, Tereza ; Babula, Petr (referee) ; Provazník, Ivo (advisor)
The thesis deals with cancer disease and its genesis. The prostate cancer is the second most dangerous disease in male population. It is very important to detect it in early stage. This thesis deals with the proteins which are or could be used as potential cancer markers. For example in current commonly used the prostate-specific antigen and newly the metallothionein. Metallothionein is a protein occuring in fauna and flora in four basic isoforms. There is a hypothesis that the isoform content or/and their ratio may have a diagnostic implication and could be used in clinical practise. For this reason they are studied and developed methods for specific and sensitive analysis of individual isoforms. In this work, capillary electrophoresis is used for the separation of the isoforms. The influence of the parameters such as type of background electrolyte, its pH and separation voltage etc. on the final separation are discussed.
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Analysis of cow milk caseins by capillary electrophoresis
Dvořáková, Irena ; Zemanová, Jana (referee) ; Vespalcová, Milena (advisor)
The diploma thesis is dealing with the determination of caseins in lyophilised samples of milk using the method of capillary electrophoresis (CE). This method was optimised and verified by Ivana Micikova in her diploma thesis in 2009. The theoretical part of the diploma thesis gives information about the chemical compound of cow's milk. Caseins and their usage in the food and non-food industries are described in a separate unit. Impact of both the nutritional and non-nutritional factors on the composition of the milk is discussed further. The description and the principles of the CE and a list of the methods of CE used in the last several years are stated at the end of this part. The experimental part is focused on the preparation of the solutions, process of casein isolation and its analysis. It summarises the setting of the experiment, sampling and sample processing and the composition of the feeding mixtures. The diploma thesis has arisen on the basis of the cooperation of the Institute of Food Science and Biotechnology, the Faculty of Chemistry, Brno University of Technology and the Research Institute for Cattle Breeding, Ltd., Department of Animal Nutrition Physiology in Pohorelice.
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Evaluation of glutathione content in plants as a marker of heavy metals environmental contamination
Borková, Marie ; Stoupalová, Michaela (referee) ; Opatřilová,, Radka (advisor)
Dependence of glutathione concentration on the amount of thallium in the plant was studied. Observed plant was maize (Zea mays) which was divided to two parts – root and overground. Two culture procedures were elaborated where seeds and young seedlings were cultivated in a solution of thallium of concentration 0, 1, 3, 5, 8, a 10 µmol/l. Extraction agents used during extraction were phosphate buffer and solution of ascorbic acid. Determination of glutathione was realized by capillary electrophoresis (CE) and high performance liquid chromatography (HPLC). Diode array detector (DAD) was used in both methods. Quantification of the thallium amount in the plant was done by method of inductively coupled plasma-atomic emission spectrometry (ICP-AES).
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