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Utilization of micropreparative isoelectric focusing in the analysis of biological samples
Vlčková, Zuzana ; Flodrová, Dana (referee) ; Šalplachta, Jiří (advisor)
The aim of this Master Thesis is the optimization and verification of conditions of separation of proteins from biological materials. For the separation a developed technique called micropreparative isoelectric focusing is being used. The theoretical part describes analytical methods of biological materials with a focus on proteins which were used for the optimization of the developing system in general. This part of the Master Thesis also presents this method itself. Furthermore, basic characteristics of other techniques used to control the efficiency of the micropreparative isoelectric focusing are introduced. The experimental part depicts the individual steps of optimization of the micropreparative isoelectric focusing. The ideal separation procedure was firstly found out on the individual standard proteins, secondly on their mixture and lastly the optimised procedure was applied to a real biological sample. The efficiency of the micropreparative isoelectric focusing was verified by SDS-PAGE and MALDI-TOF mass spectrometry.
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Analysis of selected substances contained in the extract from elderberry twigs
Škeřík, Jan ; Flodrová, Dana (referee) ; Šalplachta, Jiří (advisor)
My diploma thesis deals with the optimization and verification of conditions for separation of proteins, which are contained in the twigs of elder (Sambucus nigra L.) using the technique RP-HPLC. The measurements were made with the use of a HPLC system with a UV-VIS detector. Column Zorbax 300SB-C18 300 4,6 x 250 mm and particles the size of 5 microns were used. The theoretical part describes the attributes and usage of elder and especially its proteins. The basic characteristic of the used HPLC technique is introduced. The possibilities of how to identify proteins are described. The practical part demonstrates the individual steps of optimization of the HPLC method applied to the mixture of standard proteins. The application of this method in the real sample is also reported. The final part of the diploma thesis is focused on the comparison of analysed samples taken in different seasons.
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Determination of manganese in food supplements by atomic absorption spectrometry
Melicharová, Zuzana ; Hraníček, Jakub (advisor) ; Šíma, Jan (referee)
EN The main content of this thesis was the determination of manganese in the form of manganese ions in food supplements by the atomic absorption spectrometry. It was necessary to find the optimum conditions for this method, to conduct interference studies, to find a suitable way of transferring samples into solution and to determin manganese to achieve this goal. Following parameters were examined in optimalization study: beam height above burner edge, burner horizontal position, acetylene flow rate, torch angle, and spectral interval width. Copper, iron, sodium and calcium ions were investigated as interferents as they were present at the highest concentration in selected food supplements. The conversion of solid samples into solution was carried out in water and nitric acid at a room temperature as well as elevated temperature. These food supplements were selected for analysis: Calibrum 50 Plus, Caltrate Plus, Center A to Iron, DAS Gesunde Plus A-Z Multispectrum, GS Extra Strong Multivitamin, Spectrum 50+ and Supradyn Energy Complex. The manganese assay was performed on a GBC 933 AA spectrophotometer. Standard solutions were measured to determine the detection limit and the limit of determination of the instrument. The detection limit was 0.001 mg/l and the limit of determination was 0.004 mg/l...
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Utilization of micropreparative isoelectric focusing in the analysis of biological samples
Vlčková, Zuzana ; Flodrová, Dana (referee) ; Šalplachta, Jiří (advisor)
The aim of this Master Thesis is the optimization and verification of conditions of separation of proteins from biological materials. For the separation a developed technique called micropreparative isoelectric focusing is being used. The theoretical part describes analytical methods of biological materials with a focus on proteins which were used for the optimization of the developing system in general. This part of the Master Thesis also presents this method itself. Furthermore, basic characteristics of other techniques used to control the efficiency of the micropreparative isoelectric focusing are introduced. The experimental part depicts the individual steps of optimization of the micropreparative isoelectric focusing. The ideal separation procedure was firstly found out on the individual standard proteins, secondly on their mixture and lastly the optimised procedure was applied to a real biological sample. The efficiency of the micropreparative isoelectric focusing was verified by SDS-PAGE and MALDI-TOF mass spectrometry.
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Analysis of selected substances contained in the extract from elderberry twigs
Škeřík, Jan ; Flodrová, Dana (referee) ; Šalplachta, Jiří (advisor)
My diploma thesis deals with the optimization and verification of conditions for separation of proteins, which are contained in the twigs of elder (Sambucus nigra L.) using the technique RP-HPLC. The measurements were made with the use of a HPLC system with a UV-VIS detector. Column Zorbax 300SB-C18 300 4,6 x 250 mm and particles the size of 5 microns were used. The theoretical part describes the attributes and usage of elder and especially its proteins. The basic characteristic of the used HPLC technique is introduced. The possibilities of how to identify proteins are described. The practical part demonstrates the individual steps of optimization of the HPLC method applied to the mixture of standard proteins. The application of this method in the real sample is also reported. The final part of the diploma thesis is focused on the comparison of analysed samples taken in different seasons.
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