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Preconcentration of ion compounds from sample by epitachophoresis
Kocianová, Vanda ; Dvořák, Miloš (referee) ; Voráčová,, Ivona (advisor)
Epitachophoresis is newly developed method, which allows to separate and concentrate analytes from large volumes of complex samples. Principle is based on isotachophoresis and utilize discontinuous electrolyte system. Different design of device is used, its round shaped and due to that acceleration of analytes zone migration occurs during separation towards centre of device and at the same time widening of zones happens. Focusation, concentration and separation of analytes between leading and terminating electrolyte progress from outer line of device towards its centre. It is necessary to stabilize this boundary to prevent mixing of leading and terminating electrolyte. Main target of this diploma thesis is optimization of epitachophoretic conditions, mainly electrolyte boundary stabilization options. Tested stabilizing media were agarose gels, foamed polymer disks and PDMS columns. All leading and terminating electrolyte boundary stabilizing media undergo several optimizations so the best option can be selected for concreate applications. DNA was used as a model analyte and the media with the highest recovery were selected. The best results offered NEEO and PFEG agarose gels and Dyneema foamed polymer with recovery over 70%. Another interesting alternative are PDMS columns, with their open structure that allow concentration of large volume analytes such as microorganisms.
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Epitachophoretic preconcentration of proteins
Hrušková, Helena ; Voráčová, Ivona ; Foret, František
In this work, we present a developed method for preconcentration of standards of proteins cytochrome c, myoglobin, and hemoglobin. Firstly, the electrolyte system was selected and optimized to provide low analysis time, narrow shape of the protein zone, and eliminate overheating at the center of the device. In the next step, the developed method was tested on the preconcentration of proteins: cytochrome c, myoglobin, and hemoglobin. After several adjustments myoglobin and hemoglobin were also uccessfully preconcentrated. After evaluation of pilot results, this technique provides up to fiftyfold preconcentration and recovery of over 70 % for each protein. The next plan is to preconcentrate proteins from complex biological matrices such as urine or blood plasma.
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Preconcentration of ion compounds from sample by epitachophoresis
Kocianová, Vanda ; Dvořák, Miloš (referee) ; Voráčová,, Ivona (advisor)
Epitachophoresis is newly developed method, which allows to separate and concentrate analytes from large volumes of complex samples. Principle is based on isotachophoresis and utilize discontinuous electrolyte system. Different design of device is used, its round shaped and due to that acceleration of analytes zone migration occurs during separation towards centre of device and at the same time widening of zones happens. Focusation, concentration and separation of analytes between leading and terminating electrolyte progress from outer line of device towards its centre. It is necessary to stabilize this boundary to prevent mixing of leading and terminating electrolyte. Main target of this diploma thesis is optimization of epitachophoretic conditions, mainly electrolyte boundary stabilization options. Tested stabilizing media were agarose gels, foamed polymer disks and PDMS columns. All leading and terminating electrolyte boundary stabilizing media undergo several optimizations so the best option can be selected for concreate applications. DNA was used as a model analyte and the media with the highest recovery were selected. The best results offered NEEO and PFEG agarose gels and Dyneema foamed polymer with recovery over 70%. Another interesting alternative are PDMS columns, with their open structure that allow concentration of large volume analytes such as microorganisms.
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