Preparation and testing of capillary packed column sephadex G-10

Sephadex je v biochemických experimentech hojně využívaným sorbentem k separacím molekul v širokém rozmezí molekulových hmotností. Využívá se také k purifikacím nebo odsolování izolovaných proteinů. Dosud byly chromatografické aplikace na tomto principu prováděny jen v kolonách velkých rozměrů, čemuž musela předcházet účinná izolace proteinů, aby bylo na kolonu naneseno dostatečné množství vzorku. Naplnění sephadexu do mikrokolony by snížilo potřebné množství vzorku a mohly by tak být dále zpracovávány obtížně izolovatelné proteiny, které se získávají v malém množství. V této bakalářské práci je s použitím suspenzního plnění mikrokolona opravdu naplněna. K vytvoření gelu jako takového je použito principu přípravy monolitických kolon, tedy naplnění a následné polymerizace, v našem případě hydratace a zgelovatěním.Sephadex is a common sorbent in biochemistry for separation of molecules of a wide range of molecular masses. Sephadex is also used for purification or desalination of isolated proteins. Up to now, the gel chromatography has been performed in wide separation columns. Therefore, the isolation techniques for proteins had to be very effective to isolate enough proteins as a sufficient amount of the sample. The preparation and application of a capillary separation column packed with sephadex could decrease the needed sample amount and enable work with hardly isolable proteins obtained only in small quantities. In this bachelor project, the capillary column was packed with sephadex using the slurry packing procedure. The principle of monolithic columns was used to create a gel in the column. The column was packed in organic solvent firs, and then hydrated when flushing with water to generate the gel.Department of Analytical ChemistryKatedra analytické chemieFaculty of SciencePřírodovědecká fakult

Preparation and testing of capillary packed column sephadex G-10

Sephadex is a common sorbent in biochemistry for separation of molecules of a wide range of molecular masses. Sephadex is also used for purification or desalination of isolated proteins. Up to now, the gel chromatography has been performed in wide separation columns. Therefore, the isolation techniques for proteins had to be very effective to isolate enough proteins as a sufficient amount of the sample. The preparation and application of a capillary separation column packed with sephadex could decrease the needed sample amount and enable work with hardly isolable proteins obtained only in small quantities. In this bachelor project, the capillary column was packed with sephadex using the slurry packing procedure. The principle of monolithic columns was used to create a gel in the column. The column was packed in organic solvent firs, and then hydrated when flushing with water to generate the gel

Preparation and testing of capillary packed column sephadex G-10

Sephadex is a common sorbent in biochemistry for separation of molecules of a wide range of molecular masses. Sephadex is also used for purification or desalination of isolated proteins. Up to now, the gel chromatography has been performed in wide separation columns. Therefore, the isolation techniques for proteins had to be very effective to isolate enough proteins as a sufficient amount of the sample. The preparation and application of a capillary separation column packed with sephadex could decrease the needed sample amount and enable work with hardly isolable proteins obtained only in small quantities. In this bachelor project, the capillary column was packed with sephadex using the slurry packing procedure. The principle of monolithic columns was used to create a gel in the column. The column was packed in organic solvent firs, and then hydrated when flushing with water to generate the gel

Various methods of preparation of capillary columns packed with Sephadex for gel chromatography

In this diploma thesis, capillary columns with an inner diameter of 530 microns filled with gel particles were prepared. These columns are intended for gel chromatography. Three various methods of filling were used for their preparation. The first method was based on filling of wet particles in suspension with the organic solvent, followed by drying and swelling of particles with water into the form of gel. The second method used filling of wet particles in the suspension form with the aqueous phase. The last method started with suction of dry gel particles by air and continued with their subsequent swelling with water into the form of the gel. The prepared columns have been compared analysing hyaluronic acid and thiourea. The basic parameters specified for the gel column chromatography were determined by using phenylalanine. The pressure characteristics depending on the flow rate of mobile phase were used to compare the quality of packing of the columns prepared by various methods

Voluntary submission to criminal responsibility in Polish Fiscal Penal Code

Celem niniejszej pracy magisterskiej jest przedstawienie instytucji dobrowolnego poddania się odpowiedzialności z Kodeksu Karnego Skarbowego, jej historii, wad i zalet oraz praktyki jej stosowania na podstawie danych z krakowskich Urzędów Skarbowych.This master's thesis aim is to present the institution of voluntary submission to criminal liability from Polish Fiscal Penal Code, it's history, pros and cons and practice of using based on data from Krakow's tax offices

A Study of Methylene Blue Dye Interaction and Adsorption by Monolayer Graphene Oxide

The graphene oxide (GO) interaction with methylene blue (MB) cationic dye was studied in an aqueous solution at different pH during MB adsorption. The mutual interaction of MB with GO surface was studied and evaluated by Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD). The π-π and electrostatic interaction of MB with GO surface are the main types of interactions, and the XRD data show the monomeric arrangement of MB cation with GO. The GO surface functional groups and point of zero charge (PZC) were determined by acid-base titration. Suitability of zeta-potential measurement and acid-base titration method was briefly discussed. The quality of prepared GO was evaluated by Raman spectroscopy, XRD, and atomic force microscope (AFM). The experimental adsorption equilibrium data were analyzed using Langmuir, Langmuir-Freundlich, Freundlich, and Temkin isotherms. The GO maximum adsorption capacity increases with higher pH, that is ascribed to the facile interaction of negatively charged GO with positively charged MB structure

The effects of formation and functionalization of graphene-based membranes on their gas and water vapor permeation properties

The gas and water vapor permeabilities of graphene-based membranes can be affected by the presence of different functional groups directly bound to the graphene network. In this work, one type of carboxylated graphene oxide (GO-COOH) and two types of graphene oxide synthesized i) under strong oxidative conditions directly from graphite (GO-1) and ii) under mild oxidative conditions from exfoliated graphene (GO-2) were used as precursors of self-standing membranes prepared with thicknesses in the range of 12–55 μm via slow-vacuum filtration preparation method. It was observed that the permeabilities for all tested gases decreased in order GO-2 > GO-1 > GO-COOH and depended on both the arrangement of graphene sheets and their functionalization. The GO-1 membrane with a high content of oxygen-containing groups showed the best performance for water vapor permeability. The GO-2 membrane with a thickness of 43 μm exhibited a disordered GO sheet morphology and, therefore, unique gas-separation performance towards H2/CO2 gas pair, showing high hydrogen permeability while keeping extremely high H2/CO2 ideal selectivity that exceeds the Robeson 2008 upper bound of polymer membranes

Effect of Multiply Twinned Ag⁽⁰⁾ Nanoparticles on Photocatalytic Properties of TiO₂ Nanosheets and TiO₂ Nanostructured Thin Films

Ag-decorated TiO2 nanostructured materials are promising photocatalysts. We used non-standard cryo-lyophilization and ArF laser ablation methods to produce TiO2 nanosheets and TiO2 nanostructured thin films decorated with Ag nanoparticles. Both methods have a common advantage in that they provide a single multiply twinned Ag(0) characterized by {111} twin boundaries. Advanced microscopy techniques and electron diffraction patterns revealed the formation of multiply twinned Ag(0) structures at elevated temperatures (500 °C and 800 °C). The photocatalytic activity was demonstrated by the efficient degradation of 4-chlorophenol and Total Organic Carbon removal using Ag-TiO2 nanosheets, because the multiply twinned Ag(0) served as an immobilized photocatalytically active center. Ag-TiO2 nanostructured thin films decorated with multiply twinned Ag(0) achieved improved photoelectrochemical water splitting due to the additional induction of a plasmonic effect. The photocatalytic properties of TiO2 nanosheets and TiO2 nanostructured thin films were correlated with the presence of defect-twinned structures formed from Ag(0) nanoparticles with a narrow size distribution, tuned to between 10 and 20 nm. This work opens up new possibilities for understanding the defects generated in Ag-TiO2 nanostructured materials and paves the way for connecting their morphology with their photocatalytic activity