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Use of advanced fluorescence methods in the study of aggregation
Uhrínová, Alžbeta ; Enev, Vojtěch (referee) ; Venerová, Tereza (advisor)
The diploma thesis deals with the use of advanced fluorescence techniques in the study of aggregation of selected aggregation models (casein and sodium dodecyl sulfate). At the beginning of the research, fluorescent probes Atto 488 and Rhodamine 6G (RH6G) were characterized using a spectrofluorometry, UV-VIS spectrometry, time-resolved fluorescence, and fluorescence correlation spectroscopy (FCS). Aggregation models with the addition of fluorescent probes were characterized using the same advanced fluorescence techniques. The interaction between the fluorescent probe Atto 488 and the aggregation models was not confirmed. The excitation maximum of Atto 488 was at 500 nm and the emission maximum at 520 nm. The lifetime of the fluorescent probe Atto 488 was from 4,20 to 4,82 ns and the hydrodynamic radius was determined to 0,6 nm. Rhodamine 6G had an excitation maximum at 525 nm and an emission maximum at 555 nm. The lifetime of the fluorescent probe RH6G was from 3,99 to 5,27 ns. The intensity of RH6G was found to decrease upon addition of a higher concentration of casein, resulting in quenching. The hydrodynamic radius of the RH6G probe was determined to 0,6 nm. With the addition of casein using the maximum entropy method (MEM) evaluation was from 0,5 to 534 nm. Stable SDS micelles had a hydrodynamic radius by MEM evaluation of 2,4 nm.
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Sorption of charged organic compounds on humic acids
Krist, Tomáš ; Doskočil, Leoš (referee) ; Enev, Vojtěch (advisor)
Bachelor thesis deals with a sorption properties and physical-chemical characterization of humic acids before and after the adsorption. Sorption experiments with a methylene blue and a rhodamin 6G were performed. Adsorption isotherms were obtain via UV/VIS spectrometry. The Langmuir isotherm was used to evaluate the experimental data. Standard of HA isolated from Leonardite was characterized by an elemental and thermogravimetric analysis, by a Fourier-transform infrared spectroscopy, a surface area analysis and also by a potentiometric titration.
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Transport processes in hydrogels
Sárová, Michaela ; Kalina, Michal (referee) ; Klučáková, Martina (advisor)
This master's thesis is focused on study of transport processes in hydrogels based on humic acids. For this purpose is used methods unsteady diffusion in cuvettes, which was studied the transport of organic dyes, specifically methylene blue and rhodamine 6G, in agarose hydrogel without the addition and with the addition of individual standards humic acids (Leonardite, Elliott Soil, Suwannee River II and Pahokee Peat). This method is based on spectrophotometric monitoring of concentration changes of dyes depending on space of the cuvette and on time. The aim of this thesis was to investigate the effects of interactions between diffusing dye and the particular type of gel to the resultant effective diffusion coefficient of dye. The experiments indicate that the presence of humic acid in the hydrogel greatly affects the transport of selected dyes.
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Inverse FCS in colloidal systems research
Richterová, Veronika ; Venerová, Tereza (referee) ; Mravec, Filip (advisor)
This diploma thesis is focused on the study of inverse fluorescence correlation spectroscopy, especially with the regard for the usage of different fluorescent probes and different sized analysed particles. At first, the proper concentration of fluorescent probes was determined. In this concentration is the probe considered as a medium surrounding the analysed particles. Based on this concentration, which was determined as 400 M, several sets of samples were prepared. This samples contained different concentration of polystyrene particles of 100 and 500 nm diameter and multilamellar liposomes. Then, the FCS curves of samples with different fluorescent probes were measured. Fluorescein, rhodamine 6G and Atto 488 were used as fluorescent probes. As a result from experiments, it was found, that particles with 100 nm diameter cannot be analysed with none of the fluorescent probes. Inverse FCS method can be applied to systems, that contains particles with 500 nm diameter and fluorescein. Systems with rhodamine 6G have the same behaviour as typical FCS measurement. It is caused by dimerization of this probe and it cannot be used for 500 nm particles. Liposome samples can be established with iFCS method, but the results are biased by random distribution of liposomes size.
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Transport properties of humic gels
Sárová, Michaela ; Pospíšilová, Ľubica (referee) ; Klučáková, Martina (advisor)
This bachelor’s thesis is focused on study of transport properties of humic gels. For research of these properties was used diffusion method of organic dyes in diffusion cells which is based on spectrophotometric monitoring of concentration changes depending on time. Dyes used in the experiment were specifically methylene blue and rhodamine 6G and diffusion experiments were performed on an agarose hydrogel without the addition of humic acid, with the addition of unmethylated humic acid and then with the addition of methylated humic acid. The aim of this thesis was to investigate the effects of interactions between diffusing dye and the particular type of gel to the resultant effective diffusion coefficient of dye. It has been found that the presence of humic acid in agarose hydrogel greatly influences the transport of dyes.
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Effect of chemical modification of humic acids on their interactions with organic ions
Fryšová, Eva ; Weidlich, Tomáš (referee) ; Sedláček, Petr (advisor)
This master thesis deals with the interaction of humic acids with organic ions and effect of chemical modification of humic acids on this interaction. Rhodamine 6G was used as a model organic ion. The interaction were studied by fluorescence quenching, dialysis in diffusion cells and differential UV-VIS spectroscopy. Results showed that the bond between the reactants was not primarily dependent on carboxylic functional groups of humic acids. Fluorescence experiments indicated static quenching mechanism and differential UV-VIS spectroscopy demonstrated the existence of - interactions.
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Time-resolved fluorescence study of liquid and condensed systems based on biopolymer-surfactant interactions.
Černá, Ladislava ; Žitňan, Michal (referee) ; Mravec, Filip (advisor)
This thesis studies properties of hydrogel, which arises on the basis of electrostatic and hydrophobic interactions between hyaluronan chain and micelles of cationic surfactant. A native sodium hyaluronan at molecular weight 750–1 000 kDa and a cationic surfactant CTAB (cetyltrimethylammonium bromide) were used. This hydrogel was assessed as a material for drug delivery systems. The hydrogels were made by mixing 200mM CTAB with 0.5% hyaluronan, both dissolved in 0.15M aqueous solution of NaCl simulating physiological solution. Methods used in this study were steady-state and time-resolved fluorescence spectroscopy, more accurately time-resolved emission spectra (TRES) and deconvolution of steady-state emission spectra of a whole sample by means of parameters gained from fluorescence intensity decays at a set of wavelenghts. Selected systems were investigated by three fluorescent probes, prodan, laurdan and rhodamine 6G. The first two mentioned probes were in hydrogel localized only within micelles in three different microenvironments. Rhodamine 6G pointed out that in hydrogel the aqueous environment is significantly restricted in comparison to purely micellar solution. In addition, rhodamine informed about less available micelle surfaces, caused by hyaluronan chains occupation. There were no interactions between the probes and hyaluronan chains. Freshly made hydrogels showed almost the same results as after a week of maturation under its supernatant.
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Transport properties of agarose hydrogels
Vlasák, Jan ; Kalina, Michal (referee) ; Klučáková, Martina (advisor)
This thesis deals with the transportation processes of electrolytes using simple organic dyes as diffuse probes. The main goal of the work concerns a study of a diffusion mobility of the selected probes being in a form of simple organic dyes in a hydro-gel matrix based on a thermo-reversible linear polysaccharide, i.e., agaroses, functioning as semi-permeable membranes. The thesis describe diffusion processes used for determination of the basic diffusion parameters such as an efficient diffusion coefficient, reputed diffusion coefficient, separating coefficient, concentration of the dye in a gel and the time needed for the diffusion going throughout the entire layer of the gel. Attention is paid also to the environment in which the diffusion processes are implemented (in particular in view of the gel-like samples), porousness of the structure and its mechanical properties.
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Anisotropy and time-resolved anisotropy techniques in colloidal systems research
Holínková, Petra ; Táborský,, Petr (referee) ; Mravec, Filip (advisor)
In this diploma thesis were investigated in terms of microviscosity liquid and condensed systems composed of hyaluronan (Hya) and cationic surfactant cetyltrimethylammonium bromide (CTAB). The excitation and emission spectra, lifetime, steady-state fluorescence anisotropy and time-resolved fluorescence anisotropy of the samples were measured. First, was studied the formation of hydrophobic domains in the system Hya-CTAB at concentration of CTAB lower than its critical micelle concentration in an aqueous solution and 0.15M NaCl. It was found that in an aqueous solution small hydrophobic domains linked to chains Hya are formed. Then an increasing concentration of CTAB leads to phase separation and formation of gel. Due to the addition of NaCl then leads to the reorganization of this system and probably the formation of free micelles in the solution. Were also studied condensed phase of system Hya-CTAB-NaCl at high concentrations of surfactant during fourteen days of ageing. It was found that the microviscosity of hydrophobic domains is constant, but the microviscosity of hydrophilic parts gradually decreases.
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