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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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Transport of pharmaceuticals in model hydrogels
Horváth, Michal ; Enev, Vojtěch (referee) ; Klučáková, Martina (advisor)
This thesis was devoted to the transport properties of diclofenac in model hydrogels. Gels were prepared from agarose and some were enriched with IHSS Elliott Soil humic acid. The addition of humic acid to the hydrogel should mimic the soil environment. And it was the soil environment that was chosen to observe the transport and transfer of diclofenac. Diffusion was observed in diffusion cells, where one part contained diclofenac solution and the other MQ water. And there was a hydrogel between the cells. Samples were taken from both cells and measured spectrophotometrically at different time intervals until equilibrium was established. These measurements were finally used to describe the movement of diclofenac in the hydrogels.
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Humic substances in cosmetics: characterization and stability studies of lip balm
Dostálová, Tereza ; Venerová, Tereza (referee) ; Enev, Vojtěch (advisor)
This diploma thesis deals with the creation of lip balm containing humate to achieve an antiviral effect against the Herpes simplex virus as one of the other possible treatment options for this disease. To visually enhance the product, a dye was prepared from beetroot, the characterization of which is also part of the work. The dye was separated into its individual components by thin-layer chromatography and analyzed by FTIR to describe the structural molecular units it contains. The lip balm itself was prepared in several variants, containing both humate and dye, containing only humate or dye, and in the form of a "blank" sample. These samples were subjected to a long-term four-month stability study, where two different storage conditions were used. The analysis consisted in the assessment of organoleptic properties, spreading properties, determination of the linear viscoelastic area, flow point, melting point and bioburden. For greater specificity, the effect of dye and humate was studied using diffusion and antimicrobial tests. At the end of the study, a sensory analysis was carried out to determine the possible success of the product among consumers.
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Studz of direct and indirect plasma application on onion seeding bulbs
Krejsová, Lenka ; Enev, Vojtěch (referee) ; Krčma, František (advisor)
Nowadays, agriculture is based on conventional methods that involve the application of pesticides, agricultural chemicals and fertilizers. These substances have a negative impact on nature and human health, so more environmentally friendly methods are being sought. This thesis deals with the analysis of fragrances in onions treated with plasma before planting. The treatment was carried out to see if it would improve onion growth while maintaining or increasing the fragrance content. The theoretical part deals with the soil and it is characterization methods, plasma, quantitative and qualitative methods for the determination of volatile substances. It contains the principles of UV-VIS spectrometry, atomic absorption spectrometry (AAS), mass spectrometry (MS) as well as tandem connection of gas chromatography with mass spectrometry (GC–MS). The experimental part was carried out over a period of three years at 22 sites. The preparation and analysis of soil samples are described, which was also evaluated. Furthermore, the procedures for treating the bulbs with corona discharge and plasma-activated water before planting were described. In total, four treatment variants were carried out and bulbs from the fifth variant were not additionally treated. Each treatment variant was planted in four replications of ten bulbs each. Twenty selected grown onions from one site were individually converted to liquid samples and the saturated vapor was analysed by proton transfer reaction time of flight mass spectrometry (PTR–TOF–MS). Thanks to this analysis, the concentrations of volatile substances were determined. Some samples were analyzed by gas chromatography with mass spectrometer as detector to identify the compounds. Subsequently, the data were processed and evaluated. From the results obtained, it is evident that plasma treatment can be useful for the treatment of onions in agriculture, because it confirmed harvest increase without decrease of fragrances concentrations. Thus, it is necessary to carry out experiments on a larger scale in the future.
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Soil solution - supramolecular architecture and stability of dissolved organic matter
Trubková, Eliška ; Doskočil, Leoš (referee) ; Enev, Vojtěch (advisor)
This final thesis is focused on the complex physico-chemical characterization of soil solutions, especially with a focus on the study of the stability and molecular architecture of dissolved organic matter. The subject of the study was six soil solutions obtained from lysimeters in the Závišín locality. Three soil solution samples were taken in December 2022 and the remaining three samples were taken one month later in January 2023, at horizons at 40, 60 and 80 cm below the soil surface. Characterization of soil solutions included measurements of their pH and conductivity. Mass and optical emission spectrometry with inductively coupled plasma was used to determine the concentration of cations. The determination of the total dissolved organic matter content was carried out according to the technical standard ČSN 75 7536 and the values of absorption coefficients were calculated using UV/Vis spectrometry. Functional groups and basic structures of soil solutions were identified based on the evaluated spectra from infrared spectrometry with Fourier transformation. The main contribution was fluorescence spectrometry, which was used to determine the fluorescence intensities of A–fulvic-like and C–humic-like fluorophores. Particle size was also measured using dynamic light scattering and zeta potential, which was a suitable indicator of the stability of selected soil solution samples. The obtained results very well characterized both individual soil solutions and dissolved organic matter.
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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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Use of thin layer chromatography for fractionation of humic substances isolated from oxyhumolite
Hegrová, Nela ; Doskočil, Leoš (referee) ; Enev, Vojtěch (advisor)
The main goal of this thesis was to optimize the fractionation of humic substances (HS) using the method of Thin-layer chromatography, so that it can be used for physico-chemical characterization of the obtained individual organic fractions. Due to the high content of HS in the natural oxyhumolite matrix this oxidized brown coal was selected as a source of these biocolloid compounds which were isolated according to the international IHSS procedure. The sample of native oxyhumolite and oxyhumolite after extraction was characterized by thermal techniques such as thermogravimetric (TGA) and elemental analysis (EA). The above-mentioned oxyhumolite samples were further analyzed by Fourier transform infrared spectroscopy, mainly for deeper understanding of their structure in terms of an organic matrix containing HS. Characterization of isolated HSs was performed by the following thermochemical and spectrometric techniques, elemental analysis, thermogravimetry, ultraviolet and visible electromagnetic absorption (UV/Vis) molecular spectroscopy, Fourier transform infrared spectroscopy and steady-state fluorescence spectroscopy. Molecular absorption spectroscopy (UV/Vis) and steady-state fluorescence spectroscopy were used for physicochemical characterization of individual organic fractions.
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Influence of pH on Extraction of Matter from Biochar-Soil Complex
Jakubčeková, Kristína ; Enev, Vojtěch (referee) ; Kalina, Michal (advisor)
Biochar is one of the possible solutions to improve the soil fertility, therefore it is necessary to investigate both its composition and the impact on the soil under various conditions. The aim of this bachelor thesis is to focus on the description of the influence of pH on the amount and type of substances (molecules, ions) that are leached from the biochar into the soil and thus consequently influence its properties. As a soil model it was chosen cambisol which is the most widespread soil in the Czech Republic. The selected pH values have been defined in such a way to cover the pH range that is common for various soil types in natural conditions. In the experimental part of this thesis the extracts from the used sample of biochar and soil were characterized first. These characteristics obtained in this way were subsequently compared with the data obtained for the biochar-soil complex extracts that was obtained after a defined period of cultivation of the mixture. For the characterization of extracts the basic physico-chemical methods (pH, conductivity, elemental and thermogravimetric analysis, UV-VIS) and also the analysis of leached ions and molecules by means of ICP/OES, GC-MS were used. Evaluated experimental data show that under the condition of cultivation experiments, the influence of biochar was observed only on the basic physico-chemical characteristics of the soil (mainly pH). On the other hand, there was no significant tendency of the impact of biochar on the representation of selected elements and on the content of organic matter in the soil. Long-term biochar-soil cultivation experiments could be a suitable solution.
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Raman spectroscopy as a tool for analysis of microbial cells
Pokorný, Petr ; Enev, Vojtěch (referee) ; Samek, Ota (advisor)
This bachelor thesis is focused on the use of Raman spectroscopy for analysis of ectoine and hydroxyectoine presence in bacterial strain Halomonas elongata. Theoretical part compile characteristics of extremophiles with closer look on halophilic organisms and one of their main osmolytes, ectoines. Following by description of Raman spectroscopy method and its uses. Experimental part deals with cultivation of bacteria on different combinations of substrate and salt concentration in order to reach optimal production of PHA and for achieving the highest possible yield of biomass to be analysed by Raman spektroskopy. As the best substrate for PHA production turned out to be glukose along with salt concentration 30 g/l NaCl, where percentage representation of PHB makes 30,5229 %. As an ideal substrate for the highest yield of biomass proved to be sacharose. Bacteria that grew on sacharose with three different salt concentrations, 40, 70 and 100 g/l NaCl was measured along with Halomonas salina and Halomonas organivorans for comparison. Raman spectroscopy unfortunately turned out to be inadequate for measurement of this type of sample, thus we were not able to measure ectoine of hydroxyectoine presence in cells.
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Experimental study on the release of active ingredients from gel matrices
Volhinava, Anastasiya ; Enev, Vojtěch (referee) ; Sedláček, Petr (advisor)
This thesis deals with the characterization of hydrogels containing humic substances and salicylic acid. Experimental study of the transport of active substances was performed from the gel matrix over the skin and synthetic membranes. During this work was writing literature review focused on the use of gels with humic substances in cosmetic, medical and pharmaceutical industries. The model humic hydrogels was prepared with salicylic acid and their characterization by basic methods of material analysis. These basic methods were and visual assessment of consistency during the preparation and rheology. The study of the release of active and humic substances was performed due to the vertical diffusion cells. The amount of released humic substance was characterized by UV-VIS method and the amount of released salicylic acid from gel matrix was determined by HPLC.
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