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Preparation and characterization of triple-IPA for the formation of vesicular systems
Novotná, Ludmila ; Pekař, Miloslav (referee) ; Mravec, Filip (advisor)
This thesis deals with the preparation and characterization of vesicular systems consisting of triple-IPA (ion pair amphiphile with three hydrophobic chains). For the preparation of tripleIPA were used double-chain cationic surfactants dimethyldimyristylammonium bromide (DMSAB), dimethyldipalmitylammonium bromide (DPAB), dimethyldioctadecylammonium bromide (DODAB) and dimethyldioctadecylammonium chloride (DODAC) and the singlechain anionic surfactant sodium dodecyl sulfate (SDS). Among the cationic surfactants, DMSAB and DPAB were selected to be studied more thoroughly. Triple-IPAs were prepared by mixing solutions of oppositely charged surfactants in equimolar ratio, and the resulting triple-IPA precipitate was filtered and dried. From the obtained powder, vesicular systems were prepared by rehydration and sonication. Cholesterol and cationic surfactants, from which the triple-IPAs were prepared, were used to stabilize the cationic vesicles. The properties of the vesicular systems were evaluated by dynamic and electrophoretic light scattering measurements and turbidimetry. The dispersion solutions with the addition of cholesterol were less turbid; therefore, it can be assumed that they supported the formation of cationic vesicles. The optimum cholesterol content is 20 mol. %. At this concentration, the effect of cholesterol on vesicle membrane reorganization becomes apparent and the cationic system remains monodisperse. A positive zeta potential in the stable region was determined for all prepared solutions, which was further increased by the addition of a positive surfactant. The most suitable cationic surfactant for the preparation of triple-IPA is DPAB, which is easy to work with, produces the most monodisperse systems and has the lowest measurement uncertainties.
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Comparison of microbial metabolic production from waste and powder whey
Elefantová, Petra ; Vítová, Eva (referee) ; Babák, Libor (advisor)
The master’s thesis discusses the comparison of microbial metabolic production from waste and powder whey. Whey is obtained as a by-product of cheese production. Lactose (preferably whey) using lactic acid bacteria (eg. Lactobacillus) under suitable temperature conditions is converted to lactic acid. Effect of temperature, effect of salts and effect of yeast extract on lactic acid production by L. casei were investigated. HPLC metod was determined lactid acid. In the practical part were used bacteria of the genus Lactobacillus. It was found that for dried and waste whey is the optimal temperature of 35 °C. At this temperature is the greatest gain of lactic acid. The highest concentration of lactic acid was obtained by using 20 g of yeast extract for dried whey and for waste whey were used 24 g of yeast extract. When monitoring the effect of salt concentration on the production of lactic acid, it was found that using only MnSO4·H2O gain most of lactic acid.
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Utilization of various methods of porosimetry in investigation of internal structure of biopolymer hydrogels
Zahrádka, Jan ; Pekař, Miloslav (referee) ; Sedláček, Petr (advisor)
The main aim of the thesis was to select, optimize and test suitable methods for studying inner structure of hydrogel materials. The study of the size of pores with potential use in fundamental and applied research of these materials is the central focus of the thesis. Firstly, a literature search was done in order to collect basic information about usefulness of the individual methods. On the basis of this literature search, the experimental part of the work was divided into several steps. First, selected methods of conventional porosimetry (mercury intrusion porosimetry, BET analysis and DSC thermoporometry) were applied on reference porous materials and the results obtained by the individual methods were compared. The methods optimized with the use of these reference materials were then utilized in the analysis of porosity of model hydrogels based on polyvinyl alcohol. Based on the results of this analysis, the usability and benefits of the individual methods were discussed with respect to the sample preparation, time and financial demands, and also from the viewpoint of compliance of the determined pore sizes with the range of sizes declared for the physical PVA gels.
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Analytical methods for qualitative and quantitative determination of PHA in cyanobacteria
Černayová, Diana ; Samek, Ota (referee) ; Sedláček, Petr (advisor)
The diploma thesis is confused to verify the applicability of selected physicochemical and spectroscopic methods for characterization of cyanobacteria, with special emphasis on possibilities of qualitative and quantitative analysis of polyhydroxyalkanoates (specifically polyhydroxybutyrate (PHB)) accumulated in cyanobacterial cells. The sample basis of the work was formed by cultures of cyanobacterial strains of Synechocystis sp. PCC 6803 and Synechocystis salina CCALA 192. The cultures were were cultivated in several ways to cover the widest possible range of physiological conditions and PHB contents, in particular using an autotrophic way of cultivation on shakers and multicultural culture method in a basic culture medium,and in media enriched with 2% salt (NaCl ) as well as mixotrophic culture media with different types of the carbon substrate. After few weeks of cultivation, cyanobacterial cultures were obtained and complexly analyzed by following techniques- cell suspensions were analyzed by flow cytometry and UV-VIS spectrometry (transmission and diffusion transmission mode), dry cell biomass was characterised by gas chromatography to obtain a exact amount of PHB, and then FT-IR spectrometry and thermogravimetric analysis. The work aimed to assess whether any of these methods can be a quick and affordable alternative to the determination of PHB content to the most commonly used method of gas chromatography, but also to assess what additional information about the physiological state of cyanobacterial cells can provide test methods. The highest correlation on PHB content was determined for the parameters determined by infrared spectroscopy, in which specific peaks from the characteristic wavelengths for polyhydroxybutyrate were important. Weak correlations on PHB content were achieved in thermogravimetric analysis and cytometry, using the hydrophobic fluorescent probe BODIPY 439/503, which bound to lipophilic parts of cells. In addition to the determination of PHB, it was possible to determine pigments present in cyanobacteria (such as chlorophyll, phycocyanin and carotenoids) by flow cytometry and UV-VIS diffusion transmission spectrometry. In the end, results from all used techniques were compared by PCA analysis to determine the similarity of all analyzed samples.
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Interactions between hyaluronan and cetrimide
Adamcová, Zuzana ; Sedláček, Petr (referee) ; Mravec, Filip (advisor)
Interactions between hyaluronan and cationic Cetrimide were studied using fluorescence spectroscopy and turbidimetry in model physiological solution (0,15 M NaCl) and water. Results obtained in water showed strong electrostatic interactions before reaching CMC of the surfactant, which were screened in presence of salt due to high ionic strenght of the solution. Behavior of Cetrimide-hyaluronan systems was compared with the pure TTAB in equivalent system. It was found, that changes in Cetrimide-HA system occure at lower concentrations of the surfactant than in TTAB-HA system. This was probably caused by presence of small amount of CTAB in Cetrimide.
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Analytical methods for determination of polyhydroxyalcanoates content in bacterial cells
Černayová, Diana ; Enev, Vojtěch (referee) ; Sedláček, Petr (advisor)
This bachelor thesis deals with methods that are used for determination of polyhydroxyalkanoates (PHA) in cells of bacteria Cupriavidus necator H16. To the applied methods belong Fourier transform infrared spectroscopy, Raman spectoscopy, turbidimetry and thermal analysis (thermogravimetric analysis and differential scanning calorimetry). The results of each method were compared to amount of PHA determinated by gas chromatography. Fourier transform infrared spectroscopy and Raman spectroscopy revealed a linear relation between ratio of characteristic peaks and amount of PHA. FTIR enabled to determine not only the quantitative amount of PHA, but the quality of the sample (cristallinity, amorphous state) as well. Another method was turbidimetry, which has shown correlation between dispersion of UV-VIS light to amount of PHA in bacterial cells. Thermogravimetric analysis was the most effective technique for determination of PHA. The dependence of weight loss of PHA on its content (found by gas chromatography) was determined with the highest accuracy to gas chromatography. Tha last method- DSC was not alternative replacement for quantitative PHA determination. However, DSC indicated crystalline and amorphous state of the polymer.
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Preparation and characterization of catanionic complexes
Mušková, Alexandra ; Kalina, Michal (referee) ; Mravec, Filip (advisor)
The diploma thesis deals with the preparation and characterization of catanionic complexes and their possible application to pharmaceutics and medicine. The catanionic complexes were prepared by mixing two oppositely charged surfactants with the concentration of 20 mmol·dm3 in various volume ratios. Two systems were analyzed – CTAB + SDS and Septonex + SDS. Both systems were prepared in an aqueous and physiological environment. The turbidimetric analyses provided results of the intensity of turbidity of the prepared mixtures. The size and stability of the prepared particles were determined by DLS and ELS measurements. CTAB-rich samples showed significant viscosity changes and they were therefore characterized by rheological measurements. The results of this work show that the asymmetry of alkyl chains, surfactant selection, temperature, and ionic strength have a significant influence on the properties and self-assembly of surfactants in catanionic complexes.
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Investigation of Polyelectrolytes Interactions with Cationic Aminogroups-containing Amphiphiles
Zeman, Jan ; Běťák, Jiří (referee) ; Chytil, Martin (advisor)
The study deals with interactions of polyelectrolytes polystyrene sulfonate and hyaluronic acid with nitrogenic amphiphilic substances, represented by lysine and albumine. To study the interactions pH-metry, conductance, viscositic and turbidity measurement, DLS and reometry were used. All mixtures of different concentrations were measured and the data were compered with data obtained from measurement of samples with amphiphilic sumstances without polyelectrolytes. Observed interactions occured in the aminoacid concentrations between 0 to 20 mmoldm-3, then the PSS interaction groups were fully bonded by lysine and no more interactions were recognized. The same behaviour were observed in albumine solutions with concentration under 2 gdm-3.
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Characterization of hyaluronan interactions with albumin
Valentová, Kristýna ; Kratochvílová, Romana (referee) ; Chytil, Martin (advisor)
This bachelor thesis focuses on the research of physical interactions between hyaluronic acid and a protein albumin. In order to investigate these interactions following methods were used: pH and conductivity measurement, viscosimetry, turbidimetry and rheometry. The interactions were observed in solutions of fixed albumin concentration 1.0 gdm-3 and varying concentrations of hyaluronic acid from 0.1 to 2.0 gdm-3. Results were compared with a solution of hyaluronic acid without the addition of albumin. The results show that there are interactions between hyaluronic acid and albumin solutions, especially for those containing high molecular weight hyaluronic acid. The interactions exhibited a change in viscosity of solutions. We can see a reduction in viscosity after adding albumin to the reference solutions. The reduction is noticeable even in the low molecular weight HA solutions, but is not so significant.
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Preparation and characterization of quatsomes
Havlíčková, Anna ; Szabová, Jana (referee) ; Mravec, Filip (advisor)
This bachelor thesis deals with preparing and characterizing quatsomes and similar vesicular systems. The preparation consisted of weighing an equimolar amount of sterol and surfactant, hydration with deionized water, and subsequent sonication with an ultrasonic probe with an energy of 5, 10 and 15 kJ. In this work, two types of quatsomes were prepared, the first contained equimolar amounts of cholesterol and cetyltrimethylammonium bromide (CTAB), and the second consisted of cholesterol carbethopendecinium bromide (Septonex). The optimal dispersion energy of 10 kJ was selected for both types based on turbidity measurements, visual observation and dynamic and electrophoretic light scattering analysis. Furthermore, the observed properties were found to be constant from day 7 after sonication. In addition to quatsomes, similar vesicles were prepared, which contained a surfactant in the form of a sulfate salt. Once again, two types were prepared, the first containing sodium dodecyl sulfate (SDS) and cholesterol, the second sodium tetradecyl sulfate (STS) and cholesterol. Based on the same measurements as for quatsomes, optimal dispersion energy of 10 kJ was selected. Furthermore, mixtures with different ratios of cholesterol and a given surfactant were prepared, in which the effect of cholesterol addition was monitored. The analysis of these mixtures was performed by the same methods as the analysis of quatsomes.
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