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Agregation of palmitoyl-modified hyaluronan
Lehocká, Nikola ; Mravec, Filip (referee) ; Pekař, Miloslav (advisor)
This thesis deals with the aggregation behaviour of palmitoylhyaluronan in two degrees of substitution, namely 10 % and 16 %. Using a fluorescence spectroscopy method with pyrene as a fluorescence probe, we found a critical micellar concentration. The results were confirmed by measuring the dynamic light scattering, which also showed an increase in the size of aggre-gates with an increasing concentration. System stability is reduced by increased ionic strength as evidenced by zeta potential measurement. Experiments found that 16 % substitution sample can form a gel. The gel is very stiff and has excellent properties, which was confirmed by rhe-ology. We also managed to incorporate pyrene in the gel, which was demonstrated by the presence of highly solvated domains that could be polymer micelles. Based on these results, 16% substitution sample was subjected the MTT assay to cytotoxicity. The results confirmed that the examined sample was not toxic.
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Molecular organization of humic acids in solutions
Chytilová, Aneta ; Enev, Vojtěch (referee) ; Klučáková, Martina (advisor)
The diverse structure and content of the highly varied functional groups of humic acids determine these substances as a worthwhile study material for a number of scientific institutions. Above all, the structure causes a wide range of their interesting properties, especially spectral, colloidal, electrochemical, ion-exchange and adsorption properties. Humic acids have variety of utilization in the industry while they have been the subject of a range of further research and development. The character of these complex heterogeneous polydisperse substances is mainly cyclic and aromatic, which according to some studies results in their brown color. Generally, there are multiple theories assuming their potential secondary structure. The aim of this study is the analysis of structure and conformation of humic acids in aqueous solutions. For the purposes mentioned above a concentration series of humic acids have been prepared in four different mediums: 0,1 M NaOH, humic acids in water with pH modified to 12 (basic medium), 0,1 M NaOH + 0,1 M HCl, 0,1 M NaCl (neutral medium). In order to study conformations several analytic methods have been used: ultraviolet and visible spectroscopy, dynamic light scattering, electrophoretic light scattering, potentiometric determination of pH and direct conductimetry. Throughout the experimentation the emphasis was mainly on the purity of humic acids extracted from lignite.
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The Study of Aggregation of Humic Acids in Different Xenobiotic Environments
Hronová, Aneta ; Smilek, Jiří (referee) ; Kalina, Michal (advisor)
Theme of the presented bachelor thesis is the study of the aggregation behavior of the humic acids (HA) in the environment of extraneous substances. As extraneous substances, three cationic surfactants were selected (CTAB, TTAB and Septonex). In the experimental part of the thesis, the humic acids were characterized in detail using different methods (elemental analysis, ash content determination, titration acidity determination). The experimental part of the thesis is dealing with the structure of created HA-surfactant systems. These systems were characterized using the results of the elemental analysis and FT-IR spectroscopy. The key part of the experimental part of the study is represented by the selection of proper methods and by determination of the critical aggregation concentration of the humic acids in the surfactant environment. For these purposes, two key methods were selected in the frame of the presented bachelor thesis: isothermal titration calorimetry and dynamic light scattering. The first method is sensitive owing to the energetic fluctuations, in the case of the dynamic light scattering we are able to detect well the changes in the particle size caused by mutual interactions of particular substances in the system.
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Conformation and molecular organization of humic acids in aqueous solutions
Věžníková, Kateřina ; Pospíšilová, Ľubica (referee) ; Klučáková, Martina (advisor)
This diploma thesis deals with the conformation and molecular organization of humic acids in aqueous solutions. Humic substances have heterogeneous and polydisperse nature, therefore their secondary chemical structure has not yet been defined properly, neither has their conformational arrangement. The conformation of humic substances in the solutions is mainly stabilized by weak disperse forces, such as Van der Waals, -, CH- interactions and hydrogen bonds. Humic substances in the solutions tend to form aggregates that are held together mostly by the intermolecular hydrophobic interactions. Concentration series of humic acids were prepared in three different mediums of constant ionic strength: NaOH and NaCl (prepared either by neutralization NaOH by HCl or direct dilution by solution of NaCl). Several analytical methods have been used to determine conformation and molecular organization of humic acids: potentiometric determination of pH, direct conductometry, ultraviolet and visible spectroscopy, density measurement, dynamic light scattering, laser Doppler velocimetry and high resolution ultrasound spectrometry. It was confirmed that the conformation and molecular organization of humic acids in aqueous solutions depend not only on pH of medium, but they also depend on chemical nature and concentration with the same pH. Results showed that hydrodynamic diameter of particles is significantly increasing in NaCl medium prepared by neutralization NaOH by HCl, particularly at low concentrations, which corresponds to higher values of zeta potential obtained. Concentration dependencies of ultrasonic velocity and compressibility also indicate the changes in conformation and molecular organization corresponding with results from other methods used.
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Nanocarriers based on hydrophobized biopolymer
Velcer, Tomáš ; Hnyluchová, Zuzana (referee) ; Venerová, Tereza (advisor)
This work studies the properties of hydrophobically modified biopolymers for potential use in targeted drug delivery. Two samples of hydrophobically modified hyaluronan were selected. The critical micelle concentration (CMC) of these samples was measured using fluorescence spectroscopy, wherein it was found that micelle formation occurs over a wide concentration region. Environment of 0,15 M NaCl causes the decrease of CMC only slightly or not at all. Using the method of dynamic light scattering, particle size was measured. One of the samples exhibited a double size distribution. DLS results are consistent with the results of fluorescence spectroscopy at issue in aggregates forming near the CMC point. Zeta potential measurements provide information about stability of the samples. The results of this study confirm that one of the studied samples seems to be a suitable candidate for the post of drug carrier for targeted delivery.
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Phospholipids as the basis of biodegradable delivery systems
Burdíková, Jana ; Čeppan, Michal (referee) ; Mravec, Filip (advisor)
This thesis is focused on investigation of phospholipid-hyaluronan system. First, appropriate method for preparation of bulk solution of phospholipid/lipid and suitable fluorescence probe were chosen. Sonification was selected as a method for preparation of bulk solution and pyrene was chosen as a fluorescence probe. From the group of phospholipids lecithin was selected. Next to phospholipid, lipid with no phosphate group (DPTAP) was utilized for comparison, alternatively a mixture of lipid (DPTAP) and phospholipid (DPPC). Instead of hyaluronan another polyelectrolytes (sodium polystyrene sulfonate, sodium alginate) were used too. Measurements were performed in water environment and in phosphate buffer saline (PBS). All investigation was accomplished by fluorescence spectroscopy and dynamic light scattering.
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Fractionation and molecular organization of humic acids
Chytilová, Aneta ; Enev, Vojtěch (referee) ; Klučáková, Martina (advisor)
Humic acids are part of the natural organic matter occurring all around us. The aim of this thesis is to study the molecular organization, conformation of humic acids in aqueous solutions, that always raise a number of questions. For a long time, the scientists all over the world argue, if humic acid are polymers, micelles or supramolecules. Over time, thanks to new technologies, their opinions are moving away from a polymer model and tend rather to supramolecular arrangement of humic acids. Studying humic complex systems is not easy, because they are polydisperse and heterogeneous, which significantly complicates any characterization. Moreover, its molecular organization is affected by many factors such as e.g. pH, ionic strength and etc. For the study of the conformation of humic acids, concentration series of IHSS (International humic substances asociation) Leonardite humic acids stamdards in four different mediums were prepared: 0,1 M NaOH, humic acids in water with pH modified to 12 (basic medium), 0,1 M NaOH + 0,1 M HCl, 0,1 M NaCl (neutral environment). Furthermore it has been performed the fractionation of humic acids for the purpose of simplifying the complicated structure. Prepared concentration series were characterized with several analytical methods such as ultraviolet and visible spectroscopy, dynamic light scattering, electrophoretic light scattering, microrheology, gel permeation chromatography, potentiometric pH determination and direct conductometry. Diploma thesis is built on the previous bachelor thesis in which different sample of humic acids was studied. The measured results indicate that the studied systems show supramolecular behavior and in some cases are subject to aggregation into larger units (micelles).
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Study on interactions of protonated aminoacids with low-molecular weight hyaluronan
Chlumská, Jana ; Kalina, Michal (referee) ; Chytil, Martin (advisor)
This bachelor thesis deals with interactions between hyaluronic acid (HA) of low molecular weight (110-130 kDa) and protonated aminoacids lysine and 6-aminocaproic acid. For investigation of these interactions, methods such as pH-metry, conductance measurement, viscometry and dynamic light scattering were used. The electrostacic interactions between carboxylic group of HA and the aminogroup of the aminoacids are presumed. Proving these interactions would allow us to physically modify HA and further more, using such a system as a carrier of pharmaceuticals. The interactions of protonated lysine with HA were studied in pure water or in the solution of NaCl (c = 15 mmol•dm-3). Interactions of 6-aminocaproic acid were studied only in pure water. Both aminoacids were in the concentration range 0.9-20 mmol•dm-3. The decrease in the viscosity of the solutions indicates the interactions for lysine; for 6-aminocaproic acid the interactions also occured, but not as signifciantly as for the system containing lysine.
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The Utilization of Light Scattering Techniques for the Study on Hydrogel Gelation
Candráková, Simona ; Smilek, Jiří (referee) ; Kalina, Michal (advisor)
This bachelor thesis focuses on the study of hydrogel gelation using light scattering methods. For these purposes two different biopolymers (agarose and sodium alginate), with different sol to gelation phase transition, were selected. In the case of agarose, the gelation is caused by temperature change. On the other side, the gelation of sodium alginate is initiated by addition of polyvalent cations. In the experimental part of the work, agarose gelation was studied by DLS, temperature gradients of agarose solutions (40 – 30 °C) were measured. During the measurement the particle size distributions in the solutions having different concentrations were monitored as well as the temperature influence on the diffusion coefficient. The DLS method provided the values of the gelling temperatures of the solutions at a concentration of 0,5 wt.% and 1,0 wt.%. Furthermore, the DLS microrheology method was used, where temperature dependences of viscoelastic characteristics of agarose solutions (40 – 30 °C) were also measured, from which the gelling temperatures of agarose solutions of 0,1 wt.% and 0,5 wt.% were evaluated. The classical rheology was also used in the work to compare the viscoelastic behaviour of the samples and to determine the gelation point of the agarose solutions. By this method were determined gelling temperatures for all studied concentration of agarose (0,1 wt.%, 0,5 wt.% and 1,0 wt.%). The DLS method was also used to monitor the interactions of sodium alginate with the addition of Ca2+ ions, these interactions were then also evaluated and discussed in the experimental part.
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Colloidal particles marked with biopolymer
Pihíková, Dominika ; Víteček,, Jan (referee) ; Mravec, Filip (advisor)
The effect of hydrophobically modified hyaluronan on surfactants aggregation has been studied in this master’s thesis. The value of critical micelle concentration of anionic surfactant SDBS (sodium dodecylbenzensulfonate), cationic surfactant CTAB (cetyltrimethylamonnium bromide) and nonionic surfactant Triton X-100 (octylphenol ethoxylate) was determined by fluorescence spectroscopy using pyrene probe. Aggregation behavior of surfactants was performed with addition of hydrophobically modified hyaluronan of two molecular weights (17 kDa, 206 kDa) in aqueous solution. The greatest influence of hydrophobized hyaluronan on aggregation behavior was observed in system with cationic surfactant CTAB. Stability of system containing cationic surfactant and hydrophobically modified hyaluronan was established through zeta potential. Last part of thesis deals with size determination using dynamic light scattering.
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