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Dialysis study of hyaluronan-surfactant interactions
Šejnohová, Michaela ; Venerová, Tereza (referee) ; Pekař, Miloslav (advisor)
This diploma thesis is concentrated on the interactions between polyelectrolyte (hyaluronan) and cationic surfactant (CTAB). The experiments were performed in an aqueous solution and in an environment of physiological ionic strength (0,15mmoldm-3 NaCl). The determination of the surfactant concentration in solutions was based on the formation of colored complexes of CTAB and picric acid in chloroform. The concentrations of surfactant were measured by UV-VIS spectroscopy. The stability of CTAB+HyA was examined by a dialysis method. The results showed that, regardless of the environment, the presence of HyA in solution reduces the number of free molecules of CTAB which can be determined in the sample. It has been proved that there is an interaction between HyA and surfactant and that CTAB has greater affinity for HyA then for the picric acid. The stability of CTAB+HyA was determined by dialysis of 120 hours. After that time, the concentrations of the retentate and permeate were settled. The results showed that in the membrane remains a certain amount of CTAB bounded to hyaluronan. The system can be suitable for the preparation of targeted carriers of biologically active substances.
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Study of physical gels with hydrophobic domains
Kovářová, Lenka ; Márová, Ivana (referee) ; Mravec, Filip (advisor)
The thesis is focused on physical hyaluronan gels. The object of study is the interaction of hyaluronan (HyA) with oppositely charged surfactants in physiologic solution (0.15 M NaCl), leading to the formation of gel. In the first part of work have been determined the solids´ contents (X) in gels and their supernatants in percentage and their correlation with molecular weight concentration of original HyA solution and the ratio of binding sites on hyaluronan chain and surfactant CTAB. To conclude, decrease in HyA concentration results in higher values of X and vice versa. On the other hand, increase in the value of X with increasing molecular weight of HyA is not so significant. Analogous conclusions have been made for supernatants and the amount of solids in gel. Drying process has been recorded by drying curves. Swelling process has been used for the characterization of gels. The percentage of water that can be absorbed by dried gel, was determined. The results are in agreement with the measurements of solids´ content in gels. In the next part, the correlation between rheological properties of gels and HyA concentration, HyA molecular weight and concentration of CTAB have been studied by the oscillation and flow tests. The samples with the highest molecular weight and concentration have the most viscoelastic character. The flow test confirmed the assumed pseudoplastic behavior of gels. A very interesting trend arose while comparing HyA concentrations and viscosity in stock solutions and gels. Whereas in stock solution viscosity (at low shear rate) is lower with increasing of HyA concentration, the situation was exactly the opposite in gels. The results are in agreement with frequency tests and observed character of gels.
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Influence of the preparing of a biopolymer-surfactant complex on its stability and solubilization properties.
Pilgrová, Tereza ; Burgert,, Ladislav (referee) ; Mravec, Filip (advisor)
Influence of the preparing of a biopolymer-surfactant system on its solubilization properties and stability was investigated by using fluorescence spectroscopy and dynamic light scattering methods. Investigation was made on complex of native hyaluronan with cationic surfactant cetyltrimethylammonium bromide (CTAB). System has been studied in aqueous and in saline solutions. The effect of temperature of stock solutions and freezing effect on subsequent properties were investigated. Further was examinated, what effect has a way of introducing fluorescent probe into the system on subsequent solubilization properties. It was found that the conditions of preparing of biopolymer-surfactant system have a significant effect on the solubilization properies and stability of complex.
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Study of interaction in biopolymer-surfactant system
Sovová, Šárka ; Mondek, Jakub (referee) ; Pilgrová, Tereza (advisor)
Interaction between biopolymer-surfactant and stability of arising complexes were investigated using dynamic light scattering method Stability of the system and interactions between native hyaluronan and cationic surfactant cetyltromethyl-amonium bromide in aqueous solution depending on preparation method were investigated. System was prepared either adding CTAB to hyaluronan causing sequential micelle’s formation, or adding hyaluronan to already formed micelles. It was found out that preparing method influences properties of arisen complexes. Molecular weight of hyaluronan has an inconsiderable effect on interaction against to concentration of hyaluronan, which influences interaction of component significantly.
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Kinetic study of excited state proton transfer in polymer-surfactant system
Bačová, Jana ; Vala, Martin (referee) ; Mravec, Filip (advisor)
In this bachelor thesis were studied interactions between hyaluronic acid and cationic septonex by steady-state and time-resolved fluorescence with fluorescence probes pyrene and 1-naphthol. At first was determined critical micelle concentration of septonex. In the case of both fluorescence probes was studied area below and above critical micelle concentration. It was found, that 1-naphthol undergo very fast deprotonation in aqueous solution, while in the present of micelles the rate of deprotonation is c significantly retarded. In the system of hyaluronic acid – septonex it was observed, that formation of aggregates occurs at lower concentrations than in the system without hyaluronic acid. Molecules of probe are probably distributed between hyaluronic acid – septonex aggregates and free micelles above critical micelle concentration.
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Aggregation processes in amphiphil-polymer system
Šejnohová, Michaela ; Márová, Ivana (referee) ; Pekař, Miloslav (advisor)
This Bachelor´s thesis is focused on the exploration of polyelectrolyte-surfactant system by fluorescence spectroscopy with pyrene as fluorescent probe. Cationic cetyltrimethylammonium bromide (CTAB) was used as a surfactant. Native hyaluronan was used as polyelectrolyte. Hyaluronan was used in one constant molecular weight and one constant concentration. The whole system was prepared in an aqueous environment. Interactions were studied in area between critical aggregation concentration (CAC) and critical micelle concentration (CMC). The influences of the concentration of pyrene and the effect of the way of the samples preparation on aggregation behavior were also studied. It was found that above the critical aggregation concentration exists an area where stable properties of aggregates can be found. The concentration of pyrene has a major effect on the fluorescence properties of the system. Preparation of samples affects the aggregation of the system. At higher surfactant concentrations, above the stable area, was observed phase separation and gel formation.
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Dynamic surface tension in study of hyaluronan-surfactant interactions
Herzog, Milan ; Žídek, Jan (referee) ; Pekař, Miloslav (advisor)
The main task was to characterize interactions between negatively charged biopolymer sodium hyaluronate and two cationic surfactants: cetyltrimethylammonium bromide (CTAB) and tetradecyltrimethylammonium bromide (TTAB). Interactions between sodium hyaluronate and these surfactants are based on their different charge and can be detected by changes of surface tension. The influence of constant addition of sodium hyaluronate (MW = 90 and 1400 kDa) on interface tension values at increasing bubble lifetime was observed. For this purpose, the maximum bubble pressure method was choosen. The measurements were performed in water and in 0,15 mol•l-1 NaCl solution. The obtained results can be used as for description of dynamic properties of hyaluronate-surfactant systems as, e.g., in targeting drug delivery.
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Preparation and application of physical hyaluronan gel
Kovářová, Lenka ; Krouská, Jitka (referee) ; Venerová, Tereza (advisor)
Bachelor's work is concerning by research in physical hyaluronan gel area. Research is focusing on its preparation, especially by interaction of hyaluronan (HyA) solution with opposite charged surfactant in physiological solution (0,15 M NaCl). In the first part of work I found out influence of molecular weight, concentration of original hyaluronan solution and the ratio of binding sites on hyaluronan chain and surfactant on amount of solids in gel, expressed in percent. Finally we can say, that this value is not influenced by molecular weight of HyA neither relation of binding sides. On the other side, concentration of original HyA solution has significant influence on amount of solids in gel. There was also researched process of gel swelling, where was set amount of water (in percent) able to take dried gel back. Analogical conclusion has been reached like amount of solids in gel. In other part were researched influence of hyaluronan molecular weight and the ratio of binding sites on hyaluronan chain and surfactant on formation of excimers and spectrum change by fluorescence spectroscopy. Pyrene, 1,3-bis(pyren-1-yl)propane (P3P) and prodan were used as fluorescence probes. Excimer emission showed at P3P, while there was no growth of intensity at 470 nm for pyrene. It was found that molecular weight does not affect the value of the ratio of excimer and monomer probe P3P, or the ratio of the first and third peaks of pyrene. The ratio of binding sites HyA and surfactant influence only relation of excimer to monomer (P3P probe), while influence to relation 1:3 hasn't been noticed for pyrene. I have compared spectrum of developed gel for prodan probe, relevant solid and swelled gel by return. It also served for drying and swelling process visualisation.
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Influence of the preparing of a biopolymer-surfactant system on its aggregation behavior.
Pilgrová, Tereza ; Sedláček, Petr (referee) ; Pekař, Miloslav (advisor)
To study the effect of the method of preparation of biopolymer-surfactant on the aggregation properties several experiments were designed. Closer investigation was made on native hyaluronan-cetyltrimethylammonium bromide (CTAB) system. The system has been studied in the phosphate buffer in aqueous and saline solutions. The effect of the order of substances addition in the samples, the influence of different methods of preparation of certain concentrations of the solutions or the effect of dissolving powdered biopolymer and surfactant on the contrary, the resulting system were investigated. Fluorescence intensity was measured for all prepared samples and its changes were investigated. Pyrene and Nile red were used as fluorescent probes. Some samples were also measured by tensiometer. Measurement of surface tension was based on the method of divorcing the ring from the surface. It was found that the order of the mixing of solutions has a significant effect on the aggregation properties of the system and its appearance. Further effects were detected on the prepared samples by dissolving powdered biopolymer surfactants and vice versa, and environmental influences of the polarity of the system. It was found that sodium chloride abolished the effects of interactions in the system. Effect of method of preparation of certain concentration of CTAB in phosphate buffer is reflected only slightly.
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