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Fluorescence probes in investigation of associative colloid domains.
Mischingerová, Monika ; Krouská, Jitka (referee) ; Mravec, Filip (advisor)
The aim of this thesis was to determine how the concentration of the fluorescent probe (pyrene) affects the critical micelle concentration. In this thesis were used all types of surfactants (anionic, cationic and nonionic). As representatives of the anionic surfactants were chosen sodium dodecylsulfate (SDS) and sodium dodecylbenzenesulfate (SDBS), as the cationic surfactant was chosen cetyltrimethylamonium bromide (CTAB) and as the nonionic polyoxyethylene(9.5)octylphenol (Triton X–100). The critical micelle concentration of these surfactants were measured with a pyrene at three different concentrations. All data obtained by fluorescence spectroscopy were fitted by a Boltzmann-type sigmoid. It was found out that in the case of the anionic surfactants there is some correlation between the critical micelle concentration of the surfactant and the concentration of the fluorescent probe. In both cases (SDS, SDBS) the value of the critical micelle concentration increased with increasing concentration of the fluorescent probe. For the other surfactants (cationic and nonionic) was observed no change in the critical micelle concentration values depending on the concentration of pyrene.
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Quality of hydrophobic domains in polyelectrolyte-surfactant system
Holínková, Petra ; Vala, Martin (referee) ; Mravec, Filip (advisor)
In this bachelor thesis were studied the photophysical properties of four fluorescent probes depending on the viscosity of their surroundings by the fluorescence spectroscopy. For probes DPH, DPB a perylene was determinated dependence of fluorescence anisotropy and for P3P dependence of ratio excimer/monomer on the viscosity of the sample. Anisotropy probes DPH and perylene were also used to study interaction between hyaluronan and cationic surfactant CTAB (concentration below CMC) in aqueous solution and 0,15 M NaCl. Information about forming hydrophobic domains were obtained by measurement of fluorescence intensity and anisotropy depending on the concentration of CTAB in the sample. It was found that in an aqueuos solution, after the initial addition of CTAB to hyaluronan formed small hydrophobic domains linked to chain of hyaluronan. Increasing concentration of CTAB leads to the formation of gel and phase separation. After addition of NaCl leads to reorganization of this system and probably the formation of free micelles in the solution with higher concentration of CTAB.
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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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Influence of pH on polyectrolyte-surfactant system
Mourycová, Jana ; Omelka, Ladislav (referee) ; Pekař, Miloslav (advisor)
In this bachelor thesis, was studied influence of pH on aggregation polyelectrolyte-surfactant system using fluorescence spectroscopy and pyrene as fluorescent probe was used. As polyelectrolyte, hyaluronan with different molecular weight was used. Two surfactants cationic surfactant cetyltrimethylamonium bromide (CTAB) and nonionic surfactants polyoxyethylene (20) (TWEEN 20) were selected for interaction with hyaluronan. In the pH range from 3 to 10, no signifact changes in the aggregate non-polar core was observed. This mean system is stable against changes in the observed pH range. At system with higher concentrations of cationic surfactant (CTAB) and higher molecular weight hyaluronan occurred phase separation and gel exclusion was observed.
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Hyaluronan ionocomplexes for cosmetic and farmaceutical applications
Cimalová, Jana ; Obruča, Stanislav (referee) ; Pekař, Miloslav (advisor)
This bachelor thesis is focused on the study of a system of cationic surfactant – hyaluronan. Using fluorescent spectroscopy the critical micellar concentration of Carbethopendecinium bromide (Septonex) was determined in three environments – water, 0,15 M NaCl, and buffer PBS, with three fluorescent probes – pyrene, perylene, and nile red. It was found that the physiological environments (buffer and NaCl) reduce the value of CMC about one order. During the experiments with native hyaluronan, precipitate and gel formation were observed. The following part dealt with aggregate behaviour of the new biologically acceptable amphoteric surfactant tetradecylfosfocholine (TPC) in the environment of water and NaCl according to the increasing molar molecular weight of hyaluronan. As a fluorescent probe perylene was used. These measurements showed that the molar molecular weight or a change of ionic force has no effect on the value of CMC and therefore there are no interactions with hyaluronan. The last surfactant to study was 1,2-dihexadecanoyl-3-dimetylammonium-propan (DPTAP), which is water insoluble. DPTAP is soluble in chloroform, but after evaporation of chloroform and after addition of hyaluronan, it precipitated. The results so far, have confirmed that in terms of research of aggregation by fluorescence spectroscopy the new cationic surfactants TPC and DPTAP are not suitable for complexation with hyaluronan.
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Interaction in ionic surfactant/non-ionic surfactant/polyelectrolyte system
Pihíková, Dominika ; Sedláček, Petr (referee) ; Mravec, Filip (advisor)
The interactions in an ionic surfactant/non-ionic surfactant/polyelectrolyte system have been studied in this bachelor thesis. The value of critical micelle concentration of cationic didecyldimethylammonium chloride (DDAC) has been determined in three different mediums by fluorescence spectroscopy using Pyrene, Perylene and Nile red as probes. The CMC value of non-ionic octylphenol ethoxylate (Triton X-100) has been determined using Pyrene and Perylene as probes. The aggregation behavior of non-ionic and ionic surfactant with addition of polyelectrolyte poly(sodium4-styrensulfonate) (PSS) and native hyaluronan in aqueous solution has been investigated using Pyrene as a probe. Aggregation number in system of ionic/non-ionic surfactant has been determined by a fluorescence quenching with cetylpyridium (CPC) as a quencher and Pyrene as a fluorescence probe. Interactions in ionic surfactant/non-ionic surfactant/polyelectrolyte system were observed. This result was proved by double sigmoid Boltzman´s type curve. The addition of low molecular as well as high molecular hyaluronan had no significant effect on CMC value of the system.
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Support of physical interaction between hyaluronan and selected hydrophobic solutes
Michalicová, Petra ; Kislinger, Jiří (referee) ; Pekař, Miloslav (advisor)
The method of fluorescence spectroscopy was applied for the studying of the way of the hyaluronan-fluorescence probe system`s preparation on mutual interaction in water. Several experiments with fluorescence probes pyrene, prodan, perylene and diphenylhexatriene (DPH) had been submitted. The first part of the experiments deals with the verification of the results of T. Brown`s work and it was focused on the study of simple interaction between hyaluronan and hydrophobic compounds. In the second part of the work hyaluronan was dried at higher temperature followed by adding of the fluorescence probe. The aim of this method was the distraction of hyaluronan`s moisture packaging and the opening up the hydrophobic parts of the chain for the fluorophore. Although wide concentration ranges of fluorescence probes had been tested in the first experiments, the interactions hadn`t been observed. The similar results were obtained in the second part of the experiments.
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Microviscosity probes in study of aggregation in a biopolymer-surfactant system.
Vašíčková, Kamila ; Vala, Martin (referee) ; Mravec, Filip (advisor)
The effect of cationic surfactant concentration and ionic strength on anisotropy of fluorescence of probes diphenylhexatrien and fluorescein has been investigated in the system of cationic surfactant and in the system of cationic surfactant and hyaluronan. The investigation has been done by fluorescence emission spectroscopy. Obtained anisotropy gives information about microviscosity of investigated systems. Subsequently the system of cationic surfactant and hyaluronan has been investigated by 9-(2-carboxy-2cyanovinyl)julolidine, 4-(dicyanovinyl)julolidine and 1,3-bispyrenylpropane probes. The information about the microviscosity of the system gives the integral under the emission curve of 9-(2-carboxy2-cyanovinyl)julolidine and 4-(dicyanovinyl)julolidine and the ratio between excimer and monomer of 1,3-bispyrenylpropane. It has been discovered that the ionic strength influences the anisotropy of diphenylhexatrien and fluorescein only in the fist addition of salt (concentration 0,025 mol dm-3) and that addition of hyaluronan influences the anisotropy of diphenylhexatrien and fluorescein only in samples without addition of salt. Results of measurment with 9-(2-carboxy2-cyanovinyl)julolidine and 4(dicyanovinyl)julolidine describe the formation of aggregates of catinoc surfactant with hyaluronan and characterize these aggregates from the point of microviscosity.
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Influence of the temperature on hydrophobic domain quality in a biopolymer-surfactant system.
Hnyluchová, Zuzana ; Kopuletá, Ema (referee) ; Mravec, Filip (advisor)
The temperature dependence on polarity has been investigated by fluorescence spectroscopy with pyrene as a probe. Pyrene was chosen thanks to its unique sensitivity to polarity of the medium. As a biopolymer was chosen 73 kDa and 1,4 MDa hyaluronic acid. Emission spectra of pyrene were measured as a function of temperature in aqous solution of CTAB in a presence and absence of sodium chloride. The results show, that the addition of specific concentration of hyaluronic acid to the sample with CTAB, mostly doesn´t have effect to change increasing/decreasing slope of polarity with the temperature.
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