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Interaction between amphiphilic fluorescent probes and hyaluronan
Střondalová, Hana ; Vala, Martin (referee) ; Mravec, Filip (advisor)
This bachelor thesis dealt with studying the interaction of amphiphilic fluorescent probes with hyaluronan because these probes could significantly help in clarifying the links between hyaluronan and surfactants. This complex could be a great benefit in targeting drug distribution. This thesis was focused on the interaction between hyaluronan and amphiphilic fluorescent probes – specifically with dodecylacridine orange. First explored was the interaction with hyaluronan, and then with the hyaluronan – surfactant system. Cetyltrimethylamonium bromide was selected as surfactant. Samples were examined using fluorescence spectroscopy. Emission and absorption spectrum of dodecyl acridine orange was measured. This experiment was based on the hypothesis that dodecylacridine orange forms dimers as acridine orange and nonylacridine orange. It was found that in a system of hyaluronan-probe, dodecyl acridine orange was pushed out of the complex by cetyltrimethylamonium bromide, which was bounded instead. Surfactant interacts with hyaluronan binding positions - with carboxyl groups.
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Ionogenic fluorescence probes in research of colloidal system
Střondalová, Hana ; Burgert,, Ladislav (referee) ; Mravec, Filip (advisor)
This diploma thesis studied the interaction of ionic and amphiphilic fluorescent probes with polyelectrolytes. The research of such colloidal system could be helpful in deeper understanding of interactions between polyelectrolytes and surfactants. The aim of this thesis was to determine whether ionic or amphiphilic probes are suitable for this research. Two polyelectrolytes, sodium polystyrene sulfonate and hyaluronan were used in this study. Interactions of polyelectrolytes with fluorescent probes (4-Di-2-Asp, methylene blue, dodecyl acridine orange, DiO and DiA) were studied in this work. Then cationic surfactant cetyltrimethylamonium bromide was added to this system. Samples were studied using fluorescence and UV/VIS spectroscopy. Emission and absorption spectra of these probes were recorded. This experiment was based on hypothesis that due to positive charge these probes will bind to the negatively charged polyelectrolyte. Formation of dimers or other aggegates was presumed for fluorescent probes dodecyl acridine orange and methylene blue. When 4-Di-2-Asp was used,changes in fluorescence intensity and absorption spectra in connection with possible formation of aggregates was supposed to be observed. It should be noted that formation of dimers for 4-Di-2-Asp is not described in literature. The formation of dimers wasn't clearly demonstrated during the measurement. Formation of dimers was observed only with dodecyl acridine orange. Probes DiO and DiA were used only in the study of the interaction with polyelectrolytes. These fluorescent probes showed up as inappropriate, because they adsorbed on tube walls. When cationic surfactant was added to hyaluronan-fluorescent probe system, surfactant pushed away fluorescent probe and bonded electrostatically to negatively charged groups of polyelectrolyte.
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Fluorescence study of hydrophilic domains of associating colloids
Londinová, Monika ; Knotková,, Kateřina (referee) ; Pekař, Miloslav (advisor)
The properties of the hyaluronan were investigated by using different fluorescence probes, because hyaluronan is a hopeful carrier of an active matter in medicine and cosmetics. Selected fluorescence probes were: cationic acridine orange, Nile Blue A, methylene blue, amphiphilic 4-Di-2-ASP and anionic fluorescein. Except from fluorescence and absorption spectra of the probes were observed electrostatic and hydrophobic interactions as well. The probes in solvents with different polarity (MeOH, EtOH, DMSO) showed the bathochromic shift in the emission maximum and quenching of the fluorescence with the increasing polarity of the solvents. The influence of the ionic strength on fluorescence properties of the probe acridine orange and 4-Di-2-ASP was investigated in aqueous solutions of chlorides. The formation of acridine orange dimer is inhibited with increasing ionic strength. CaCl2 increased the ionic strength the most, then prevented repulsion of carboxylate groups, so it means the expansion of hyaluronan cluster into the solution. However, the emission of the probe 4-Di-2-ASP was quenched with the addition of CaCl2 the most. The first additions of COO– groups cause the formation of dimers of AO shown as decreasing in extinction coefficient and fluorescence intensity. Next addition of the hyaluronan caused a depolymerization of formed dimers and the increase of the emission intensity. The repolymerization caused the decrease and then again the increase. In case of 4-Di-2-ASP was the pattern of the fluorescence (the intensity and the position of the emission) firstly the same, but at the concentration of 1 g dm-3 the emission intensity increased. The probes MB and F were used for spectroscopic studies of the interaction between methylene blue-fluorescein complex and anionic and cationic surfactants. The absorbance of separate MB and F changed only with the addition of surfactants with the opposite electric charge. Absorbance of the mixture MB-F changed with the addition of the CTAC surfactant, while the addition of SDS into the mixture caused only the change of MB absorption spectra.
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Ionogenic fluorescence probes in research of colloidal system
Střondalová, Hana ; Burgert,, Ladislav (referee) ; Mravec, Filip (advisor)
This diploma thesis studied the interaction of ionic and amphiphilic fluorescent probes with polyelectrolytes. The research of such colloidal system could be helpful in deeper understanding of interactions between polyelectrolytes and surfactants. The aim of this thesis was to determine whether ionic or amphiphilic probes are suitable for this research. Two polyelectrolytes, sodium polystyrene sulfonate and hyaluronan were used in this study. Interactions of polyelectrolytes with fluorescent probes (4-Di-2-Asp, methylene blue, dodecyl acridine orange, DiO and DiA) were studied in this work. Then cationic surfactant cetyltrimethylamonium bromide was added to this system. Samples were studied using fluorescence and UV/VIS spectroscopy. Emission and absorption spectra of these probes were recorded. This experiment was based on hypothesis that due to positive charge these probes will bind to the negatively charged polyelectrolyte. Formation of dimers or other aggegates was presumed for fluorescent probes dodecyl acridine orange and methylene blue. When 4-Di-2-Asp was used,changes in fluorescence intensity and absorption spectra in connection with possible formation of aggregates was supposed to be observed. It should be noted that formation of dimers for 4-Di-2-Asp is not described in literature. The formation of dimers wasn't clearly demonstrated during the measurement. Formation of dimers was observed only with dodecyl acridine orange. Probes DiO and DiA were used only in the study of the interaction with polyelectrolytes. These fluorescent probes showed up as inappropriate, because they adsorbed on tube walls. When cationic surfactant was added to hyaluronan-fluorescent probe system, surfactant pushed away fluorescent probe and bonded electrostatically to negatively charged groups of polyelectrolyte.
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Fluorescence study of hydrophilic domains of associating colloids
Londinová, Monika ; Knotková,, Kateřina (referee) ; Pekař, Miloslav (advisor)
The properties of the hyaluronan were investigated by using different fluorescence probes, because hyaluronan is a hopeful carrier of an active matter in medicine and cosmetics. Selected fluorescence probes were: cationic acridine orange, Nile Blue A, methylene blue, amphiphilic 4-Di-2-ASP and anionic fluorescein. Except from fluorescence and absorption spectra of the probes were observed electrostatic and hydrophobic interactions as well. The probes in solvents with different polarity (MeOH, EtOH, DMSO) showed the bathochromic shift in the emission maximum and quenching of the fluorescence with the increasing polarity of the solvents. The influence of the ionic strength on fluorescence properties of the probe acridine orange and 4-Di-2-ASP was investigated in aqueous solutions of chlorides. The formation of acridine orange dimer is inhibited with increasing ionic strength. CaCl2 increased the ionic strength the most, then prevented repulsion of carboxylate groups, so it means the expansion of hyaluronan cluster into the solution. However, the emission of the probe 4-Di-2-ASP was quenched with the addition of CaCl2 the most. The first additions of COO– groups cause the formation of dimers of AO shown as decreasing in extinction coefficient and fluorescence intensity. Next addition of the hyaluronan caused a depolymerization of formed dimers and the increase of the emission intensity. The repolymerization caused the decrease and then again the increase. In case of 4-Di-2-ASP was the pattern of the fluorescence (the intensity and the position of the emission) firstly the same, but at the concentration of 1 g dm-3 the emission intensity increased. The probes MB and F were used for spectroscopic studies of the interaction between methylene blue-fluorescein complex and anionic and cationic surfactants. The absorbance of separate MB and F changed only with the addition of surfactants with the opposite electric charge. Absorbance of the mixture MB-F changed with the addition of the CTAC surfactant, while the addition of SDS into the mixture caused only the change of MB absorption spectra.
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Interaction between amphiphilic fluorescent probes and hyaluronan
Střondalová, Hana ; Vala, Martin (referee) ; Mravec, Filip (advisor)
This bachelor thesis dealt with studying the interaction of amphiphilic fluorescent probes with hyaluronan because these probes could significantly help in clarifying the links between hyaluronan and surfactants. This complex could be a great benefit in targeting drug distribution. This thesis was focused on the interaction between hyaluronan and amphiphilic fluorescent probes – specifically with dodecylacridine orange. First explored was the interaction with hyaluronan, and then with the hyaluronan – surfactant system. Cetyltrimethylamonium bromide was selected as surfactant. Samples were examined using fluorescence spectroscopy. Emission and absorption spectrum of dodecyl acridine orange was measured. This experiment was based on the hypothesis that dodecylacridine orange forms dimers as acridine orange and nonylacridine orange. It was found that in a system of hyaluronan-probe, dodecyl acridine orange was pushed out of the complex by cetyltrimethylamonium bromide, which was bounded instead. Surfactant interacts with hyaluronan binding positions - with carboxyl groups.
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