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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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Time-resolved fluorescence in investigation of colloid system
Černá, Ladislava ; Ouzzane, Imad (referee) ; Mravec, Filip (advisor)
This thesis is focused on determination of the basic spectral characteristics, such as fluorescence maximum and lifetime, of fluorescent probes prodan [6-propionyl-2-(dimethylamino)naphthalene] and perylene by steady-state and time-resolved fluorescence spectroscopy. Time-resolved measurements were realized through TCSPC method (time-correlated single photon counting). The characteristics of prodan were found in water, acetone, n-heptane and in mixtures of these, so that a polar scale of environments would arise. Both of the mentioned probes were further investigated in systems of aqueous solutions of hyaluronic acid (300 kDa) at the concentration of 10 mg•l-1 together with cationic surfactant CTAB (cetyltrimethylammonium bromide) at the concentration range from 0.01 to 0.20 mmol•l-1 (before CMC). The aim was to confirm the reach and the range of critical aggregation concentration of the surfactant (CAC), to uncover the basic spectral characteristics of the mentioned probes in this concentration range and to verify the stability of this system in the presence of NaCl at the concentration of 0.15 mol•l-1. Furthermore, by TCSPC method were obtained time-resolved emission spectra of prodan from the system of hyaluronic acid and CTAB at one concentration of CTAB in CAC interval both without salt and with salt at the concentration of 0.15 mol•l-1. The obtained time-resolved emission spectra correlated with the results of the CAC concentration range measurement. After addition of salt the system of CAC proved to be unstable.
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Study of interaction between hyaluronan and phospholipid in a presence of biocompatible non-ionic surfactant
Burdíková, Jana ; Klučáková, Martina (referee) ; Mravec, Filip (advisor)
This work is based on investigation of aggregation behavior of sugar surfactants and sn-glycero-dipalmitoylphosphatidylcholin (DPPC) in aqueous environment and on study of the interactions in nonionic surfactant-DPPC and DPPC-nonionic surfactant-hyaluronan systems. Sugar surfactants were used from non-ionic surfactants. The behavior of each systems was studied by fluorescence spectroscopy. Values of critical micellar concentrations (CMC) of sugar surfactants, begining of aggregation of DPPC, DPPC aggregation effect of sugar surfactants and the effect of hyaluronan on the system non-ionic surfactant-DPPC has been investigated. Also CMC of the CPC (cetylpyrimidium chloride), which was used for fluorescence quenching in the determination of aggregate numbers of sugar surfactants, has been determined. Fluorescent probes pyrene, perylene, nile red, acridine orange and hydrophobic dye sudan red were used for measurements.
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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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Fluorescenec spectroscopy in study of aerosol-forming substances
Lippay, Josef ; Jedelský, Jan (referee) ; Mravec, Filip (advisor)
To study the spectral characteristics of a substance DEHS several experiments were suggested. Closer investigation was made on study of the effect of different environments on the spectral properties of the substance DEHS. It was found that the highest positive effect on the fluorescence of the substance DEHS have a medium with hydroxyl groups. IPA was evaluated as the best medium for further experiments. It was found that the medium xylene decreases intensity of the fluorescence to the detection limit. Spectral characterization of the nondegraded and degraded DEHS was performed. It was found that for the degraded DEHS shift to higher wavelengths occurs due to degradation. Possible contamination of the DEHS with substances causing luminescence has not been proven.
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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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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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Study of aggregation in a biopolymer-surfactant system at low surfactant concentration.
Stiborský, Filip ; Vojtová, Lucy (referee) ; Mravec, Filip (advisor)
In this bachelor thesis, the interaction between hyaluronate and surfactant using fluorescent spectroscopy and pyrene as fluorescent probe was studied. A cationic surfactant cetyltrimethylammonium bromide and a native hyaluronate with different molecular size was used. Before a detected critical micelle concentracion, there were no interactions observed in phosphate buffer. However, we could observe electrostatical interactions in the water systém measuring. At low concentration of surfactant (between critical aggregation concentration and critical micelle concentration), the production of gel phase appeared.
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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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