|
|
Characterization of coloid particles by excited-state proton transfer with advanced fluorescence techniques
Kotouček, Jan ; Mravec, Filip (referee) ; Pekař, Miloslav (advisor)
The deprotonation characteristics of fluorescent probes -naphthol and 8-hydroxypyrene-1,3,6-trisulphonic acid (HPTS) were studied in this diploma thesis, using steady-state and time-resolved fluorescence spectroscopy. Two cationic surfactants, Septonex and cetyltrimethylammonium bromide (CTAB), were studied. These surfactants were measured in the complex with hyaluronan (1.75 MDa, 1 MDa and 300 kDa). Steady-state fluorescence was used for determination of critical aggregation concentration of each surfactant and pKa*. Time-resolved fluorescence decays were used to calculate the average lifetimes and the deprotonation constants of naphthol and HPTS. The measurement with hyaluronan were compared with the polystyrenesulfonate (PSS) – surfactant system. The effect of hydration shell of hyaluronan on hyaluronan – surfactant complex formation results from the comparison of above mentioned systems. Large differences were found in the deprotonation characteristic between surfactants and even between individual molecular weights of hyaluronan. The measurement shows that the hydration shell is located near to the dissociated carboxyl groups of hyaluronan chain, where the interaction with the positively charged surfactants occurs. Furthermore, the aggregation number of Septonex was determined by quenching of pyrene using cetylpyridinium chloride (CPC) as a quencher. The aggregation number for 20 mM Septonex solution was determined as a value of 104 molecules. CPC was used for confirmation of the localization of -naphthol in the micelles of CTAB and polymer – CTAB, respectively.
|
|
|
Fluorescence probes to study the properties of hydrogel systems
Máčala, Jakub ; Venerová, Tereza (referee) ; Mravec, Filip (advisor)
This bachelors thesis is focused on possibility of selected fluorescent probes for study of properties of hydrogel systems. Chosen probes are Prodan for polarity study, HPTS for measurement of pH of the probe environment and platinum complex [2,6-Bis(2-pyridyl)fenyl-C,N,N]chloroplatinum(II) for its unique fluorescence properties and potential of energy transfer studies in hydrogel systems. Spectral properties of fluorophores were defined in selected set of solvents with different polarity with steady state fluorescence spectroscopy method. Properties of HPTS probe were also tested in buffers with different pH. Subsequently, hydrogels based on hyaluronan-septonex and dextran-SDS with incorporated fluorescence probes were prepared. Steady state fluorescence measurement of fluorophores from hydrogel were made using fluorimeter optical fibres. It was found, that probes Prodan and HPTS are suitable for study of polarity and pH of hydrogels and probably they will be able to be used for study of hydrogel with other techniques, like fluorescence microscopy, however platinum complex is, due to the low fluorescence intensity and possible quenching by polar water environment, probably not suitable for other measurements.
|
|
|
Fluorescence probes to study the properties of hydrogel systems
Máčala, Jakub ; Venerová, Tereza (referee) ; Mravec, Filip (advisor)
This bachelors thesis is focused on possibility of selected fluorescent probes for study of properties of hydrogel systems. Chosen probes are Prodan for polarity study, HPTS for measurement of pH of the probe environment and platinum complex [2,6-Bis(2-pyridyl)fenyl-C,N,N]chloroplatinum(II) for its unique fluorescence properties and potential of energy transfer studies in hydrogel systems. Spectral properties of fluorophores were defined in selected set of solvents with different polarity with steady state fluorescence spectroscopy method. Properties of HPTS probe were also tested in buffers with different pH. Subsequently, hydrogels based on hyaluronan-septonex and dextran-SDS with incorporated fluorescence probes were prepared. Steady state fluorescence measurement of fluorophores from hydrogel were made using fluorimeter optical fibres. It was found, that probes Prodan and HPTS are suitable for study of polarity and pH of hydrogels and probably they will be able to be used for study of hydrogel with other techniques, like fluorescence microscopy, however platinum complex is, due to the low fluorescence intensity and possible quenching by polar water environment, probably not suitable for other measurements.
|
|
|
Characterization of coloid particles by excited-state proton transfer with advanced fluorescence techniques
Kotouček, Jan ; Mravec, Filip (referee) ; Pekař, Miloslav (advisor)
The deprotonation characteristics of fluorescent probes -naphthol and 8-hydroxypyrene-1,3,6-trisulphonic acid (HPTS) were studied in this diploma thesis, using steady-state and time-resolved fluorescence spectroscopy. Two cationic surfactants, Septonex and cetyltrimethylammonium bromide (CTAB), were studied. These surfactants were measured in the complex with hyaluronan (1.75 MDa, 1 MDa and 300 kDa). Steady-state fluorescence was used for determination of critical aggregation concentration of each surfactant and pKa*. Time-resolved fluorescence decays were used to calculate the average lifetimes and the deprotonation constants of naphthol and HPTS. The measurement with hyaluronan were compared with the polystyrenesulfonate (PSS) – surfactant system. The effect of hydration shell of hyaluronan on hyaluronan – surfactant complex formation results from the comparison of above mentioned systems. Large differences were found in the deprotonation characteristic between surfactants and even between individual molecular weights of hyaluronan. The measurement shows that the hydration shell is located near to the dissociated carboxyl groups of hyaluronan chain, where the interaction with the positively charged surfactants occurs. Furthermore, the aggregation number of Septonex was determined by quenching of pyrene using cetylpyridinium chloride (CPC) as a quencher. The aggregation number for 20 mM Septonex solution was determined as a value of 104 molecules. CPC was used for confirmation of the localization of -naphthol in the micelles of CTAB and polymer – CTAB, respectively.
|
guest ::
