This work is focused on physical interactions of negatively charged polymers with small ionogenic fluorescent molecules. Trying to verify the presence of these interactions using fluorescence correlation spectroscopy (FCS) and provides a comprehensive view of the problem. The aim of this work is to observe the effect of concentration on the diffusion properties. P/D ratio, where P represents number of polymer binding sites and D number of dye binding sites, was chosen for this issue. Hyaluronate, sodium chondroitin sulfate and sodium polystyrene sulfonace were used as polymers and Acridine Orange, and Rhodamine 6G were chosen as fluorescent probes. The reason why this experiment uses these probes, was the assumption, that the positive charge occuring on the fluorescent probe will lead to the electrostatic interaction with the negatively charged polymer. As a result, the bond between acridine orange and polyelectrolyte was not clearly demonstrated, but the interaction with Rhodamine 6G have been proved.

This master thesis is dealing with the use of light scattering techniques in the study on formation of polyelectrolyte complexes in the system biopolymer-surfactant. Sodium hyaluronate was chosen as biopolymer and cetyltrimethylammonium bromide and karbethopendecinium bromide were selected as surfactants. The first precipitation experiments were performed to determine the optimal concentration range of surfactants for subsequent titration measurements performed on Zetasizer Nano ZS and for SEC-MALS analysis. The key parameters obtained from these measurements were the values of critical aggregation concentrations in hyaluronate-surfactant systems. It was found out that the increasing molecular weight of hyaluronan in these systems is causing reduction of obtained values of critical aggregation concentrations. Changes of Z-average particle size, zeta potential, polydispersity index, conformation plot, radius of gyration and molecular weight in the hyaluronan-surfactant system were monitored during experiments.

This diploma thesis deals with study of hydrogels formed by phase separation of hyaluronan with oppositely charged surfactants cetyltrimethylammonium bromide (CTAB) and Septonex. It follows the bachelor thesis and extends the knowledge about the detailed characterisation of the inner environment of the hydrogel by determining the diffusion behaviour of the fluorescent probes Atto 488 and Nile Red using fluorescence correlation spectroscopy (FCS) technique and its modified version dual-focus fluorescence correlation spectroscopy (2f-FCS). Compared results showed that both methods show similar values and probes specifically interact with CTAB but Atto 488 shows only weak interaction with Septonex compared to Nile Red. Additionally, these interactions were not affected by the molecular weight of hyaluronan. In conclusion, it was recommended to measure this type of hydrogel in a small depth of gel using a conventional method.

Biopolymer surfactant systems have been studied by using different physico chemical methods. As the biopolymer, it was chosen high (1400–1600 kDa), medium (250–450 kDa) and low (8–15 kDa) molecular weight sodium hyaluronate. Two cationic surfactants – cetyltrimethylammonium bromide (CTAB) and carbethopendecinium bromide (Septonex) were selected to provide polyelectrolyte complexes with oppositely charged hyaluronan. The critical aggregation concentration of the surfactant in the system was monitored by dynamic light scattering (Zetasizer Nano ZS). SEC-MALS method was used for the description of the conformation of sodium hyaluronate of different molecular weights and polyelectrolytes complexes hyaluronan surfactant after reaching the aggregation point. Isothermal titration calorimetry (ITC) was used to determine the critical aggregation concentration of the surfactant by monitoring the thermal changes accompanying the aggregation of the system. It was found out that the increasing molecular weight of hyaluronan in these systems decreases the critical aggregation concentrations and conversely.