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The hydration of hyaluronic acid
Průšová, Alena ; Šimon, Peter (referee) ; Kučerík, Jiří (advisor)
Hydratace patří mezi nejdůležitější faktory ovlivňující sekundární strukturu a tím i funkci molekul v živých systémech. Díky vysoké afinitě tvoří molekuly vody specifické struktury jejichž složení a fyzikální vlastnosti jsou ovlivněny přítomností studované látky. Hyaluronan patří mezi biomolekuly s obrovskou schopností vázat a zadržovat vodu. Cílem této práce bylo prozkoumat hydratační vlastnosti hyaluronanu o různé molekulové hmotnosti a vyčíslit množství molekul vody v jednotlivých hydratačních vrstvách. V první části práce byla využita metoda diferenční kompenzační kalorimetrie. V druhé části diplomové práce, na základě rozdílné kompresibility, byla vázaná voda studována metodou vysoko rozlišovací ultrazvukové spektroskopie.
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Determination of additives in multicomponent electrospinning systems
Gajdová, Barbora ; Petr, Knotek (referee) ; Knotková,, Kateřina (advisor)
This bachelor thesis deals with methods of producing nanofibres from mixture of different polymers. Different methods of preparation and production nanofibres by 4SPIN® are described in theoretical part. Experimental part deals with incorporation of additives into fibred layer. There were four different dyes used in this thesis: brilliant green, erythrosine, riboflavin and eriochrom black. UV – VIS spectrophotometry was used to measure volume of additives in nanofiber layer.
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Preparation of hyaluronan oligosaccharides with N-acetylglucosamine at both ends of the chain using enzymatic catalysis
Gadová, Martina ; Hermannová,, Martina (referee) ; Šílová, Tereza (advisor)
This bachelor thesis deals with preparation of odd-numbered hyaluronan oligosaccharides with N-acetylglucosamine at both ends of the chain using enzymatic catalysis by -Glucuronidase. The theoretical part focuses on present knowledge of hyaluronic acid and it fragments. It focuses on properties, enzymatic hydrolysis, application in different branches of science, but also analysis of oligosaccharides. One chapter is devoted to properties of enzyme -Glucuronidase. The experimental part deals with preparation of oligosaccharides type HANN. The reaction conditions for preparation of HA oligosacharides were optimized (pH, temperature) and the progress of enzymatic hydrolysis and the mixture composition after digestion were monitored by method UHPLC.. After optimization the mixture of oligosaccharides HANN was prepared and consenquently it was analysed by methods UHPLC and LC-MS. All the data were acquired in company Contipro Pharma, a.s., Dolní Dobrouč, Czech republic.
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Effect of pH on quantitative determination of additives in nanofibers
Grufíková, Jana ; Marek, Pokorný (referee) ; Knotková,, Kateřina (advisor)
The object of my thesis was to prepare a nanofiber layer with dye additives and to quantify the additive content in dependence on pH. The nanofiber layers with additives were prepared by means of electrospinning. The analysis of the additive content in samples was performed by UV-VIS spectrophotometry. Concretely Erythrosine, Eriochrome Black, Riboflavin and Brilliant Blue were analysed. Each of this colors has different chemical structure and charge. The pH was defined by use of buffers, namely acetate buffer (pH 5), phosphate buffer (pH 7,4) and carbonate buffer (pH 10). For Erythrosine and Eriochrome Black the research was extended to study the effect of ionic strength on the quantitative determination of these dyes. It was found that the quantity of the determined additive depends on the pH and also on the ionic strength. However, this dependency is different for each additive. This confirms the assumption of influenced chemical structure and charge by interaction with polymers and buffers used in this experiment. The accuracy of evaluation is influenced by the way of data processing.
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Polysaccharide-protein gels
Hájovská, Pavla ; Smilek, Jiří (referee) ; Chytil, Martin (advisor)
This bachelor thesis deals with gelation of albumin solutions and mixed solutions of albumin and the high molecular weight hyaluronic acid due to heating. Study was carried out by measuring rheological properties of the solutions, formed gels and gelation process. For the protein samples in water, the effect of concentration, ionic strength and pH were tested. For mixed solutions, the effect of concentration and a composition of the polymeric component were tested. With increasing concentration of the polymeric component, the gel point is shifted towards lower temperature values, with the concentration difference of 2 % (w/v) up to 5°C. The resulting gels exhibit higher values of loss and storage moduli. Similar behavior was observed also in the samples influenced by the ionic strength as, a result of alternation of the electrostatic interactions between the charged groups of the polymer chains and as a result of change of this charge due to the pH reduced below the pI value of the protein.
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Kinetic study of excited state proton transfer in polymer-surfactant system
Bačová, Jana ; Vala, Martin (referee) ; Mravec, Filip (advisor)
In this bachelor thesis were studied interactions between hyaluronic acid and cationic septonex by steady-state and time-resolved fluorescence with fluorescence probes pyrene and 1-naphthol. At first was determined critical micelle concentration of septonex. In the case of both fluorescence probes was studied area below and above critical micelle concentration. It was found, that 1-naphthol undergo very fast deprotonation in aqueous solution, while in the present of micelles the rate of deprotonation is c significantly retarded. In the system of hyaluronic acid – septonex it was observed, that formation of aggregates occurs at lower concentrations than in the system without hyaluronic acid. Molecules of probe are probably distributed between hyaluronic acid – septonex aggregates and free micelles above critical micelle concentration.
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Hyaluronan-albumin system dynamics
Jarábková, Sabína ; Smilek, Jiří (referee) ; Chytil, Martin (advisor)
The bachelor thesis deals with the hyaluronan and albumin system dynamics. In this study was to use these methods: interfacial rheology, tensiometry, method of maximum pressure in the bubble and UV-VIS spectrophotometry. The measurements were used with various concentrations of albumin and a constant concentration of hyaluronan( high molecular and low molecular too) and then conversely. All solutions were measured and the obtained data were compared with the results of BSA in buffer without HA. Solutions were prepared with citrate-phosphate buffer of pH about 4. Were observed interaction between HA and BSA.
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Characterization of hyaluronan interactions with albumin
Valentová, Kristýna ; Kratochvílová, Romana (referee) ; Chytil, Martin (advisor)
This bachelor thesis focuses on the research of physical interactions between hyaluronic acid and a protein albumin. In order to investigate these interactions following methods were used: pH and conductivity measurement, viscosimetry, turbidimetry and rheometry. The interactions were observed in solutions of fixed albumin concentration 1.0 gdm-3 and varying concentrations of hyaluronic acid from 0.1 to 2.0 gdm-3. Results were compared with a solution of hyaluronic acid without the addition of albumin. The results show that there are interactions between hyaluronic acid and albumin solutions, especially for those containing high molecular weight hyaluronic acid. The interactions exhibited a change in viscosity of solutions. We can see a reduction in viscosity after adding albumin to the reference solutions. The reduction is noticeable even in the low molecular weight HA solutions, but is not so significant.
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Electrolytic properties of hyaluronan solutions
Suchá, Šárka ; Smilek, Jiří (referee) ; Chytil, Martin (advisor)
This batchelor thesis deals with electrolytic properties of hyaluronan in aqueous solutions. In this study measuring of conductance, acid-base and conductrometric titration, and UV-VIS spectroscopy were used. The measuring of conductance was made on the solutions of different concentrations of hyaluronic acid and of different ionic strength. Acid-base titrations were performed in water solution and conductometric titrations were performed in 0,3mmol.dm–3 solution of sodium chloride. UV-VIS spectra of hyaluronic acid were measured in solution with addition of acid, base and in buffer solution. Values of pKa obtained from the above mentioned measurements was between 2,8–6,5.
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