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Hyaluronane interactions with hydrophobic solutes
Slezáková, Dagmar ; Knotková,, Kateřina (referee) ; Pekař, Miloslav (advisor)
The diploma thesis is based on the study of hydrophobic interactions of the native hyaluronan with selected solutes. On the basis of a literature search were chosen fluorescent probes and fluorescing biologically active substances, which are useful for investigation of colloids as 6-(p-toluidino)-2-naphthalenesulfonic acid (polarity probe), lipophilic vitamin (±)-alpha-tocopherol, pyrene (polarity probe) and finally hydrophilic vitamin riboflavin. In the experimental part of this thesis was studied the influence of solvents with different polarities, or more precisely dielectric constant, on the emission spectra, as well. There were investigated interactions of native hyaluronan with TNS and then interactions, which were influenced by the ionic strength. Such influenced interactions were not observed, that was probably due to the strong solvation´s wrapping of the hyaluronan. Interactions were observed after the process of lyophilisation followed-up by the rehydratation of the samples. For the next study of interactions the riboflavin was chosen and was investigated the REES effect in the native hyaluronan in different concentrations of its different molecular weights. In this case were not observed any shifts in the emission maximum with the excitation wavelenght shift and that is why the interactions of hyaluronan with riboflavin were not demonstrated in the field of chosen concentrations. By using another probe alpha-tocopherol was investigated the associative behaviour of hyaluronan and moreover was observed anisotropy of alpha-tocopherol in different concentrations of different molecular weights of native hyaluronan. The anisotropy reached high values in contrast to the reference solute that was the mixture of glycerol and ethanol. The anisotropy depended more on the molecular weight than on the concentration of hyaluronan. Interactions of hyaluronan were also studied by using the polarity probe pyrene in different concentrations of different molecular weights of the hyaluronan. The pyrene 1:3 ratio did not show the concentration dependence within the chosen concentrations except for the molecular weight 253.9 kg mol–1. Both probes alpha-tocopherol and pyrene were performed by the process of lyophilisation followed-up by the rehydratation, which improved interactions of these probes with hyaluronan.
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PCP sorption of lignite
Šnédarová, Gabriela ; Taraba, Boleslav (referee) ; Pekař, Miloslav (advisor)
Within the framework of this diploma thesis, the sorptive capability of a lignite as a natural adsorbent was applied on an aqueous solution of pentachlorophenol. The aqueous solution of this substance, which is very dangerous for the environment, was prepared in various concentration ranges according to reached solubility. The solubility is noticed in different literatures variously and then is not applicable. That is why it was necessary to find out the ”real“ solubility. The aqueous solution of pentachlorophenol of given concentration was subsequently put to adsorption with exactly defined quantity of the lignite and as a result the adsorptive isotherms were obtained. These isotherms represent the adsorption capability in dependence on the adsorption duration, quantity of used lignite and concentration of pentachlorophenol solution. By the adsorption with duration longer than one hour, the quantity of adsorbed PCP does not increase markedly.
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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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Solubilizattion properties of polysaccharides
Lenartová, Radka ; Foglarová, Marcela (referee) ; Pekař, Miloslav (advisor)
In this diploma thesis were studied solubilization properties of polysaccharides by using hydrophobic solutes (Sudan Orange G, Sudan Red G, (±)-alpha-Tocopherol, Pyrene, Perylene, Nile red), which were represented by alkyl derivates of hyaluronan. At first, a behaviour of individual hydrophobic solutes was investigated in variously polar solvents (Methanol, 1 Propanol, Chloroforme, Cyklohexane, n Heptane) and in the environment of varying ionic strength (water, 0.1 M and 0.4 M NaCl). Afterwards, solubilization properties of Sodium Dodecyl Sulfate model solubilizated the hydrophobic solutes into a core of micelles was examinate. We were interested in the solubilization capacity as the mol of solubilized molecules per mol micelles of surfactant corresponding with a state of micelles saturation. In the case of the solubilization of (±)-alpha-Tocopherol into the core of micelles, it was not possible to determine the solubilization capacity. So we changed the determination of universally solubilization power. The solubilization power is defined as mol of molecules solubilized per mol surfactant relative to the quantity solubilizate at the micelles saturation. Model system of Sodium Dodecyl Sulfate as a simple surfactant carrying a negative charge as the alkyl derivates of hyaluronan was selected bacause of its characteristics.The surfactant forms unimolar micelles and its critical micelle concentrations and aggregation numbers are tabelated for the investigated microenvironment. The main aim of the study was investigating of hydrophobic domains of alkyl derivates of hyaluronan as free places for incorporation hydropbobic solutes in the microenvironment of varying ionic strength. The critical aggregation concentrations were determined by the Pyrene 1:3 ratio method. For the research of micropolarity of alkyl derivates hyaluronan’s domains were selected two concentrations of derivates for the next research of solubilization experiments - the first concentration near the critical aggregation concentration and the second concentration above it. The effect of concentration of Pyrene on a core polarity of derivates was investigated. We discovered the influence of the concentration and the other we found a stationary area of the concentration. In the end we investigated the influence of preparation of solutions of derivates of hyaluronan on the core polarity by the concentration of pyrene which corresponds to the stationary area. The study of solubilization properties of alkyl derivates of hyaluronan is not a simple case as we assumed. When we measured spectra of the absorbance, higher concentration of derivates of hyaluronan belittle absorbance of solubilizates. At the experiment of solubilization with Sudan Red G we found out that Sudan Red G is not able to solubilizate into the hydrophobic core of micelles of hyaluronan’s derivates because of lipophilic or steric effects. We had to change Perylene as a new solubilizate. From the measured emission spectra we found saturation micelles. We can express the solubilization power of hyaluronan’s derivates for the concentration of Perylene. The main aim of the diploma thesis was to determine optimal way of the preparation of hyaluronan’s derivates solutions with required degree of solubilization.
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Colloid properties of hyaluronane sols
Krouská, Jitka ; Mašek, David (referee) ; Pekař, Miloslav (advisor)
Předmětem této práce bylo popsat agregační vlastnosti kyseliny hyaluronové (HA) a jejích hydrofobně modifikovaných derivátů. Za tímto účelem byly připraveny vodné roztoky kyseliny a dvou derivátů, lišící se stupněm substituce a druhem anorganické soli jako prostředím. Koncentrační řady čisté kyseliny hyaluronové a jejích derivátů byly studovány dvěma metodami, tensiometricky a spektrometricky. Povrchové vlastnosti roztoků byly měřeny s použitím kroužku Du Noüy. Zkoumáno bylo také solubilizační chování ve vodě a v chloridu sodném pomocí dvou sond: Coomassie brilantní modři a sudanu III. Amfifilní deriváty hyaluronanu mohou skutečně tvořit hydrofobní agregáty v roztoku a adsorbovat se na rozhraní vzduch–voda. Tato vlastnost je důležitá v souvislosti s transportními systémy léčiv, protože právě hydrofobní domény agregátů hyaluronanu mohou být použity jako nosiče hydrofobních léčiv. Byly zjištěny kritické agregační koncentrace roztoků derivátů hyaluronanu (tensiometricky a spektrometricky), které odpovídají právě těm koncentracím, při kterých vznikají hydrofobní domény.
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Dynamic tensiometry of biocolloids
Kulilová, Pavlína ; Mašek, David (referee) ; Pekař, Miloslav (advisor)
Hyaluronová kyselina je v současné době velmi významná biomolekula používaná v medicíně a kosmetice, a její výzkum je důležitý pro další budoucí použití. Zaměření této práce je studování povrchových vlastností hyaluronové kyseliny, jejích hydrofobních derivátů a roztoků SDS pro srovnání, za použití tenziometrie. Tyto sloučeniny byly zkoumány ve vodě a v roztocích ve formě sodné soli. Sledované vzorky byly měřeny dvěmi metodami v různých koncentračních rozmezích při laboratorní teplotě pomocí nového BPA-800P bublinového tenziometeru. Byly navrženy takové experimenty, aby se zjistily využitelné možnosti tohoto přístroje pro další výzkum. Výsledky práce ukazují rozdílnosti jednotlivých povrchově aktivních látek v závislosti na jejich koncentraci a použitém prostředí. Hyaluronová kyselina nevykazuje povrchovou aktivitu, zatímco její deriváty a SDS ano.
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Rheology of hyaluronane solutions
Hlisnikovská, Kristýna ; Soukupová, Lenka (referee) ; Pekař, Miloslav (advisor)
Předmětem tohoto studia bylo prozkoumat reologické chování vodných roztoků vysokomolekulárního hyaluronanu. Byl studován vliv zvyšující se koncentrace biopolymeru v roztoku, která se pohybovala v rozmezí od 1 do 3 hmotnostních procent, a také vliv vzrůstající iontové síly rozpouštědla, způsobené přídavkem chloridu sodného, na viskoelasticitu a stabilitu těchto roztoků. Pro obsáhlejší popis viskoelasických vlastností roztoků byla použita, vedle běžných oscilačních měření, také metoda ceepových testů, ze které bylo možno určit důležité veličity, jako je procentuální poměr viskozní a elastické složky vzorku, rovnovážná poddajnost, viskozita při nulovém smykovém napětí a retardační čas. Ty byly následně porovnávány s výstupy z jiných typů měření, jako jsou právě oscilační a tokové křivky, nebo nesly dopňující informace důležité pro detailnější popis viskoelastických vlastností těchto roztoků. Ke studiu stability vzorků během namáhání pak byla použita metoda peak-hold, která ukázala na velmi dobou mechanickou i časovou odolnost roztoků hyaluronanu a naznačila hranice, za kterými už dochází k trvalému poškození struktury a degradaci řetězců hyaluronanu a je s němi proto potřeba při manipulaci s roztoky tohoto biopolymeru pro jejich další použití v aplikacích počítat.
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Isolation and characterization of the soil gels
Kundera, Ondřej ; Pekař, Miloslav (referee) ; Sedláček, Petr (advisor)
The aim of this study are the gel-creative components in soil, mainly the isolation and characterization using electron microscopy and available analytical methods like FT-IR spectrometry, elementary and thermogravimetric analysis. The knowledge from this study may help to define function and significance of this gels to soils and nature. The theoretical part of this thesis deals with humic substances which are, according to published articles so far, expected to play an important role in the formation of the soil gels. The experimental part solves especially the optimization of the method of isolation soil gels and his further characterization.
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Interactions of aminoclays with natural polyelectrolytes
Jančík Procházková, Anna ; Mravec, Filip (referee) ; Pekař, Miloslav (advisor)
This thesis deals with the preparation of products of interactions of aminoclay with natural polyelectrolytes. Sodium hyaluronate was chosen as the natural polyelectrolyte. The concentration and volume ratio of solutions of aminoclay and natural polyelectrolyte which led to the formation of any type of product were studied. The resulting coagulates were characterized by microscopy, FTIR spectroscopy and rheology. Furthermore the possibility of interactions with fluorescent probes was studied.
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Aminoclays - optimization of preparation and characterization
Plotěná, Michaela ; Sedláček, Petr (referee) ; Pekař, Miloslav (advisor)
The objektive of this paper is to give an overview of synthetis, properities and application of magnesium phyllosilicate with 3-aminopropyl functional groups (aminoclay). The research in this paper focuses on preparing aminoclay, specifically to prove the prescribed procedure and its modifications. The structure of the final product was verified by X-ray analysis and scanning electron microscope. The main motivation is the synthesis of aminoclay opportunities in the pharmaceutical industry. Aminoclay would serve as an adsorbent that would ensure greater bioavailability of the drug.
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