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Development and characterization of hydrogel carriers for modern agricultural applications
Mai, Thuy Ha ; Kalina, Michal (referee) ; Sedláček, Petr (advisor)
The bachelor thesis is focused on characterization of hydrogel carriers and their application in agriculture. Entrapment of active phase within the hydrogel matrix would allow release of its content in controlled rate. The theoretical part describes encapsulation techniques and suitable biocompatible materials involved in encapsulation. For the experimental part, the release of soil bacteria Cupriavidus necator H16 from alginate matrix was examined. The focus was on the release rate of bacteria via diffusion and later, on the release rate of bacteria encouraged by chemical or enzymatic reagents. Further modification of carrier was suggested by adding hydrophobic barrier composed of polyhydroxybutyrate (PHB). The modified alginate beads seemed to be more resistant toward enzymatic or chemical agents, thus the process of the release was to some extent suppressed. This fact might suggest PHB as a possible vessel for optimization of controlled release system of hydrogel carriers.
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Encapsulation of soil bacteria in hydrogel carriers
Orišková, Sofia ; Smilek, Jiří (referee) ; Sedláček, Petr (advisor)
The goal of this bachelor thesis was both to review existing literature regarding the topic of inoculation of soil bacteria and test a relevant encapsulation method and optimize it. The evaluation process involved the study of various encapsulation techniques that involve hydrogel carriers suitable for agronomic purposes. Encapsulation allows controlled release of soil bacteria, and protects and stabilizes it, while ensuring longer shelf life. For the practical testing, Cupriavidus necator H16 was chosen as a nitrogen fixing bacteria for the inoculation. Through an ionic gelation method, it was encapsulated into alginate carriers, forming matrix microcapsules. Sodium alginate reacts with the cross-linking agent calcium chloride to form the capsules. The impact on the quality of the product was tested through several variables. What was revealed was that molecular weight of alginate was proven to have a significant impact. Alginate with higher molecular weight was shown to be suitable for the given system. The most desirable environment was distilled water or a TRIS-HCl buffer. However, the TRIS-HCl was unsuitable in cases of retrieving a dry product by lyophilization. Eventually, the viability of encapsulated cells was examined, and it was proven that encapsulated bacteria endure the process of encapsulation in the above-mentioned environment and they were gradually released from the carrier.
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The Utilization of Light Scattering Techniques for the Study on Hydrogel Gelation
Candráková, Simona ; Smilek, Jiří (referee) ; Kalina, Michal (advisor)
This bachelor thesis focuses on the study of hydrogel gelation using light scattering methods. For these purposes two different biopolymers (agarose and sodium alginate), with different sol to gelation phase transition, were selected. In the case of agarose, the gelation is caused by temperature change. On the other side, the gelation of sodium alginate is initiated by addition of polyvalent cations. In the experimental part of the work, agarose gelation was studied by DLS, temperature gradients of agarose solutions (40 – 30 °C) were measured. During the measurement the particle size distributions in the solutions having different concentrations were monitored as well as the temperature influence on the diffusion coefficient. The DLS method provided the values of the gelling temperatures of the solutions at a concentration of 0,5 wt.% and 1,0 wt.%. Furthermore, the DLS microrheology method was used, where temperature dependences of viscoelastic characteristics of agarose solutions (40 – 30 °C) were also measured, from which the gelling temperatures of agarose solutions of 0,1 wt.% and 0,5 wt.% were evaluated. The classical rheology was also used in the work to compare the viscoelastic behaviour of the samples and to determine the gelation point of the agarose solutions. By this method were determined gelling temperatures for all studied concentration of agarose (0,1 wt.%, 0,5 wt.% and 1,0 wt.%). The DLS method was also used to monitor the interactions of sodium alginate with the addition of Ca2+ ions, these interactions were then also evaluated and discussed in the experimental part.
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Encapsulation of probiotics and prebiotics for use in nutritional products for children
Šnajdarová, Karolína ; Němcová, Andrea (referee) ; Skoumalová, Petra (advisor)
The Diploma thesis deals with designing of probiotic dietary supplement for children with strains Lactobacillus acidophilus and Bifidobacterium breve and with prebiotics. Used prebiotics were Inulin, Chia fiber, Bamboo fiber, Chlorella + Spirulina and Yakon syrup. The theoretical part is focused on probiotics, prebiotics and their biological influence. In experimental part the possibilities of encapsulation into alginate particle and lyophilisation of probiotic cells were observed to find their good form to final nutritional product for children. Several types of probiotic with addition of prebiotics were tested in model conditions of gastrointestinal tract. It was found that addition of prebiotic highly increases viability of probiotic cells and their resistance to model conditions of gastrointestinal tract. In this case, the best prebiotic was found in Yakon syrup. The prebiotics were also characterised in terms of nutritional composition, amount of total and reducing sugars, oligosaccharides, proteins, lipids, polyphenols and chlorophyll were obtained. Finally, Chia fiber, Chlorella + Spirulina and Yakon syrup were chosen as prebiotics with best characterisation/properties. In conclusion, a dietary supplement with lyophilized alginate particles containing probiotic cells and with the most appropriate prebiotics were designed. Forms of the product were powder and gummy-bear.
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Use of microcalorimetry in the study of hydration of biopolymers
Bola, Tomáš ; Smilek, Jiří (referee) ; Krouská, Jitka (advisor)
This master thesis deals with the using of microcalorimetry in the study of hydration of biopolymers. Lactose has been selected together with the other biopolymers although it is not among biopolymers but disaccharides. Selected biopolymers are alginate, dextrane, chitosan and hyaluronan of two molecular weights. Lactose has been selected for these purposes mainly because it is a model example to determine whether or not the reaction to moisture between the other samples and the saturated salt solution occurs. The biopolymer hydration study, as opposed to the commonly used perfusion calorimetry method using the possibility of measuring with adjustable moisture has been used an isothermal microcalorimetry method where at two constant temperatures the reaction of the sample to the different moisture released by the saturated salt solution was monitored.
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Study of barrier and transport properties of polyelectrolytes using diffusion techniques in hydrogels
Valentová, Kristýna ; Kalina, Michal (referee) ; Smilek, Jiří (advisor)
This diploma thesis was focused on study of barrier and transport properties of selected polyelectrolytes in hydrogel matrices by using diffusion techniques. The study of these properties was performed in horizontal diffusion cells where is observed the change in diffusion probe concentration over time. Diffusion experiments were performed on an agarose hydrogel with the addition of alginate, hyaluronic acid, polystyrene sulfonate, humic acids and as a model probe rhodamine 6G was used. Important parts of this thesis are also the methods which characterize the substances and hydrogel matrices such as rheology and potentiometric titration. The main aim of this diploma thesis was to investigate the effect of interactions between passing model dye (rhodamine 6G) and the appropriate gel (agarose + polyelectrolyte) on the fundamental diffusion parameters (effective diffusion coefficient, lag time, etc.).
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Development of a new type of energy drinks containing natural active components
Vysoká, Marie ; Němcová, Andrea (referee) ; Skoumalová, Petra (advisor)
The aim of this thesis is the development and preparation of a new energy drink. The theoretical part of the thesis focuses on the comparison of the drinks available in market, the presentation of the main active ingredients and the proposal of enriching the new energy drink. It also describes the possibility of using an encapsulation and the method used to determine specific compound in energy drink or encapsulated particles. In the experimental part of the work, at the beginning, the main content of energy drinks were characterized, for instance caffeine, sugar, polyphenol and vitamin C. Subsequently the energy drink based on selected types of loose teas that were enriched with the encapsulated extract of acai, acerola, maca, goji and ginseng was designed. All extracts contained large amounts of polyphenols and antioxidants. In addition, acerola extracts contained very high amounts of vitamin C. Two types of particles, alginate particles and liposomes were prepared for encapsulation. For all particles, encapsulation efficiency, size, stability, and antioxidant activity were determined. Long-term stability was then monitored within three months. The particles were also exposed to artificial gastric, pancreatic and bile juice. All tested particles showed very good encapsulation efficiency. In terms of long-term storage stability in an aqueous environment. Particularly liposome particles, which also showed high stability by zeta potential measurement, were suitable. On the contrary, in case of alginate particles, when stored in an aqueous medium, the release of active substances occurred. Finally, an energy drink was created and subjected to sensory analysis. The best energy drink was based on black or green tea with alginate particles acai, goji and ginseng.
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Encapsulation of selected natural extract for food application.
Vyskočilová, Terezie ; Kočí, Radka (referee) ; Márová, Ivana (advisor)
This diploma thesis deals with encapsulation of natural extracts. In the theoretical part the methods of encapsulation, materials for particle preparation, as well as application of encapsulation techniques in food industry were described. In experimental part selected natural extracts of propolis, green barley and probiotics were characterized. There substances were encapsulated into alginate and chitosan. In the total of 25 types of prepared particles long-term stability in some model physiological conditions as well as in four different model foods was evaluated. Additionally, stability of selected particles in several real milk-based products was followed too. The stability of particles was determined spectroptohometrically. In natural extract a content of polyphenols, proteins, chlorophylls, as well as total antioxidant activity were analysed. To analysis of probiotics optical and fluorescence microscopy were used. In propolis and green barley antimicrobial activity was tested too. Moreover, in the sample of propolis also cytotoxic assay was applied. Agar-chitosan was chosen as the best shell material for propolis due to its optimal stability in model physiological conditions as well as model foods. Liposomes were evaluated as unstable and were not recommended for further application. As the suitable shell material for powdered green barley starch-alginate (rate 1:4) and agar-chitosan were proposed, while the second one showed better stability for released proteins. Agar-chitosan shell material was usable for fresh green barley too. For probiotics encapsulation alginate or alginate-starch were chosen because of their porosity and possibility of nutrients diffusion. In real foods the best results were reached with application of probiotic particles into milk. Coencapsulation of powdered barley and probiotics did not confirm inhibition of culture growth. Neither the antimicrobial effect of propolis and barley nor the cytotoxic effect of propolis were confirmed.
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Phospholipids as the basis of biodegradable delivery systems
Burdíková, Jana ; Čeppan, Michal (referee) ; Mravec, Filip (advisor)
This thesis is focused on investigation of phospholipid-hyaluronan system. First, appropriate method for preparation of bulk solution of phospholipid/lipid and suitable fluorescence probe were chosen. Sonification was selected as a method for preparation of bulk solution and pyrene was chosen as a fluorescence probe. From the group of phospholipids lecithin was selected. Next to phospholipid, lipid with no phosphate group (DPTAP) was utilized for comparison, alternatively a mixture of lipid (DPTAP) and phospholipid (DPPC). Instead of hyaluronan another polyelectrolytes (sodium polystyrene sulfonate, sodium alginate) were used too. Measurements were performed in water environment and in phosphate buffer saline (PBS). All investigation was accomplished by fluorescence spectroscopy and dynamic light scattering.
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Magnetic modification of microbial cells
BALDÍKOVÁ, Eva
Baker´s yeast (Saccharomyces cerevisiae) were magnetically modified by three different methods, namely, surface modification by magnetic fluid, entrapment of cells into alginate and covalent immobilization on particles of magnetic chitosan. The ability of H2O2 decomposition was tested for all types of modification. It is apparent that the most amount of hydrogen peroxid was degraded by magnetic fluid - modified cells (84-95%), while the efficiency of cell which were modified by other methods was much lower (40-60%). Thanks to immobilization on particles of magnetic chitosan, we made completely new type of magnetic material, which was tested for adsorption of Crystal violet and Safranin O. It was founded that magnetic chitosan adsorbs no dyes, so all adsorption belongs to immobilized yeast. The maximum adsorption capacities were determined using Langmuire isotherm at 69,4 mg/g for Crystal violet and 99,0 mg/g for Safranin O.
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