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Inverse FCS in colloidal systems research
Richterová, Veronika ; Venerová, Tereza (referee) ; Mravec, Filip (advisor)
This diploma thesis is focused on the study of inverse fluorescence correlation spectroscopy, especially with the regard for the usage of different fluorescent probes and different sized analysed particles. At first, the proper concentration of fluorescent probes was determined. In this concentration is the probe considered as a medium surrounding the analysed particles. Based on this concentration, which was determined as 400 M, several sets of samples were prepared. This samples contained different concentration of polystyrene particles of 100 and 500 nm diameter and multilamellar liposomes. Then, the FCS curves of samples with different fluorescent probes were measured. Fluorescein, rhodamine 6G and Atto 488 were used as fluorescent probes. As a result from experiments, it was found, that particles with 100 nm diameter cannot be analysed with none of the fluorescent probes. Inverse FCS method can be applied to systems, that contains particles with 500 nm diameter and fluorescein. Systems with rhodamine 6G have the same behaviour as typical FCS measurement. It is caused by dimerization of this probe and it cannot be used for 500 nm particles. Liposome samples can be established with iFCS method, but the results are biased by random distribution of liposomes size.
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Preparation and characterization of foods and food supplements containing selected vitamins
Krupičková, Lucie ; Hoová, Julie (referee) ; Skoumalová, Petra (advisor)
The thesis is focused on preparation of protected form of vitamins for aplication to food and diatery supplemets. The theoretical part characterizes vitamins and describes their role in human organism. Furthermore, this chapter summarizes different methods of determination vitamins soluble in water and encapsulation of substances into liposomes. The last sections are focused on functional foods and dietary supplements. In experimental part, an optimalization of determination method was made. Selected vitamins were encapsulated into liposomes. Size, stability, encapsulation efficiency, long-term stability and long-term encapsulation efficiency after two weeks was determined there. All prepared liposomes were found to be stable enough. Furthermore, the vitamin solutions, liposomes and lyophilized particles were analyzed during the process of model digestion. The active forms of vitamins were degraded during the model digestion of vitamin solutions. Vitamins were gradually released from the liposomes and lyophilized particles into the digestive juice. The analysis of content declared by producer was executed for functional foods and dietary supplements. Finally, applications of protected form of vitamins into the food and dietary supplemets were created.
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PREPARATION AND APPLICATION OF SOME BIOPOLYMERS, NANOPARTICLES AND NANOFIBRES FOR COSMETICS AND FOOD
Bokrová, Jitka ; Pekař, Miloslav (referee) ; Kráčmar, Stanislav (referee) ; Márová, Ivana (advisor)
The presented doctoral thesis is focused on preparation of nanoparticles and nanofibers with natural active ingredient and testing their biological effects. Modern types of application forms were prepared from biomaterials based on one or more natural polymers. Chitosan particles were prepared from cross-linked polymer using ultrasonication. A mixture of soy lecithin and cholesterol was used for preparation of liposomes. Poly-3-hydroxybutyrate was used for preparation of combined liposomes, too. All liposome particles were prepared by ultrasonication. Nanofibers were obtained from polyhydroxybutyrate using electrospinning. Mixtures of low-molecular antioxidants obtained by extraction from natural sources were used as active ingredients. Different types of teas, barks, herbs, spices, fruits and vegetables were selected as sources of natural antioxidants. Total phenolic and flavonoid content and total antioxidant activity of extracts were determined using spectrophotometrical methods. Obtained natural extracts were subsequently used for encapsulation. Prepared application forms were characterized in terms of their physicochemical properties. Particle size was monitored by dynamic light scattering. Colloidal stability of particles in suspension was determined using zeta potential. Spectrophotometry was used to evaluate the efficiency of encapsulation of active compounds into particles. The morphology of the new type of combined PHB liposomes was monitored by electron microscopy. Chromatography was used for quantification of individual components of particles. Morphology of nanofibers and incorporation of active agent into their structure were monitored using FTIR-ATR spectroscopy and electron microscopy. Afterwards, antimicrobial, cytotoxic and genotoxic effects of preparations were evaluated. It was found that the most suitable types of extracts for liposome preparation are aqueous and lipid extracts of natural antioxidants. Prepared particles showed excellent stability and good encapsulation efficiency. The study confirmed that incorporation of polydroxybutyrate into liposome structure does not reduce neither the colloidal stability of the particle, nor the efficiency of encapsulation process. Antimicrobial and antimycotic effect of preparations against model microorganisms Micrococcus lutues, Serratia marcescens and Candida glabrata was detected. It was found that process of encapsulation increases the inhibitory effect of natural extracts of antioxidants. The safety of preparations was assessed using two human cell cultures: epidermal keratinocytes and HaCaT cell line. Assays of cell viability and plasma membrane integrity were used to determine cytotoxicity of preparations. Low toxicity of liposome particles was confirmed by a series of cytotoxic tests. Obtained data showed that association of phospholipid with PHB polymer does not cause a significant increase in cytotoxicity in human skin cells. Genotoxicity testing on model procaryotic organism confirmed zero genotoxic potential of preparations. The new type of combined particles and polymeric fibers cant thus be used as a carrier for active ingredients, complex natural extracts, antimicrobial agents and many others.
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Characterization of beverages and food supplements containig caffeine
Patočková, Klára ; Pekař, Miloslav (referee) ; Márová, Ivana (advisor)
This master thesis is focused on characterization of some energy drinks and formulation of their modified composition. The theoretical part was focused on summary of energy drinks and some of their natural components - guarana, goji as well as on techniques of encapsulation. In the experimental part concentrations of caffeine, vitamin C, polyphenols and carbohydrates in commercially purchased drinks was evaluated. Subsequently, three kinds of particles (particles with caffeine, particles with guarana and particles with guarana and goji) by 6 different techniques were prepared. The efficiency of encapsulation was determined by HPLC/UV-VIS (caffeine, vitamin C) and by spectrophotometry (phenolics). Stability of the particles was followed in real food and in a model physiological environment of artificial digestive juices. Size of prepared particles was determined by dynamic light scattering. In this work it was found that particles with best activity and stability were obtained by encapsulator. For manual particles the best results were found in the liposome particles. Liposomes exhibited the smallest particle size, the highest efficiency encapsulation, excellent stability in real food and juices and the best value of zeta potential.
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Development of new type of energy drinks based on coffee with active substances
Dudrová, Markéta ; Skoumalová, Petra (referee) ; Márová, Ivana (advisor)
This bachelor thesis deals with development and preparation of new energy drinks with active plant component in free and encapsulated form. The theoretical part is focused on characterization of the energy drinks, their components and properties. The experimental part deals with the preparation of plant extracts, their characterization and subsequent encapsulation into liposome particles. All particles were determined for size, colloidal stability, long-term stability, antioxidant activity and encapsulation efficiency. Energy drinks based on various types of coffee were prepared and enriched with free or encapsulated guarana, goji, matcha or cocoa extract. In prepared energy drinks the content of caffeine, sugar, phenolics and antioxidant aktivity was monitored. Finally, selected energy drinks were subjected to sensory analysis, where appearance, color, smell and taste were evaluated.
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Use of algae extracts in cosmetics
Plášková, Anna ; Rapta, Marek (referee) ; Márová, Ivana (advisor)
This thesis is focused on a preparation of extracts from selected species of algal and cyanobacterial strains, their characterization, a cytotoxicity testing and a use in cosmetic products. The theoretical part contains the review of a taxonomic classification and parameters of algae and cyanobacteria and basic information about used organisms. There is a description of the most significant factors influencing the algae cultivation and production of biomass. Further, active substances of algae and cyanobacteria, which could be used in cosmetic products as UV protective substances, were described. An overview of extraction techniques, encapsulation of active substance into liposomes and particles characterization, antioxidant activity analysis, and cytotoxicity tests is given too. In experimental part, antioxidant activity of algal extracts and chlorophyll a and carotenoid content were determined spectrophotometrically. The long-term stability of extracts was monitored for three weeks. Liposomes were prepared by sonification and encapsulation activity was analysed. The Dynamic Light Scattering method (DLS) was used for analysis of polydispersity and size of liposomes. Colloid stability of particles was determined by zeta potential. Cytotoxicity tests were performed using MTT assay. The prepared extracts, particles and lyophilized biomass of algal and cyanobacterial strain were further used as active substances in the formulation of cosmetic products (type o/w). These creams were tested on human skin.
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Preparation and characterization of biomaterials with natural UV filtres
Pavelková, Renata ; Čarnecká, Martina (referee) ; Márová, Ivana (advisor)
This presented diploma thesis is focuses on preparation and characterization of biomaterials with natural UV filters. The theoretical part is aimed at effects of sunlight on the skin and how to avoid these effects by using cosmetic products. The materials and methods for extraction of lipid part of the selected vegetable and microorganism species and methods for preparation and characterization of nanoparticles and emulsions were characterized. The practical part deals with extraction of lipid part of selected vegetable and microorganism species. The liposome particles with encapsulated active compounds were prepared. The particle size, polydispersity index, colloidal stability, encapsulation efficiency, SPF, antioxidant capacity, phospholipid content and cytotoxic effect on human keratinocytes were observed. These particles were used for preparation of the resulting emulsions. Their protection level on UV light were set and stability were tested by using analytical centrifugation.
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Preparation of edible antimicrobial packaging
Kozubová, Petra ; Dzurická, Lucia (referee) ; Skoumalová, Petra (advisor)
This bachelor’s thesis is focused on preparation and characterization of eatable packaging. Eatable packaging consists of two components, antimicrobial component and base of packaging. The task of eatable packaging is to protect product against antimicrobial attack and to extend durability of the product. As antimicrobial components were used water and oil extracts of several herbs (mint, nettle, lavender, cinnamon, cloves, vanilla, ginger and rhinoceros). Oil extracts were also encapsulated into liposomes for better application. Firstly, concentrations of polyphenoles and antioxidants of extracts were determined. Oil extract of clove contained the highest concentration of both determined substances. Encapsulation efficiency, stability and size of liposomes were tested too. All prepared liposomes were stable and relatively of the same size. Also high encapsulation efficiency was observed. Next, antimicrobial activity of prepared extracts and liposome particles against yeast Candida glabrata and two bacteria strains Escherichia coli and Staphycococcus epidermidis was tested and the high antimicrobial activity reported mainly all tested liposomes. Then alginate, chitosan and carboxymethylcellulose and their combinations respectively were used as bases of packaging. Created film had to meet several parameters, especially suitable sensory properties such as pleasant taste and smell, as well as invisibility, gloss, affordability and suitable manipulability. Combination of 2% carboxymethylcellulose and 1% alginate in a ration of 2:1 with liposomes of mint as antimicrobial component achieved the best results.
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Preparation and use of selected nanoparticles in cosmetics
Hoová, Julie ; Němcová, Andrea (referee) ; Márová, Ivana (advisor)
The Bachelor thesis is focused on preparation and use of selected nanoparticles in cosmetics. The theoretical part deals with general characterization and use of caffeine, phenolics and vitamin C in cosmetics. Further, characterization of liposomes and possibilities of encapsulation of active substances gained from natural sources were introduced. In experimental part liposomes with encapsulated active substances were prepared by three different techniques. Encapsulation efficiency was determined by HPLC/PDA, phenolics spectrophotometrically and vitamin C by titration. Prepared particles were monitored for size and stability by dynamic light scattering. Furthermore, long-term stability of liposomes in model conditions was evaluated as the amount of the active substances released during incubation. In this work was found out that from all tested samples co-encapsulated liposomes are probably the best application form which can be used in face lotion or gel. Co-encapsulated liposomes exhibited relatively high encapsulation efficiency, good stability and thanks to quality of active substances they meet customer’s requirements of cosmetic industry.
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Isolation and characterization of natural antimycotics and their potential applications
Dvořáková, Eva ; Hoová, Julie (referee) ; Skoumalová, Petra (advisor)
This bachelor thesis was focused on preparation and characterization of products containing natural antimycotic ingredients. The theoretical part summarizes the current issue of increased presence of mycoses and yeast diseases in connection with the extension of the problem of reduced immunity. Furthermore, the problem of synthetic drugs and antimycotics compared to alternatives of natural origin was described. In the experimental part, selected extracts (aqueous, ethanolic and lipidic) were prepared, characterized and subsequently encapsulated into liposomes. The content of polyphenols, flavonoids and the total antioxidant activity of extracts was measured. In liposomes, the encapsulation efficiency, size, stability and long-term stability were measured. In addition, the antimycotic activity of individual extracts and liposomes against Candida glabrata was studied. For comparison, the antifungal activity of the commercial drug clotrimazole was also tested. Lipidic extracts showed highest antimycotic activity from all of the prepared extracts, which is why these extracts were used to prepare antifungal gels and suppositories. More specifically, lipidic extracts of nasturcium, cinnamon, ginger and liposomes containing them were used. Their antimycotic activity was also tested and confirmed for these preparations. Finally, the cytotoxicity test on the HaCaT human cell line showed that used extracts and liposome particles are safe and that the prepared products could thus serve as an alternative to synthetically prepared antimycotics in the future.
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