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Preservative systems for natural cosmetics
Koblasová, Dana ; Veselá, Mária (referee) ; Zemanová, Jana (advisor)
The aim of this bachelor thesis is to evaluate the antimicrobial activity of naturally derived preservatives against selected microorganisms. Theoretical part is focused on description of the most used natural, naturally derived and synthetic preservatives, on the safety of preservatives and finally, the issue of contamination of cosmetic products by microorganisms. In experimental part, antimicrobial activity of 4 naturally derived preservatives was evaluated against selected microorganisms, which were bacteria Micrococcus luteus, Kocuria rosea, Bacillus cereus, Bacillus subtilis, Serratia marcescens and one yeast Candida vini. The tested preservatives were from MinaSolve company - E-LEEN Green A, which contains pentylene glycol and phenylpropanol, E-LEEN Green OR, which is composed of citrus extracts, citric acid, ascorbic acid and pentylene glycol, Green B, which contains benzoic acid, sodium benzoate and pentylene glycol and Hexam+, which is a mixture of pentylene glycol and hexamidin diisethionate. Antimicrobial activity was evaluated by agar diffusion method. The highest antimicrobial activity showed Hexam+, the lowest E-LEEN Green OR. E-LEEN Green A and Green B did not show any antimicrobial activity against any microorganism in the 1% tested concentration. The measured values of the inhibition zones show that the most resistant microorganisms from all the tested in this thesis are Bacillus cereus and Bacillus subtilis and the least resistant are Micrococcus luteus and Kocuria rosea.
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Use of some encapsulation techniques to controlled release of active substances in food and cosmetics products.
Skoumalová, Petra ; Rittich, Bohuslav (referee) ; Kráčmar, Stanislav (referee) ; Márová, Ivana (advisor)
The presented doctoral thesis is focused on preparation, characterization and application of organic micro- and nanoparticles as transport systems for active components and some their complex natural sources. Active component were packed into liposomes and polysaccharide particles. As active components were used caffeine, some drugs – clotrimazole and ibuprofen, further antioxidants and vitamins. Antimicrobial herbs and spices extract, antimicrobial peptides lysozyme, nisin and other antimicrobial ingredients were encapsulated too. Encapsulation of selected hydrolytic enzymes was tested, too. Particles were also used for encapsulation of probiotic strains Bifidobacterium breve and Lactobacillus acidophilus and prebiotic components. These prebiotics were co-encapsulated into capsules with probiotic cells. Natural extracts were encapsulated e.g. extracts of guarana, ginseng, goji, green barley, propolis, black, green and white tea, coffee, fruit and vegetable extracts. The efficiency of encapsulation was determined by HPLC/PDA and by spectrophotometry. Long-term stability of particles and amount of released component in model/real foods, in model cosmetic conditions and in a model physiological environment were monitored too. Size of prepared liposomes and polysaccharide particles was determined by dynamic light scattering and by light microscopy and electron microscopy, respectively. Stability of the particles was measured using a zeta potential. Also, analytical centrifugation was used to measurement of sedimentation velocity and stability of the prepared particles. The antimicrobial activity were tested using two Gram-positive (Bacillus subtilis, Micrococcus luteus), two Gram-negative (Escherichia coli, Serratia marcescens) bacteria and one fungal strains (Candida glabrata). For determining the antimicrobial properties of active component and prepared particles two the most widely used methods were used - agar diffusion method and broth dilution method. The viability of probiotic strains were performed using flow cytometry and fluorescence microscopy. Encapsulation of active component was successful in all types of particles. Liposome showed a very good long-term stability mainly in water conditions with neutral pH and polysaccharide particles were stable in acidic conditions. Prepared particles showed a very good stability in model stomach environment, while in model intestines environments particles were disintegrated and active component were released. Prepared particles with encapsulated caffeine as well as other tested antioxidants and vitamins could be used to modern types of energy drinks, food supplements and also for some cosmetics applications. Encapsulated antimicrobial components could be used for food application as well as for cosmetics and pharmaceutical application like antimicrobial wound formulation. Encapsulated enzymes can be used for controlled release of proteases in wound healing, as delivery systems in digestive tract and as a part of pharmaceutical preparative and food supplements for enzyme therapy. The study revealed that encapsulation of probiotics and also co-encapsulation of probiotics with prebiotics exhibited longer stability of particles and survival bacterial cells. So, prepared particles are suitable for use to food product with beneficial effects on the human body.
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Antimicrobial materials based on nanostructures with enzymes
Jurová, Bronislava ; Vysoká, Marie (referee) ; Skoumalová, Petra (advisor)
This bachelor thesis is focused on the preparation and characterization of antimicrobial gels and coatings containing antimicrobial components and enzymes. The theoretical part deals with a brief description of antimicrobial substances, their structure and mainly enzymes with antimicrobial effect. The methods used in the testing are also listed here. The practical part deals with testing the antimicrobial effects of extracts from cinnamon and cloves, clove essential oil, lysozyme and bromelain against Micrococcus luteus, Serratia marcescens and Candida glabrata. Various types of extracts were prepared from the selected spices, namely aqueous and ethanol extracts in different percentages. The individual extracts were characterized and, according to the total proportion of phenolic substances, the best ones were selected for encapsulation in liposome particles and gels. Extracts and enzymes were also added to the hydrogel and non-fibrous covers. A solution of sodium alginate and chitosan was used to prepare non-fibrous covers. A broth dilution method was used to test the antimicrobial effects on the given microorganisms. Furthermore, nanofibers based on PHB were prepared using the forcespinning method. These nanofibers were enriched with clove essential oil and their antioxidant activity was monitored.
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Biologically active substances of chokeberry (Aronia spp.)
Palkovská, Kateřina ; Pořízka, Jaromír (referee) ; Vítová, Eva (advisor)
This master thesis deals with preparation of extracts from fruits of aronia, optimization of conditions for extraction and subsequent characteristics of the obtained optimal extract. In the theoretical part information about the botanical characteristics of Aronia melanocarpa and about its utilization are mentioned. Furthermore, the chemical composition of aronia fruits is described and the main biologically active substances contained in the plant are described. At the end of the theoretical part extraction and separation techniques, determination methods of antioxidant, antimicrobial activity and determination of aromatic active compounds are described. The experimental part describes the optimization of maceration so that the macerate contains as much polyphenolic substances as possible, as well as aromatic active substances. 40% ethanol, at a temperature of 60 ° C, with a weight solvent ratio of 1: 5, for 40 minutes proved to be optimal for the maceration of aronia fruits. The total polyphenol content of the optimal extract was determined to be 5237.2 ± 40.7 mgGAEkg-1 and the antioxidant activity by TEAC to be 512.5 ± 5.04 gml-1. 24 aromatic active compounds were identified in the extract, with the largest percentage being monoterpenes and aldehydes. Furthermore, it was found that the extract did not show antimicrobial activity against the microorganisms used (Bacillus cereus, Escherichia coli and Candida glabrata). The obtained results confirm that aronia extracts are an important source of many biologically active compounds and can be used in food as a natural source of nutritionally and sensory valuable substances.
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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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Bioactive compounds of fennel
Nentvichová, Aneta ; Vespalcová, Milena (referee) ; Veselá, Mária (advisor)
The aim of this bachelor thesis is to study the bioactive substances contained in fennel, to determine the content of polyphenols, flavonoids and to investigate the antioxidant and antimicrobial activity in aqueous and ethanol extracts of fennel. The theoretical part discusses the characteristics of fennel, its classification in the system, its use and especially the bioactive substances contained in it. The experimental part is divided into two sections, where the first part focuses on spectrophotometric determination of polyphenols, flavonoids and antioxidant activities. In the second part the antimicrobial effects of prepared extracts of fennel were examined. Two microorganisms Serratia marcescens and Bacillus cereus were exposed to the said extracts. Antioxidant activity and relatively high content of polyphenols and flavonoids were found in the extracts in the first experimental part. The second experimental part proved that the antimicrobial effect of the extracts was very low, in most cases almost negligible. Ethanol extracts showed higher antimicrobial activity, while hot water extracts found to support the growth of Bacillus cereus.
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Ion-selective membrane with antimicrobial effect
Gablech, Evelína ; Farka,, Zdeněk (referee) ; Soukupová,, Jan (referee) ; Drbohlavová, Jana (advisor)
Tato práce se zabývá zlepšením antimikrobiálních vlastností komerčních polymerních membrán používaných v mlékárenském průmyslu pro filtraci mléčných výrobků jako je syrovátka. Tyto membrány trpí kontaminací způsobenou mikroorganismy přítomnými v mléce během procesu filtrace. Toto biologické znečištění ovlivňuje životnost membrány i samotný filtrační proces. Pro ochranu membrány před vytvořením biofilmu z mikroorganismů byly tyto membrány modifikovány pomocí plazmových metod tak, aby došlo k funkcionalizaci aminovými skupinami pro následnou imobilizaci nanočástic stříbra a selenu. Antimikrobiální aktivita obou typů nanočástic a membrán s imobilizovanými nanočásticemi byla stanovena proti buněčným kmenům Staphylococcus aureus a Escherichia coli.
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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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Characterization of plasma activated water for biomedical applications
Šindelková, Kateřina ; Dzik, Petr (referee) ; Kozáková, Zdenka (advisor)
The aim of this master thesis is to characterize plasma activated water (PAW) prepared in various plasma systems using direct and indirect interaction of plasma with liquid or its surface. Plasma was generated by electric discharges at atmospheric pressure. The prepared plasma-activated water was studied with respect to its use in biomedical applications. The theoretical part deals with plasma activated water. It contains a description of the PAW preparation, its characterization in terms of produced particles or its physico-chemical properties. Also, methods for antimicrobial activity estimation or colorimetric determination of produced particles are described in this section. In the experimental part of this work, the types of discharges that were used for the PAW preparation and their comparison are presented. Two types of liquids were selected for the activation, namely distilled and tap water. Subsequently, concentrations of selected reactive particles in PAW (hydrogen peroxide, nitrates and nitrites) produced in three different plasma systems were determined. Besides the comparison of reactive particles production in various systems, their time stability was studied as well. Over time after the PAW preparation, the values of active particles concentrations have changed due to mutual interactions between the particles. Mainly, nitrites were oxidized to nitrates or vice versa depending on the PAW type and other conditions. Part of the work was also monitoring selected physical properties of PAW, namely pH value and specific conductivity. It has been found that PAW does not retain its activated properties for too long, depending on the type of preparation and methods of storage. Therefore, one of the important goals of this work was to determine and compare suitable storage conditions for PAW with respect to its potential biomedical applications.
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Preparation and testing of application forms for natural antimicrobial substances
Vajglová, Klára ; Bokrová, Jitka (referee) ; Márová, Ivana (advisor)
The goal of this bachelor thesis was the study of antimicrobial and antioxidant effects of some herbal extracts: cloves, nettle, sage, chamomile, rosemary, elderberry, lavender and also lysozyme. Further, herbal extracts effect on Gram positive and Gram negative bacteria were compared. In the theoretical part were briefly characterized lysozyme, polyphenols, flavonoids and selected strains of microorganisms. In the experimental section characterization of herbs with antimicrobial and antioxidant effect was proved. Particles containing herbal extracts were prepared and encapsulation efficiency was determined. In some of tested herbs encapsulation efficiency of up to 80% was reached. Stability of the prepared liposomes was tested in the model digestive conditions and after 1 week and 1 month incubation period in model foods. Antimicrobial activity was tested on selected representatives of Gram negative and Gram positive bacteria. Gram positive bacteria were more sensitive to herbal extracts and liposomes containing encapsulated herbs, while lysozyme exhibited higher effect to Gram negative bacteria. Prepared particles with encapsulated antimicrobial extracts are especially suitable for use in the food industry.
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