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Study of properties and effect of some natural antimicrobial substances
Horňáková, Nikola ; Kostovová, Iveta (referee) ; Márová, Ivana (advisor)
This bachelor thesis is focused on the testing of natural antimicrobial substances of plant origin, dealing with their characterisation, their effect against the microorganisms, optimization of antimicrobial testing and increasing of efficiency of their active compounds. Theoretical part of this thesis describes a characterisation of used natural species, their important properties, spectrophotometrical methods applied in determination of active compounds, opportunities of stabilization of natural substances and various techniques of antimicrobial testing. In experimental part 8 kinds of tea and non-tea plants (Pai Mu Tan, Formosa Gunpowder, Gunpowder Black, Honeybush, Lapacho, Aspalathus linearis and Paullinia cupana) were used and combined with 6 herbal species (Syzygium aromaticum, Urtica dioica, Hippophae rhamnoides, Matricaria recutita, Plantago lanceolata and cinnamon), extracted by different solvents (water, ethanol and dimethyl sulfoxide). In these extracts amount of antioxidants and effect to three bacterial cultures (Escherichia coli, Micrococcus luteus and Serratia marcescens) were determined. To improve antimicrobial, antioxidant activity and stability aqueous tea extracts were encapsulated into liposomal particles.
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Study of natural antimicrobial mixtures and their stabilization
Vejrostová, Petra ; Veselá, Mária (referee) ; Márová, Ivana (advisor)
Present diploma thesis is focused on natural antimicrobial substances. The aim of the experimental part of thesis was testing of antimicrobial activity of selected natural substances in free and encapsulated forms, preparation of particles and monitoring of their long-term stability. Various teas extracts (aqueous, ethanoic and DMSO) and aqueous herbs extracts were used as plant antimicrobial substances. Liposomes and chitosan particles with encapsulated aqueous teas extracts were prepared by ultrasonication. Encapsulation efficiency, size, stability and antioxidant activity for all particles were specified. Consequently, within a two months period, long-term stability for these particles was monitored. Spectrophotometric method for determining total polyphenols was used for evaluating encapsulation efficiency as well as for monitoring of long-term stability. The DLS method was used for determination of size and stability of particles. Antioxidation activity was monitored by using spectrophotometric method using ABTS. For testing antimicrobial activity of various types of teas, herbs extracts and their mixtures and combianitons with lysozyme and antibiotics, broth dilution methods were used while using both Gram positive and Gram negative bacterial test strains. Antimicrobial activity of prepared particles was also monitored, the antimicrobial activity was especially significant with the chitosan particles. Suitable combinations of natural substances whatever with antimicrobial or antioxidation effect either in free or encapsulated form could be used for applications for example in cosmetics industry, food processing or in medicine.
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Determination of Natural Compounds in Select Plant Materials and Less Obvious Kinds of Fruit
Hohnová, Barbora ; Márová, Ivana (referee) ; Ventura, Karel (referee) ; Omelková, Jiřina (advisor)
Flavonoids are natural compounds widely distributed in plant kingdom. They are inseparable from human diet because they showed a protective effect against cancer, stroke and coronary heart diseases related to their antioxidant properties. Therefore, rapid and efficient extraction procedure prior to chromatographic analysis is required. The liquid extraction at elevated temperature and pressure – Pressurized fluid extraction (PFE) and Pressurized hot water extraction (PHWE), present fast, effective and environmentally friendly extraction methods for the determination of flavonoids in plant materials. PFE and PHWE followed by reversed phase high-performance liquid chromatography with UV-visible detection have been utilized for the determination of a group of flavonoids (rutin, myricetin, quercetin, luteolin, apigenin and kaempferol) in the leaves and berries of less common plants. The matrices were extracted by methanol, ethanol and water at higher temperature 40-120 oC and pressure 15 MPa during 15 minutes. The obtained results were compared with conventional Soxhlet extraction and ultrasound-assisted extraction, the same solvents were used. The most effective extraction of selected flavonoids was achieved by PHWE. PFE showed the extraction yields comparable to those of the Soxhlet extraction, and the lowest extraction power was displayed by ultrasound-assisted extraction.
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Isolation of compounds from Cannabis sativa and their anti-inflammatory effect
Baranová, Zuzana ; Vespalcová, Milena (referee) ; PharmDr.Daniela Veselá (advisor)
The object of this thesis is a traditional and also controversial plant Cannabis sativa. The theoretical part is focused on ubiquitous polyphenolic compounds – flavonoids, also present in the studied plant. All previously researched and confirmed effects on human body are described in detail. The experimental part of this thesis describes the isolation and identification of substances of the chloroform and hexane yields of extract obtained from this plant. Chromatographic methods were used for isolation – thin layer and column chromatography and semipreparative HPLC. Obtained substances were characterized using UV, IR spectrophotometry and NMR. Then the anti-inflammatory activity of obtained substances and also of the ethanolic extract and its yields was observed. The result of the experimental part was the isolation of one pure substance which was identified as the cannabidiolic acid. Using the THP 1 assay, we demonstrated mild anti-inflammatory effect of the non-polar yields and isolated CBDA.
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Encapsulation of natural antioxidants
Štindlová, Jitka ; Obruča, Stanislav (referee) ; Márová, Ivana (advisor)
In theoretical part of this diploma thesis the basic properties of antioxidants, especially anthocyanins and phenolic compounds are described, as well as the basic features and principles of free radicals formation. The theoretical part also describes some possible ways of encapsulation of antioxidants into polysaccharide and lipid particles. In the experimental part basic characteristics of extracts from selected lyophilized fruits and vegetables (carrots, apples and mixed berries) are described. As group parameters of plant extracts the total antioxidant activity, content of flavonoids and phenolics, carotenoids, anthocyanins and ascorbic acid were determined. In experimental part also various encapsulation techniques were tested, encapsulation effectiveness of each technique was evaluated and the stability and size of the created particles were determined. As the best encapsulation method in terms of encapsulation efficiency in most of samples/parameters ethanol injection was found. On the other hand the particles prepared by ethanol injection are relatively unstable in terms of zeta potential, which is followed by their tendency to aggregate. As the most stable particles prepared by thin layer evaporation (TLE) and reverse phase thin layer evaporation (RP-TLE) were evaluated. Particles prepared by TLE, RP-TLE and chitosan-alginate particles exhibited a negative charge, while particles prepared by ethanol injection stayed uncharged and chitosan particles have a positive charge.
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Determination of some nutrition factors in selected kinds of fruit
Smělá, Margita ; Vítová, Eva (referee) ; Vespalcová, Milena (advisor)
Diploma thesis is concerned with determination of some nutrion factors and vitamin C in various cultivars of apples (Pirus Malus, L.) and quinces (Cydonia oblonga, Mill.). In theoretical part there are described general characterization in selected kinds of fruits, their medically significant sunstances and utilization of these fruits. Main focus is paid to vitamin C. There are described its chemical properties and effect for human health. Various methods for determination of some nutrion factors with focus on determination of vitamin C by high-performance liquid chromatography. In experimental part, total of 27 various cultivars of aplles and 16 various cultivars of quinces were analyzed. These cultivars are from grower station Faculty of garden of Mendel university. For determitation vitamin C was used by high-performance liquid chromatography. Vitamin C was determination as an ascorbic acid. Some nutrion factors were analyzed in experimental part. There were analyzed dry matter, total flavonoids, total phenolics and titratable acid in various cultivars of apples and quinces. There were compared of some various cultivars after representation and content of significant substances as result. The last part of thesis was sensory analysis of apples.
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Changes in selected local fruits during freezing.
Matějková, Markéta ; Melounová, Jitka (referee) ; Márová, Ivana (advisor)
This diploma thesis was focused on study of low-molecular antioxidants present in various kinds of fruits, namely berries. In theoretical part detailed characterization of antioxidants studied in presented work and their biological effects are introduced. Further, principals of processing and storage of specified species of fruits are discussed. Changes observed during storage of specific fruits are described too. In practical part changes in content of selected antioxidants in berries were followed. As studied material strawberries, raspberries, blueberries, red and black currants were used. All kind of these fruits was stored in fridge for six months using several different kind of processing. Berries were stored as whole fruits, whole osmotically processed fruits by sucrose, raw pulp and sweetened pulp. At first, starting values of selected antioxidant parameters were measured in fresh fruits (ascorbate, total antioxidant activity, total phenolics, catechins and individual flavonoids). After freezing the same parameters were measured regularly in two month periods. As the most suitable method of fruit processing, with regard to antioxidant content, storage of the whole fruits or pulp can be recommended. Content of ascorbate decreased in all forms and species of the processed fruits during all the time of storage. Oppositely, the content of other studied antioxidants exhibited fluctuations or increasing trend. Sensory analysis was proved for the whole fruits and also for whole sweetened fruits.
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Pressurized solvent extraction of plant material for food industry use
Holasová, Petra ; Vítová, Eva (referee) ; Vespalcová, Milena (advisor)
Flavonoids were identified and quantified in samples of dry leaves of plant Stevia rebaudiana Bertoni. Content of flavonols (kaempherol, quercetin, myricetin, rutin) and flavons (apigenin, luteolin) were compared in three samples (standard leaves of Stevia rebaudiana Bertoni, Stevia rebaudiana Bertoni origin from Ukraine and from Czech republic), that were extracted by three extraction methods (PSE, ultrasonic extraction, Soxhlet extraction) with two polar solvents, methanol and ethanol, in combination with non-polar hexane. Extracts were analyzed by HPLC with detection UV-VIS. Antioxidant activity and total content of flavonoids in extracts were measured by spectrometry. Stable free radical DPPH. was used for determination of scavenging effect in extracts. PSE was the most effective extraction method with this conditions – 120 °C, dynamic mode and with methanol like solvent.
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Determination of total antioxidation activity of tea pickles in selected sorts of tea
Bartošová, Jitka ; Vitoulová, Eva (referee) ; RNDr.Jana Skopalová, Ph.D. (advisor)
This thesis deals with the total antioxidant activity (TAA) evaluation of various sorts of tea (green, black, half-fermented, white, mate, rooibos and herbal) depending on the process and time of their treatment. The main emphasis is put on testing a new method of measuring the total antioxidant activity by flow coulometry. The results acquired by this method are compared to the results of the chemiluminescent Trolox equivalent antioxidant capacity (TEAC) method which is a standard for measuring the total antioxidant capacity in various materials and raw materials. The work discusses the reasons for the differences in the values found by both methods.
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Preparation of some application formula of antimicrobial substances
Krempaský, Kamil ; Němcová, Andrea (referee) ; Márová, Ivana (advisor)
The main aim of this study is to determine antimicrobial activity of herbal extracts and also encapsulate these extracts and lysozyme to liposomes. The extracts were prepared from above ground part of wormwood (Artemisia absinthium), aerial part of breckland thyme (Thymus serpyllum), the root of purple coneflower (Echinacea purpurea), liquorice (Glycyrrhiza glabra), corolla of pot marigold (Calendula officinalis) and berries of sea buckthorn (Hippophae rhamnoides). In all extracts antioxidant activity and the amount of total polyphenols and flavonoids was determined. All of these extracts and also lysozyme were used for encapsulation to liposomes. Prepared liposomes were then tested for their stability. Further, encapsulated as well as free extracts were tested for their antimicrobial activity on Escherichia coli, Serratia marcescens, Micrococcus luteus and Bacillus subtilis. Extract from Echinacea purpurea showed the highest antimicrobial activity.
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