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Separation and determination of possible products of enzymatic cleavage of 4-nitrophenyl-N,N'-diacetyl-β-D-chitobioside using capillary electrophoresis
Velvarská, Romana ; Křížek, Tomáš (advisor) ; Kalíková, Květa (referee)
This work deals with the development and optimization of conditions of a method that can be used to compare the activity of the enzyme β-N-acetylhexosaminidase in hydrolysis of a natural substrate and a chromogenic substrate, which is often used in the study of enzyme kinetics. As a substrate, 4-nitrophenyl-N,N'-diacetyl-β-D-chitobioside was selected for cleavage. This oligosaccharide contains bond, which the enzyme cleaves in the natural substrate, and the bond that occurs in the chromogenic substrate. To determine the products arising from enzymatic hydrolysis of 4-nitrophenyl-N,N'-diacetyl-β-D-chitobioside, capillary zone electrophoresis was used. First, it was necessary to find the optimal composition of the electrolyte, its pH and concentration. The optimal background electrolyte was a solution of sodium tetraborate at a concentration of 25 mmol/l and a pH of 10.25. Subsequently, repeatability, calibration curves and linearity, limit of detection and limit of quantification were investigated. Repeatability of migration times ranged up to 0.6%, the repeatability of peak areas between 2.5 and 6.3%. Limits of detection were ranging from 0.005 to 0.120 mmol/l. Finally, the optimized method was successfully used to monitor the actual enzyme cleavage.
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Educational Project Saccharides Topic Work in Chemistry for high school
Šedivá, Renata ; Šulcová, Renata (advisor) ; Součková, Danuše (referee)
Charles University in Prague - Faculty of Science Department of Teaching and Didactics of Chemistry Albertov 3, 128 40 Praha 2, Czech Republic Educational project Saccharides Topic Work in Chemistry for high School Bc. Renata Šedivá sedivarenata@gmail.com This diploma thesis is a suggestion of educational project Saccharides classifiable into education of Chemistry in 3rd year of four-year grammar school. In the first part there is a research of literature sources concerning the issue project education. The Following part is a suggestion of educational project Saccharides, its product are lessons about the topic saccharides, which are designed, prepared and taught by the students. The suggestion of the project contains a timetable of the project, instructions and methodology for individual parts of the project (motivation, preparation of lessons, laboratory works, presentation of lessons, final practice, test and reflection), methodological notes and documentation to the implementation of the project at Grammar school in Dvůr Králové nad Labem.
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Determination of macroelements, organic acids and other parameters in ketchups
Popelová, Nikola ; Divišová, Radka (referee) ; Diviš, Pavel (advisor)
The aim of this master’s thesis is perform basic analyses of samples ketchups. The theoretical part describes the basic characteristics of tomato, tomatoes processing and production of puree and ketchup. Then there are described analytical methods which were used for analysing the selected parameters in ketchups. The experimental part deals with the determination of pH, total solids, organic acids, saccharides, macroelements and lycopene in ketchups. Citrate and acetate were analysed by IC. The amount of acetate in ketchups was about 5 times greater than the amount of citrate. Glucose, fructose and sucrose were analysed by HPLC. Glucose and fructose were determined in all samples but sucrose not. The ICP-OES method was used to determine four elements. Magnesium was the least represented element and sodium was the most represented element in the analysis. The amount of sodium was recalculated on the amount of salt. Lycopene was analysed by UV-VIS spectrometry. Its amount in ketchups is influenced by the variety of used tomatoes and plenty of other factors.
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Evaluation of Physical and Chemical Parameters of Still Less Exploited Species of Small Fruit and Proposal of a New Soft Drink from this Fruit
Cetkovská, Jitka ; Krška,, Boris (referee) ; Barták,, Petr (referee) ; Omelková, Jiřina (advisor)
This thesis deals with still less expolited species of small fruit, evaluation of its physical and chemical parameters and proposal of a new soft drink from one of the studied species of furit. Firstly, for the characterisation of basic nutrition parameters (dry matter, content of organic acids and saccharides) and biologicaly active substances (vitamin C, anthocyanins and phenolic compounds) suitable titration, spectrophotometric, chromatographic and electrophoretic methods were chosen, optimized and validated. The sample preparation procedure was optimized as well. For three years (2010–2012) the content of listed parameters was monitored in some cultivars of five species of small fruit: in seven cultivars of sea buckthorn, in ten cultivars of cornelian cherry, in twelve cultivars of rowanberry, in two cultivars of chokeberry and in seventeen cultivars of elderberry. By means of Duncan’s test, analysis of principal components (PCA) and cluster analysis (CLU) the main differences between studied cultivars were observed mainly in sea buckthorn and rowanberry, where single cultivars were easily distinguishable from others on the basis of genetic origin. On the basis of comparison of determined parameters in studied species of fruit and with collaboration with fruit processing company the suitable species of fruit for commercial utilization was chosen – elderberry, as a important source of anthocyanins and phenolic compounds. A new soft drink as a proposal of utilization of this fruit was created – fruit juice or nectar with portion of elderberry juice. Mixed juices and nectars with various composition were prepared from elderberry and grape juice and the sensory analysis of these drinks was performed. The best evaluated were 100% fruit juices prepared with 30–50% portion of elderberry juice. The composition was jurally protected. On the basis of this protection the product consisted from 100% fruit juice with 10% of elderberry component mixed with apple and grape juice was introduced to the market by the fruit processing company.
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Determination of the basic physical and chemical parameters of fruit juice from aronia
Zídková, Anežka ; Jurečková, Zuzana (referee) ; Diviš, Pavel (advisor)
Theoretical part of bachelor´s thesis contains basic information about aronia (Aronia melanocarpa) and examples of their use. There is a description of analysed substances, which are contained in aronia berries, and their importance for human organism. Experimental part is focused on determination of minerals, saccharides and vitamin C in aronia juices, syrups, nectar and aronia wine. Elements were determined by inductively coupled plasma optical emission spectrometry. Liquid chromatography was chosen for analysis of saccharides and vitamin C. All of analysed products are significant sources of macroelements. The highest concentrations of elements were determined in the sample named Aroniový skorosirup, which was sugared by vaporized apple juice. Fructose and glucose were contained in all products. Sucrose was only in products which were sugared. The amount of vitamin C was lower in consequence of pasteurization and storage.
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Cereals - Active Substances, Biological Effects and Selected Applications in Food Industry
Lichnová, Andrea ; Tremlová, Bohuslava (referee) ; Vávrová, Milada (referee) ; Márová, Ivana (advisor)
The presented doctoral thesis is focused on the study of the biological effects of active compounds found in cereals, on the development of methods of analysis these effects, on the determination of the content of active substances and characterization of the relationship between composition and biological effects of cereals and cereal products. For the analysis several kinds of raw cereal samples (flakes, flour, germ, bran), flavored extruded cereal products and also samples of paddy and husked rice were chosen. To major types of analysed active compounds belong mainly phenolic compounds in the form of glycosides and aglycones, and also saccharides. Group parameters such as total polyphenols, flavonoids and total and reducing saccharides were determined spectrophotometrically, individual phenolics and saccharides were determined by high performance liquid chromatography (HPLC). TEAC, DPPH and CLAMS methods were used to determine the antioxidant activity. Indirect methods of determination of substances with antioxidant effect were used as well. Results of the total antioxidant activity were compared with values of antimutagenic/genotoxic activity obtained by several microbial test systems. Antimutagenic effect was expressed as a percentage of inhibition of effect of standard mutagen and could be considered as a potential preventive effect of cereals to DNA arising primarily by free radicals effect. The highest values of group and individual phenolics, antioxidant and antimutagenic activity were found in germs, bran, in buckwheat products and in coloured and raw rice. In flavoured cereal products addition of chocolate or fruit positively influences content of active phenolic substances as well as sugars, antixidant and antimutagenic activity. In a representative sample of Czech population, questionnairy study was performed to monitor interest in cereals and consumer preferences. The most of consumers consider cereals with chocolate flavour as less healthy than confirmed results of laboratory analyses. In this study some new food products were developed. Several types of model cereal products containing plant (fruit, vegetables) extracts were proposed. Extracts were added to cereals in freeze-dried and encapsulated form. The highest positive effect exhibited addition of local forrest fruit extract. Within preparation of encapsulated extracts several methods of preparing lipid or saccharides particles were also tested. The encapsulation efficiency of the methods and stability and size of particles were analysed. Optimal type of fortified cereal foods could be suggested based on the acquired results. Selected cereals were used as alternative carbon substrates (processed or raw) for the cultivation of microorganisms to produce enriched biomass usable in the feed industry. We can conclude that cereals in raw as well as processed form belong to universal foods and rich sources of biologically active substances. They can be processed by many ways. They can be used for direct consumption, as a part of new products and also undirectly as a substrate for feedstock.
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Determination of selected chemical parameters of fruits of cultivated elderberry varieties
Hamalová, Veronika ; Vránová, Dana (referee) ; Vespalcová, Milena (advisor)
In this thesis was determinated selected chemical and nutritional parameters in the wild elderberry and 17 cultivated varieties of elderberry. The fruits was provided by the Research and Breeding Institute of Pomology Holovousy Ltd. The theoretical part deals describes elderberry, chemical composition of elderberry, especially biological active substances and its using not only food industry. Closer attention has been focused on effect of saccharides, anthocyanins and methods of their determination, specially determination by HPLC. The experimental part describes the various methods of determination of selected chemical and nutritional parameters. Total dry matter, content of total polyphenolic compounds, monomeric anthocyanin pigment and antioxidant activity was defined in all varieties of elderberry fruits. For some varieties was defined total soluble dry matter, content of organic acids and saccharides by various methods. In the end was compared the values from different methods varieties.
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Fortification of selected cereal products
Hurtová, Jana ; RNDr.Renata Mikulíková, Ph.D. (referee) ; Márová, Ivana (advisor)
Present diploma thesis describes the study of bioactive compounds in the cereals. The work is focused on possible fortification of cereal products in order to increase nutritional value of final products which is reduced during the food processing and storage. Cereal grains are rich in several phenolic compounds, vitamins and minerals that reduce the risk of chronic diseases. The aim of this study was the preparation and analysis of model fortified cereals using lyophilized fruit originating from Czech Republic. A comparative analysis of content of bioactive substances in commercial and fortified products for children was performed too. The experimental part deals with a content of individual flavonoids by RP-HPLC/UV/VIS method, spectrophotometric determination of total and reducing saccharides and analysis of mono- and disaccharides by the HPLC/RI method as well. Differences in the content of individual flavonoids and saccharides after the application of acid hydrolysis of samples were investigated. The content of these compounds increased in most products because of their release from glycosidic forms. Presence of phenolics was confirmed in all analyzed cereals. Addition of 10% of lyophilized fruit/vegetable preparative (carrot, berries, apples) caused several fold increase of phenolics content in model mixtures. In commercially available cereal products there are relatively high differences in total phenolics content, proportionally to fruit component amount. The work included the spectrophotometric analysis of total polyphenols and flavonoids in a model physiological environment (artificial stomach juice) in which the hydrolysis of bound phenolic compounds occurred. The results of this study show that addition of lyophilized fruit seems to be a good way to increase the nutritional value and maintain the content of bioactive compounds in cereal products.
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Analysis of saccharides and markers of wood combustion in atmospheric aerosols
Kubátková, Nela ; Mravcová, Ludmila (referee) ; Mikuška,, Pavel (advisor)
Diploma thesis deals with the analysis of saccharides and markers of biomass and wood combustion in atmospheric aerosol, PM2,5. Theoretic part is focused on general characterisation of atmospheric aerosols and on the properties, sources and analysis methods of selected analysed organic compounds. The experimental part is focused on the optimization of method for simultaneous analysis of saccharides and tracers from biomass and wood combustion. Optimization of method includes selection of solvent for the extraction of compounds and optimization of derivatization process and GC/MS analysis. The optimized method was then applied for the analysis of selected compounds in real aerosol samples in the size fraction PM2.5. The concentrations of analysed compounds were compared in term of sampling seasons.
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Analysis of some active substances in different products from rice
Valentová, Radka ; Melounová, Jitka (referee) ; Márová, Ivana (advisor)
Presented diploma thesis is focused on analysis of selected biologically active substances in different kinds of natural and peeled rice. In rice samples phenolic compounds, antioxidant activity, content of saccharides and vitamin C were analysed. Active substances and saccharides were analysed by UV-VIS spectrophotometry and RP-HPLC-UV/VIS. Content of phenolic compounds and saccharides was measured before and after acid hydrolysis. The content of these compounds increased several times because of release of active substances from glycosides. As a part of diploma thesis basic sensory analysis was performed and consumer questionnaires were evaluated. The highest level of antioxidant activity was detected in Indian Wild rice. The highest content of total polyphenols and flavonoids was detected in Red Rice Natural before acid hydrolysis. After acid hydrolysis the highest content was detected in Indian Wild rice. The highest content of individual flavonoids, phenolic acids and catechins was detected in natural rice Indian Wild, Rice Three colours and in two samples of rice parboiled. The highest content of total and reducing saccharides was found in rice Indian Wild before hydrolysis, while after hydrolysis the content was very similar in all rice samples. The amount of individual monosaccharides after acid hydrolysis increased, while total content of disaccharides decreased. The highest value of simple sugars was detected in rice parboiled and Indian Wild rice. The content of vitamin C was detected only in four rice kinds - in two samples of rice parboiled, Rice Three colours and rice Indian Wild.
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