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Use of Kluyveromyces marxianus to bioethanol produce from waste paper
Tomečková, Andrea ; Hrstka, Miroslav (referee) ; Babák, Libor (advisor)
The diploma thesis is focused on production possibilities of bioethanol from waste paper by yeast Kluyveromyces marxianus. Waste cardboard was used as a potential substrate for bioethanol production. Several methods for cardboard preparation were introduced and compared as well as methods of fermentation. Simultaneous sacharification and fermentation and separate hydrolysis and fermentation of preprepared cardboard paper were performed in different pH buffer (4,8-7). Simultaneous sacharification and fermentation was held at a temperature of 45°C. Hydrolysis in separate hydrolysis and fermentation was performed at 50°C and fermentation at 25°C. Procedures outputs were obtained by sampling in specific time intervals and samples were analyzed by HPLC for presence and concentration glucose and ethanol. The results of the analysis have shown that the highest concentration of glucose produced by enzymatic hydrolysis was achieved by using microwaves, 2% H2SO4 and 2% NaOH pretreated paperboard at pH 4,8. The highest yield of ethanol was obtained by separate hydrolysis and fermentation of pulp pretreated by microwaves, 2% H2SO4 and 2% NaOH in pH 5,4 buffer. The method SHF proved to be more effective for the production of ethanol than SSF.
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Alcohol-free beers with herbal extracts
Vyskočilová, Terezie ; Lichnová, Andrea (referee) ; Márová, Ivana (advisor)
This bachelor thesis deals with alcohol-free beers and herbal extracts, which are the beers flavoured with. In the theoretical part the methods of non-alcoholic beer production and the representation of the active ingredients in herbs are described. A total of 7 herbal extracts were prepared and added into 3 non-alcoholic Czech beers. Each variant was examined in order to detect changes of the content of active substances before and after the addition of the herbal extract. The content of polyphenols, flavonoids, bitter substances and iso-bitter acids as well as antioxidant activity were analyzed spectrophotometrically. Using HPLC/UV-VIS analysis a content of catechins was assayed. The two selected flavours were encapsulated into liposome; the particles were tested of size and stability. Sensory analysis of all herbal extracts including the encapsulated ones was performed to determine the response of consumers and flavour preferences.
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Analytical Evaluation of Biologically Active Substances by Chromatographic Methods
Pasáková, Ivana ; Sochor, Jaroslav (advisor) ; Nobilis, Milan (referee) ; Holčapek, Michal (referee)
Introduced dissertation thesis deals with the development and validation of the chromatographic methods for analytical evaluation of selected biologically active substances. HPLC coupled with UV and MS detection was chosen for determination of analytes, because of the dominant role of the HPLC in pharmaceutical analysis. The theoretical part is focused on the theory of chromatographic methods and topics of the experimental work. At first, summary of all the chromatographic methods is briefly introduced and subsequently, the most common analytical method HPLC is described in details. The next part deals with the mass spectrometry. Thank to its high sensitivity and ability to provide structural information about the analytes, MS became an indispensable tool not only in modern pharmaceutical analysis. Besides MS theory and instrumentation, its applications and new trends are also mentioned. Last chapters deal with the transdermal application of drugs, specifications of antiretroviral therapy and especially they provide basic information about the physical - chemical and biological properties of analysed substances. The experimental part is consisted of the original research papers with appropriate comments divided into two thematic sections. The first one is composed of three papers focused on analytical...
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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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HPLC Analysis of Anthelmintics Using Core-shell Columns
Marková, Jitka ; Kubíček, Vladimír (advisor) ; Lázníčková, Alice (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biophysics and Physical Chemistry Candidate: Jitka Marková Consultant: Ing. Vladimír Kubíček, CSc. Title of Thesis: HPLC Analysis of Anthelmintics Using Core-shell Columns This diploma thesis deals with HPLC analysis of three anthelmintics included into AAD group (i. e. amino-acetonitrile derivatives). Suitable separation conditions were studied using three core-shell columns (i. e. Ascentis Express RP - Amide, Ascentis Express F5 and Kinetex PFP). Finally, an efficiency of two preextraction methods (SPE, LLE) was tested. The obtained results show the Ascentis Express RP-Amide column to be the suitable choice for the required purpose. The proposed mobile phases are MS compatible. SPE column STRATA X, exhibiting very high recoveries, is the best choice for SPE. The LLE recovery ratio can be taken as satisfactory.
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Methods for the extraction of selected toxins from solid matrices and their subsequent determination by HPLC/MS
Herman, David ; Zlámalová Gargošová, Helena (referee) ; Čáslavský, Josef (advisor)
This diploma thesis is focused on the analysis of toxins and their extraction from sandyloamy soil. Particularly, saxitoxin and trichothecene mycotoxins deoxynivalenol, HT-2 and T-2 toxins are in the centre of interest of this work. Their occurrence, toxic properties and influence on living organisms are described in theoretical part of this thesis. In next chapters, currently published extraction methods for individual toxins and analytical approaches for their quantitative evaluation are summarized. In experimental part of this thesis, optimized process of sample pretreatment based on extraction of toxins from soil using 1mM HCOONH4 in 84% acetonitrile was proposed as the best option. Simultaneous determination of toxins was performed by liquid chromatography on a CN column (3.0 x 150 mm, 3 m, 100 ) in gradient elution mode. Mass spectrometer with electrospray as ion source and linear ion trap as analyzer was used as detector. Recovery of designed method was over 80% for trichothecene mycotoxins and 51% for saxitoxin.
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HPLC determination of selected incestisides in cosmetic
Kameníčková, Daniela ; Šatínský, Dalibor (advisor) ; Chocholouš, Petr (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Daniela Kameníčková Supervisor: Doc. RNDr. Dalibor Šatínský, Ph.D. Title of Diploma Thesis: HPLC determination of selected incestisides in cosmetic Active ingredients fenoxycarb and permethrin were determined in cosmetic anti- parasitic product Arpalit® Neo shampoo against parasites with bamboo extract. Analysis was performed by HPLC using RP-Amide column 100 x 3 mm with a particle size of 2.7 micron and detection by UV at a wavelength of 225 nm. Separation proceeded by isocratic elution with mobile phase of acetonitrile - water (55:45) at a flow rate 1.0 ml/min. Temperature during measurement was 60 řC. For evaluation method was used an internal standard method, as the internal standard was used dye sudan II. For separation was needed time interval of 6 min. The retention time of fenoxycarb was 1.01, sudan II 2.87 and permethrin 4.42 min. Resolution of chromatographic peaks fenoxycarb and sudan II was 10.612 and sudan II and permethrin 6.524. Keywords: fenoxycarb, permethrin, HPLC.
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Analytical evaluation of drugs using chromatographic methods III.
Strýčková, Kristýna ; Mokrý, Milan (advisor) ; Sochor, Jaroslav (referee)
Analytical evaluation of drugs using Chromatographic Methods III. Thesis Kristýna Strýčková Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Chemistry and Drug Control The thesis draws up the optimum chromatographic conditions for the HPLC analysis of fumagillin in honey and the procedure of the isolation of the drug from honey. The optimum results were provided by the analysis on the Tessek column with the Separon SGX NH2 filling, while using the mobile phase composed of methanol : ammonium acetate (aqueous solution 0.005 mol/l, v/v) in the ratio of 60:40, at a flow rate of 1 ml/min and pressure of 25 to 26 bar. The detection was performed at 335 nm using the UV detector. The internal standard method was drawn up for the determination of fumagillin in honey. Nimesulid suited the selected conditions most. As far as the validation parameters are concerned, the linearity was verified on the basis of calibration curve.
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Electrophoretic determination of 8-hydroxy-2-deosyguanosine as the marker of oxidative stress
Bydžovská, Lucie ; Polášek, Miroslav (advisor) ; Pospíšilová, Marie (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Anylytical Chemistry Candidate: Lucie Bydžovská Supervisor: . RNDr. Miroslav Polášek, CSc. Title of diploma thesis: Electrophoretic determination of 8-hydroxy-2'-deoxyguanosine as a marker of oxidative damage Abstract 8-Hydroxy-2'-deoxyguanosine (8-OHdG) is probably the most important and well-documented product of oxidative DNA damage in biological systems. The study of oxidative DNA destruction is valuable in biomedical research and in diagnostics concerning cancer and ageing. Capillary electrophoresis (CE) has been tested as a highly efficient separation method for the analysis of 8-OHdG as an alternative to conventionally used HPLC. The aim of this work was to develop CE/UV method for the determination of 8-OHdG in urine samples. Optimal CE separation of 8-OHdG from other urine components was achieved with background electrolyte based on 150mM borate buffer of pH* 9.2 (adjusted with NaOH) containing 10% (v/v) of acetonitrile; 2'-deoxyguanosine was selected as optimal internal standard (IS). The detection wavelength was 282 nm. The samples analysed were standard aqueous solutions of 8-OHdG + IS and human urine spiked with 8-OHdG + IS. For increasing the sensitivity of the CE special separation capillary equipped with a...
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