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HPLC in quality control of food supplements based on goldenrod and containing rutin and chlorogenic acid
Kotyková, Klára ; Šatínský, Dalibor (advisor) ; Chocholoušová Havlíková, Lucie (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Klára Kotyková Supervisor: prof. RNDr. Dalibor Šatínský, Ph.D. Title of the diploma thesis: HPLC in quality control of food supplements based on goldenrod and containing rutin and chlorogenic acid In this diploma thesis, a high-performance liquid chromatography (HPLC) method was developed and validated for the separation and determination of chlorogenic acid, 3,5- dicaffeoylquinic acid, rutin, quercetin, and kaempferol. This method was applied to the analysis of food supplements based on goldenrod: drops Celík zlatobýl (Valdemar Grešík), Celík obecný BIO (Bylinné kapky), Celík obecný (Bylinné kapky), Zlatobýl (Green Idea), Zlatobýl - tinktura z pupenů (Serafin), Zlatobýl - tinktura z bylin (Serafin), Zlatobýlová směs (Dědek kořenář), Zlatobýl (Inca Botanica), and capsules Celík (Ex Herbis). Furthermore, two food supplements containing rutin (Solgar and Walmark®) and a food supplement Zelená káva (Botanic) containing chlorogenic acid were analyzed. An Ascentis® Express 90 Å AQ-C18 150 × 4.6 mm column with 2.7 µm particles was selected for analysis. A gradient elution with a mobile phase of acetonitrile/0.085% phosphoric acid was used. Detection was performed with a diode array detector at...
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Synthesis of novel quinazoline derivatives as a potential multipotent therapeutics against Alzheimer's disease
Tošnerová, Daniela ; Karabanovich, Galina (advisor) ; Matouš, Petr (referee)
1 ABSTRACT Charles University, Faculty of Pharmacy in Hradec Králové Department of: Organic and Bioorganic Chemistry Mentor: Ing. Galina Karabanovich, Ph.D. Consultant: Ing. Barbora Svobodová, Ph.D. Student: Daniela Tošnerová Title of Thesis: Synthesis of novel quinazoline derivatives as a potential multipotent therapeutics against Alzheimer's disease Alzheimer's disease (AD) is a severe neurologic disorder. Current treatments only temporarily delay the progression of disease. The main limiting factor for developing of new therapeutic compounds is the simultaneous presence of multiple pathologies. Thus, a new group of potential drugs called multi-target directed ligands (MTDLs) emerged as an alternative option to combat AD. To this date, dozens of MTDLs have been published. As part of the experimetnal section of this thesis, a series of small molecules based on quinazoline scaffold was rationally designed with the intention to influence the activity of acetylcholinesterase, butyrylcholinesterase, monoamine oxidase A and B (MAO-A/MAO-B), and the GluN1/GluN2B subunit of N-methyl-D-aspartate receptor (NMDAR). Subsequently twenty-four new quinazoline derivatives were synthesized, and biologically evaluated. The overall results highlighted compound II-6h ((5E)-8-chlor-N-cyklohexyl-3-methyl-1-...
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The genus Neotinea Rchb.f. in Europe
Vavřina, Štěpán ; Chumová, Zuzana (advisor) ; Vosolsobě, Stanislav (referee)
The thesis deals with the genus Neotinea from the phylogenetic, molecular and ecologic points of view. Most of its species have been classified in the genus Orchis until the use of molecular methods. Within the Orchidoideae, Neotinea forms a sister clade to the genera Anacamptis, Serapias, Ophrys and Himantoglossum. Six species are accepted now. The current taxonomic circumscription calls for a revision under the influence of new knowledge. Especially in N. tridentata, which contains several well-differentiated lineages and is the ancestor of the tetraploid N. commutata. Also the relationship between N. lactea and N. conica remains unresolved. The genome size within the genus ranges from 6.48 pg in N. maculata to 31.14 pg in N. lactea. All species except N. commutata, whose tetraploid origin is debated, are diploid (2n = 42). Partial endoreplication is present in the genus. The range of endoreplicated DNA is relatively narrow regardless of differences in genome size. Species prefer open, slightly basic habitats. In mycorrhizal interactions they appear generalistic with a preference for Ceratobasidiaceae. At the level of reproductive strategies, the genus Neotinea prefers the generative mode over the vegetative one. N. maculata is autogamous, the other species are allogamous, food deceptive, with...
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Derivatives of boronic acids as potential drugs I.
Koucká, Kateřina ; Kučerová, Marta (advisor) ; Zitko, Jan (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Pharmaceutical Analysis Student: Kateřina Koucká Supervisor: PharmDr. Marta Kučerová, Ph.D. Consultant: PharmDr. Petr Šlechta Title of diploma thesis: Derivatives of boronic acids as potential drugs I. The thesis includes the design, synthesis, and biological evaluation of boronic acid derivatives. These derivatives were prepared as hybrid compounds containing an amide linker, combining mainly pyrazinoic acids with 4-aminophenylboronic acid. Pyrazinamide is used as 1st line antituberculosis drug and 4-aminophenylboronic acid is a bioisoster of 4-aminobenzoic acid, which is a crucial precursor in the folate pathway. Bioisosteric replacement of the carboxylic group with boronic acid could afford the ability of the compounds to form a reversible covalent bond toward a potential biological target. The presented compounds were synthesized in a two-step reaction. The first step was the condensation of 4-aminophenylboronic acid pinacol ester with different derivatives of (hetero)aryl carboxylic acids, that underwent previous activation. The second step was the deprotection of boronic acid pinacol ester to obtain free boronic acids derivatives. The obtained compounds were screened for their in vitro...
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Effect of dysbiosis on proportion of particular neutrophil subsets and their functional capacities
Sklenářová, Lydie ; Hrdý, Jiří (advisor) ; Dobeš, Jan (referee)
The gut microbiota is crucial for maintaining physiological balance and influences metabolic processes, immune responses, and intestinal barrier function. Dysbiosis, or the imbalance of microbial composition, is associated with a range of health complications, including chronic inflammatory conditions such as non-specific intestinal inflammations. Inflammatory processes associated with dysbiosis and changes in microbial metabolites can directly affect the activation of neutrophils, impacting the pathogenesis of various diseases. Probiotics, defined as live microorganisms which, when administered in adequate amounts, confer a health benefit on the host, offer the potential for positive modulation of these inflammatory conditions The aim of this thesis was to explore how experimentally induced intestinal dysbiosis affects the heterogeneity of neutrophils in the bone marrow. Dysbiosis was induced by administering antibiotics to mice, which were subsequently treated with the probiotic strain Escherichia coli O83:K24:H31 (EcO83). Neutrophil phenotypes were assessed using flow cytometry based on the expression of surface markers CD11b, Ly6G, CD62L, and CXCR2. Meanwhile, gene expression related to their antimicrobial functions and the inflammatory environment was analyzed by quantitative PCR. The results...
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Development of LC-MS method for compounds used for determination of cytochrome activity
Smolík, Martin ; Lochman, Lukáš (advisor) ; Horáček, Ondřej (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Pharmaceutical Analysis Candidate: Bc. Martin Smolík Supervisor: PharmDr. Lukáš Lochman, Ph.D. Consultant: prof. PharmDr. Petr Pávek, Ph.D. Title of thesis: Development of LC-MS method for compounds used for determination of cytochrome activity. Cytochromes P450 (CYP450) are enzymes that play a key role in the metabolism of xenobiotics and some endobiotics (cholesterol, steroid hormones, and eicosanoids). They are present in the liver, gastrointestinal tract, lungs, and kidneys. Determination of CYP450 activity is a valuable tool to detect drug interactions resulting from inhibition or induction of various isoforms of this enzyme. This thesis is devoted to the development and validation of a new analytical method using liquid chromatography coupled with tandem mass spectrometry for the determination of activity CYP450 isoforms 3A4, 2C9, and 2B6, which are crucial for the metabolism of many drugs and biologically active substances. In the framework of this thesis, a new method was developed providing a fast, accurate, and reliable measurement of the activity of these enzymes based on the quantification of specific CYP450 substrates, namely midazolam (3A4), diclofenac (2C9), bupropion (2B6) and their...
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Food composition and feeding selectivity of zooplankton
Lis, Jonáš ; Škaloud, Pavel (advisor) ; Čablová, Radka (referee)
Zooplankton are an important link in the food web between phytoplankton and higher tro- phic levels, and understanding their feeding preferences is the key to unraveling the functions and dynamics of aquatic ecosystems. Current knowledge of zooplankton food composition is limited, consisting mainly of laboratory experiments that have revealed different hunting strategies and selection of prey based on size and nutritional value. Recently, publications have emerged that, through the use of DNA metabarcoding, provide a completely new and comprehensive insight into the diversity of zooplankton diet composition, especially copepods, cladocerans and rotifers. These methods reveal that the feeding niches of copepods and cladocerans are distinct and do not overlap. In general, both groups are said to prefer mainly diatoms as their food, when available. The food niches of rotifers are so diverse that no clear trend can be established for the entire phylum. Cyanobacteria appear to be the main source of energy for zooplankton and are proba- bly consumed by metazooplankton largely as secondary prey when consuming ciliates and other heterotrophic prey. Furthermore, differences in feeding niches between species are known and can be partly explained by their vertical distribution, and different levels of feeding...
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Development of sensitive determination of diclofenac using fluorescent properties of complexes with cyclodextrins
Pekařová, Ivana ; Sklenářová, Hana (advisor) ; Chocholouš, Petr (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of analytical chemistry Candidate: Ivana Pekařová Supervisor: doc. PharmDr. Hana Sklenářová, Ph.D. Title of diploma thesis: Development of sensitive determination of diclofenac using fluorescent properties of complexes with cyclodextrins This experimental thesis deals with the development of a sensitive determination using the formation and detection of a complex consisting of an active substance and cyclodextrin (CD) using sequential injection analysis (SIA). A spectrofluorometer was used for subsequent detection. The tested biologically active substance was diclofenac in the form of sodium salt. In the beginning the conditions for setting the spectrofluorometer, namely the appropriate excitation and emission wavelengths, which were 364,5 and 290 nm, slit size and detection sensitivity were found. After that, 13 CDs were used in order to find the appropriate type and ratio of CD with diclofenac so that the measured fluorescence intensity differs from the intensity of CD alone. This testing divided the CDs into 4 groups. When testing the stability of selected complexes, the increase or decrease in fluorescence intensity over time was monitored to ensure the formation of complexes with CD. SIA was used to automate the determination,...
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Political trials of the 1950s: a case study
Svoboda, David ; Kuklík, Jan (advisor) ; Blažek, Lukáš (referee)
81 Political trials of the 1950s - a concrete case study (The Babice Case) Abstract The subject of the thesis "Political Trials of the 1950s - a Specific Case Study", subtitled The Babice Case, was a description of the application of the general tools of political persecution used in the Czechoslovak Republic in the 1950s to the specific case of the shooting at the Babice school and the subsequent political trials. Also significant is the aim of placing the political trials in Czechoslovakia in the context of the times and the international political situation, which played an important role in the Babice case. The first chapter of the thesis formulates the aims of the thesis and the background on which it is based. The second chapter of the thesis describes the significance of the political processes and the general background of their organisation. An important part of Chapter 2 of the thesis is a description of the legal framework for the conduct of political trials, focusing both on substantive aspects, such as the newly defined offences, and on procedural issues, especially the limitations on the rights of the accused. The legal biennium, an initiative aimed at changing the legal system to reflect the needs and ideology of the Communist Party, is also described as a significant phenomenon. The third...
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Interference in the photochemical generation of volatile Se species using TiO2 photocatalyst
Nentvich, Daniel ; Nováková, Eliška (advisor) ; Hraníček, Jakub (referee)
The diploma thesis deals with the study of interferences in the method of UV-photochemical vapour generation of volatile Se species in formic acid media catalysed by nanocrystalline titanium dioxide. High resolution continuum source atomic absorption spectrometry was used for detection. It was observed that SeVI was less resistant to pH change relative to SeIV , while at pH = 2.3, a 20% increase in response was observed for this specie. It was further observed that both the generated volatile SeIV and SeVI species are resistant to UV radiation in the gas phase. The Cu2+ and Ag+ ions were particularly significant interferents in the SeIV determination. Interference behaviour was also observed for Cd2+ ions and silver nanoparticles. In the case of Cd2+ ions, the generation of cadmium selenide was likely observed. For Zn2+ , Fe2+ , Co2+ ions, there was an increase in response of more than 30% and 40%, in case of Ni2+ ions. The Co2+ and Ni2+ ions were chosen as reaction modifiers. For the UV-photochemical vapour generation of selenium species with photocatalyst and reaction modifiers, there was a 4-fold improvement in detection limits and limits of quantification compared to the method without these modifiers. In addition, the overall generation efficiencies for SeIV (64.7 %) and SeVI (68.6 %) were...
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