Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Botany Candidate: Ivana Lakatošová Supervisor: PharmDr. Anna Hošťálková, Ph.D. Title of Master's thesis: Bioactive metabolites of representative members in the genus Magnolia 2019, 72 p. Key words: Magnolia, secondary metabolites, traditional medicine, CNS bioactivity The aim of this Master's thesis was to summarise current knowledge of significant secondary metabolites of species Magnolia officinalis and Magnolia obovata, and their biological activity. Botanical features of genus Magnolia were defined and described, followed by the classification of bioactive compounds isolated from species Magnolia officinalis and Magnolia obovata. The Master's thesis focused predominantly on the effects of biphenol compounds of magnolol and honokiol, which have been subjected to many studies. The main attention was given to the immunosuppressive effect and pharmacologic effects on the nervous system. Essential oils were observed to have an anti-inflammatory and a myorelaxant effect. Alkaloids and neolignans, which had been known to relax spasms of skeletal muscles, expressed neuroprotective activity. Neolignans were shown to have the inhibitory effect on bacteria, viruses and pathogenic fungi. They also proved themselves to be...

Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Simona Rudišarová Supervisor: PharmDr. Marie Vopršalová, CSc. Title of diploma thesis: The effect of quercetin metabolite on the rat isolated arteries This study is focused on the research of the effects of flavonol quercetin and its selected metabolites (3-(3-hydroxyphenyl)propionic acid and 3-hydroxybenzoic acid) on blood vessels in vitro. A standardized method of an isolated rat aorta was used. The aim of this study was to find out how substances act on the isolated aorta and if the effect of the effective metabolite is dependent on the presence of the endothelium in the vessel. After precontracting with noradrenaline, individual concentrations of the test substances were added to the aortic ring bath (final concentrations in the bath ranging from 10-7 to 10-3 M). From the measured values EC50 values (quercetin = 3,63 . 10-5 M, 3-(3-hydroxyphenyl)propionic acid = 3,21 . 10-6 M, 3-hydroxybenzoic acid = 4,38 . 10-5 M) were obtained and the corresponding DRC curves were constructed using S.P.E.L. Advanced Kymograph Software. The experiment was performed once again with quercetin and 3-(3-hydroxyphenyl)propionic acid on the endothelium-denuded vessel. According to the results is...

Klíčová A.: Alkaloids from wood of the species Liriodendron tulipifera L. and their biological activity. Diploma thesis, Charles University, Faculty of Pharmacy in Hradec Králové, Department: Pharmaceutical Botany, Hradec Králové, 2018. Supervisor: PharmDr. Anna Hošťálková, Ph.D. Key words: Liriodendron tulipifera, secondary metabolites, alkaloids, biological activity Alzheimer's disease (AD) is the most common type of dementia. It is neurodegenerative disorder. Patients suffer from cognitive disorder in combination with aphasia, apraxia or disorders of executive functions, which leads to exacerbation of working or social skills. Nowadays, there exist no casual treatment for AD, and that is why the intense research is still waging. Currently there are available remedies, which can slow down progress of AD. These drugs includes also natural substances. An alkaloid extract of Liriodendron tulipifera L. wood showed promising inhibition of human cholinesterases in a preliminary testing. These results were the reason why this extract has been chosen for further isolation of single alkaloids and identification of their biological activity. Isolation was made by column chromatography with step gradient elution. Then was used preparative TLC to isolate single alkaloids. Identification of alkaloids was...

The object of this thesis was the study on the pharmacokinetics and biotransformation of selected abused new synthetic drugs, specifically belonging to cathinone and aminoindane groups, in experimental rats. This work consisted of two parts: a biotransformation study of mephedrone, methylone, 5,6-methylenedioxy-2- aminoindane (MDAI) and a distribution profile study of mephedrone, methylone, MDAI and naphyrone. This document is presented as a commentary to the original papers, which were published in peer reviewed journals. The samples of blood and tissue (brain, lungs, liver) were taken at specific times after a subcutaneous single bolus dose of the studied drugs. The urine samples for the biotransformation studies were collected over a 24-hour period following the sc. dose. The LC/HRMS method was used for metabolite detection and quantitative analysis. Our findings not only included testing metabolites of the aforementioneddrugs against the synthesized reference standards, but also discovering previously unknown metabolites of these drugs. Ten phase I and five phase II metabolites aside from the parent drug were identified in urine after mephedrone administration by comparison of their spectra against those of reference standards and/or MS2 spectra of previously identified metabolites. The main...

Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Botany and Ecology Candidate: Jana Puzyrevská Supervisor: PharmDr. Anna Hošťálková, Ph.D. Title of Thesis: Biological activity of plant metabolites XXXIV. Alkaloids from the herb of Glaucium flavum CRANTZ and their impact on human cholinesterases. Key words: Glaucium flavum Crantz, cataline, N-methyllaurotetanine, norchelidonine, protopine, AChE, BuChE. Alzheimer's disease, the most widespread neurodegenerative disease, causes decrease of cognitive functions and dementia. The most effective therapeutic approach is the application of central cholinesterase inhibitors, which alleviate cholinergic deficit in brain and thus improve memory. Currently, intensive investigation of new active compounds including natural substances is carried on. Within the preliminary testing, alkaloid extract from Glaucium flavum Crantz herb showed promising inhibition of human cholinesterases, so it was selected for further examination. The primary alkaloid extract was acquired from dried flowering herb by extraction with ethanol and subsequent liquid extraction at different pH. This extract was treated by preparative thin layer chromatography. The structure of alkaloids was determined by spectrometric methods (MS, NMR) and their optical...

Baková I.: Isolation of alkaloids of the species Magnolia soulangeana Soul.-Bod. and study of their biological activity. Diploma thesis, Charles University, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Botany and Ecology, Hradec Králové, 2017. Key words: Magnolia solulangeana, secondary metabolites, alkaloids, biological activity. Secondary metabolites of plants are responsible for various biological activities. Alkaloids were described as a potentially suitable for Alzheimer's disease therapy (AD) through their inhibition activities against cholinesterases. Nowadays, these substances are important medicine for AD, therefore a screening of herbal drugs is still a current topic. An alkaloid extract of Magnolia × soulangeana flowers was tested in a preliminary testing on anticholinesterase activity. Because of the promising results, it was chosen for an isolation and identification possible effective alkaloids. The extract was separated by a column chromatography using aluminium oxide and a step gradient elution. Alkaloids were isolated by a repeated preparative thin-layer chromatography. Individual alkaloids were identified by a structural analysis (NMR, MS) and then their optical activity was measured. Substances were tested for an inhibition activity against human...

Lipases are enzymes catalyzing primarily the hydrolytic cleavage of triacylglycerol bonds. The production of lipolytic enzymes is known in many microorganisms, especially those who are able to utilize a fatty carbon substrate. Some genera of carotenogenic yeasts are characterized by this ability. Carotenogenic yeasts are characterized primarily by the formation of intracellular carotenoids, lipids and lipid-soluble substances. In addition to these metabolites, they may also produce some biosurfactants. This work deals with the production of extracellular lipolytic enzymes and biosurfactants by carotenogenic yeasts Rhodotorula glutinis, Cystofilobasidium macerans, Rhodotorula mucilaginosa and Sporidiobolus pararoseus cultivated mainly on animal waste fat at various C/N ratios (13, 25, 50, 100). Lipase activity was detected in all strains studied. Enzyme activities were measured by spectrophotometric method. Lipase induction has also been observed during cell growth, where several peaks of lipase activity have been reported, suggesting cell-associated lipase and lipase secreted into the environment. Lipase activities have also been found in cultures on glucose and glycerol carbon substrates. Further, the molecular characterization of lipolytic enzymes was performed using polyacrylamide gel electrophoresis. The formation of biosurfactants is to some extent formed by all strains. In particular, the biosurfactants of C. macerans and S. pararoseus yeast have emulsifying and solubilizing properties. Simultaneously with the production of lipase and biosurfactants, the production of characteristic high value added intracellular metabolites in S. pararoseus and R. mucilaginosa was evaluated too.

The subject of the thesis is the issue of pesticides in the environment and the possibility of their removal through activated carbon. The thesis is divided into two parts, the search part and the practical part. The research part is focused on description of activated carbon. It also deals with the distribution of pesticides and their degradation in the environment. In the practical part, a laboratory experiment was carried out to determine the effectiveness of removing pesticides and metabolites from the test water on two types of activated carbon.

Plants are an important source of secondary metabolites, such as substances that have a beneficial biological effect on the health of humans and they are irreplaceable in the modern medicine. They may operate in prevention and in the treatment of civilization diseases. In terms of content substances, the phenolic come to fore, especially polyphenols, which have been related to a number of overview studies due to their wide distribution and high concentration in plants. They also represent an important part of substances with redox effects present in the human diet and have an important role for the plant itself. Currently, many laboratories are dedicated to pharmaceutical and biological testing of plants. Individual polyphenols in plant matrices are determined primarily by chromatography, electrophoresis methods, mass spectrometry and nuclear magnetic resonance. Valuable information on the content of polyphenols in plant extract can be obtained also by using spectrophotometric methods. The subject of this thesis is to provide information, characterization and comparison of conventional and modern techniques for the determination of natural substances – phenolic compounds using spectral techniques.

(EN) This dissertation thesis contains scientific results achieved in the field of analytical chemistry, particularly liquid chromatography. The major part of the results has been published in prestigious international journals in five papers. In addition to that, relevant yet unpublished results have been included as well. In general terms, the work presented here contributed to the concerted efforts to tackle the current lack of novel antibiotics. Specifically, high-performance liquid chromatography (HPLC) and ultra high-performance liquid chromatography (UHPLC) techniques coupled to a variety of detection systems have been employed for analysis of antibiotics and actinomycete secondary metabolites. The first thematic part describes the development of liquid chromatography methods for analysis of lincomycin precursors, lincomycin precursor analogues, and lincomycin derivatives. The methods have been applied to study lincomycin biosynthetic pathway and obtain improved lincomycin derivatives by mutasynthesis. The second thematic part aims at investigating alternative approaches for analysis of antibiotics. Firstly, the core-shell particle and the sub-2 μm particle chromatographic columns were compared. The core-shell particle columns compatible with HPLC proved to be a convenient alternative to the...