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Preparation of a method for detection of the extent of chelation of calcium and magnesium ions
Konečný, Lukáš ; Mladěnka, Přemysl (advisor) ; Jáč, Pavel (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Candidate: Lukáš Konečný Supervisor: prof. Přemysl Mladěnka, Pharm.D., Ph.D. Title of Thesis: Preparation of a method for detection of the extent of chelation of calcium and magnesium ions Calcium and magnesium are biogenic elements belonging to the second group of the periodic table. Calcium is the fifth most abundant element in the human body. In addition to its important role in bone homeostasis, calcium is involved in other processes such as blood clotting, muscle contraction and regulation of nerve excitability Magnesium is the cofactor of many enzymatic reactions essential for a row of physiological functions. The aim of this work was to propose a novel screening method for determination of the degree of calcium and magnesium chelation. In the first step the UV-vis spectra of the selected indicator o-cresolphthalein complexone and its complexes with calcium and magnesium were measured. This was followed by verification of the linearity and the determination of the sensitivity of the method. Concurrently the stability of the indicator and its complexes with both ions was measured. In the last step, the suitability of the method was verified on a standard chelator ethylenediaminetetraacetic acid...
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Search of effective cobalt chelators - 8-hydroxyquinolines
Pelnářová, Karolína ; Mladěnka, Přemysl (advisor) ; Carazo Fernández, Alejandro (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Karolína Pelnářová Supervisor: Prof. Přemysl Mladěnka, PharmD., Ph.D. Consultant: Václav Tvrdý, MSc.; Marcel Hrubša, MSc. Title of diploma thesis: Searching of effective cobalt chelators - 8-hydroxyquinolines Cobalt is one of the essential trace elements present in the human body. It forms a part of the organometallic complex of vitamin B12, which is essential for many physiological functions. Both overload and lack of cobalt in the body is associated with pathological conditions. Manifestations of deficiency can lead up to pernicious anemia or hypofunction of the thyroid gland. Cobalt intoxication can occur in an industrial environment, such as by cobalt metal dust during heavy metal processing, or by its release due to corrosion from orthopedic prostheses. Systemic toxicity is manifested by a number of endocrine, cardiovascular and neurological symptoms. The main aim of this diploma thesis was to find effective chelators of cobalt from the group of substances derived from 8-hydroxyquinoline. In vitro spectrophotometric measurement was used to determine the degree of chelation. The effect of chelation was also monitored ex vivo in rat erythrocytes. All tested chelators - 8-hydroxyquinoline,...
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Studium vlivu požití zeleného čaje na antioxidační stav organismu
Papanová, Veronika
This bachelors thesis Study of green tea ingestion on antioxidant status of the organism is divided into two parts theoretical and practical. The theoretical part deals with materials for the production of green tea and its production, chemical composition, health effect and the impact on the human organism during use. The practical part is about experiment with capillary blood samples using Ellmans method, Bradford assay and High performance liquid chromatography with electrochemical detection. The practical part also including the sensory questionnaire. The results show that green tea is able to increase the substances with antioxidant effect in blood. Values of Ellmans method and HPLC ED analysis are comparable and thiol groups and reduced glutathione growth is observed. According the result of the Bradford assay it seems that green tea has no effect on the protein content in blood. The overall impression of the sensory analysis is satisfactory 73,8 points.
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Technologie výroby a jakost hroznového moštu
Floriánová, Jana
The thesis deals with the production of grapes musts, their quality and sensory profile. The theoretical part describes the chemical composition of grapes, production technology and methods of increasing the shelf life of musts. The practical part is divided into thee larger units. The first part deals with the two varieties used and their origin. It also deals with the technology of grape must production. The second part is devoted to the analysis of must, such as pH, spectrophotometric color measurement and content of ascorbic acid. The last part of the practical part is devoted to sensory analysis of grape musts. There was no difference in pH values that had different pasteurization temperatures. The highest content of ascorbic acid was measured in fresh must, followed by must, which was heated to 80 ℃. The lowest ascorbic acid content was measured in the must, which was heated to 65 ℃. The sensory analysis revealed a difference in colour assessment ant he presence of foreign particles the varieties.
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Izolace, separace a studium významných kov-vazných proteinů
Skaličková, Sylvie
The metal binding proteins are important group of biomolecules effects in living organisms. They represent series of vital important functions such as homeostasis of essential metals, they are involved in heavy metals detoxification, helps antioxidant system with free radical neutralization to prevent oxidation stress. Lot of them belongs to group of important enzymes catalyzing biochemical reactions. Last, but not least they are part of innate immunity system against pathogenes and could be utilized as a carrier cages in target drug therapy due to their unique transporter properties. A number of methods have been developed to study and observe the interactions between metalloproteins and specific biomolecules. First of all, the liquid chromatography enables isolation of required proteins from biological matrices. Further experiments utilized immunochemical, spectrophotometrical and electrochemical methods to clarify their properties and behaviour in various environments. The aim of the thesis was optimized analytical methods for separation and studying of the metal binding proteins.
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Preparation of a novel method for screeing of cobalt chelators
Moravcová, Monika ; Mladěnka, Přemysl (advisor) ; Karlíčková, Jana (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Monika Moravcová Supervisor: Assoc. Prof. Přemysl Mladěnka, Pharm.D., Ph.D. Title of thesis: Preparation of a Novel Method for Screening of Cobalt Chelators Cobalt as a structural part of the vitamin B12 is an essential microelement for living organisms including humans. However, its excess is associated with pathological conditions. Cobalt poisoning can be caused for example by exposure to cobalt metal dust during the production of hard metals or follow the corrosion of metal hip prosthesis. Patients intoxicated by cobalt can develop different manifestations including neurological impairment, hypothyroidism or cardiomyopathy. The aim of this work is to prepare a standardized, rapid, cheap but precise method for the screening of cobalt chelators. For this purpose, spectrophotometric detection using 1-nitroso-2-nafphhol-3,6-disulfonic acid disodium salt as the indicator was used. Firstly, it was found that the addition of cobalt ions led to a clear bathochromic shift of the maximum absorbance of the indicator. The relationship between the absorbance and cobalt concentration was highly linear from 470 to 560 nm at all 4 tested pH conditions (4.5, 5.5, 6.8 and 7.5). The sensitivity of the method...
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Adaptogens in the Shizandra chinensis plant
Pospíchalová, Lucie ; Půčková, Helena (referee) ; Kotlík, Josef (advisor)
This diploma thesis deals with plant adaptogens (bioactive substances) in the plant Schisandra chinensis. The theoretical part is focused on characterization of the Schisandra genus, characterization of the Schisandra chinensis as a plant, its properties, occurrence and growing possibilities in the climate zone. Furthermore, The effects of these adaptogens on the organism and the prediction of possible use in food supplements are described. The experimental part deals with the extraction of fruits and leaves from plant Schisandra chinensis. Further, chlorophyll a, chlorophyll b in the leaves extract was determined by UV – VIS spectrophotometry, whereby the optimal amount 3:1 of chlorophyll was confirmed for the plants growing in optimal conditions. Finally, the concentration of the adaptogen schisandrin A in the dried fruits was from different kind of extraction measured by HPLC. In all fruits, schisandrin A was confirmed, but the amount is different in each sample.
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Control and monitoring of parameters in microfluidic systems
Meloun, Jan ; Sekora, Jiří (referee) ; Čmiel, Vratislav (advisor)
This thesis discusses the construction of Microfluidic systems for a microscope. The theoretical part of this thesis deals with light, light’s behaviour and optical spectral analysis in some methods are discussed. Also, the use of the development platform Arduino with wireless data transfer for data visualization is mentioned. Methods for measuring pH are mentioned as well. The practical part is dedicated to the construction of the Microfluidic system with the use of Arduino, smart mobile devices and a developed mobile application including controlling liquid flow with the measured data visualization.
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Interference of reducing agents with the hematoxylin assay for determination of cupric chelation
Zsáková, Emily ; Mladěnka, Přemysl (advisor) ; Vorčáková, Katarína (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Emily Zsáková Supervisor: Assoc. Prof. Přemysl Mladěnka, PharmD., Ph.D. Title of diploma thesis: Interference of reducing agents with the hematoxylin assay for determination of cupric chelation Copper is an essential trace element of living organisms. Disorders of its homeostasis affect various pathological conditions. In excessive amounts, it is toxic due to its ability to form free radicals. Vitamin C, trolox (a water soluble form of vitamin E), glutathione and hydroxylamine possess reducing properties that protect our body from oxidative tissue damage. Oxidized glutathione (GSSG) was also tested for comparison. The object of study was to test the chelating ability of these substances. A simple and fast spectrophotometric method was used. The combination of reducing properties of copper and chelating agents could lead to an improvement in chelation therapy either in heavy metal poisoning, treatment of Wilson's disease, or other diseases. We found that all substances were chelators/copper-chelating agents in vitro in a slightly competitive environment, using hematoxylin as an indicator. However, by using the bathocuproine assays to verify chelating properties, none of the substances...
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Content of biologically active phenolic compounds in cultivated of species of Vaccinium genus.
CARDOVÁ, Michaela
This bachelor´s thesis includes a knowledge about active biologic compounds from the most common grown species of cranberry (Vaccinium) in the Czech Republic. Active substances have been analyzed in varieties of Vaccinium vitis-idaea like Koralle, Runo bielawskie, Sanna, Sussi, Linnea and Ida. Polyphenols are biomolecules, which are founded in products of plants and there have been proved an antioxidant and anti-inflammatory effects in vivo and in vitro (Chiva-Blanch, 2017). In during of evolution, in plants evolved an ability to produce of secondary polyphenolic metabolites, which weren´t intervened to their growth and development but they are very important for environment, for reproduction strategy and mechanism of protection (Cheynier, 2013). Between polyphenols belong to for examples flavonoids and anthocyanins, which are co-pigment of their property. Next analyzed compound was vitamin C alias ascorbic acid. This substance is simple low-molecular-weight carbohydrate. Synthesis of vitamin C is localized of plants and some species of animal, (Lykkesfeldt, 2014). Ascorbic acid is a good antioxidant, which save organism before ROS (reactive oxygen form) and stimulated immune system. The aim of this study was analyzed the content of quercetin, ascorbic acid and the content of total anthocyanin in six selected varieties of cranberry. The determination as such was performed using high performance liquid chromatography (HPLC) and determination of content of anthocyanin were analyzed by use spectrophotometer for define concentration from absorbance of valid sample. We analyzed all of three mentioned kinds of compounds. The higher results of anthocyanin were analyzed from kind Sussi. Concentration was 617 ? 8.7 mg/kg. In kind of Sanna, the ascorbic acid amount to 110 ? 1.4 mg/kg and there were measured the content of quercetin 1019 ? 36 mg/kg in freeze-dryied material.
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