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HPLC evaluation of tyrosine and its metabolites
Michailellis, Panagiotis ; Kastner, Petr (advisor) ; Kučera, Radim (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Pharmaceutical Analysis Author: Panagiotis Michailellis Supervisor: PharmDr. Petr Kastner, PhD. Title of Diploma Thesis: HPLC evaluation of tyrosine and its metabolites Tyrosine is an important precursor of catecholamines, which are dopamine, norepinephrine (noradrenaline), and epinephrine (adrenaline). These are neurotransmitters and hormones, crucial for every living organism. Therefore, their identification and evaluation in biological material would aim to understand their function and behavior more accurately. This Diploma Thesis is a review on how to evaluate catecholamines and their metabolites in biological matter with analytical methods used in Pharmaceutical Analysis, especially with HPLC. First chapters present theoretical knowledge about metabolism of tyrosine and its metabolites, how a sample from an organism should be treated in order to be examined, information about HPLC, CE, GC apparatuses and the main detectors used in analysis of these compounds. In literature review, tables are presented, concerning different sample preparation methods, HPLC details and characteristics, and finally a sum of all the analytical methods studied for this Diploma Thesis. All the articles and...
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Molecular mechanism of DNA regulatory segment recognition by MADS box family transcription factors
Profantová, Barbora ; Štěpánek, Josef (advisor) ; Jelínek, Otakar (referee) ; Manfait, Michel (referee)
The thesis deals with physico-chemical properties of the MADS box, binding domain of transcription factors, which are important for the formation of complexes with the DNA regulatory segment bearing the CArG box. The study was performed also on model oligopeptides, selected segments of the MADS box and their analogues with a point mutation. A wide range of spectroscopic techniques was employed, namely absorption, circular dichroism, fluorescence and Raman spectroscopies. Advanced approaches including multivariate methods were used for data processing. The three tyrosines of the MADS box located in amino-acid vicinities of different charge and hydrophobicity, were used as intrinsic spectroscopic probes. The obtained characteristics of the MADS box and its segments structural arrangement, flexibility and acid-base equilibria are the main results of the work.
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Biogenic amins in beer
Zima, Matěj ; Orsák, Matyáš (advisor) ; Braný, Pavel (referee)
One of the most popular and widely used alcoholic beverages is beer. There is a long-standing tradition of beer production in the Czech Republic and it is our significant export commodity. Hops, specifically the so called Saaz Late, is a typical ingredient in the production of bottom fermented a Pilsen kind of beer that has made us famous in the world.
Beer is a complicated beverage and its production is not an easy process, although it may seem differently at first glance. Apart from the convenient ingredients, for the production of beer are also needed technological facilities and most importantly the knowledge. The procedure itself consists of grinding malt, mixing it with water and mashing process, which includes a shortage of polysaccharide chains by influence of malt enzymes. The extracted product is called hopped wort and it is further boiled together with hops.
This process is in Czech language known as chmelovar (hereinafter referred to as wort). During the wort, there are some bitter substances coming to work (this is how mixture of ingredients and water is generally called during the whole process of production), the bitter substances are specifically alfa bitter acids and beta bitter acids.
The extracted product is called hopped wort. After fermenting the hopped wort, there comes a stage of main and secondary fermentation. During this phase two crucial ingredients of beer are produced. Alcohol and carbon dioxide. Traditional for Czech regions is the bottom fermentation in which yeasts are descending to the base of a container.
Biogenic amines are one of many components of the beer. However, they can be found not only in the beer, but also in almost all basic foods. We can find them in meat, fish, vegetables and fruits, and even in different concentrations. Their production is often associated with the fermentation process. Regarding the content of biogenic amines in beer, due to the research it is obvious that a part of them gets into the beer from the original raw materials. Another part arises during the fermentation process. Tyramine and histamine (among others), discovered in the beer, can have a serious impact on human health. During the consumption of large quantities of such a beer, but even other foods that contain a lot of these biogenic amines, it may affect the circulatory system, breathing, cause a facial flushing and in extreme cases even cerebral haemorrhage. The intoxication by histamine is often manifested as food poisoning or allergic reactions.
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