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Detekce nukleových bazí pomocí kapilární elektroforézy
Hendrychová, Kateřina
Capillary electrophoresis (CE) is one of the most important techniques used for high-throughput separations in modern analytical chemistry. Its main advantages include the possibility of using different detection techniques, speed and efficiency of separation as well as reproducibility. These properties are used in the separation of a wide variety of analytes ranging from inorganic ions to nanoparticles and were also verified in this work, in which the basic building blocks of DNA and RNA, nucleic bases, which serve as biomarkers for many disorders, diseases and food safety, were measured using capillary electrophoresis with a UV detector. The bachelor thesis is focused on the development and optimization of a method to detect and separate 5 nucleobases and purine. The separation was achieved by using 40 mM borate buffer pH 9,8 with the addition of 60 mM SDS as a detergent which forms micelles.
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Totální syntéza [15N4] cytokininů a jejich využití
BUČEK, Jan
Cytokinins (CKs) and their metabolites and derivatives are essential for cell division, plant growth regulation and development. They are typically found at minute concentrations in plant tissues containing very complicated biological matrices. Therefore, defined standards labelled with stable isotopes are required for precise metabolic profiling and quantification of CKs, as well as in vivo elucidation of CK biosynthesis in various plant species. In this work, eleven [15N]-labelled C6-purine derivatives were prepared, among them five aromatic (4, 5, 6, 7, 8) and three isoprenoid (9, 10, 11) CKs. Compared to current methods, optimized syntheses of 6-amino-9H-[15N5]-purine (adenine, 1) and 6-chloro-9H-[15N4]-purine (6-chloropurine, 3) were performed to achieve more effective, selective and generally easier approaches. The chemical identity and purity of prepared compounds were confirmed by physico-chemical analyses (TLC; HRMS; HPLC-MS; 1H, 13C, 15N NMR). The presented approach is applicable for the synthesis of any other desired [15N4]-labelled C6-substituted purine derivatives.
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HPLC and LC-MS/MS methods for diagnosis of inherited metabolic diseases
Bártl, Josef
This dissertation thesis is focused on development and optimization of high- performance liquid chromatography (HPLC) and tandem mass spectrometry (LC-MS/MS) methods, and its utility for diagnosis of inherited metabolic diseases. The first thematic part describes a comprehensive laboratory approach to diagnostics of patients with hereditary xanthinuria by determination of specific markers and enzyme activity. For this purpose HPLC method with diode array detection for measurement of hypoxanthine, xanthine, allopurinol and oxypurinol in urine and plasma and HPLC method with fluorimetric detection for analysis of pterin and isoxanthopterin in plasma were employed. These methods were successfully applied in clinical practice to ascertain two patients with hereditary xanthinuria type I. The second thematic part aims at developing and clinical application of new LC-MS/MS method for simultaneous determination of total homocysteine (tHcy), methionine (Met) and cystathionine (Cysta) in dried blood spots (DBS) and plasma. The results demonstrated the clinical utility of this method for detection of patients with homocystinuria and possibility to distinguish between defects in the remethylation and transsulfuration pathways of homocysteine metabolism. Due to ease of DBS collection and sample transportation...
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Gout diet
KOZÁK, Martin
This Bachelor's thesis, that is concerned with the disease gout, is divided into 3 parts. Theoretical part of this thesis is dedicated to the description of this disease, including its treatment and related diet measures, which have to be followed. Part of this study is a research of respondents with gout disease. This research was made as an interview with the respondents and it discovers necessary information including their diet. Results of this research are described at the end of this thesis and it shows that gout disease cannot be treated only with pharmaceutical means, but it is important to follow appropriate diet including an overall change of a life style
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HPLC and LC-MS/MS methods for diagnosis of inherited metabolic diseases
Bártl, Josef
This dissertation thesis is focused on development and optimization of high- performance liquid chromatography (HPLC) and tandem mass spectrometry (LC-MS/MS) methods, and its utility for diagnosis of inherited metabolic diseases. The first thematic part describes a comprehensive laboratory approach to diagnostics of patients with hereditary xanthinuria by determination of specific markers and enzyme activity. For this purpose HPLC method with diode array detection for measurement of hypoxanthine, xanthine, allopurinol and oxypurinol in urine and plasma and HPLC method with fluorimetric detection for analysis of pterin and isoxanthopterin in plasma were employed. These methods were successfully applied in clinical practice to ascertain two patients with hereditary xanthinuria type I. The second thematic part aims at developing and clinical application of new LC-MS/MS method for simultaneous determination of total homocysteine (tHcy), methionine (Met) and cystathionine (Cysta) in dried blood spots (DBS) and plasma. The results demonstrated the clinical utility of this method for detection of patients with homocystinuria and possibility to distinguish between defects in the remethylation and transsulfuration pathways of homocysteine metabolism. Due to ease of DBS collection and sample transportation...
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HPLC and LC-MS/MS methods for diagnosis of inherited metabolic diseases
Bártl, Josef ; Jelínek, Ivan (advisor) ; Čabala, Radomír (referee) ; Cvačka, Josef (referee)
This dissertation thesis is focused on development and optimization of high- performance liquid chromatography (HPLC) and tandem mass spectrometry (LC-MS/MS) methods, and its utility for diagnosis of inherited metabolic diseases. The first thematic part describes a comprehensive laboratory approach to diagnostics of patients with hereditary xanthinuria by determination of specific markers and enzyme activity. For this purpose HPLC method with diode array detection for measurement of hypoxanthine, xanthine, allopurinol and oxypurinol in urine and plasma and HPLC method with fluorimetric detection for analysis of pterin and isoxanthopterin in plasma were employed. These methods were successfully applied in clinical practice to ascertain two patients with hereditary xanthinuria type I. The second thematic part aims at developing and clinical application of new LC-MS/MS method for simultaneous determination of total homocysteine (tHcy), methionine (Met) and cystathionine (Cysta) in dried blood spots (DBS) and plasma. The results demonstrated the clinical utility of this method for detection of patients with homocystinuria and possibility to distinguish between defects in the remethylation and transsulfuration pathways of homocysteine metabolism. Due to ease of DBS collection and sample transportation...
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Možnosti využití hmyzu jako potraviny v podmínkách České republiky
Bednářová, Martina
The growing interest in edible insects in the Czech Republic is confronted with a minimal amount of information available on this subject. Therefore, the main aim of this work was to map the possibilities of using insects as food in the Czech Republic. To meet the objectives, two hypotheses were defined: 1. Nutritional value of insects bred in the Czech Republic is comparable with the nutritional value of common foods and insects is the alternative in human nutrition. 2. The professionals and laymen in the Czech Republic are ready for including insects as food in the consumer basket. Methodically the objectives were addressed through: a) analyzes of selected nutritional parameters of seven insect species: migratory locust (Locusta migratoria), mealworm (Tenebrio molitor), superworm (Zophobas atratus), honey bee (Apis mellifera), wax moth (Galleria mellonella), silkworm (Bombyx mori), field cricket (Gryllus assimilis); b) analyzes of purine derivates in three species of insects: mealworm (Tenebrio molitor), superworm (Zophobas atratus), field cricket (Gryllus assimilis); c) a survey regarding the use and insect breeding among the consumers, breeders and processors; d) gathering information of the legislative framework from legal authorities of the Czech Republic and the Novel Food Working Group of European Union in Brussels. Analyses proved a high nutritional value of selected species, but also confirmed statistically significant differences in the nutrient content of nutrientss and purines between the species. The survey confirmed the different public attittude to the species, the best score was gained by mealworm (Tenebrio molitor), the worst by silkworm (Bombyx mori). Breeders are willing to produce edible insects for entomophagy. Analysis of legislation and questioning showed that the legislative framework for entomophagy does not yet exist in the Czech Republic. Aims of the study were met, Hypothesis 1 was confirmed completely, Hypothesis 2 only partially. While consumers, farmers and processors are prepared for the edible insects to enter the market in the Czech Republic, the legislative framework is yet to be created.
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