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New procedures in metabolomic analysis of biofluids
Řimnáčová, Lucie ; Čabala, Radomír (advisor) ; Feltl, Ladislav (referee) ; Sýkora, David (referee)
The aim of this thesis has been the research on reactivity of protic metabolites with chloroformates and its application for GC-MS analysis of biofluids. The research was conducted in three separate studies and the results are three new, original methods for GC-MS determination of low- molecular protic metabolites in biological material especially in biofluids. The first study explores the discovery of fast derivatization of alicyclic hydroxyl groups by fluoroalkyl chloroformates (FCFs) under anhydrous conditions [1]. FCF fully converts the hydroxyl group into a corresponding carbonate and the step can easily be coupled with liquid-liquid microextraction (LLME) of the arising derivatives into organic phase. The reaction of the alicyclic OH group with FCFs was tested on 12 clinically relevant steroids and 4 tocopherols. The analytical properties of determined analytes were described and the method was validated for the GC-MS determination of 6 diagnostic sterols and 4 tocopherols in human serum and amniotic fluid. The new method was further successfully used for determination of sterols and tocopherols in tissues of the bug Pyrrhocoris apterus [2]. The second study was focused on the reactions of protic, particularly urinary metabolites with FCFs, mainly heptafluorobutyl chloroformate (HFBCF)....
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Inhibitory effects of B. velezensis strain on plant pathogens of the genus Xanthomonas
Švecová, Magdaléna ; Palyzová, Andrea (advisor) ; Jelínková, Markéta (referee)
Biological control is a method of applied ecology that uses one organism, or its product, to inhibit another pathogenic organism. This method in plant protection should be more environmentally friendly than commonly used pesticides. The rhizobacterium Bacillus velezensis FZB42, which can suppress the growth of other microorganisms, can be used as a biological control agent. Xanthomonas campestris pv. campestris is a phytopathogen that causes significant losses in agricultural production. This diploma thesis proved an antagonistic effect of the bacterial strain B. velezensis FZB42 on the phytopathogenic bacteria Xanthomonas campestris pv. campestris SU in mixed culture. The study confirmed that the antibiotic effect of B. velezensis strain FZB42 against X. campestris pv. campestris SU in mixed culture depends on the inoculation ratios of both strains. The interaction of the two bacterial strains was characterized by metabolomics analysis, it was confirmed that B. velezensis FZB42 produces four dominant secondary metabolites: lipopeptides surfactin, fengycin, and bacillomycin and siderophore bacillibactin, which show antifungal and antibiotic activity. The inhibitory activity of B. velezensis strain FZB42 was further monitored by SEM analysis, which showed damage to X. campesris pv. campestris SU...
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New procedures in metabolomic analysis of biofluids
Řimnáčová, Lucie ; Čabala, Radomír (advisor) ; Feltl, Ladislav (referee) ; Sýkora, David (referee)
The aim of this thesis has been the research on reactivity of protic metabolites with chloroformates and its application for GC-MS analysis of biofluids. The research was conducted in three separate studies and the results are three new, original methods for GC-MS determination of low- molecular protic metabolites in biological material especially in biofluids. The first study explores the discovery of fast derivatization of alicyclic hydroxyl groups by fluoroalkyl chloroformates (FCFs) under anhydrous conditions [1]. FCF fully converts the hydroxyl group into a corresponding carbonate and the step can easily be coupled with liquid-liquid microextraction (LLME) of the arising derivatives into organic phase. The reaction of the alicyclic OH group with FCFs was tested on 12 clinically relevant steroids and 4 tocopherols. The analytical properties of determined analytes were described and the method was validated for the GC-MS determination of 6 diagnostic sterols and 4 tocopherols in human serum and amniotic fluid. The new method was further successfully used for determination of sterols and tocopherols in tissues of the bug Pyrrhocoris apterus [2]. The second study was focused on the reactions of protic, particularly urinary metabolites with FCFs, mainly heptafluorobutyl chloroformate (HFBCF)....
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