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Development of mass spectrometric methods for structural analysis of fatty acids and estolides
Cudlman, Lukáš ; Cvačka, Josef (advisor) ; Kozlík, Petr (referee) ; Lísa, Miroslav (referee)
(EN) This doctoral thesis summarizes the use of modern mass spectrometric methods for the structural identification of fatty acids and triacylglycerol estolides (TG-EST) and their subsequent characterization in the lipidome of vernix caseosa, i.e., the biofilm covering the skin of the human fetus during the last trimester of in-utero development. Vernix caseosa has mainly antimicrobial properties and the ability to heal wounds and burns. The first part of the doctoral thesis focuses on the diversity of fatty acids in the lipidome of vernix caseosa. These acids form the structural subunits of complex lipids, such as the abundant triacylglycerols and wax esters. By studying fatty acids and their structural features, this thesis has provided valuable insights into the structural variability of the vernix caseosa lipidome. Without a full understanding of the structure of these key subunits, it would be impossible to complete the analysis of this lipidome in the future. In this thesis, fatty acids using high-performance liquid chromatography (HPLC) and modern mass spectrometry methods, namely ultraviolet photodissociation (UVPD) and ozone-induced dissociation (OzID), were characterized. The coupling of HPLC and mass spectrometry (HPLC-MS) provided unambiguous information on the positions and geometry of...
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Sample preparation for analyzing oligomeric forms of amyloid beta by mass spectrometry
Mrkvičková, Simona ; Cvačka, Josef (advisor) ; Kozlík, Petr (referee)
Alzheimer's disease is the most widespread chronic and progressive neurodegenerative disease. The main problem of the disease is the reduction and damage of nerve cells. This leads to loss of brain mass and memory impairment. The disease mainly affects the elderly, for whom it is the most common cause of dementia. Around 50 million people suffer from this disease worldwide, and the number is growing. Alzheimer's disease thus represents a serious public health problem. The pathogenesis of this disease is controlled by the production and deposition of amyloid beta peptide. This peptide tends to aggregate. Its lower oligomeric forms are water-soluble and have a toxic effect on nerve cells. Amyloid beta oligomers therefore represent a possible target for the diagnosis of Alzheimer's disease. Oligomeric forms of beta amyloids have a non-covalent character, which can negatively affect their stability during analyzes in model and biological samples. In this bachelor thesis, suitable conditions for the preparation of model amyloid beta samples were sought. It was observed how the distribution of oligomeric forms changes under the influence of solvents, salts and other additives. The effect of pH on the incubation of amyloid beta in ammonium acetate and water was also investigated. Finally, a method for the...
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