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Optimization of metabolomics workflow: critical evaluation of sample clean-up step
Syřínek, Jakub Eduard ; Nováková, Lucie (advisor) ; Kočová Vlčková, Hana (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Jakub Eduard Syřínek Supervisor: Assoc. Prof. PharmDr. Lucie Nováková, Ph.D. Title of Diploma Thesis: Optimization of metabolomics workflow: critical evaluation of sample clean-up step Metabolomics workflow involves following steps: experiment planning, sampling, sample preparation, sample analysis and both data processing and data interpretation. Metabolomics is focused on small molecules with mass lower than 1500 Da. This is reason, why sample preparation of biological samples is critical. This work deals with the optimization of sample preparation method for polyethylene glycol (PEG) elimination from sample. PEG can interfere with metabolites during the ionization in the ion source of mass spectrometer. Solid phase microextraction in pipette tips (SPE-PT) was used in this work for PEG elimination. Porous titanium dioxide was employed as a sorbent. Vaginal swabs were collected from patients suffering from vulvovaginal discomfort caused by Candida albicans. Samples were first dissolved in phosphate buffer saline solution. In SPE-PT method 5 different composition of binding solution, number of aspiration/expelling repeats in sample loading step, 5 different composition of washing buffer and...
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