| Summary: | Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of analytical chemistry Candidate: Hana Plečková Supervisor: Doc. PharmDr. Lucie Nováková Ph.D. Title of Doctoral Thesis: Development of method for the identification of alkaloids in plant extracts from Amaryllidaceae using UHPLC-MS/MS. The purpose of this thesis is the development of method for identification of alkaloids from the family Amaryllidaceae in plant extracts using coupling of two methods UHPLC and MS/MS. The combination of methods UHPLC and MS/MS enables to work with the very small particles (< 2 µm) thereby fast separation with high efficiency and reduction of analysis time is achieved. MS is used for determination of atomic weight, molecular weight after the ionization of neutral species. The development of methods was first conducted on UHPLC with PDA detection which served for the choice of optimal stationary and mobile phase. The tested columns were following: BEH PHENYL, BEH C18, BEH SHIELD RP18, HSS T3. The possibilities of separation and the behavior of the monitored substances at different pH (3 and 9), retention time and flow rate were studied. The column BEH SHIELD RP18 provided the best results of these columns, using buffer pH 3 and methanol as an organic modifier, and then flow rate 0.4 ml/min....
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