| Summary: | Charles University, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Tereza Lacmanová Supervisor: Assoc. Prof. PharmDr. Lucie Nováková, Ph.D. Title of Diploma Thesis: Application of supercritical fluid chromatography in analysis of natural compounds The goal of this diploma thesis is to verify the suitability of UHPSFC method with PDA and MS detection for analysis of natural products. For the analyzes, two representatives of natural compounds - the alkaloids of the family Amaryllidaceae with effect in the treatment of Alzheimer's disease and phenolic compounds with antioxidant activity - flavonoid rutin, quercetin and their metabolites were chosen. Separations were carried out on the UHPSFC system Acquity UPC2 with PDA and MS detector. The basic screening was performed on 10 stationary phases: BEH 2-EP, HSS SB C18, BEH, Torus DIOL, Torus 2-PIC, Torus DEA, CSH PFP, Torus 1-AA, Cortecs HILIC and BEH Amide. Four mobile phases were tested: 1. CO2 + MeOH, 2. CO2 + MeOH + 0.1 % NH4OH, 3. CO2 + MeOH + 10 mM ammonium formate, and 4. CO2 + MeOH + 10 mM ammonium formate + 2 % H2O. Separation conditions were as follows: flow rate 1.5 ml/min, temperature 40 řC, pressure 140 bar, gradient elution from 2 % to 40 % within 4 minutes, with two isocratic steps at the beginning and...
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