Influence of sample preparation method on occurence of matrix effects in UHPSFC-MS analysis of vitamin E

Charles University, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Lucia Vasilisková Supervisor: doc. PharmDr. Lucie Nováková, Ph.D. Title of Diploma Thesis: Influence of sample preparation method on occurence of matrix effects in UHPSFC-MS analysis of vitamin E The goal of this thesis was to optimize the chromatographic method for the determination of 8 different isomers of vitamin E that would provide better analysis results than two existing methods; high speed (HS) and high resolution (HR). New Torus stationary phases (SP) were tested (diethylamine, 1-aminoantracene, 2-picolylamine and diol under conditions: CO2:MeOH 98:2, flow rate 2 ml/min, injection volume 2 µl, column temperature 40řC, BPR 12.9 MPa, make-up solvent 0.1% NH4OH in MeOH. The modifiers (ethanol, isopropanol and a 1:1 mixture of acetonitrile/methanol) were tested on the 2-PIC column. The effect of additives in the mobile phase was also compared (0.1% NH4OH, 10mM ammonium formate, 10mM ammonium acetate and 2% H2O). The next part of the thesis describes the development of the sample preparation method to reduce or eliminate the matrix effects. Dilution method was tested using solvents - H2O, 50% acetonitrile (ACN) and 75% ACN. Urine samples were 10, 20, 50 and 100 times diluted by the most...