Using of separation methods in clinical research

Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Mgr. Jiří Plíšek Supervisor: prof. RNDr. Petr Solich, CSc. Supervisor specialist: assoc. prof. RNDr. Dagmar Solichová, Ph.D. Title of dissertation thesis: Using of separation methods in clinical research This thesis is devoted to development and validation of new LC methods for the determination of biologically active substances, specifically a fat-soluble vitamins (vitamin A, E and D) and theirs metabolites. Vitamins are essential micronutrients which cannot be synthesized by human organism with the exception of steroid hormone precursor of vitamin D. Vitamins are involved in biochemical processes of organism directly or as coenzymes. Interest in the clinical monitoring of these substances is increasing constantly over the last years. It needs to use modern instrumental techniques such as the UHPLC coupled with MS detection for the determination of vitamin D metabolites (25-hydroxy vitamin D3 and 25-hydroxy vitamin D2) due to their low concentrations in serum. The use of classical liquid chromatography with diode array detection is suitable in case of monitoring of levels of vitamin A (retinol) and E (α-tocopherol) in breast milk and vitamin E (α- tocopherol) in red blood cell..