Vývoj UHPLC metody pro separaci osmi strukturně blízkých derivátů vitamínu E

Pružinská, Klára

Charles University, Faculty of Pharmacy in Hradec Králové Department of analytical chemistry Candidate: Klára Pružinská Supervisor: Assoc. prof. PharmDr. Lucie Nováková, Ph.D. Title od the diploma thesis: Development of UHPLC method for separation of eight structurally related derivatives of vitamin E The aim of this diploma thesis was to develop UHLPC method for the separation of eight derivates of vitamin E. They include a group of tocopherols and tocotrienols. To achieve the aim ultra-high performance liquid chromatography was used. In process of optimalization the most important thing was the choice of stationary phase, consistence of mobile phase, flow speed, temperature of column and the time of separation. The Ascentis Express F5 Column (100 x 3.0 mm, 2.7 μm), Sigma-Aldrich was the best stationary phase because the separation of all eight derivatives on this column including two problematic pairs: β-T3 with γ-T3 and the pair β-T with γ-T was performed. The mobile phase was a mixture of a methanol with acetonitrile (1:1) and water in the ratio 75:25 with a flow rate 1 ml/min, isocratic elution and PDA and FLR detection. In PDA detection the wavelength was set 290 nm. In more sensitive FLR detection the emission wavelenght was 325 nm and the exciting wavelength of exciting was set on 280 nm....

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