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Development of HPLC method for determination of coumarins in Tokaj wines
 Sudová, Tereza ; Šatínský, Dalibor (advisor) ; Sklenářová, Hana (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Bc. Tereza Sudová Supervisor: Doc. RNDr. Dalibor Šatínský, Ph.D. Title of the diploma thesis: Development of HPLC method for determination of coumarins in Tokaj wines A high performance chromatography method was used for simultaneous determination of coumarins: umbelliferone (7-hydroxycoumarin), coumarin and 7-methoxycoumarin (herniarin) in 60 samples of Tokaj wines and 12 samples of wines and spirits. Before the analysis, HPLC parameters were optimized: detector wavelength, analytical column, mobile phase and gradient elution. The suitability of the HPLC method for the determination of coumarins was confirmed. Then the method was validated by a validation process. The analysis was performed under gradient elution conditions with mobile phase methanol (organic phase) and 0.1% phosphoric acid (aqueous phase) at a flow rate of 1 ml/min. Ascentis® Express F5 - 150  4.6 mm reversed phase column at a temperature of 30 řC was used. The particle size of the stationary phase was 5 µm. Coumarins were detected by a DAD detector at wavelenghts 276 nm and 323 nm. Untreated samples were injected in a volume of 5 µl. The total analysis was 15.51 min.
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