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Comparison and optimization of P-glycoprotein inhibitors separation using different stationary phases in the SIC system
Vrábelová, Dóra ; Sklenářová, Hana (advisor) ; Horstkotte Šrámková, Ivana (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Dóra Vrábelová Supervisor: doc. PharmDr. Hana Sklenářová, Ph.D Title of the diploma thesis: Comparison and optimization of P-glycoprotein inhibitors separation using different stationary phases in the SIC system The diploma thesis was focused on development, comparison and optimization of method for separation of selected inhibitors of P-glycoprotein - verapamil, quinidine and rhodamine 123 using sequential injection chromatography system. In the optimization step several types of stationary phases - chromatographic columns based on monolithic columns (C18, Phenyl, CN and DIOL), and core-shell particle columns (C18, RP-Amide, Phenyl-Hexyl, and F5), and several types and combinations of mobile phases were tested. Following conditions of separation for verapamil and rhodamine 123 were chosen after optimization: Analysis using monolithic column Chromolith Flash RP-18e 25 x 4.6 mm (Merck, Germany). Spectrophotometric detection was performed at wavelenghts of 229 nm, 278 nm, 504 nm, (600 nm reference), isocratic elution was used. Mobile phase was acetonitrile/5 mM ammonium acetate buffer 50/50 (v/v) pH was modified to 7. The flow rate of mobile phase was 15 ɥl/s and the volume was 3 ml. Injection...
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