Optimalization of sample preparation step for UHPLC-MS/MS analysis of atorvastatin, rosuvastatin and their metabolites

Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of analytical chemistry Candidate: Veronika Pilařová Supervisor: Doc. PharmDr. Lucie Nováková Ph.D. Title of GraduationThesis: Optimalization of sample preparation step for UHPLC-MS/MS analysis of atorvastatin, rosuvastatin and their metabolites The purpose of this graduation thesis was the optimalization of UHPLC-MS/MS method for the determination of concentrations of atorvastatin, rosuvastatin and their metabolites in biological material, and then development and optimalization of a MEPS (microextraction by packed sorbent) method for sample preparation of biological material and validation of this method. Acquity BEH C18 column (50 x 2.1 mm, 1.7 µm, Waters) was used for the separation of the analytes. Electrospray ionization was performed in both negative and positive ion mode. Triple quadrupole mass analyser was used for detection. Precursor ions and fragment ions were chosen for each statin. Collision energy and cone voltage were optimized for all analytes individually. Quantification of analytes was performed using the SRM (selected reaction monitoring). Protonated molecule [M+H]+ , which was measured in the positive ion mode, was chosen as a precursor ion for rosuvastatin lactone. Ion [M-H]- , which was measured in..