Development of Stage tips-UHPLC-MS/MS method for determination of statins in human serum

Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of analytical chemistry Candidate: Lukáš Otoupal Supervisior: RNDr. Hana Vlčková, Ph.D. Title of Thesis: Development of Stage tips-UHPLC-MS/MS method for determination of statins in human serum This diploma thesis deals with development of a method for determination of statins in human serum. Extraction by Stage tips pipette tips was chosen and optimized for their extraction. The measurement was performed by the ultra-high performance liquid chromatography coupled to triple quadrupole mass spectrometer. The extraction procedure and important mass spectrometer parameters were optimized. Previously developed LC method for determination of statins was used for the analysis. The optimization of the mass spectrometer parameters was started by the selection of precursor ions, consequently the ion source parameters were tuned and finally appropriate ionization mode, SRM transition and collision energy for each analyte were chosen. The suitability of the LC-MS method was verified by the repeatability method (RSD ≤ 1 for the retention time and RSD ≤ 10 for the peak area), linearity (correlation coefficient ≥ 0,997) and sensitivity (LOQ in the range of 5 x 10-10 - 1 x 10- 9 g/ml). Firstly, the optimization employing mixed standard..

Development of Stage tips-UHPLC-MS/MS method for determination of statins in human serum

Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of analytical chemistry Candidate: Lukáš Otoupal Supervisior: RNDr. Hana Vlčková, Ph.D. Title of Thesis: Development of Stage tips-UHPLC-MS/MS method for determination of statins in human serum This diploma thesis deals with development of a method for determination of statins in human serum. Extraction by Stage tips pipette tips was chosen and optimized for their extraction. The measurement was performed by the ultra-high performance liquid chromatography coupled to triple quadrupole mass spectrometer. The extraction procedure and important mass spectrometer parameters were optimized. Previously developed LC method for determination of statins was used for the analysis. The optimization of the mass spectrometer parameters was started by the selection of precursor ions, consequently the ion source parameters were tuned and finally appropriate ionization mode, SRM transition and collision energy for each analyte were chosen. The suitability of the LC-MS method was verified by the repeatability method (RSD ≤ 1 for the retention time and RSD ≤ 10 for the peak area), linearity (correlation coefficient ≥ 0,997) and sensitivity (LOQ in the range of 5 x 10-10 - 1 x 10- 9 g/ml). Firstly, the optimization employing mixed standard..