Optimization and validation of UHPSFC-UV methods for quality control of agomelatin and atorvastatin in drug substance and in tablets

Charles University, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Zdeňka Němcová Supervisor: Assoc. Prof. PharmDr. Lucie Nováková, Ph.D. Title of Diploma Thesis: Optimization and validation of UHPSFC-UV methods for quality control of agomelatin and atorvastatin in drug substance and in tablets The aim of this master thesis was to develop two ultra-high performance supercritical fluid chromatography (UHPSFC) methods with PDA detection for determination of agomelatine and atorvastatin and its potential impurities. UHPSFC system Acquity UPC2 with PDA detector was used for measurement. The separation of agomelatine was performed by Torus Diol column (100 × 3.0 mm, 1.7 µm). Gradient elution was performed using CO2 with the mixture of methanol/acetonitrile (1:1) and 31.5 mM (0,1 %) ammonium hydroxide as a mobile phase. Column temperature was set at 40 řC and BPR (back pressure regulator) pressure at 2000 psi. The detection wavelength was set at 225 nm. The method was developed for measurment of API (active pharmaceutical ingredient) and subsequently for tablets. The separation of atorvastatin was performed using Acquity UPLC HSS C18 SB column (100 × 3.0 mm, 1.8 µm). Gradient elution was performed using CO2 with the mixture of methanol/ acetonitrile (2:1), 15 mM..