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Development and validation of method for determination of of Cyclosporine in biorelevant mediums
Tichoň, Matej ; Matysová, Ludmila (advisor) ; Kujovská Krčmová, Lenka (referee)
Charles university Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Matej Tichoň Supervisor: assoc. prof. PharmDr. Ludmila Matysová, Ph.D. Title of diploma thesis: Development and validation of method for determination of cyclosporine A in biorelevant mediums For the determination of cyclosporine A in biorelevant mediums, a new analytical method was developed by using of ultra-high performance liquid chromatography. The analysis was performed on a Kinetex™ 1.7 µm, C18 column, 50 x 2.1 mm (Phenomenex, USA). The mobile phase consisted of ultrapure water acidified with phosphoric acid to pH = 2.40 and mixture of methanol : acetonitrile (90 : 10) in a total ratio of 30 : 70 aqueous to organic phases. The analysis was carried out at 60 řC for 5 minutes with a constant flow rate of 0.5 ml/min. Gradient elution was used in the analysis, starting at 70 % organic phase at 0 minutes and increasing to 100 % at 5 minutes. Cyclosporine A was detected at 210 nm. The retention time of the substance was 3.08 min. Having the optimized conditions in hands, the method was fully validated. Keywords: cyclosporine A, UHPLC, biorelevant mediums
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