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The Development of SPE Extraction Procedure for Isolation of Pravastatin and its Metabolite from Biological Matrix
Rabatinová, Martina ; Nováková, Lucie (advisor) ; Nobilis, Milan (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Mgr. Martina Rabatinová Supervisor: Doc. PharmDr. Lucie Nováková, Ph.D. Title of Rigorous Thesis: The development of SPE extraction procedure for isolation of pravastatin and its metabolite from biological matrix Statins are a class of drugs used to lower cholesterol levels. Statins have an inhibitory activity against 3-hydroxy-3-methylglutaryl coenzyme A reductase. The main purpose of this work was to develop and validate a quantitative method for sensitive analysis of pravastatin and pravastatin lactone. The analytes were extracted from rat urine by a solid phase extraction method. In this method urine samples were prepared on C18 Discovery DSC-18. Washing and elution agents consisted of a mixture of acetonitrile and 0.01 M ammonium acetate buffer, pH 4.5. Number of extraction steps were optimized to eliminate most of matrix effects. The method involved use of ultra high performance liquid chromatography tandem mass spectrometry. Chromatography was carried out on a BEH C18 1.7 µm, 2.1×100 mm column. A mobile phase consisted of a mixture of acetonitrile and 0.5 mM ammonium acetate solution, pH 4.0. Linear gradient conditions were used. Total run time of this anal- ysis was 6 minutes...

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