title: Structural characterization of electrochemically and in vitro biologically generated oxidation products of atorvastatin using UHPLC/MS/MS.
abstract: Ultrahigh-performance liquid chromatography coupled with high-mass-accuracy tandem mass spectrometry (UHPLC-MS-MS) has been used for elucidation of the structures of oxidation products of atorvastatin (AT), one of the most popular commercially available drugs. The purpose of the study was identification of AT metabolites in rat hepatocytes and comparison with electrochemically generated oxidation products. AT was incubated with rat hepatocytes for 24 h. Electrochemical oxidation of AT was performed by use of a three-electrode off-line system with a glassy carbon working electrode. Three supporting electrolytes (0.1 mol L(-1) H2SO4, 0.1 mol L(-1) HCl, and 0.1 mol L(-1) NaCl) were tested, and dependence on pH was also investigated. AT undergoes oxidation by a single irreversible process at approximately +1.0 V vs. Ag/AgCl electrode. The results obtained revealed a simple and relatively fast way of determining the type of oxidation and its position, on the basis of characteristic neutral losses (NLs) and fragment ions. Unfortunately, different products were obtained by electrochemical oxidation and biotransformation of AT. High-mass-accuracy measurement combined with different UHPLC-MS-MS scans, for example reconstructed ion-current chromatograms, constant neutral loss chromatograms, or exact mass filtering, enable rapid identification of drug-related compounds. β-Oxidation, aromatic hydroxylation of the phenylaminocarbonyl group, sulfation, AT lactone and glycol formation were observed in rat biotransformation samples. In contrast, a variety of oxidation reactions on the conjugated skeleton of isopropyl substituent of AT were identified as products of electrolysis.
authors.affiliation: Department of Analytical Chemistry, Faculty of Chemical Technology, University of Pardubice, Studentská 573, 53210, Pardubice, Czech Republic. Robert.Jirasko@upce.cz
authors.forename: Robert
authors.initials: R
authors.lastname: Jirásko
authors.forename: Tomáš
authors.initials: T
authors.lastname: Mikysek
authors.forename: Vitaliy
authors.initials: V
authors.lastname: Chagovets
authors.forename: Ivan
authors.initials: I
authors.lastname: Vokřál
authors.forename: Michal
authors.initials: M
authors.lastname: Holčapek
mesh_terms: D000818:Animals; D000069059:Atorvastatin; D001692:Biological Transport; D001711:Biotransformation; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004572:Electrolysis; D006018:Glycols / Q000737:chemistry* / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D006538:Heptanoic Acids / Q000737:chemistry* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D006900:Hydroxylation; D007783:Lactones / Q000737:chemistry* / Q000378:metabolism; D008297:Male; D008970:Molecular Weight; D010084:Oxidation-Reduction; D011758:Pyrroles / Q000737:chemistry* / Q000378:metabolism; D051381:Rats; D053719:Tandem Mass Spectrometry
chemical_list: D006018:Glycols; D006538:Heptanoic Acids; D007783:Lactones; D011758:Pyrroles; D000069059:Atorvastatin