title: Application of polarity switching in the identification of the metabolites of RO9237.
abstract: Polarity switching mass spectrometry is an efficient way to collect structural data on drug metabolites. The value of this approach is illustrated with the in vitro metabolism of RO9237. Metabolites are identified by positive and negative electrospray ionization (ESI) full scan mass spectrometry, MS/MS and MS(3) using unlabelled and (14)C-radiolabelled versions of the drug. Comparison of the relative detectability of these metabolites by +ESI and -ESI shows that neither ESI mode is universal. It is advantageous to screen for metabolites using both positive and negative ionization modes. This is especially true for phase II metabolism which tends to make molecules more polar and often more acidic. Identification of phase II metabolites also benefits greatly from MS(3) experiments because the conjugating groups typically are cleaved in MS/MS and information on the core structure is only obtained in MS(3). A special case of phase II metabolism is the generation of glutathione (GSH) conjugates from reactive metabolites. The detection of GSH conjugates also benefits from generating both positive and negative ESI mass spectral data.
authors.affiliation: Drug Metabolism and Pharmacokinetics Department, Roche Palo Alto, LLC, 3431 Hillview Ave., Palo Alto, CA 94304, USA. Bill.fitch@roche.com
authors.forename: William L
authors.initials: WL
authors.lastname: Fitch
authors.forename: Limin
authors.initials: L
authors.lastname: He
authors.forename: Ya-Ping
authors.initials: YP
authors.lastname: Tu
authors.forename: Ludmila
authors.initials: L
authors.lastname: Alexandrova
mesh_terms: D000818:Animals; D001711:Biotransformation; D002851:Chromatography, High Pressure Liquid; D000882:Haplorhini; D022781:Hepatocytes / Q000378:metabolism; D006801:Humans; D007202:Indicators and Reagents; D008862:Microsomes, Liver / Q000378:metabolism; D011985:Receptors, Serotonin / Q000187:drug effects; D012702:Serotonin Antagonists / Q000493:pharmacokinetics*; D021241:Spectrometry, Mass, Electrospray Ionization; D013450:Sulfones / Q000493:pharmacokinetics*
chemical_list: D007202:Indicators and Reagents; C522134:RO9237; D011985:Receptors, Serotonin; D012702:Serotonin Antagonists; D013450:Sulfones; C097216:serotonin 6 receptor