title: Bio-generation of stable isotope labeled internal standards for absolute and relative quantitation of drug metabolites in plasma samples by LC-MS/MS.
abstract: In order to achieve a better understanding of the toxicity of drug candidates, quantitative characterization of circulatory drug metabolites has been of increasing interest in current pharmaceutical research. Stable isotope labeled (STIL) internal standards (IS) are ideally used to simplify drug metabolite quantitation via liquid chromatography and tandem mass spectrometry (LC-MS/MS) analysis, primarily due to their capability to compensate matrix effects, thereby leading to faster method establishment by using generic assay conditions. However, chemical synthesis of STIL metabolites can often be resource intensive, requiring lengthy exploratory synthesis route development and/or extensive optimization to achieve the required stability for some metabolites. To overcome these challenges, we developed a general method that could generate STIL metabolites in a matter of hours from STIL parent drugs through the utilization of an appropriate in vitro metabolic incubation. This methodology can potentially save valuable synthesis resources, as well as provide timely availability of STIL IS. The following work demonstrates the proof-of-concept that multiple STIL metabolites can be generated simultaneously to provide satisfactory performance for both absolute quantitation of drug metabolites and for potential use in assessment of relative exposure coverage across species in safety tests of drug metabolites (MIST).
authors.affiliation: Department of Pharmaceutical and Biomedical Sciences, College of Pharmacy, University of Georgia, Athens, GA 30602, USA.
authors.forename: Pei
authors.initials: P
authors.lastname: Li
authors.forename: Yong
authors.initials: Y
authors.lastname: Gong
authors.forename: Heng-Keang
authors.initials: HK
authors.lastname: Lim
authors.forename: Wenying
authors.initials: W
authors.lastname: Jian
authors.forename: Richard W
authors.initials: RW
authors.lastname: Edom
authors.forename: Rhys
authors.initials: R
authors.lastname: Salter
authors.forename: Jose
authors.initials: J
authors.lastname: Silva
authors.forename: Naidong
authors.initials: N
authors.lastname: Weng
mesh_terms: D002853:Chromatography, Liquid / Q000379:methods*; D006801:Humans; D007554:Isotopes; D057230:Limit of Detection; D008861:Microsomes / Q000378:metabolism; D004364:Pharmaceutical Preparations / Q000097:blood* / Q000378:metabolism; D012015:Reference Standards; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D007554:Isotopes; D004364:Pharmaceutical Preparations