title: Liquid chromatography and tandem mass spectrometry method for the quantitative determination of saxagliptin and its major pharmacologically active 5-monohydroxy metabolite in human plasma: method validation and overcoming specific and non-specific binding at low concentrations.
abstract: A liquid chromatography and tandem mass spectrometry (LC-MS/MS) method was developed and validated to simultaneously determine the concentrations of saxagliptin (Onglyza™, BMS-477118) and its major active metabolite, 5-hydroxy saxagliptin to support pharmacokinetic analyses in clinical studies. The dynamic range of the assay was 0.1-50 ng/mL for saxagliptin and 0.2-100 ng/mL for 5-hydroxy saxagliptin. Protein precipitation (PPT) with acetonitrile was used to extract the analytes from plasma matrix before injecting on an Atlantis(®) dC18 column (50 mm × 2.1 mm, 5 μm) for LC-MS/MS analysis. The sample pre-treatment process was carefully controlled to disrupt DPP4-specific binding and non-specific binding observed at lower concentrations. The recoveries for both analytes were >90%. The assay was selective, rugged and reproducible; storage stability of at least 401 days at -20°C was demonstrated. Under these chromatographic conditions, the isomers of saxagliptin and 5-hydroxy saxagliptin were chromatographically separated from saxagliptin and 5-hydroxy saxagliptin. The assay has been used to support multiple clinical studies and regulatory approvals.
authors.affiliation: Research & Development, Bristol-Myers Squibb, Princeton, NJ 08543, USA. xiaohui.xu@bms.com
authors.forename: Xiaohui Sophia
authors.initials: XS
authors.lastname: Xu
authors.forename: Roger
authors.initials: R
authors.lastname: Demers
authors.forename: Huidong
authors.initials: H
authors.lastname: Gu
authors.forename: Lisa J
authors.initials: LJ
authors.lastname: Christopher
authors.forename: Hong
authors.initials: H
authors.lastname: Su
authors.forename: Laura
authors.initials: L
authors.lastname: Cojocaru
authors.forename: David W
authors.initials: DW
authors.lastname: Boulton
authors.forename: Mark
authors.initials: M
authors.lastname: Kirby
authors.forename: Bruce
authors.initials: B
authors.lastname: Stouffer
authors.forename: William G
authors.initials: WG
authors.lastname: Humphreys
authors.forename: Mark E
authors.initials: ME
authors.lastname: Arnold
mesh_terms: D000218:Adamantane / Q000031:analogs & derivatives* / Q000097:blood / Q000737:chemistry / Q000493:pharmacokinetics; D002853:Chromatography, Liquid / Q000379:methods*; D004151:Dipeptides / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D004355:Drug Stability; D006801:Humans; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013237:Stereoisomerism; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: C558749:5-hydroxysaxagliptin; D004151:Dipeptides; C502994:saxagliptin; D000218:Adamantane