title: A supported liquid extraction LC-MS/MS method for determination of concentrations of GDC-0425, a small molecule Checkpoint kinase 1 inhibitor, in human plasma.
abstract: A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the determination of GDC-0425 concentrations in human plasma has been developed and validated. Supported liquid extraction was used to extract plasma samples (50 μL) and the resulting samples were analyzed using reverse-phase chromatography and mass spectrometry coupled with a turbo-ionspray interface. The mass analysis of GDC-0425 was performed using multiple reaction monitoring transitions in positive ionization mode. The method was validated over the calibration curve range of 1.00-1000 ng/mL using linear regression and 1/x2weighting. Within-run relative standard deviation ranged from 0.8 to 5.1%, while between-run RSD varied from 1.9 to 4.7% for QCs. The accuracy ranged from 90.0 to 101.0% of nominal for within-run and from 94.0 to 100.0% of nominal for between-run. Overall extraction recovery was 87.4% for GDC-0425 and 87.9% for GDC-0425-d9. Stability of GDC-0425 was established in human plasma for 374 days at -20 and -70 °C and established in reconstituted sample extracts for 88 h when stored at 2-8 °C. Stable-labeled internal standard was used to minimize matrix effects. This assay was used to characterize the pharmacokinetics of GDC-0425 in cancer patients.
authors.affiliation: Genentech, Drug Metabolism and Pharmacokinetics, 1 DNA Way, South San Francisco, CA, USA.
authors.forename: Xiao
authors.initials: X
authors.lastname: Ding
authors.affiliation: Genentech, Drug Metabolism and Pharmacokinetics, 1 DNA Way, South San Francisco, CA, USA.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Genentech, Small Molecule Clinical Pharmacology, 1 DNA Way, South San Francisco, CA, USA.
authors.forename: Srikumar
authors.initials: S
authors.lastname: Sahasranaman
authors.affiliation: Covance Laboratories, 3301 Kinsman Blvd., Madison, WI, USA.
authors.forename: Yifan
authors.initials: Y
authors.lastname: Shi
authors.affiliation: Covance Laboratories, 3301 Kinsman Blvd., Madison, WI, USA.
authors.forename: Janine
authors.initials: J
authors.lastname: McKnight
authors.affiliation: Genentech, Drug Metabolism and Pharmacokinetics, 1 DNA Way, South San Francisco, CA, USA.
authors.forename: Brian
authors.initials: B
authors.lastname: Dean
mesh_terms: D000071877:Checkpoint Kinase 1 / Q000037:antagonists & inhibitors*; D002853:Chromatography, Liquid / Q000379:methods*; D006461:Hemolysis; D006575:Heterocyclic Compounds, 3-Ring / Q000097:blood* / Q000493:pharmacokinetics; D006801:Humans; D057230:Limit of Detection; D010880:Piperidines / Q000097:blood* / Q000493:pharmacokinetics; D047428:Protein Kinase Inhibitors / Q000097:blood*; D012015:Reference Standards; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D006575:Heterocyclic Compounds, 3-Ring; D010880:Piperidines; D047428:Protein Kinase Inhibitors; C000614032:GDC-0425; D000071877:Checkpoint Kinase 1
keywords: GDC-0425; LC-MS/MS; checkpoint kinase 1 (ChK1); human plasma; supported liquid extraction