title: LC-MS/MS assay for the quantitation of the tyrosine kinase inhibitor neratinib in human plasma.
abstract: Neratinib is an orally available tyrosine kinase inhibitor targeting HER2 (ERBB2) and EGFR (ERBB). It is being clinically evaluated for the treatment of breast and other solid tumors types as a single agent or in combination with other chemotherapies. In support of several phase I/II clinical trials investigating neratinib combinations, we developed and validated a novel LC-MS/MS assay for the quantification of neratinib in 100μL of human plasma with a stable isotopic internal standard. Analytes were extracted from plasma using protein precipitation and evaporation of the resulting supernatant followed by resuspension. Chromatographic separation was achieved using an Acquity UPLC BEH Shield RP18 column and a gradient methanol-water mobile phase containing 10% ammonium acetate. An ABI 4000 mass spectrometer and electrospray positive mode ionization were used for detection. The assay was linear from 2 to 1,000ng/mL and proved to be accurate (98.9-106.5%) and precise (<6.2%CV), and met the FDA guidance for bioanalytical method validation. This LC-MS/MS assay will be an essential tool to further define the pharmacokinetics of neratinib.
authors.affiliation: Cancer Therapeutics Program, University of Pittsburgh Cancer Institute, Pittsburgh, PA, USA; Department of Pharmaceutical Sciences, School of Pharmacy, University of Pittsburgh, Pittsburgh, PA, USA.
authors.forename: Brian F
authors.initials: BF
authors.lastname: Kiesel
authors.affiliation: Cancer Therapeutics Program, University of Pittsburgh Cancer Institute, Pittsburgh, PA, USA.
authors.forename: Robert A
authors.initials: RA
authors.lastname: Parise
authors.affiliation: PUMA Biotechnology Inc., Los Angeles CA, USA.
authors.forename: Alvin
authors.initials: A
authors.lastname: Wong
authors.affiliation: PUMA Biotechnology Inc., Los Angeles CA, USA.
authors.forename: Kiana
authors.initials: K
authors.lastname: Keyvanjah
authors.affiliation: NSABP Foundation, Inc., Pittsburgh, PA, USA.
authors.forename: Samuel
authors.initials: S
authors.lastname: Jacobs
authors.affiliation: Cancer Therapeutics Program, University of Pittsburgh Cancer Institute, Pittsburgh, PA, USA; Department of Pharmaceutical Sciences, School of Pharmacy, University of Pittsburgh, Pittsburgh, PA, USA; Division of Hematology-Oncology, Department of Medicine, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA. Electronic address: beumerj@gmail.com.
authors.forename: Jan H
authors.initials: JH
authors.lastname: Beumer
mesh_terms: D002853:Chromatography, Liquid / Q000379:methods; D005260:Female; D006801:Humans; D047428:Protein Kinase Inhibitors / Q000032:analysis / Q000097:blood*; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors*; D011804:Quinolines / Q000032:analysis / Q000097:blood*; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D047428:Protein Kinase Inhibitors; D011804:Quinolines; D011505:Protein-Tyrosine Kinases; C487932:neratinib
keywords: Assay; Neratinib; Tandem mass spectrometry; Validation