Development, validation, and application of an LC-MS/MS method for the determination of the AXL/FLT3 inhibitor gilteritinib in mouse plasma

J Chromatogr B Analyt Technol Biomed Life Sci. 2021 Aug 1:1179:122882. doi: 10.1016/j.jchromb.2021.122882. Epub 2021 Jul 30.

Abstract

A simple, fast and precise LC-MS/MS method for the quantitation of the tyrosine kinase inhibitor gilteritinib was developed and validated for micro-volumes of mouse plasma. The assay procedure involved a one-step extraction of gilteritinib and the internal standard [2H5]-gilteritinib with acetonitrile. An Accucore aQ column was used to separate analytes using a gradient elution delivered at a flow rate of 0.4 mL/min, and a total run time of 2.5 min. Validation studies with quality control samples processed on consecutive days revealed that values for intra-day and inter-day precision were <7.04%, with an accuracy of 101-108%. Linear responses were observed over the entire calibration curve range (up to 500 ng/mL), and the lower limit of quantification was 5 ng/mL. The developed method was successfully used to examine the pharmacokinetics of oral gilteritinib in wild-type mice and mice lacking the organic cation transporters OCT1, OCT2, and MATE1 to further understand mechanisms contributing to drug-drug interactions and causes of inter-individual pharmacokinetic variability.

Keywords: Gilteritinib; LC-MS/MS; Mouse plasma; Pharmacokinetics.

MeSH terms

  • Aniline Compounds / blood*
  • Aniline Compounds / chemistry
  • Aniline Compounds / pharmacokinetics
  • Animals
  • Chromatography, Liquid / methods*
  • Female
  • HEK293 Cells
  • Humans
  • Limit of Detection
  • Linear Models
  • Mice
  • Pyrazines / blood*
  • Pyrazines / chemistry
  • Pyrazines / pharmacokinetics
  • Reproducibility of Results
  • Tandem Mass Spectrometry / methods*

Substances

  • Aniline Compounds
  • Pyrazines
  • gilteritinib