A validated LC-MS/MS assay for the simultaneous determination of the anti-leukemic agent dasatinib and two pharmacologically active metabolites in human plasma: application to a clinical pharmacokinetic study

J Pharm Biomed Anal. 2012 Jan 25;58:130-5. doi: 10.1016/j.jpba.2011.09.008. Epub 2011 Sep 16.

Abstract

Dasatinib (Sprycel) is a potent antitumor agent prescribed for patients with chronic myeloid leukemia (CML). To enable reliable quantification of dasatinib and its pharmacologically active metabolites in human plasma during clinical testing, a sensitive and reliable liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated. Samples were prepared using solid phase extraction on Oasis HLB 96-well plates. Chromatographic separation was achieved isocratically on a Luna phenyl-hexyl analytical column. Analytes and the stable labeled internal standards were detected by positive ion electrospray tandem mass spectrometry. The assay was validated over a concentration range of 1.00-1000 ng/mL for dasatinib and its two active metabolites. Intra- and inter-assay precision values for replicate QC control samples were within 5.3% for all analytes during the assay validation. Mean QC control accuracy values were within ± 9.0% of nominal values for all analytes. Assay recoveries were high (>79%) and internal standard normalized matrix effects were minimal. The three analytes were stable in human plasma for at least 22 h at room temperature, for at least 123 days at -20°C, and following at least six freeze-thaw cycles. The validated method was successfully applied to the quantification of dasatinib and two active metabolites in a human pharmacokinetic study.

Publication types

  • Clinical Trial

MeSH terms

  • Antineoplastic Agents / blood*
  • Antineoplastic Agents / chemistry
  • Antineoplastic Agents / pharmacokinetics*
  • Calibration
  • Chromatography, Liquid / methods*
  • Dasatinib
  • Humans
  • Leukemia / drug therapy
  • Pyrimidines / blood*
  • Pyrimidines / chemistry
  • Pyrimidines / pharmacokinetics*
  • Reference Standards
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Solid Phase Extraction / methods
  • Spectrometry, Mass, Electrospray Ionization / methods
  • Tandem Mass Spectrometry / methods*
  • Thiazoles / blood*
  • Thiazoles / chemistry
  • Thiazoles / pharmacokinetics*

Substances

  • Antineoplastic Agents
  • Pyrimidines
  • Thiazoles
  • Dasatinib