Simultaneous quantitative detection of afatinib, erlotinib, gefitinib, icotinib, osimertinib and their metabolites in plasma samples of patients with non-small cell lung cancer using liquid chromatography-tandem mass spectrometry

Clin Chim Acta. 2022 Feb 15:527:1-10. doi: 10.1016/j.cca.2021.12.028. Epub 2022 Jan 6.

Abstract

Background and aims: As numerous studies have reported the concentration-exposure relationships of epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs), therapeutic drug monitoring is a promising approach in lung cancer treatment, aiming to avoid treatment failure or toxicity. A new method for the simultaneous analysis of five EGFR-TKIs (afatinib, erlotinib, gefitinib, icotinib and osimertinib) and their metabolites in human plasma samples was developed and validated using liquid chromatography-tandem mass spectrometry (LC-MS/MS).

Materials and methods: Afatinib-d6, erlotinib-d6, OSI-420-d4, gefitinib-d6 and osimertinib-C13,d3 were used as internal standards (ISs). The samples were prepared by liquid-liquid extraction using tert-butyl methyl ether. Chromatographic separation was undertaken on an XBridge C18 column using a linear gradient elution. LC-MS/MS was conducted in positive ionization mode with multiple reaction monitoring.

Results: The proposed method showed satisfactory results in terms of linearity, sensitivity, specificity, precision (intra- and inter-day coefficients of variation ranged from 1.1 to 13.9%), and accuracy (from 93.3 to 111.1%). The IS-normalized matrix factors were below 15%. The sensitivity and linearity were highly appropriate for the expected concentrations according to the analysis of samples from non-small cell lung caner (NSCLC) patients who received EGFR-TKIs.

Conclusions: The proposed method showed an acceptable reproducibility, high sensitivity and selectivity, and low matrix effects. This method could be significant for monitoring plasma concentrations of the mentioned EGFR-TKIs in NSCLC patients, aiming to improve the efficacy and safety of targeted therapies.

Keywords: EGFR-TKIs; LC–MS/MS; Metabolites; Non-small cell Lung cancer; Therapeutic drug monitoring.

MeSH terms

  • Acrylamides
  • Afatinib
  • Aniline Compounds
  • Carcinoma, Non-Small-Cell Lung* / drug therapy
  • Chromatography, Liquid
  • Crown Ethers
  • Erlotinib Hydrochloride
  • Gefitinib
  • Humans
  • Lung Neoplasms* / drug therapy
  • Mutation
  • Protein Kinase Inhibitors / therapeutic use
  • Quinazolines
  • Reproducibility of Results
  • Tandem Mass Spectrometry

Substances

  • Acrylamides
  • Aniline Compounds
  • Crown Ethers
  • Protein Kinase Inhibitors
  • Quinazolines
  • osimertinib
  • Afatinib
  • icotinib
  • Erlotinib Hydrochloride
  • Gefitinib