Development and validation of a new liquid chromatography-tandem mass spectrometry assay for the simultaneous quantification of afatinib, dacomitinib, osimertinib, and the active metabolites of osimertinib in human serum

• Main Authors: Chishima, H. (Author), Hakamata, J. (Author), Ikemura, S. (Author), Ishikawa, E. (Author), Jibiki, A. (Author), Kawazoe, H. (Author), Kimura, M. (Author), Kuniyoshi, O. (Author), Muramatsu, H. (Author), Nakada, H. (Author), Nakajima, H. (Author), Nakamura, T. (Author), Nakaya, N. (Author), Sato, I. (Author), Suehiro, N. (Author), Suzuki, S. (Author), Yokoyama, Y. (Author)
• Format: Article
• Language: English
• Published: Elsevier B.V. 2022
• Subjects: Active metabolites; Amino acids; Biological organs; Biomolecules; Diseases; Drug products; Enzymes; Epidermal growth factor receptors; Epidermal growth factor receptor-tyrosine kinase inhibitor; Epidermal growth factor receptor-tyrosine kinase inhibitors; Human serum; Lc.ms/ms; LC-MS/MS; LC-MS-MS; Liquid chromatography; Mass spectrometry; Metabolites; Non small cell lung cancer; Non-small cell lung cancer; Patient monitoring; Patient treatment; Precipitation (chemical); Protein precipitation; Receptor-tyrosine kinase; Safety factor; Tyrosine kinase inhibitor
• Online Access: View Fulltext in Publisher

 Reports on the therapeutic drug monitoring (TDM) of second- and third-generation epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) in non-small cell lung cancer patients are limited and are required to improve the safety of EGFR-TKI therapy. Some EGFR-TKIs have active metabolites with similar or higher potency compared with the parent compounds; thus, monitoring the parent compound as well as its active metabolites is essential for truly effective TDM. In this study, we developed and validated a method that simultaneously quantifies second- and third-generation EGFR-TKIs (afatinib, dacomitinib, and osimertinib) and the active metabolites of osimertinib, AZ5104 and AZ7550, in the human serum using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The clinical application of the method was also evaluated. The analytes were extracted from a 100 μL serum sample using a simple protein precipitation method and analyzed using LC-MS/MS. Excellent linearity of calibration curves was observed at ranges of 2.5–125.0 ng/mL for afatinib, 2.5–125.0 ng/mL for dacomitinib, 4.0–800.0 ng/mL for osimertinib, 1.0–125.0 ng/mL for AZ5104, and 2.5–125.0 ng/mL for AZ7550. The precision and accuracy were below 14.9% and within ± 14.9% of the nominal concentrations, respectively. The mean recovery was higher than 94.7% and the coefficient of variation (CV) was lower than 8.3%. The mean internal-standard normalized matrix factors ranged from 94.6 to 111.9%, and the CVs were lower than 9.7%. This analytical method met the acceptance criteria of the U.S. Food and Drug Administration guidelines. The method was also successfully applied to the analysis of 45 clinical samples; it supports the efficient and valuable analysis for TDM investigations of EGFR-TKIs. © 2022 The Authors