Comprehensive identification, fragmentation pattern, and metabolic pathways of gefitinib metabolites via UHPLC-Q-TOF-MS/MS: in vivo study of rat plasma, urine, bile, and faeces

Abstract

1. Gefitinib, the first approved inhibitor for oral epidermal growth factor receptor (EGFR), has been proved to be effective in non-small cell lung cancer with EGFR mutation. However, there are many metabolites of gefitinib that have not been identified in vivo.

2. This study aims to identify the metabolites of gefitinib and its metabolic pathways in rats using ultra-high-performance liquid chromatography coupled with a quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS/MS) detector. Protein precipitation, solid-phase and ultrasonic extraction were used for the pre-treatment of plasma, urine, bile and faeces samples.

3. In this study, a total of 28 compounds were identified in rat plasma, 29 in bile, 20 in urine and 16 in faeces. 20 new compounds were firstly reported as metabolites of gefitinib. Reduction, hydroxylation, dealkylation and dehalogenation were the major metabolic pathways in phase I. For phase II, the main pathways were sulphate and glucuronide conjugation. The fragment ions of gefitinib and its metabolites were usually generated via the fracture of C₁–O bond of propoxy on the C₆ position of aniline quinazoline ring.

4. The results may be valuable and important for understanding the metabolic process of gefitinib in clinical application and drug safety.

Keywords:

• Gefitinib
• metabolisms identification
• fragmentation pattern
• metabolic pathways
• UHPLC-Q-TOF-MS/MS

Disclosure statement

The authors report no declarations of interest.

Additional information

Funding

The present work was financially supported by Priority Academic Program Development of Jiangsu Higher Education Institutions of China and Open-End Funds of Jiangsu Key Laboratory of Marine Pharmaceutical Compound Screening.