Premium
Determination of intracellular anlotinib, osimertinib, afatinib and gefitinib accumulations in human brain microvascular endothelial cells by liquid chromatography/tandem mass spectrometry
Author(s) -
Ma Zhiyuan,
Lu Shuanghui,
Zhou Hui,
Zhang Shirong,
Wang Yuqing,
Lin Nengming
Publication year - 2020
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.8955
Subject(s) - afatinib , gefitinib , chemistry , osimertinib , erlotinib , electrospray ionization , chromatography , tandem mass spectrometry , orbitrap , mass spectrometry , liquid chromatography–mass spectrometry , epidermal growth factor receptor , receptor , biochemistry
Rationale Brain metastases are a common complication in patients with non‐small‐cell lung cancer (NSCLC). Anlotinib hydrochloride is a novel multi‐target tyrosine kinase inhibitor (TKI) exhibiting a superior overall response rate for brain metastases from NSCLC. The penetrability of anlotinib and three generations of epidermal growth factor receptor (EGFR) TKIs (osimertinib, afatinib and gefitinib) into brain microvascular endothelial cells (HBMECs) was compared. Methods A sensitive quantification method for the four TKIs was developed using liquid chromatography coupled to tandem mass spectrometry (LC/MS/MS). Anlotinib and the three EGFR TKIs were separated on an ACQUITY BEH C18 column after a direct protein precipitation, and then analyzed using electrospray ionization in positive ion mode. The linearity, accuracy, precision, limit of quantification, specificity and stability were assessed. Results The four analytes could be efficiently quantified in a single run of 3.8 min. The validation parameters of all analytes satisfy the acceptance criteria of bioanalytical method guidelines. The calibration range was 0.2–200 ng mL −1 for anlotinib and gefitinib, 1–500 ng mL −1 for osimertinib and 1–200 ng mL −1 for afatinib. The penetration of anlotinib across HBMECs was comparable with that of afatinib and gefitinib but less than that of osimertinib. Conclusions A sensitive LC/MS/MS method to simultaneously measure anlotinib, osimertinib, afatinib and gefitinib in cell extracts was successfully validated and applied to determine their uptake inside HBMECs, which could pave the way for future research on the role of anlotinib in NSCLC brain metastases.