Highly sensitive HPLC‐DAD method for the assay of gefitinib in patient plasma and cerebrospinal fluid: application to a blood‐brain barrier penetration study

Abstract

The quantification of intracranial gefitinib (GEF) exposure is limited owing to the sensitivity of analytical equipment. Although mass spectrometry (MS) is the preferred method because of its high sensitivity, the equipment is not available in many laboratories, especially in developing Asian countries. In this paper, we developed a highly sensitive high performance liquid chromatography-diode array detector (HPLC-DAD) method for the assay of GEF in human cerebrospinal fluid (CSF) and plasma. GEF was extracted from CSF and plasma by solid-phase extraction and liquid-liquid extraction, respectively. The chromatographic separation was performed on a C18 column with gradient elution of 0.1% triethylamine solution and acetonitrile, then finally detected at 344 nm. This method was validated and proved to be highly sensitive with a lower limit of quantitation value of 0.11 ng/mL in CSF and 11 ng/mL in plasma. The blood-brain barrier penetration ratio of GEF ranged from 1.48 to 2.41%. This method provides a reliable MS-independent solution for the quantitation of GEF in patients' CSF and plasma.

Recent developments in the chromatographic bioanalysis of approved kinase inhibitor drugs in oncology

“…To assess the blood-brain barrier (BBB) permeability of TKIs, cerebrospinal fluid (CSF) concentrations have been compared to blood-concentrations [50]. A number of TKI assays for the analysis and subsequent calculation of the CSF-penetration rate have been published for erlotinib, gefitinib, sorafenib and vandetanib [8,[51][52][53][54][55]. CSF is a relatively clean ultrafiltrate, with a low protein level of approximately 0.15 to 0.60 g/L (blood-plasma contains ca.…”

“…One of the problems that arises when validating an assay for TKIs in CSF is acquiring drug-free CSF. A way to overcome this is to use artificial CSF [53], if justifiable [18,19]. To incorporate CSF bioanalysis in plasma-assays, CSF is sometimes diluted with drug-free plasma.…”

“…melanoma B-Raf-V600E No [148] dasatinib Sprycel BMS354825 Ph+CML (Bcr-Abl) BCR-ABL, PDGFR, Src, c-Kit, … No [7,15,23,31,47,62,83,95,99,114,119,124,126,130,149] erlotinib Tarceva OSI774 NSCLC EGFR, JAK2V617F No [15,23,51,61,67,84,91,105,107,114,118,124,125,128,129,135,1 50,151] gefitinib Iressa ZD1839 NSCLC EGFR No [15,52,53,107,114,124,125,152] ibrutinib Imbruvica PCI32765 MCL, CLL, WM Brutons TK (BTK) Yes, Cys481 [72,76,98,113,145] imatinib Gleevec, Glivec ST1571 Ph+CML (Bcr-Abl), GIST BCR-ABL, PDGFR, SCF, c-KIT No [7,8,10,15,23,26,27,31,33,36,47,60,94,97,99,100,102,…”

An Exploration of Advancement in Analytical Methodology for Quantification of Anticancer Drugs in Biomatrices

“…Total chromatographic run time was 10 min. 94 Wang et al developed a method for quantification of ponatinib in rat feces using the LLE technique and separation was achieved using a Waters Cortecs column kept at 40 C. The mobile phase used was water (having 0.1% formic acid and 2 mM ammonium formate) and methanol in gradient elution. The mass detection was performed in MRM with ESI in positive ion mode.…”