Quantification of henatinib maleate,a novel potent inhibitor of VEGF receptors,in rat plasma by LC‐MS/MS

Henatinib maleate (R,Z)‐2‐(5‐fluoro‐1,2‐dihydro‐2‐oxo‐3H‐indol‐3‐ylidene) methyl]‐5‐(2‐hydroxy‐3‐morpholinopropyl)‐3‐methyl‐5,6,7,8‐tetrahydro‐1H‐pyrrolo3,2‐c] azepin‐4‐ketone maleate is a potent inhibitor of vascular endothelial growth factor receptors, and is currently under preclinical evaluation as an anticancer drug. A novel method for the quantification of henatinib maleate in rat plasma using high performance liquid chromatography–tandem mass spectrometry has been developed. The analyte (henatinib maleate) and internal standard (papaverine hydrochloride) were extracted from 50 μL of rat plasma by protein precipitation and separated on a C₁₈ column using a mixture of 25 mm ammonium acetate buffer : methanol : acetonitrile (35 : 50 : 15, v/v/v) as mobile phase with a run time of 4.5 min. The detection was performed by means of triple quadrupole mass spectrometer equipped with an ESI interface operating in the multiple‐reaction monitoring mode. A linear response was observed over the concentration range 5.0–1000 ng/mL. The limit of quantification was 5.0 ng/mL. Both intra‐ and inter‐day precision, defined as relative standard deviation, were within 9.7%. Accuracy, defined as relative error, was within ± 3.1%. The developed method was successfully applied to preclinical pharmacokinetic studies of henatinib maleate in rat after a single oral administration of the drug. Copyright © 2009 John Wiley & Sons, Ltd.