Liquid chromatography tandem mass spectrometry method for the quantification of vandetanib in human plasma and rat liver microsomes matrices: metabolic stability investigation

Vandetanib (VNT) is a new oral tyrosine kinase inhibitor that acts mainly by inhibiting vascular endothelial growth factor receptor (VEGFR). Fast, specific, sensitive and validated LC-MS/MS was established for the determination of VNT in two various matrices including rat liver microsomes (RLMs) and human plasma. This method was applied in metabolic stability investigation of VNT. Resolution of two analytes was performed using C18 column and isocratic mobile phase composed of binary system of 10 mM ammonium formate (pH 4.1) and acetonitrile in a ratio of 1:1. The flow rate was set at 0.25 mL/min and total run time was 4 min with injection volume of 5 mu L. Ions were generated by ESI source and analyzed by multiple reaction monitoring mode (basis for quantification) in the Agilent 6410 QqQ analyzer. The linearity of the established method ranged from 5 to 500 ng/mL (r(2) >= 0.9996) in human plasma and RLMs. LOQ and LOD were 2.48 and 7.52 ng/mL, and 2.14 and 6.49 in human plasma and RLMs matrices. The intra-day and inter-day precision and accuracy were 0.66-2.66% and 95.05-99.17% in human plasma matrix-while in RLMs matrix, ranged from 0.97 to 3.08% and 95.8 to 100.09%, respectively. In vitro half-life was 39.85 min and intrinsic clearance was 3.92 +/- 0.28 mL/min/kg.