Abstract
A simple, rapid and sensitive liquid chromatography–tandem mass spectrometric (LC/MS–MS) method has been developed and validated for the quantitative determination of pazopanib in mouse plasma and brain tissue homogenate. Single liquid–liquid extraction step with ethyl acetate was employed for analysis of pazopanib and the internal standard (IS); vandetanib. HPLC separation was performed on an XTerra® MS C18 column 50mm×4.6mm, 5.0μm. The mobile phase consisted of 70% acetonitrile and 30% water with 0.1% formic acid, pumped at a flow rate of 0.25ml/min. Analysis time was 3.5min per run and both the analyte and IS eluted within 1.8–2.0min. Multiple reactions monitoring (MRM) mode was utilized to detect the compounds of interest. The mass spectrometer was operated in the positive ion mode for detection. The precursor to product ions (Q1→Q3) selected for pazopanib and internal standard during quantitative optimization were (m/z) 438.1→357.2 and 475.0→112.2 respectively. The calibration curves were linear over the range of 3.9–1000ng/ml in both biological matrices. Lower limit of quantification (LLOQ) for mouse plasma and brain tissue was 3.9ng/ml. The values for inter and intra day precision and accuracy were well within the ranges acceptable for analytical assessment (<15%). This method was applied to determine brain to plasma concentration ratio and relevant pharmacokinetic parameters of pazopanib after a single intravenous dose of 5mg/kg in FVB wild type mice.
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