A high‐throughput, fully automated liquid/liquid extraction liquid chromatography/mass spectrometry method for the quantitation of a new investigational drug ABT‐869 and its metabolite A‐849529 in human plasma samples

Abstract

ABT‐869 is a novel ATP‐competitive inhibitor for all the vascular endothelial growth factor (VEGF) and platelet‐derived growth factor (PDGF) receptor tyrosine kinases (RTKs). It is one of the oncology drugs in development at Abbott Laboratories and has great potential for enhanced anti‐tumor efficacy as well as activity in a broad range of human cancers. We report here an accurate, precise and rugged liquid chromatography/mass spectrometry (LC/MS/MS) assay for the quantitative measurement of ABT‐869 and its acid metabolite A‐849529. A fully automated 96‐well liquid/liquid extraction method was achieved utilizing a Hamilton liquid handler. The only manual intervention required prior to LC/MS/MS injection is to transfer the 96‐well plate to a drying rack to dry the extracts then transfer the plate back to the Hamilton for robotic reconstitution. The linear dynamic ranges were from 1.1 to 598.8 ng/mL for ABT‐869 and from 1.1 to 605.8 ng/mL for A‐849529. The coefficient of determination (r²) for all analytes was greater than 0.9995. For the drug ABT‐869, the intra‐assay coefficient of variance (CV) was between 0.4% and 3.7% and the inter‐assay CV was between 0.9% and 2.8%. The inter‐assay mean accuracy, expressed as percent of theoretical, was between 96.8% and 102.2%. For the metabolite A‐849529, the intra‐assay CV was between 0.5% and 5.1% and the inter‐assay CV was between 0.8% and 4.9%. The inter‐assay mean accuracy, expressed as percent of theoretical, was between 96.9% and 100.6%. Copyright © 2006 John Wiley & Sons, Ltd.