title: 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%.
authors.affiliation: Department of Drug Analysis, Abbott Laboratories, Abbott Park, IL 60064, USA. Ramona.Rodila@abbott.com
authors.forename: Ramona C
authors.initials: RC
authors.lastname: Rodila
authors.forename: Joseph C
authors.initials: JC
authors.lastname: Kim
authors.forename: Qin C
authors.initials: QC
authors.lastname: Ji
authors.forename: Tawakol A
authors.initials: TA
authors.lastname: El-Shourbagy
mesh_terms: D001774:Blood Chemical Analysis / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D006801:Humans; D007191:Indazoles / Q000097:blood*; D044085:Microfluidics / Q000379:methods; D010671:Phenylurea Compounds / Q000097:blood*; D015203:Reproducibility of Results; D012371:Robotics / Q000379:methods*; D012680:Sensitivity and Specificity; D052617:Solid Phase Microextraction / Q000379:methods*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
chemical_list: D007191:Indazoles; D010671:Phenylurea Compounds; C513486:linifanib