title: A high-throughput liquid chromatography/tandem mass spectrometry method for simultaneous quantification of a hydrophobic drug candidate and its hydrophilic metabolite in human urine with a fully automated liquid/liquid extraction.
abstract: ABT-869 (A-741439) is an investigational new drug candidate under development by Abbott Laboratories. ABT-869 is hydrophobic, but is oxidized in the body to A-849529, a hydrophilic metabolite that includes both carboxyl and amino groups. Poor solubility of ABT-869 in aqueous matrix causes simultaneous analysis of both ABT-869 and its metabolite within the same extraction and injection to be extremely difficult in human urine. In this paper, a high-performance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS) method has been developed and validated for high-speed simultaneous quantitation of the hydrophobic ABT-869 and its hydrophilic metabolite, A-849529, in human urine. The deuterated internal standards, A-741439D(4) and A-849529D(4), were used in this method. The disparate properties of the two analytes were mediated by treating samples with acetonitrile, adjusting pH with an extraction buffer, and optimizing the extraction solvent and mobile phase composition. For a 100 microL urine sample volume, the lower limit of quantitation was approximately 1 ng/mL for both ABT-869 and A-849529. The calibration curve was linear from 1.09 to 595.13 ng/mL for ABT-869, and 1.10 to 600.48 ng/mL for A-849529 (r2 > 0.9975 for both ABT-869 and A-849529). Because the method employs simultaneous quantification, high throughput is achieved despite the presence of both a hydrophobic analyte and its hydrophilic metabolite in human urine.
authors.affiliation: Department of Drug Analysis, Abbott Laboratories, Abbott Park, IL 60064, USA.
authors.forename: Perry G
authors.initials: PG
authors.lastname: Wang
authors.forename: Jun
authors.initials: J
authors.lastname: Zhang
authors.forename: Eric M
authors.initials: EM
authors.lastname: Gage
authors.forename: Jeffrey M
authors.initials: JM
authors.lastname: Schmidt
authors.forename: Ramona C
authors.initials: RC
authors.lastname: Rodila
authors.forename: Qin C
authors.initials: QC
authors.lastname: Ji
authors.forename: Tawakol A
authors.initials: TA
authors.lastname: El-Shourbagy
mesh_terms: D002851:Chromatography, High Pressure Liquid; D015507:Drugs, Investigational; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D007191:Indazoles / Q000652:urine*; D010671:Phenylurea Compounds / Q000652:urine*; D020794:Receptor Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors*; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry
chemical_list: D015507:Drugs, Investigational; D007191:Indazoles; D010671:Phenylurea Compounds; C513486:linifanib; D020794:Receptor Protein-Tyrosine Kinases