Liquid chromatography–tandem mass spectrometric assay for the cyclin-dependent kinase inhibitor AT7519 in mouse plasma

• Published in: Journal of pharmaceutical and biomedical analysis 2014-01, Vol.88, p.216-220
• Main Authors: Dolman, M. Emmy M., den Hartog, Ilona J.M., Molenaar, Jan J., Schellens, Jan H.M., Beijnen, Jos H., Sparidans, Rolf W.
• Format: Article
• Language: English
• Subjects: Animals; AT7519; Calibration; Chromatography, Liquid; Cyclin-dependent kinase inhibitor; Cyclin-Dependent Kinases - antagonists & inhibitors; Female; Indoles - analysis; LC–MS/MS; Mice; Mice, Nude; Mouse plasma; Neoplasm Transplantation; Neuroblastoma - drug therapy; Piperidines - blood; Protein Kinase Inhibitors - blood; Pyrazoles - blood; Reproducibility of Results; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/j.jpba.2013.08.051

•The first validated bioanalytical assay for AT7519 has been reported.•The assay has successfully been validated in the 5–10,000ng/ml range.•The drug is sufficiently stable under all conditions relevant for the assay.•Pharmacokinetics in mice with neuroblastoma xenografts have been reported. A quantitative bioanalytical liquid chromatography–tandem mass spectrometric (LC–MS/MS) assay for the cyclin-dependent kinase inhibitor AT7519 in mouse plasma was developed and validated. Plasma samples were pre-treated using protein precipitation with acetonitrile containing rucaparib as internal standard. After dilution with water, the extract was directly injected into the reversed-phase LC system. The eluate was transferred into the electrospray interface with positive ionization and the analyte was detected in the selected reaction monitoring mode of a triple quadrupole mass spectrometer. The assay was validated in a 5–10,000ng/ml calibration range using double logarithmic calibration, 5ng/ml was the lower limit of quantification. Within day precisions (n=6) were 2.9–5.6%, between day (3 days; n=18) precisions 3.2–7.2%. Accuracies were between 95.9 and 99.0% for the whole calibration range. The drug was stable under all relevant analytical conditions. Finally, the assay was successfully used to determine plasma pharmacokinetics after intraperitoneal administration of AT7519 in mice with neuroblastoma xenografts.