Liquid chromatography-tandem mass spectrometric assay for the cyclin-dependent kinase inhibitor AT7519 in mouse plasma.
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.
Amsterdam Medical Center, University of Amsterdam, Department of Oncogenomics, Meibergdreef 15, PO Box 22700, 1105 AZ Amsterdam, The Netherlands.
M Emmy M
ME
Dolman
Ilona J M
IJ
den Hartog
Jan J
JJ
Molenaar
Jan H M
JH
Schellens
Jos H
JH
Beijnen
Rolf W
RW
Sparidans
D000818:Animals; D002138:Calibration; D002853:Chromatography, Liquid; D018844:Cyclin-Dependent Kinases / Q000037:antagonists & inhibitors*; D005260:Female; D007211:Indoles / Q000032:analysis; D051379:Mice; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D009447:Neuroblastoma / Q000188:drug therapy; D010880:Piperidines / Q000097:blood*; D047428:Protein Kinase Inhibitors / Q000097:blood*; D011720:Pyrazoles / Q000097:blood*; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry
C531230:4-(2,6-dichlorobenzoylamino)-1H-pyrazole-3-carboxylic acid piperidin-4-ylamide; D007211:Indoles; D010880:Piperidines; D047428:Protein Kinase Inhibitors; D011720:Pyrazoles; C531549:rucaparib; D018844:Cyclin-Dependent Kinases
AT7519; AUC(0−∞); C(max); CDK; Cyclin-dependent kinase inhibitor; EDTA; IS; LC–MS/MS; MYCN; Mouse plasma; T(1/2); T(max); V-myc myelocytomatosis viral related oncogene neuroblastoma derived (avian); area under the plasma concentration–time curve; cyclin-dependent kinase; elimination half-life; ethylenediaminetetraacetic acid; i.p.; internal standard; intraperitoneal; maximum plasma concentration; time to C(max)