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Development and validation of a LC‐MS assay for the quantification of ikh12 a novel anti‐tumor candidate in rat plasma and tissues and its application in a pharmacokinetic study
Author(s) -
Otaegui Dorleta,
Masdeu Carme,
Aldaba Eneko,
Vara Yosu,
Zubia Aizpea,
San Sebastian Eider,
Alcalá Maria,
Villafruela Sergio,
Cossío Fernando P.,
RodriguezGascón Alicia
Publication year - 2015
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.3414
Subject(s) - chemistry , chromatography , electrospray ionization , calibration curve , formic acid , selected reaction monitoring , pharmacokinetics , electrospray , extraction (chemistry) , methanol , tandem mass spectrometry , liquid chromatography–mass spectrometry , mass spectrometry , high performance liquid chromatography , detection limit , analytical chemistry (journal) , medicine , organic chemistry
IKH12 is a novel histone deacetylase 6 selective inhibitor. A rapid and sensitive liquid chromatography tandem mass spectrometry method was developed and validated for the quantification of IKH12 in rat plasma and tissue with kendine 91 as internal standard (IS). The samples were prepared by liquid–liquid extraction with tert ‐butyl methyl ether. The chromatographic separation was accomplished by using a Zorbax Extend C 18 4.6 × 150 mm, 5 µm column, with a mobile phase consisting of methanol and 0.1% formic acid (75:25 v/v). Multiple reaction monitoring, using electrospray ionization in positive ion mode, was employed to quantitatively detect IKH12 and IS. The monitored transitions were set at m / z 418 → 252 and 444 → 169 for IKH12 and kendine 91, respectively. The calibration curve was linear over the concentration range 2–1000 ng mL −1 . The intra‐ and inter‐assay precision and accuracy of the quality controls and the limit of quantification were satisfactory in all cases (according to European Medicines Agency guidelines). Stability studies showed that plasma samples were stable in the chromatography rack for 24 h and at −80 °C for 2 months and also after three freeze–thaw cycles. This method was successfully applied to a pharmacokinetic study of IKH12 in rat. Copyright © 2015 John Wiley & Sons, Ltd.