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Simultaneous determination of a novel oral Janus kinase inhibitor ASP015K and its sulfated metabolite in rat plasma using LC‐MS/MS
Author(s) -
Oda Kazuo,
Mera Katsumi,
Nagasaka Yasuhisa,
Tokoro Kazumi
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.3380
Subject(s) - chemistry , chromatography , electrospray ionization , metabolite , ammonium acetate , electrospray , sulfation , analyte , selected reaction monitoring , tandem mass spectrometry , mass spectrometry , extraction (chemistry) , bioanalysis , pharmacokinetics , solid phase extraction , liquid chromatography–mass spectrometry , high performance liquid chromatography , pharmacology , biochemistry , medicine
Abstract A sensitive and selective liquid chromatography with tandem mass spectrometry (LC‐MS/MS) was developed for determining the concentrations of novel Janus kinase inhibitor ASP015K and its sulfated metabolite M2 in rat plasma. This method involves solid‐phase extraction (SPE) from 25 μL of rat plasma. LC separation was performed on an Inertsil PH‐3 column (100 mm L ×4.6 mm I.D., 5 µm) with a mobile phase consisting of 10 m M ammonium acetate and methanol under linear gradient conditions. Analytes were introduced to the LC‐MS/MS through an electrospray ionization source and detected in positive‐ion mode using selected reaction monitoring. Standard curves were linear from 0.25 to 500 ng/mL ( r ≥0.9964). This assay enabled quantification of ASP015K and M2 at a concentration as low as 0.25 ng/mL in rat plasma. Validation data demonstrated that the method is selective, sensitive and accurate. Further, we also successfully applied this method to a preclinical pharmacokinetic study in rats. Copyright © 2014 John Wiley & Sons, Ltd.