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Simultaneous quantitation of IC87114, roflumilast and its active metabolite roflumilast N‐oxide in plasma by LC‐MS/MS: application for a pharmacokinetic study
Author(s) -
Thappali Satheeshmanikandan RS,
Varanasi Kanthikiran VS,
Veeraraghavan Sridhar,
Vakkalanka Swaroop Kumar VS,
K Mukkanti
Publication year - 2012
Publication title -
journal of mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 121
eISSN - 1096-9888
pISSN - 1076-5174
DOI - 10.1002/jms.3103
Subject(s) - chemistry , chromatography , analyte , ammonium acetate , tolbutamide , roflumilast , metabolite , pharmacokinetics , selected reaction monitoring , active metabolite , calibration curve , liquid chromatography–mass spectrometry , mass spectrometry , tandem mass spectrometry , high performance liquid chromatography , detection limit , pharmacology , medicine , psychology , biochemistry , psychiatry , copd , endocrinology , diabetes mellitus
A sensitive and reliable high‐performance liquid chromatography–mass spectrometry (LC–MS/MS) was developed and validated for simultaneous quantification IC87114, roflumilast (RFM), and its active metabolite roflumilast N‐oxide (RFN) using tolbutamide as an internal standard. The analytes were extracted by using liquid–liquid extraction and separated on a reverse phase C 18 column (50 mm × 3 mm i.d., 4.6 µ) using methanol: 2 mM ammonium acetate buffer, pH 4.0 as mobile phase at a flow rate 1 mL/min in gradient mode. Selective reaction monitoring was performed using the transitions m/z 398.3 > 145.9, 403.1 >186.9, 419.1 > 187.0 and 271.1 > 155.0 to quantify quantification IC87114, RFM, RFN and tolbutamide, respectively. The method was validated over the concentration range of 0.1–60 ng.mL −1 for RFM and RFN and 6 to 2980 ng.mL −1 for IC87114. Intra‐ and inter‐day accuracy and precision of validated method were within the acceptable limits of <15% at all concentrations. Coefficients of correlation ( r 2 ) for the calibration curves were >0.99 for all analytes. The quantitation method was successfully applied for simultaneous estimation of IC87114, RFM and RFN in a pharmacokinetic drug–drug interaction study in Wistar rats. Copyright © 2012 John Wiley & Sons, Ltd.