Simultaneous quantitation of IC87114, roflumilast and its active metabolite roflumilast N-oxide in plasma by LC-MS/MS: application for a pharmacokinetic study

• Published in: Journal of mass spectrometry. 2012-12, Vol.47 (12), p.1612-1619
• Main Authors: Thappali, Satheeshmanikandan RS, Varanasi, Kanthikiran VS, Veeraraghavan, Sridhar, Vakkalanka, Swaroop Kumar VS, K, Mukkanti
• Format: Article
• Language: English
• Subjects: Adenine - analogs & derivatives; Adenine - blood; Adenine - chemistry; Adenine - pharmacokinetics; Aminopyridines - blood; Aminopyridines - chemistry; Aminopyridines - pharmacokinetics; Analysis; Animals; Benzamides - blood; Benzamides - chemistry; Benzamides - pharmacokinetics; Biological and medical sciences; Chromatography, High Pressure Liquid - methods; Correlation analysis; Cyclopropanes - blood; Cyclopropanes - chemistry; Cyclopropanes - pharmacokinetics; Drug Stability; Flow rate; General pharmacology; IC87114; LC-MS/MS; Linear Models; Liquids; Male; Mass spectrometry; Medical sciences; Metabolites; Methyl alcohol; pharmacokinetics; Pharmacology. Drug treatments; plasma; Quinazolines - blood; Quinazolines - chemistry; Quinazolines - pharmacokinetics; Rats; Rats, Wistar; Reproducibility of Results; Reproduction; roflumilast; roflumilast N-oxide; Sensitivity and Specificity; Tandem Mass Spectrometry - methods
• ISSN: 1076-5174; 1096-9888
• DOI: 10.1002/jms.3103

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 C18 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 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.