title: Simultaneous quantitation of IC87114, roflumilast and its active metabolite roflumilast N-oxide in plasma by LC-MS/MS: application for a pharmacokinetic study.
abstract: 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.
authors.affiliation: Drug Metabolism and Pharmacokinetics Division, Incozen Therapeutics Private Limited, 450, Alexandria Knowledge Park, Shamirpet Hyderabad, 500078, Andhra Pradesh, India. satheeshtrs@gmail.com
authors.forename: Satheeshmanikandan R S
authors.initials: SR
authors.lastname: Thappali
authors.forename: Kanthikiran V S
authors.initials: KV
authors.lastname: Varanasi
authors.forename: Sridhar
authors.initials: S
authors.lastname: Veeraraghavan
authors.forename: Swaroop Kumar V S
authors.initials: SK
authors.lastname: Vakkalanka
authors.forename: K
authors.initials: K
authors.lastname: Mukkanti
mesh_terms: D000225:Adenine / Q000031:analogs & derivatives* / Q000097:blood / Q000737:chemistry / Q000493:pharmacokinetics; D000631:Aminopyridines / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D000818:Animals; D001549:Benzamides / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003521:Cyclopropanes / Q000097:blood / Q000737:chemistry / Q000493:pharmacokinetics; D004355:Drug Stability; D016014:Linear Models; D008297:Male; D011799:Quinazolines / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D051381:Rats; D017208:Rats, Wistar; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D000631:Aminopyridines; D001549:Benzamides; D003521:Cyclopropanes; C477872:IC 87114; D011799:Quinazolines; C424423:Roflumilast; C517734:roflumilast N-oxide; D000225:Adenine