title: Sensitive LC-MS/MS-ESI method for simultaneous determination of montelukast and fexofenadine in human plasma: application to a bioequivalence study.
abstract: A rapid, simple, sensitive and selective LC-MS/MS method was developed and validated for simultaneous quantification of montelukast (MT) and fexofenadine (FF) in human plasma (200 μL) using montelukast-d6 (MT-d6 ) and fexofenadine-d10 (FF-d10 ), respectively as an internal standard (IS) as per the US Food and Drug Administration guidelines. The chromatographic resolution was achieved on a Chromolith RP18e column using an isocratic mobile phase consisting of 20 mm ammonium formate-acetonitrile (20:80, v/v) at flow rate of 1.2 mL/min. The LC-MS/MS was operated under the multiple-reaction monitoring mode using electrospray ionization. The total run time of analysis was 4 min and elution of MT, FF, MT-d6 and FF-d10 occurred at 2.5, 1.2, 2.4 and 1.2 min, respectively. The standard curve found to be linear in the range 2.00-1000 ng/mL with a coefficient of correlation of ≥0.99 for both the drugs. The intra- and inter-day accuracy and precision values for MT and FF met the acceptance as per FDA guidelines. MT and FF were found to be stable in a battery of stability studies viz., bench-top, auto-sampler and repeated freeze-thaw cycles. The validated assay was applied to an oral bioequivalence study in humans.
authors.affiliation: Veeda Clinical Research Pvt. Ltd, Shivalik Plaza-A, Ambawadi, Ahmedabad, 380015, India; Department of Pharmacy, Annamalai University, Annamalai Nagar, 608002, India.
authors.forename: Rajendraprasad
authors.initials: R
authors.lastname: Muppavarapu
authors.forename: Swati
authors.initials: S
authors.lastname: Guttikar
authors.forename: Manavalan
authors.initials: M
authors.lastname: Rajappan
authors.forename: Kannan
authors.initials: K
authors.lastname: Kamarajan
authors.forename: Ramesh
authors.initials: R
authors.lastname: Mullangi
mesh_terms: D000085:Acetates / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D000328:Adult; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D018592:Cross-Over Studies; D003521:Cyclopropanes; D004355:Drug Stability; D006801:Humans; D016014:Linear Models; D008297:Male; D011804:Quinolines / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D016037:Single-Blind Method; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D013440:Sulfides; D053719:Tandem Mass Spectrometry / Q000379:methods*; D016593:Terfenadine / Q000031:analogs & derivatives* / Q000097:blood / Q000737:chemistry / Q000493:pharmacokinetics; D013810:Therapeutic Equivalency
chemical_list: D000085:Acetates; D003521:Cyclopropanes; D011804:Quinolines; D013440:Sulfides; D016593:Terfenadine; C093230:fexofenadine; C093875:montelukast
keywords: LC-MS/MS; bioequivalence; fexofenadine; human plasma; method validation; montelukast; pharmacokinetics