title: Development and validation of a UPLC-ESI-MS/MS method for the determination of N-butylscopolamine in human plasma: application to a bioequivalence study.
abstract: A sensitive and fast ultra performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UPLC-ESI-MS/MS) method for measurements of N-butylscopolamine in plasma was developed and validated. A single protein precipitation was proposed for the clean up of the plasma and N-methylhomatropine was added as internal standard (IS). The analyses were carried out using a C(18) column and mobile phase of acetonitrile: 5 mM ammonium acetate + 0.1% formic acid (90:10, v/v). The triple quadrupole mass spectrometer equipped with an electrospray source in positive mode, was set up in selective reaction monitoring, to detect precursor → product ion 360.0 → 194.0 m/z and 290.3 → 138.0 m/z transitions, for N-butylscopolamine and IS, respectively. The method was linear in 0.03 (lower limit of quantitation; LLOQ) - 10.00 ng/ml range for N-butylscopolamine. Satisfactory selectivity, linearity, precision, accuracy, and robustness were obtained for the UPLC-ESI-MS/MS method. The proposed method was successfully applied to a pharmacokinetic study of healthy human volunteers; the results showed that the two scopolamine butylbromide formulations tested are not bioequivalent in rate and extent of absorption.
authors.affiliation: Biocinese-Centro de Estudos Biofarmacêuticos Ltda, Av. Cirne Lima, 1541-Vila Becker, Toledo, PR, Brazil.
authors.forename: Wagner Alex Jann
authors.initials: WA
authors.lastname: Favreto
authors.forename: Ana Maria
authors.initials: AM
authors.lastname: Pugens Pinto
authors.forename: Josélia Larger
authors.initials: JL
authors.lastname: Manfio
authors.forename: Karina Graziella
authors.initials: KG
authors.lastname: Fiametti
authors.forename: Maycon Fernando
authors.initials: MF
authors.lastname: Percio
authors.forename: Mauricio Bedim dos
authors.initials: MB
authors.lastname: Santos
mesh_terms: D000328:Adult; D002086:Butylscopolammonium Bromide / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D018727:Muscarinic Antagonists / Q000097:blood*; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods; D055815:Young Adult
chemical_list: D018727:Muscarinic Antagonists; D002086:Butylscopolammonium Bromide