title: Validation of a liquid chromatographic/tandem mass spectrometric method for the determination of scopolamine butylbromide in human plasma: application of the method to a bioequivalence study.
abstract: A sensitive and specific LC/MS/MS method was developed and validated for the determination of scopolamine butylbromide in human plasma. Scopolamine butylbromide and propanolol (internal standard) were extracted from the plasma by liquid-liquid extraction with dichloromethane as the extraction solvent and separated on a C18 analytical column (50 x 4.6 mm id) maintained at 40 degrees C. The analytes were eluted at a constant flow rate of 0.45 mL/min; the mobile phase consisted of acetonitrile and a buffer of 5 mM ammonium acetate and 0.1% formic acid (60 + 40, v/v). The mass spectrometer, equipped with an electrospray source in the positive ionization mode, was set up in the multiple-reaction monitoring mode to monitor the transitions m/z 360.6 > 102.5 (scopolamine butylbromide) and m/z 259.7 > 115.6 (propanolol). The chromatographic separation was obtained within 2.0 min, and the responses were linear over the concentration range of 0.10-40.00 ng/mL. The mean extraction recoveries of scopolamine butylbromide and propanolol from plasma were 69.00 and 80.76%, respectively. Method validation parameters, such as specificity, linearity, precision, accuracy, and stability, were within the acceptable range. Moreover, when 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, Av. Cirne Lima, 1541, 97.105-900--Toledo-PR, Brazil. manfiojoselia@bol.com.br
authors.forename: Josélia Larger
authors.initials: JL
authors.lastname: Manfio
authors.forename: Mauricio Bedim
authors.initials: MB
authors.lastname: Dos Santos
authors.forename: Wagner Alex Jann
authors.initials: WA
authors.lastname: Favreto
authors.forename: Fabiane Ines
authors.initials: FI
authors.lastname: Hoffmann
authors.forename: Adriana Cristina
authors.initials: AC
authors.lastname: Mertin
mesh_terms: D000085:Acetates / Q000032:analysis; D000293:Adolescent; D000328:Adult; D002623:Chemistry Techniques, Analytical; D002626:Chemistry, Pharmaceutical / Q000379:methods*; D002853:Chromatography, Liquid / Q000379:methods*; D005260:Female; D005561:Formates / Q000032:analysis; D006801:Humans; D006842:Hydrocarbons, Brominated / Q000032:analysis* / Q000097:blood*; D008297:Male; D010949:Plasma / Q000187:drug effects; D015203:Reproducibility of Results; D012601:Scopolamine / Q000032:analysis* / Q000097:blood*; D012997:Solvents / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods*; D013696:Temperature
chemical_list: D000085:Acetates; D005561:Formates; D006842:Hydrocarbons, Brominated; D012997:Solvents; C030544:formic acid; D012601:Scopolamine; C018824:ammonium acetate; C047757:butyl bromide