title: Validation of an ultra-performance liquid chromatography-tandem mass spectrometry method for the determination of flecainide in human plasma and its clinical application.
abstract: A simple and reproducible bioanalytical method for the determination of flecainide in human plasma was developed and validated using an ultra-performance liquid chromatography with tandem mass spectrometry (UPLC-MS/MS) to obtain higher sensitivity than the current available methods. After simple protein precipitation, flecainide and a stable isotope-labeled internal standard (IS) were chromatographed on an Acquity UPLC BEH C18 column (2.1 × 100 mm, 1.7 µm) with isocratic elution of mobile phase consisting of 45% methanol containing 0.1% formic acid at a flow rate 0.25 mL/min. Detection was performed in positive electrospray ionization by monitoring the selected ion transitions at m/z 415.4/301.1 for flecainide and m/z 419.4/305.1 for the IS. The method was validated according to current bioanalytical method validation guidelines. The calibration standard curve was linear from 2.5 to 1000 ng/mL using 0.1 mL of plasma. No significant interferences were detected in blank human plasma. Accuracy and precision in the intra- and inter-batch reproducibility study were within acceptance criteria. Neither hemolysis effects nor matrix effects were observed. The UPLC-MS/MS method developed was successfully applied to determine plasma flecainide concentrations to support clinical studies and incurred sample reanalysis also ensured the reproducibility of the method.
authors: Drug Metabolism and Pharmacokinetics, Biopharmaceutical Assessment Core Function Unit, Eisai Co. Ltd, 1-3, 5-chome, Tokodai, Tsukuba-shi, Ibaraki, 300-2635, Japan.
authors: Yuji
authors: Y
authors: Mano
authors: Analysis group, Tsukuba Division, Sunplanet Co. Ltd, 1-3, 5-chome, Tokodai, Tsukuba-shi, Ibaraki, 300-2635, Japan.
authors: Yoshiki
authors: Y
authors: Asakawa
authors: Analysis group, Tsukuba Division, Sunplanet Co. Ltd, 1-3, 5-chome, Tokodai, Tsukuba-shi, Ibaraki, 300-2635, Japan.
authors: Kenji
authors: K
authors: Kita
authors: Analysis group, Tsukuba Division, Sunplanet Co. Ltd, 1-3, 5-chome, Tokodai, Tsukuba-shi, Ibaraki, 300-2635, Japan.
authors: Takuho
authors: T
authors: Ishii
authors: Drug Metabolism and Pharmacokinetics, Biopharmaceutical Assessment Core Function Unit, Eisai Co. Ltd, 1-3, 5-chome, Tokodai, Tsukuba-shi, Ibaraki, 300-2635, Japan.
authors: Koichiro
authors: K
authors: Hotta
authors: Drug Metabolism and Pharmacokinetics, Biopharmaceutical Assessment Core Function Unit, Eisai Co. Ltd, 1-3, 5-chome, Tokodai, Tsukuba-shi, Ibaraki, 300-2635, Japan.
authors: Kazutomi
authors: K
authors: Kusano
mesh_terms: D000889:Anti-Arrhythmia Agents / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005424:Flecainide / Q000097:blood*; D006801:Humans; D057230:Limit of Detection; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D000889:Anti-Arrhythmia Agents; D005424:Flecainide
keywords: ISR; UPLC-MS/MS; flecainide; hemolysis; validation