title: Development and validation a LC-MS/MS method for the simultaneous determination of agomelatine and its metabolites, 7-desmethyl-agomelatine and 3-hydroxy-agomelatine in human plasma: Application to a bioequivalence study.
abstract: A novel sensitive and selective LC-MS/MS method for the determination of agomelatine, 7-desmethyl-agomelatine and 3-hydroxy-agomelatine in human plasma was developed and validated. After simple protein precipitation, the analytes were separated on a Phenomenex ODS3 column (4.6×150 mm, 5μm, Phenomenex, USA) with mobile phase consisted of methanol and 5mM ammonium formate solution (containing 0.2% formic acid) at a ratio of 70:30 (v/v) with a flow rate of 0.8mL/min. The MS acquisition was performed in multiple reactions monitoring (MRM) mode with a positive electrospray ionization source. The mass transitions monitored were m/z 244.1→185.1, m/z 230.1→171.1, m/z 260.1→201.1 and m/z 180.1→110.1 for agomelatine, 7-desmethyl-agomelatine, 3-hydroxy-agomelatine and internal standard (phenacetin), respectively. The method was validated for specificity, linearity and lower limit of quantification, precision and accuracy, extraction recovery, matrix effect and stability. The calibration curves for agomelatine, 7-desmethyl-agomelatine and 3-hydroxy-agomelatine in human plasma were linear over concentration ranges of 0.0457-100ng/mL, 0.1372-300ng/mL and 0.4572-1000ng/mL, respectively. Intra- and inter-day precisions and accuracies data met the acceptance criteria of FDA guideline for bioanalytical method validation. The developed method has been successfully applied to a bioequivalence study in healthy Chinese volunteers.
authors.affiliation: School of Pharmaceutical Sciences, Central South University, Changsha 410013, PR China.
authors.forename: Meizhen
authors.initials: M
authors.lastname: Li
authors.affiliation: School of Pharmaceutical Sciences, Central South University, Changsha 410013, PR China.
authors.forename: Fang
authors.initials: F
authors.lastname: Tang
authors.affiliation: School of Pharmaceutical Sciences, Central South University, Changsha 410013, PR China. Electronic address: feifanxie@hotmail.com.
authors.forename: Feifan
authors.initials: F
authors.lastname: Xie
authors.affiliation: School of Pharmaceutical Sciences, Central South University, Changsha 410013, PR China.
authors.forename: Yisha
authors.initials: Y
authors.lastname: Lv
authors.affiliation: School of Pharmaceutical Sciences, Central South University, Changsha 410013, PR China.
authors.forename: Peng
authors.initials: P
authors.lastname: Yu
authors.affiliation: Hunan Tiger-Xiangya R&D Company Ltd., Changsha 410013, PR China.
authors.forename: Zhi
authors.initials: Z
authors.lastname: Liu
authors.affiliation: School of Pharmaceutical Sciences, Central South University, Changsha 410013, PR China. Electronic address: chengzn@csu.edu.cn.
authors.forename: Zeneng
authors.initials: Z
authors.lastname: Cheng
mesh_terms: D000081:Acetamides / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D000293:Adolescent; D000328:Adult; D000928:Antidepressive Agents / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005260:Female; D006801:Humans; D008297:Male; D015394:Molecular Structure; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*; D013810:Therapeutic Equivalency; D055815:Young Adult; D009281:Naphthalenes
chemical_list: D000081:Acetamides; D000928:Antidepressive Agents; C084711:agomelatine; D009281:Naphthalenes
keywords: 3-Hydroxy-agomelatine; 7-Desmethyl-agomelatine; Agomelatine; Bioequivalence; LC–MS/MS; Metabolite; Plasma