Determination of mianserin in human plasma by high performance liquid chromatography-electrospray ionization mass spectrometry (HPLC-ESI/MS): application to a bioequivalence study in Chinese volunteers

This study aims to develop a standard protocol for the bioequivalence study of mianserin hydrochloride tablets--a tetracyclic antidepressant drug. For this purpose, a rapid, convenient and selective method using high performance liquid chromatography coupled with electrospray ionization mass spectrometry (HPLC-ESI/MS) has been developed and validated to determine mianserin in human plasma. Mianserin and the internal standard (I.S.), cinnarizine were extracted from plasma by N-hexane:dimethylcarbinol (98:2, v/v) after alkalinized with sodium hydroxide. LC separation was performed on a Thermo Hypersil-Hypurity C18 (5 microm, 150 mm x 2.1 mm) with the mobile phase consisting of 10mM ammonium acetate (pH 3.4)-methanol-acetonitrile (35:50:15, v/v/v) at 0.22 ml/min. The retention time of mianserin and cinnarizine was 3.4 and 2.1 min, respectively. Quadrupole MS detection and quantitation was done by monitoring at m/z 265 [M+H]+ for mianserin and m/z 369 [M+H]+ for cinnarizine. The method was validated over the concentration ranges of 1.0-200.0 ng/ml for mianserin. The recovery was 81.3-84.1%, intra- and inter-day precision of the assay at three concentrations were 9.6-11.4% with accuracy of 97.5-101.2% and the lower limit of quantitation (LLOQ) detection was 1.0 ng/ml for mianserin. The stability of compounds was established in a battery of stability studies, i.e., short-term and long-term storage stability as well as freeze-thaw cycles. This method proved to be suitable for the bioequivalence study of mianserin hydrochloride tablets in healthy human male volunteers.