Determination of dapoxetine in rat plasma by ultra-performance liquid chromatography–tandem mass spectrometry

•UPLC–MS/MS for quantitation of dapoxetine in rat plasma was developed and validated.•The method was specific, precise, and accurate, and no matrix effect was observed.•The method was successfully applied in pharmacokinetic studies of dapoxetine in rats. In this study, we describe and validate a rapid and sensitive method for quantitation of dapoxetine in rat plasma by using ultra-performance liquid chromatography–electrospray ionization tandem mass spectrometry (UPLC–ESI/MS/MS). Plasma samples were prepared by protein precipitation with acetonitrile, and sildenafil was used as an internal standard (IS). The mobile phase consisted of 0.5% formic acid/acetonitrile (60:40, v/v); a C18 reversed-phase column (2.0×50mm, 1.7μm) was used for chromatographic separation. Multiple reaction monitoring (MRM) was used in the positive ion mode for mass spectrometric detection. The calibration curve for dapoxetine was linear (r2=0.999) in the concentration range of 1–500ng/mL. The intra- and inter-day precision was between 3.8% and 8.3%, and the intra- and inter-day accuracy was between 101.1% and 109.0%. Dapoxetine was found to be stable in various conditions with the recoveries>87.0% (RSD