LAUSR

HighlightsA simple, rapid and efficient pretreatment of AF‐HF001 is reported.A full validated LC–MS/MS method of AF‐HF001 is described for the first time.Successful application to a pharmacokinetics study of AF‐HF001 in mice plasma.First mice pharmacokinetics report of AF‐HF001 to date. Abstract Heart failure (HF) is one of the most serious health problems worldwide. A new positive inotropic compound, an isonicotinylhydrazide derivative (AF‐HF001) was designed recently. A liquid chromatography‐tandem mass spectrometry (LC–MS/MS) method was developed for the determination of the target analyte in mouse plasma. Samples were prepared by one step precipitation with ethyl acetate and stored in acetonitrile. Chromatographic analysis was carried out on a Hypersil Gold C18 column (2.1 mm × 50 mm, 3 &mgr;m) with a gradient mobile phase consisting of acetonitrile and 0.1% aqueous formic acid. The analyte was detected by selective reaction monitoring (SRM) mode with target quantitative ion pair of m/z 292.1 → 148.2, using praziquantel as the internal standard (IS) m/z 313.1 → 203.2. Good linearity (r = 0.995) was observed over a wide concentration range. The validation of method showed acceptable recovery and precision. The method has been then applied to a very first pharmacokinetic assay of AF‐HF001 in mice.