An effective separation and simultaneous determination of corynoxeine and its metabolites using high-performance liquid chromatography with tandem mass spectrometry was developed and validated. The method was applied to pharmacokinetics and in vivo distribution investigations in rats after oral (0.105 mmol kg(-1)) and intravenous (0.0105 mmol kg(-1)) doses of corynoxeine. Its brain uptake index was of 3.08 × 10(-11) mol g(-1) at 3 h and 3.75 × 10(-11) mol g(-1) at 74 min after oral and intravenous doses, respectively.