Hyperoside is one of the major components of Hypericum perforatum L. and also present in many plant species such as Abelmoschus manihot （L.） Medik., Ribes nigrum L. and Rosa agrestis Savi （Rosaceae）. Because hyperoside exhibits many biological activities, the pharmacokinetics profile of hyperoside needs to be studied for further elucidating its mechanism of action. A simple method for the determination of hyperoside in rat plasma was developed by using ultra-high performance liquid chromatography coupled with mass spectrometry （UPLC-MS）. Only 50 ~tL plasma samples were required for sample preparation. The quantitative detection of hyperoside was accomplished by selected ion monitoring （SIM） in negative ion mode. Hyperoside was analyzed in less than 10 rain. Good linearity was obtained （r2〉0.999） and the intra- and inter-day precision of the method were lower than 15%. Lower limit of quantification （LLOQ） was 4 ng/mL for hyperoside in rat plasma. Our method showed advantage in the lower LLOQ compared with the reported method; furthermore, smaller amount of plasma was needed. The method was successfully applied for the pharmacokinetics study of hyperoside in rat after intravenous administration of hyperoside.
