RATIONALE: Artemisitene shows a wide variety of pharmacological activities, such as antioxidant protection in vitro and in vivo. It has been identified as a novel Nrf2 inducer. However, there is no report on an ultra-performance liquid chromatography/tandem mass spectrometry (UPLC/MS/MS) method to quantitate artemisitene in rat plasma and its application to a pharmacokinetic profile study. METHODS: An ACQUITY UPLCTM BEH Symmetry Shield RP18 column (1.7 mu m, 2.1 mm x 100 mm) was used at a flow rate of 0.3 mL.min(-1). Mass detection was performed by electrospray ionization tandem mass spectrometry via multiple reaction monitoring (MRM) in positive mode. Plasma samples were pre-treated by a single-step extraction with 0.1% formic acid aqueous solutions-acetonitrile, and tolbutamide was used as internal standard. RESULTS: The calibration curve was from 0.98 to 1000 ng.mL(-1) (r(2) = 0.995). The extraction recoveries were 61.5-79.4% and 81.7-94.6% for artemisitene and tolbutamide, respectively. The lower limit of quantification (LLOQ) was 0.98 ng.mL(-1). The absolute bioavailability of artemisitene was 3.7% after intravenous and oral administration in rats. CONCLUSIONS: The UPLC/MS/MS assay was validated for linearity, accuracy, stability, extraction recovery, matrix effects, and intra-day and inter-day precision. The method, for the first time, achieved some pharmacokinetic parameters and was successfully applied to a pharmacokinetic study. Copyright (c) 2017 John Wiley & Sons, Ltd.