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First LC–MS/MS method for trans-ε-viniferin quantification in mouse plasma.A simple protein precipitation that allowed for efficient/reproducible recovery yields was used.The small plasma volume (10 μl) supports the ability to examine pharmacokinetics in mice.In this study, a simple and sensitive liquid chromatography-tandem mass spectrometry (LC–MS/MS) method for the quantification of trans-ε-viniferin in small volumes (10 μl) of mouse plasma using chlorpropamide as an internal standard was developed and validated. Plasma samples were precipitated with acetonitrile and separated using an Eclipse Plus C18 column (100 × 4.6 mm, 1.8-μm) with a mobile phase consisting of 0.1% formic acid in acetonitrile and 0.1% formic acid in water (60:40 v/v) at a flow rate of 0.5 ml/min. A triple quadrupole mass spectrometer operating in positive ion mode with selected reaction-monitoring mode was used to determine trans-ε-viniferin and chlorpropamide transitions of 455.10 → 215.05 and 277.00 → 111.00, respectively. The lower limit of quantification was 5 ng/ml with a linear range of 5–2500 ng/ml (r ≥ 0.9949). All validation data, including the selectivity, precision, accuracy, recovery, dilution integrity, and stability, conformed to the acceptance requirements. No matrix effects were observed. The developed method was successfully applied to pharmacokinetic studies of trans-ε-viniferin following intravenous (2.5 mg/kg), intraperitoneal (2.5, 5 and 10 mg/kg), and oral (40 mg/kg) administration in mice. This is the first report on the pharmacokinetic properties of trans-ε-viniferin. The results provide a meaningful basis for evaluating the pre-clinical or clinical applications of trans-ε-viniferin.