Pharmacokinetics of eupalinolide A, eupalinolide B and hyperoside from Eupatorium lindleyanum in rats by LC/MS/MS

•EA and EB are the cis and trans isomers.•A method of simultaneous determination of EA, EB and HYP in plasma was first developed.•The LLOQs of EA, EB and HYP were 1.28, 1.98 and 2.00 ng/mL, respectively.•PK on EA and EB isomers from Eupatorium lindleyanum was first carried out.

A simple, selective, and sensitive LC/MS/MS method was developed and validated for simultaneous determination of eupalinolide A, eupalinolide B, and hyperoside in rat plasma. Plasma samples were processed by protein precipitation with acetonitrile. The three analytes, together with internal standard (IS, lysionotin), were separated on a Venusil MP-C18 column (50 mm × 2.1 mm, 3 μm) using a mobile phase of methanol and 10 mM ammonium acetate (45:55, v/v) with isocratic elution. Mass spectrometric detection was performed by multiple-reaction monitoring mode via electrospray ionization source. Linear calibration curves were obtained for the following concentration range: 1.28–640 ng/mL for EA; 1.98–990 ng/mL for EB; and 2.00–1000 ng/mL for HYP. The intra- and inter-day precision was less than 10.25%, and the accuracy was between 89.16% and 110.63%. The extraction recovery of the analytes and IS from rat plasma was above 88.75%. The validated method has been successfully applied to pharmacokinetic studies of the three analytes following intragastric administration of Eupatorium lindleyanum extract at a single dose of 100, 250, and 625 mg/kg to Sprague-Dawley rats, respectively. The pharmacokinetic results may help to better understand the pharmacological actions of the herb E. lindleyanum.