Simultaneous determination and pharmacokinetic study of Atractylenolide I, II and III in rat plasma after intragastric administration of Baizhufuling extract and Atractylodis extract by UPLC–MS/MS

• A UPLC–MS method was developed for the determination of Atractylenolides.
• The total analysis time was 4.0 min.
• The LLOQs of Atractylenolide I, II, III were 0.8, 0.8 and 4 ng/ml, respectively.
• This method was applied to the comparative pharmacokinetic study.

A simple and rapid ultra high performance liquid chromatography-tandem mass spectrometry (UPLC–MS/MS) method was developed for the simultaneous determination of Atractylenolide I, II and III in rat plasma. Plasma samples were processed by liquid–liquid extraction with ethyl acetate, using schisandrin as internal standard (IS). Chromatographic separation was accomplished on a Thermo Hypersil GOLD C18 column (2.1 mm × 50 mm, 1.9 μm) with mobile phase consisting of acetonitrile and 0.1% formic acid–water (50:50, v/v). The detection was carried out by ESI-MS (positive ionization mode) and low-energy collision dissociation tandem mass spectrometric analyses using the multiple-reaction monitoring (MRM) scan mode. The quantification was performed using the transitions of the protonated molecule → product ion at m/z 231.0 → 185.1 for Atractylenolide I, at m/z 233.1 → 187.1 for Atractylenolide II and at m/z 249.1 → 231.1 for Atractylenolide III, respectively. Method validation revealed excellent linearity over investigated range together with satisfactory intra- and inter-day precision, accuracy, matrix effects and extraction recoveries. This method was successfully applied to the comparative pharmacokinetic study of Atractylenolide I, II and III in rat plasma after intragastric administration of Baizhufuling extract and Atractylodis extract.