Development of a hydrophilic interaction liquid chromatography–tandem mass spectrometric method for the determination of kinsenoside, an antihyperlipidemic candidate, in rat plasma and its application to pharmacokinetic studies

•HILIC–MS/MS for quantitation of kinsenoside in rat plasma was developed and validated.•The method was specific, precise, and accurate, and no matrix effect was observed.•The method was successfully applied to the pharmacokinetic study of kinsenoside in rats.

Kinsenoside is a major bioactive constituent isolated from Anoectochilus formosanus and is investigated as an antihyperlipidemic candidate. In this study, a rapid, sensitive, and reliable bioanalytical method was developed for the determination of kinsenoside in rat plasma using hydrophilic interaction liquid chromatography–tandem mass spectrometry (HILIC–MS/MS). The plasma sample was pretreated with 1% acetic acid, followed by protein precipitation with acetonitrile:methanol (70:30). Chromatographic separation was performed on a HILIC silica column (2.1 mm × 100 mm, 3 μm). The mobile phases consisted of 0.1% acetic acid in distilled water (solvent A) and 0.1% acetic acid in acetonitrile (solvent B). A gradient program was used at a flow rate of 0.2 mL/min. For mass spectrometric detection, the multiple reaction monitoring mode was used; the MRM transitions were m/z 265.2 → m/z 102.9 for kinsenoside and m/z 163.3 → m/z 132.1 for the internal standard (IS) nicotine in the positive ionization mode. A calibration curve was constructed in the range of 2–500 ng/mL. The intra- and interday precision and accuracy were within 5%. The HILIC–MS/MS method was specific, accurate, and reproducible and was successfully applied in a pharmacokinetic study of kinsenoside in rats.

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