Pharmacokinetic studies of six major 2‐(2‐phenylethyl) chromones in rat plasma using ultra high performance liquid chromatography with tandem mass spectrometry after oral administration of agarwood ethanol extract

Abstract In this study, a simple, quick, sensitive and reliable method utilizing ultra‐high performance liquid chromatography with tandem mass spectrometry method was validated for simultaneous quantification of six main 2‐(2‐phenylethyl) chromones, including agarotetrol, isoagarotetrol, (5S,6R,7R,8S)‐5,6,7,8‐tetrahydroxy‐(4‐methoxyphenethyl)‐5,6,7,8‐tetrahydro‐4H‐chromen‐4‐one, 8‐chloro‐2‐(2‐phenyl ethyl)‐5,6,7‐trihydroxy‐5,6,7,8‐tetrahydrochromone, 6,7‐dimethoxy‐2‐(2‐phenylethyl) chromone, and 2‐(2‐phenylethyl) chromone in rat plasma after oral administration of agarwood ethanol extract. Separation was performed on a Waters ACQUITY UPLC BEH C 18 column (2.1 × 100 mm, 1.7 μm) using gradient elution with mobile phase of 0.2% formic acid‐water and acetonitrile. The tandem mass was performed in the multiple reaction monitoring mode with positive ionization. The calibration curves indicated good linearity ( r 2 > 0.99) over the corresponding concentration range. The precision and accuracy were within the acceptable range. Mean absolute recoveries of all analytes were between 73.31% and 94.76%, and the relative standard deviations of matrix effects were not higher than 15%. The six analytes were proven to be stable during sample storage and analysis procedures. The validated method was successfully applied to pharmacokinetic study of six 2‐(2‐phenylethyl) chromones in rat after oral administration of agarwood ethanol extract for the first time. This study could serve as a reference and provide theoretical guidance for further pharmacodynamic research and clinical applications of agarwood.