Determination of eurycomanone in rat plasma using hydrophilic interaction liquid chromatography-tandem mass spectrometry for pharmacokinetic study

Abstract

In this study, a rapid, sensitive, and reliable hydrophilic interaction liquid chromatography-tandem mass spectrometry (HILIC-MS/MS) method for the determination of eurycomanone in rat plasma was developed and validated. Plasma samples were pretreated with a protein precipitation method and quercitrin was used as an internal standard (IS). A HILIC silica column (2.1x100mm, 3m) was used for hydrophilic-based chromatographic separation, using the mobile phase of 0.1% formic acid with acetonitrile in gradient elution at a flow rate of 0.25mL/min. Precursor-product ion pairs for multiple-reaction monitoring were m/z 409.1391.0 for eurycomanone and m/z 449.1303.0 for IS. The linear range was 2-120ng/mL. The intra- and inter-day accuracies were between 95.5 and 103.4% with a precision of <4.2%. The developed method was successfully applied to the pharmacokinetic analysis of eurycomanone in rat plasma after oral dosing with pure compound and E. longifolia extract. The C-max and AUC(0-t), respectively, were 40.43 +/- 16.08ng/mL and 161.09 +/- 37.63ngh/mL for 10mg/kg eurycomanone, and 9.90 +/- 3.97ng/mL and 37.15 +/- 6.80ngh/mL for E. longifolia extract (2mg/kg as eurycomanone). The pharmacokinetic results were comparable with each other, based on the dose as eurycomanone.