A Validated Liquid Chromatography–Tandem Mass Spectrometry Method for the Determination of Methylophiopogonanone A in Rat Plasma and Its Application to Pharmacokinetic Study

ABSTRACT Methylophiopogonanone A (MOA) is one of the homoisoflavonoids isolated from Ophiopogon japonicus , which has been demonstrated to have extensive pharmacological activities. The aim of this study was to develop and validate a liquid chromatography–tandem mass spectrometry method for the measurement of MOA in rat plasma. Methylophiopogonanone B (MOB) was used as internal standard. After precipitation with acetonitrile, the samples were separated using a Waters ACQUITY HSS T 3 column with 0.1% formic acid solution and acetonitrile as mobile phase. Mass detection was monitored using multiple reactions monitoring mode with precursor‐to‐product ion transition of m/z 343.2 > 135.1 for MOA and m/z 329.2 > 121.1 for internal standard. The assay showed good linearity over the concentration range of 1–1000 ng/mL, with correlation coefficient > 0.997. The lower limit of quantification (LLOQ) was 1 ng/mL. Acetonitrile‐mediated precipitation showed high extraction efficiency (> 80%). The accuracy and precision were within the acceptable limits. MOA was demonstrated to be stable in rat plasma under the tested storage conditions. After validation, the proposed method was applied to the pharmacokinetic study of MOA in rat plasma, and the data revealed that MOA showed rapid absorption and elimination from rat plasma. The oral bioavailability of MOA in rat was 24.5%.