Development and validation of high‐performance liquid chromatography/electrospray ionization mass spectrometry for assay of madecassoside in rat plasma and its application to pharmacokinetic study

A rapid, sensitive and specific high‐performance liquid chromatography/electrospray ionization mass spectrometric (LC‐ESI‐MS) method was developed and validated for the quantification of madecassoside, a major active constituent of Centella asiatica (L.) Urb. herbs, in rat plasma. With paeoniflorin as an internal standard (IS), a simple liquid–liquid extraction process was employed for the plasma sample preparation. Chromatographic separation was achieved within 6 min on a Shim‐pack CLC‐ODS column using acetonitrile and water (60:40, v/v) containing 0.1% (v/v) formic acid as the mobile phase. The detection was performed by MS with electrospray ionization interface in negative selected ion monitoring (SIM) mode. The linear range was 11–5500 ng/mL with the square regression coefficient ( r 2 ) of 0.9995. The lower limit of quantification was 11 ng/mL. The intra‐ and inter‐ day precision ranged from 4.99 to 9.03%, and the accuracy was between 95.82 and 111.80%. The average recoveries of madecassoside and IS from spiked plasma samples were >92%. The developed method was successfully applied to the pharmacokinetic study of madecassoside in rats after an oral administration. Copyright © 2011 John Wiley & Sons, Ltd.