Pharmacokinetic Study of Tussilagone in Rat Plasma by UPLC-MS/MS

Tussilagone was isolated from the flower of buds of Tussilago farfara (Compositae). In this work, a sensitive and selective UPLC-MS/MS method for determination of tussilagone in rat plasma was developed. After addition of segetalin A as an internal standard (IS), protein precipitation by acetonitrilemethanol (9:1, v/v) was used to prepare samples. Chromatographic separation was achieved on an UPLC BEH C18 column (2.1 x 50 mm, 1.7 mu m) with 0.1% formic acid and acetonitrile as the mobile phase with gradient elution. An electrospray ionization source was applied and operated in positive ion mode; multiple reactions monitoring (MRM) mode was used for quantification using target fragment ions m/z 413.2 -> 299.2 for tussilagone, and m/z 632.2 -> 604.3 for IS. Calibration plots were linear throughout the range 2-1000 ng/mL for tussilagone in rat plasma. Mean recoveries of tussilagone in rat plasma ranged from 87.6 to 95.4%. RSD of infra-day and inter-day precision were both less than 8%. The accuracy of the method was between 97.6 and 105.7%. The method was successfully applied to pharmacokinetic study of tussilagone after intravenous administration in rats. The main pharmacokinetic parameters after intravenous administration of tussilagone in rats were AUC((0-t)) 175.7 +/- 255 ng/mL.h, t(1/2) 0.5 +/- 0.3 h, and CL 11.5 +/- 1.8 L/h/kg.