A Quick Method for the Determination of Neomangiferin in Rat Plasma by UPLC-MS/MS

An ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method was developed to determine neomangiferin in rat plasma using midazolam as the internal standard (IS). Sample preparation was accomplished through a protein precipitation procedure with acetonitrile to 0.1 mL plasma sample. The analyte and IS were separated on an Acquity UPLC BEH C18 column (2.1 x 50 mm, 1.7 mu m) with the mobile phase of acetonitrile and 0.1% formic acid in water with gradient elution at a flow rate of 0.40 mL/min. The injection volume was 2 mu L. The detection was performed on a triple quadrupole tandem mass spectrometer equipped with electrospray ionization (ESI) by multiple reactions monitoring (MRM) of the transitions at m/z 585.2 -> 273.1 for neomangiferin and m/z 326.2 -> 291.1 for IS. The linearity of this method was found to be within the concentration range of 1-1000 ng/mL with a lower limit of quantification of 1 ng/mL. Only 3.0 min was needed for an analytical run. The matrix effect was 92.6 to 108.4% for neomangiferin. The intra- and inter-day precision (RSD%) were less than 9.4% and accuracy (RE%) was within +/- 9.1%. The recovery ranged from 84.8 to 95.1%. Neomangiferin was sufficiently stable under all relevant analytical conditions. The method was also successfully applied to the pharmacokinetic study of neomangiferin in rats. The pharmacokinetic parameters were demonstrated as followed: t(1/2) was 0.89 +/- 0.09 h, C-max was 481.20 +/- 69.47 ng/mL, and AUC(0 -> 8) was 1044.42 +/- 210.77 ng/mL.h.u