Simultaneous determination of icariin, naringin and osthole in rat plasma by UPLC-MS/MS and its application for pharmacokinetic study after oral administration of Gushudan capsules.

Abstract A rapid, sensitive and selective ultra performance liquid chromatography tandem mass spectrometry (UPLC–MS/MS) method was developed and validated for the simultaneous determination of icariin, naringin and osthole in rat plasma. Plasma samples pretreatment involved a one-step liquid–liquid extraction with a mixture of ethyl acetate-methyl tert-butyl ether (3:1, ν/ν). The separation was performed on an ACQUITY UPLC™ BEH C 18 column with a gradient mobile phase system of methanol and water. The detection was performed on a triple quadrupole tandem mass spectrometer equipped with electrospray ionization (ESI) by multiple reactions monitoring (MRM), with the transitions at m / z 513.3 → 366.8 (icariin), m / z 579.3 → 150.9 (naringin), m / z 245.1 → 189.0 (osthole) and m / z 237.1 → 194.1 (IS), respectively. A good linear response was observed over the concentration ranges of 1.06–424 ng/ml, 2.10–525 ng/ml and 1.05–1.05 × 10 3  ng/ml with lower limit of quantification (LLOQ) of 1.06, 2.10 and 1.05 ng/ml for icariin, naringin and osthole, respectively. The intra- and inter-day precisions ( R.S.D. ) were within 14.3%, and the accuracy ( R.E. ) ranged from −4.1% to 4.6% at three quality control levels. The sensitive and selective method was applied to a pharmacokinetic study of icarrin, naringin and osthole in rats after oral administration of Gushudan capsule.