Characterization and identification of saponins in Achyranthes bidentata by rapid-resolution liquid chromatography with electrospray ionization quadrupole time-of-flight tandem mass spectrometry.

A rapid-resolution liquid chromatography (RRLC) method coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (Q-TOF MS/MS) has been developed for analysis of oleanane-type triterpenoid saponins in Achyranthes bidentata. Collision-induced dissociation techniques were used to fragment the precursor molecular ions and the resulting product ions. A retro-Diels-Alder rearrangement from the oleanane aglycone skeleton in the MS/MS process yielded characteristic fragment ions in positive ion mode. These characteristic ions were helpful in predicting the aglycone structure. Losses of monosaccharide sequences, presence of sugar-chain fragment ions, and cleavage of CO(2) were observed for important information on sugar types and attachment sequences. Fragmentation rules of three major groups of saponins from A. bidentata were summarized, and the possible fragmentation pathways were proposed. A total of 22 compounds including both the target and unknown oleanane-type triterpenoid saponins were rapidly screened and predicted in the herbal extract by the developed method. The RRLC-Q-TOF MS/MS method has provided a powerful approach for rapid separation, target screening and structural elucidation of oleanane-type saponins, and also opened perspectives for similar studies on other herbal medicines.