Determination of Bioactive Markers for the Discrimination of Syneilesis palmata and Paris verticillata by High Performance Liquid Chromatography (HPLC) with Diode Array Detection (DAD) and Ion Trap Time-of-Flight Mass Spectrometry (IT-TOF-MS)

Abstract

Syneilesis palmata (S. palmata) is a common Korean therapeutic herb extensively used to treat pain. The species Paris verticillata (P. verticillata) is morphologically similar to S. palmata but with no medicinal values. This study was designed to establish a technique for the discrimination of S. palmata and P. verticillata leaves. The methanolic extracts of the leaves of both species were analyzed by high performance liquid chromatography (HPLC) with diode array detection (DAD) and ion trap time-of-flight mass spectrometry (IT-TOF-MS). Three marker peaks identified only in P. verticillata leaves were assigned to be quercetin-3-6-rhamnosylgentiobioside (peak 1), kaempferol-3-O-gentiobioside-7-O-rhamnose (peak 2), and kaempferol-3-O-gentiobioside (peak 3). These were characterized by IT-TOF-MS due to the unavailability of standards. Formulations prepared with different compositions also displayed these indicator peaks at the target retention time and same ultraviolet spectra by DAD analysis. The peak area increased with the ratio of P. verticillata leaves in the mixtures. The developed technique is rapid and efficient for authentication and discrimination of S. palmata and P. verticillata leaves.