An off-line three-dimensional liquid .....

ABSTRACT

To comprehensively elucidate the herbal metabolites is crucial in natural products research to discover new lead compounds. Ginsenosides are an important class of bioactive components from the Panax plants exerting the significant tonifying effects. However, to identify new ginsenosides by the conventional strategies trends to be more and more difficult because of the large spans of acid-base property(the neutral and acidic saponins), molecular mass (40 0–140 0 Da), and rather low content. Herein, an offline multidimensional chromatography/high-resolution mass spectrometry approach was presented: ion exchange chromatography (IEC) as the first dimension of separation, hydrophilic interaction chromatography (HILIC) in the second dimension, and reversed-phase chromatography (RPC) for the third dimension which was hyphenated to a Q Exactive Q-Orbitrap mass spectrometer. 

By applying to the flower buds of P. ginseng (PGF), P. quinquefolius (PQF), and P. notoginseng (PNF), IEC using a PhenoSphere TM SAX column could fractionate the total extracts into the neutral (unretained) and acidic (retained) fractions, while  HILIC (an XBridge Amide column) and RPC (BEH Shield RP18 column) achieved the hydrophilic interaction and hydrophobic interaction separations, respectively. Q-Orbitrap mass spectrometry offered rich structural information and complementary resolution to the co-eluting components, particular to those minor ones by including precursor ion lists in data-dependent acquisition. We could characterize 803 ginsenosides from PGF, 795 from PQF, and 833 from PNF, and 1561 thereof are potentially unknown. These results can indicate the great potential of this multidimensional approach in the ultra-deep characterization of complex herbal samples supporting the efficient discovery of potentially novel natural compounds.

Materials and methods

The reference compounds of 74 ginsenosides (Fig. S1 and Table S1) were purchased from  Chengdu Desite Biotechnology Co., Ltd. (Chengdu, China).

原文链接：https://doi.org/10.1016/j.chroma.2022.463177