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Oxytropis falcata  Bunge, known as “the King of Herbs” in Tibetan medicine, is used for treatment of hyperpyrexia, pain, wounds, inflammation and anthrax. However, it is difficult to isolate compound with high-purity from  O. falcata  because of its complexity. In this work, an efficient method was successfully established for the separation of 3-hydroxy-3-methylglutaryl (HMG) flavonoid glycosides from  O. falcata  by 2D preparative chromatography (2D-HPLC). An ODS C 18  preparative column was used for the first-dimension preparation, and the XCharge C 18  preparative column with different separation selectivity to the ODS C 18  stationary phase was used for the second-dimension preparation. Four HMG flavonoid glycosides (oxytroflavosides A–D) and two flavonoid glycosides (oxytroflavosides F and G) were separated with purities over 98%. Their structures were elucidated by nuclear magnetic resonance spectroscopy. The 2D-HPLC method used in this study is effective for preparative separation of HMG flavonoid glycosides from  O. falcata.  Additionally, this method shows great potential for the separation of flavonoid glycosides from other plant extracts.

Separation of 3-Hydroxy-3-Methylglutaryl Flavonoid Glycosides from Oxytropis falcata Using 2D Preparative Chromatography