Separation of Four Phenylpropanoid Glycosides from a Chinese Herb by HSCCC

Four phenylpropanoid glycosides (PPGs), such as echinacoside, wiedemannioside C, forsythoside B and verbascoside, were isolated and purified from the Tibetan medicinal herb Pedicularis longiflora Rudolph. var. tubiformis (Klotz) Tsoong by high-speed counter-current chromatography (HSCCC) combined with macroporous resin (MR) column separation for the first time. In the present study, the two-phase solvent system composed of ethyl acetate-n-butanol-water (10 : 6:15, v/v/v) was used for HSCCC separation. A total of 3.5 mg of echinacoside, 12.6 mg of wiedemannioside C, 22.7 mg of forsythoside B and 48.7 mg of verbascoside with the purity of 93.6, 97.9, 97.8 and 98.1%, respectively, were obtained from 120 mg of crude sample. The HSCCC fractions were analyzed by HPLC, and the chemical structures were identified by (1)H NMR and (13)C NMR. The results demonstrate that MR coupled with HSCCC is a powerful technique for separation of PPGs from natural products.