Isolation of Hyperoside and Luteolin‐Glucoside from Agrimonia pilosa Ledeb Using Stepwise Elution by High-Speed Countercurrent Chromatography

Preparative high-speed countercurrent chromatography was successfully used for isolation and purification of hyperoside and luteolin‐glucoside from Agrimonia pilosa Ledeb. The separation was performed by stepwise elution with a pair of two‐phase solvent systems composed of ethyl acetate‐methanol‐water at volume ratios of 50:1:50 and 5:1:5, which had been selected by analytical high-speed countercurrent chromatography (HSCCC). Using a preparative unit of the HSCCC centrifuge, about a 300 mg amount of the crude extract was separated, yielding 7.3 mg of hyperoside and 10.4 mg of luteolin‐glucoside at a high purity of over 97%.