Application of High-Speed Countercurrent Chromatography to the Separation of Flavanonol, Phenylcoumaran and Flavonolignans in Silybum marianum

Abstract

Separation of flavanonol, phenylcoumaran and flavonolignans in Silybum marianum was examined using high-speed countercurrent chromatography (HSCCC). In order to prepare analytical standards, a flavanonol of aromadendrin (87.4% purity, 7.8 mg), three phenylcoumarans of jatrointelignan D (84.5% purity, 13.2 mg), dehydrodiconiferyl alcohol (76.1% purity, 1.4 mg) and dihydrodehydrodiconiferyl alcohol (93.0% purity, 5.8 mg), and three flavonolignans of silybin (88.8% purity, 35.6 mg), silydianin (99.3% purity, 25.1 mg) and silychristin (96.9% purity, 12.6 mg) were separated from the seeds of S. marianum using common column chromatography and ODS-HPLC, and identified by 1H and 13C NMR spectra. Then, HSCCC with the hexane/ethyl acetate/methanol/water (3 : 7 : 4 : 6, v/v) system was applied to the separation of aromadendrin, jatrointelignan D, silydianin and silybin. In this separation, it was revealed that silybin and silydianin were successfully separated from each other. The present HSCCC system was directly applied to the ethyl acetate extract and resulted in the separation of silybin. The overall results suggested that HSCCC is useful for the separation of bioactive compounds in S. marianum.