Application of high‐performance countercurrent chromatography for the isolation of steroidal saponins from Liriope plathyphylla

High‐performance countercurrent chromatography (HPCCC) with electrospray light‐scattering detection was applied for the first time to isolate a spirostanol and a novel furostanol saponin from Liriope platyphylla . Due to the large differences in K D values between the two compounds, a two‐step HPCCC method was applied in this study. The primary HPCCC employed methylene chloride/methanol/isopropanol/water (9:6:1:4 v/v, 4 mL/min, normal‐phase mode) conditions to yield a spirostanol saponin ( 1 ). After the primary HPCCC run, the solute retained in the stationary phase (SP extract) in HPCCC column was recovered and subjected to the second HPCCC on the n ‐hexane/ n ‐butanol/water system (1:9:10 v/v, 5 mL/min, reversed‐phase mode) to yield a novel furostanol saponin ( 2 ). The isolated spirostanol saponin was determined to be 25( S )‐ruscogenin 1‐ O ‐β‐ d ‐glucopyranosyl (1→2)‐[β‐ d ‐xylopyranosyl (1→3)]‐β‐ d ‐fucopyranoside (spicatoside A), and the novel furostanol saponin was elucidated to be 26‐ O ‐β‐ d ‐glucopyranosyl‐25( S )‐furost‐5(6)‐ene‐1β‐3β‐22α‐26‐tetraol‐1‐ O ‐β‐ d ‐glucopyranosyl (1→2)‐[β‐ d ‐xylopyranosyl‐(1→3)]‐β‐ d ‐fucopyranoside (spicatoside D).