An Efficient Method to Separate the Main Constituents From Polygonatum cyrtonema Hua by a Combination of Online‐Storage Inner‐Recycling and Consecutive High‐Speed Counter‐Current Chromatography

ABSTRACT A novel strategy combining XAD‐4 macroporous resin pretreatment and online‐storage inner‐recycling high‐speed counter‐current chromatography (OS‐IRCCC) was developed for efficient separation of multi‐polarity constituents from Polygonatum cyrtonema Hua. The innovation lies in the OS‐IRCCC mode, which integrates online storage of target fractions via a six‐way valve‐equipped recycling loop and inner‐recycling separation, overcoming limitations of conventional high‐speed counter‐current chromatography in resolving structurally similar compounds. After resin fractionation, 40%, 60%, and 95% ethanol eluates were separated using methyl tert‐butyl ether/ n ‐butanol/acetonitrile/water (1:4:1:5, v/v) solvent system, chloroform/methanol/water (7:6:4, v/v) solvent system (with OS‐IRCCC for Compounds 8 and 9 ), and petroleum ether/ethyl acetate/methanol/water (5:5:4:3, v/v) solvent system, respectively. Thirteen compounds were successfully isolated, including four high‐polarity glycosides ( 1 – 4 ), five medium‐polarity phenolic constituents ( 5 – 9 ), and four low‐polarity terpenes ( 10 – 13 ). This study demonstrates that the established separation method provides an efficient, rapid, and straightforward approach for isolating active constituents from P. cyrtonema Hua.