Systematic and efficient separation of 11 compounds from Rhizoma Chuanxiong via counter-current chromatography–solid phase extraction–counter-current chromatography hyphenation

•A CCC–SPE–CCC system was developed.•Trapping conditions of SPE were optimized.•Five intractable compounds were separated by baseline through adopting recycling CCC.•11 compounds with large polarity difference were separated by running CCC only once.

A counter-current chromatography (CCC)–solid phase extraction (SPE)–CCC system with high preparative capacity was used to realize rapid one-run systematic separation of natural products, in which two six-port valves and the SPE cartridge served as the interface. In the orthogonal separation system, equal column volumes of TEB-300A and TEB-300B were employed for the first dimension (1st-D) and second dimension (2nd-D), respectively. An optimized solid-phase column (25 mm × 10 mm i.d.) packed with Oasis HLB materials acted as the trapping SPE column. The analyte-focusing effect of the trapping column associated with the considerable preparative capacities of 1st-D and 2nd-D using totally different solvent systems significantly facilitated this one-run systematic separation of natural product. Therefore, this proposed approach was successfully applied to isolate chemical compounds from the crude extract of Rhizoma Chuanxiong. As a result, 11 compounds with widely different polarities were separated by running CCC for only one time. More importantly, this hyphenated strategy could serve as a rapid and efficient systematic pathway for the separation of natural products.