Comprehensive multi-channel multi-dimensional counter-current chromatography for separation of tanshinones from Salvia miltiorrhiza Bunge

• In this work, we built a novel multi-channel multi-dimensional counter-current chromatography.
• Several 2D and 3D CCC systems were established.
• These multi-dimensional CCC systems were applied to separation of tanshinones from Salvia miltiorrhiza Bunge.
• All 2D and 3D CCC isolations were found to be successful and efficient.
• They decrease analysis time, reduce solvent consumption and increase sample throughput significantly.

Multi-dimensional chromatography offers the increased resolution and peak capacity by coupling of multiple columns with the same or different separation mechanisms. In this work, a novel multi-channel multi-dimensional counter-current chromatography (CCC) has been successfully constructed and used for several two-dimensional (2D) and three-dimensional (3D) CCC separations including 2D A × B/A × C, A × B–C and A–B × C, and 3D A × B × C systems. These 2D and 3D CCC systems were further applied to separate the bioactive tanshinones from the extract of Tanshen (or Danshen, Salvia miltiorrhiza Bunge), a famous Traditional Chinese Medicine (TCM). As a result, the developed 2D and 3D CCC methods were successful and efficient for resolving the tanshinones from complex extracts. Compared to the 1D multiple columns CCC separation, the 2D and 3D CCC decrease analysis time, reduce solvent consumption and increase sample throughput significantly. It may be widely used for current drug development, metabolomic analysis and natural product isolation.