Development of ultrasound-assisted centrifugal extraction combined with two countercurrent chromatography systems for the simultaneous extraction and isolation of phytochemicals.

We proposed a method for the extraction of medicinal herbs, called ultrasound-assisted centrifugal extraction, and an online solvent concentration method. These techniques were coupled with two countercurrent chromatography systems and applied to the continuous extraction and online isolation of chemical constituents from Inonotus obliquus. Raw plants were extracted using a two-phase petroleum-ethanol-water (2.0:1.0:2.0, v/v/v) process, and then the aqueous and organic phases were concentrated using the proposed online solvent concentrator. The countercurrent chromatography preparation prior to separation includes pumping of the two-phase solution, rotating column, and equilibrium column. Following online concentration, the extracted solution was pumped into a second countercurrent chromatography process for separation. During separation, the extraction solution and concentrated extract were prepared automatically. Upon completion of the first cycle of ultrasound-assisted centrifugal extraction/two countercurrent chromatography, the second cycle experiment starts. This process can be indefinitely repeated. In this study, six target compounds with purities above 97.71% were successfully extracted and isolated online using a two-phase solvent system consisting of n-hexane-ethyl acetate-acetonitrile (4.5:1.5:5.5, v/v/v) and n-hexane-ethyl acetate-methanol-water (0.4:3.0:1.5:2.5, v/v/v/v). Compared to conventional extraction methods, the instrumental setup of the proposed method provides enhanced automation, efficiency, purity, and systematic extraction and isolation of natural products.