Ten compounds, including three lignan glycosides and seven lignans, were purified from Justicia procumbens L. in 8 h using an efficient strategy based on high-speed counter-current chromatography (HSCCC). The two-phase solvent system composed of petroleum–ethyl acetate–methanol–H2O (1:0.7:1:0.7, v/v) was firstly employed to separate the crude extract (320 mg), from which 19.3 mg of justicidin B (f), 10.8 mg of justicidin A (g), 13.9 mg of 6′-hydroxyjusticidin C (h), 7.7 mg of justicidin E (i), 6.3 mg of lignan J1 (j) were obtained with 91.3 mg of enriched mixture of compounds a–e. The enriched mixture (91.3 mg) was further separated using the solvent system consisting of petroleum–ethyl acetate–methanol–H2O (3:3.8:3:3.8, v/v), yielding 12.1 mg of procumbenoside E (a); 7.6 mg of diphyllin-1-O-β-d-apiofuranoside (b); 7.4 mg of diphyllin (c); 8.3 mg of 6′-hydroxy justicidin B (d); and 7.9 mg of diphyllin acetyl apioside (e). The purities of the 10 components were all above 94%, and their structures were identified by NMR and ESI-MS spectra. The results demonstrated that the strategy based on HSCCC for the separation of lignans and their glycosides was efficient and rapid.

Article Highlights• The combination of ultrasonic extraction and pH-zone-refining CCC is used in the present study • Three isoquinoline alkaloids with high purities are obtained in a single run • Optimizing the concentration of retainer and eluter is an important aspect in pH-zone--refining CCC AbstractBerberidis radix, as a traditional Chinese herbal medicine, is well known as the main source of berberine. This herbal medicine has been used for the treatment of many diseases and alkaloids including jatrorrhizine, palmatine, and berberine are major bioactive constituents. Few literatures report the preparative separation of these alkaloids from Berberidis radix. In the present study, the combinative application of ultrasonic extraction and pH-zone-refining countercurrent chromatography was used to extract and purify these alkaloids. The PZRCCC separation was performed in a two-phase solvent system composed of chloroform-methanol-water (4:3:3 volume ratio), where 40 mM HCl was added in the upper aqueous stationary phase as a retainer and 10 mM triethylamine was added in the lower organic mobile phase as an eluter. From a crude sample of 1.5 g, palmatine (251.6 mg), berberine (392.5 mg), and jatrorrhizine (412.6 mg) were obtained in a single run with purities of all being over 93.0%, analyzed by HPLC. The chemical structures of the separated alkaloids were identified by electrospray ionization-mass spectrometry (ESI-MS), 1 H-NMR and 13 C-NMR.