An effective high–speed counter–current chromatography (HSCCC) method was established for semi-preparative isolation and purification of bioactive chemical constituents from the Tibetan medicinal plant Pedicularis longiflora var. tubiformis (Klotzsch) P. C. Tsoong. With a two-phase solvent system composed of chloroform–methanol–water (8:4:5, v/v), 40 mg of extract was separated to yield luteolin (12.5 mg), apigenin (9.6 mg), and chrysoeriol (4.8 mg), with purities of 99.3, 98.2, and 98.6%, respectively, as determined by high-performance liquid chromatography (HPLC). The chemical structures of these three components were identified by ¹H NMR and ¹³C NMR.

Fig. HPLC chromatograms of the crude extract from BCM (A) after cleaning up by silica-gel column chromatography and the three targeted compounds (peak fractions I, II, and III, B–D) purified by HSCCC. Conditions: column: Eclipse XDB–C18 column (5 μm, 4.6150 mm); mobile phase: methanol–water (60:40, ν/v); flow-rate: 1.0mL/min; column temperature: 25οC; detection wavelength: 254 nm.

Additional Information:
1 Author Information:Xiaohui Zhao, Fa Han, Yulin Li, Guoying Zhou & Huilan Yue
Correspondence: E-mail: hizhaoxh@163.com
2 Published:
Journal of Liquid Chromatography & Related Technologies, 36:13, 1751-1761, 2013