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