Abstract: High-speed counter-current chromatography (HSCCC) method for isolation and purification of flavonoids crystalloid, i.e. apigenin-7-O-rutinoside(I), luteolin-7-O-glucoside(Ⅱ), apigenin-7-O-glucoside(Ⅲ) and acacetin-7-O-glucoside(IV) from Chrysanthemum morifolium Ramat. was established successfully. The separation was performed in two steps with two different types of solvent systems: ethyl acetate-ethanol-water-acetic acid (4:1:5:0.2, v/v) and chloroform-methanol-water (4:3:2, v/v). In the first separation step, 100 mg of the flavonoids crystalloid yielded 11.2 mg of apigenin-7-O-rutinoside, 15.3 mg of luteolin-7-O-glucoside, and 28.2 mg of apigenin-7-O-glucoside. Then we collected tail blowing fluid and obtained 35 mg of extract by evaporation to dryness. In the second step, when we used [chloroform-methanol-water (4:3:2, v/v)] solvent system to separate the 35 mg of extract, 14.5 mg of acacetin-7-O-glucoside could be obtained. Their four compounds purities were 99.4%, 93.6%, 99.1% and 99.5%, respectively. Their structures were identified by ESI-MS, ¹H NMR, and ¹³C NMR.

Key words: flavonoid, HSCCC, Chrysanthemum morifolium Ramat