Highspeed countercurrent chromatography(HSCCC)was developed for separation and purification of astaxanthin from Haematococcus pluvialis.The optimized twophase solvent system, revolution speed, flow rate, temperature, sample concentration and sample volume were nhexaneethyl acetateethanolwater(6.5∶5∶6.5∶3, v/v/v/v), 850 r/min, 3 mL/min, 25  ℃, 10 mg/mL and 10 mL, respectively. The lower phase was used as stationary phase, while the upper phase was the mobile phase of HSCCC. Furthermore, the compound collected from HSCCC analyzed by high performance liquid chromatography(HPLC), mass spectrometry(MS)and standard and was confirmed as astaxanthin. This paper can provide technical support to efficient preparation of astaxanthin from H. pluvialis using HSCCC.