A rapid method of microwave-assisted extraction coupled with high-speed counter current chromatography and UV detector was established for the preparation of phytosterols from edible marine algae. Extraction conditions, namely microwave power, liquid/solid ratio, irradiation time and extraction temperature were optimized using orthogonal array design. The microwave-assisted extraction of Undaria pinnatifida and Sargassum fusiforme were separated and purified with a non-aqueous two-phase solvent system composed of n-hexane-acetonitrile-methanol (5:5:3, v/v/v). The isolation was done in less than 220 min, and a total of 13.0 mg fucosterol, 1.5 mg 24-methylenecholesterol and 10.7 mg phytol were obtained from 15.0 g U. pinnatifida; 4.6 mg fucosterol, 03 mg 24-methylenecholesterol and 3.5 mg phytol were obtained from 15.0 g S. fusiforme. The purities of all products were over 97% determined by high performance liquid chromatography. The results demonstrate that microwave-assisted extraction coupled with high-speed counter-current chromatography is a feasible, economical and efficient technique for rapid extraction, separation and purification of effective compounds from natural products. (C) 2012 Elsevier B.V. All rights reserved.