| Summary: | A method for batch preparation of fucoxanthin from brown algae was established, which possessed the advantages of high yield and high purity. The ultrasonic-assisted extraction method was used to obtain a crude extract from <i>Sargassum fusiforme</i> as the separation sample. Then the crude extract was separated by elution-extrusion countercurrent chromatography. The optimum preparation conditions of fucoxanthin were determined as follows: <i>n</i>-hexane-ethanol-water (20:9:11, <i>v</i>:<i>v</i>:<i>v</i>) as a two-phase solvent system, the mobile phase flow rate was 5 mL min<sup>−1</sup>, the revolution speed was 800 r min<sup>−1</sup>, the loading capacity was 60 mg 10 mL<sup>−1</sup> and the temperature was 25 °C. By this method, 12.8 mg fucoxanthin with a purity of 94.72% was obtained from the crude extract of <i>Sargassum fusiforme</i>. In addition, when the loading capacity was 50 mg 10 mL<sup>−1</sup>, the purity of fucoxanthin reached 96.01%. Two types of by-products, chlorophyll and pheophytin, could also be obtained during the process of separation. This optimal method was further applied to separate fucoxanthin from <i>Laminaria japonica</i> and <i>Undaria pinnatifida</i>, and 6.0 mg and 9.7 mg fucoxanthin with a purity of 96.24% and 92.62% were acquired, respectively. Therefore, it was demonstrated that the preparation method of fucoxanthin established in this study had an applicability to brown algae, which improved the utilization value of raw materials.
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