With the EtOAC extracts of fermentation broth of a Bacillus atrophaeus strain associated with a sponge as the material,the natural algicide bacillamide C was separated and purified by three different methods:silica gel column chromatography,centrifugal TLC and semi-preparative HPLC. By comparing injection volume, time, eluent consumption, the purity and the recovery of bacil-lamide C,both advantages and disadvantages of the three methods were evaluated comprehensively. The results showed that the semi-preparative HPLC has the potential to become the ideal method for rapid separation of bacillamide C.%以一株海绵共附生芽孢杆菌发酵液的乙酸乙酯萃取物为原料，分别用硅胶吸附柱层析、离心薄层层析和半制备HPLC三种方法对其中的天然杀藻活性物质bacillamide C进行分离提纯，通过对分离过程中进样量、时间、洗脱液消耗量和分离物中bacillamide C的纯度和回收率的分析比较，对三种方法的优缺点做出综合评定，认为半制备HPLC法有成为理想的快速分离bacillamide C方法的潜力。
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