Abstract: The reason why the strong polar compounds couldn`t be well separated by conventional J type high speed countercurrent chromatography has been studied by analyzing the total and single phase’s polarity distribution rules of commonly used 28 kinds HEMWat solvent systems in this paper.According to the results,an effective sample pre-treatment method was established to segment the Sea Cucumber polypeptides.The method is based on the polypeptides` physical and chemical properties to find a suitable modifier improving the distribution behavior of the samples in mobile phase and stationary phase.After two-step separation,eight sea cucumber polypeptide fractions with different polarity were obtained,which provided technical support for the tracking and screening of the best active substance of sea cucumber polypeptide.

Key words: high speed counter-current chromatography, HEMWat solvent systems, high polar compounds, sea cucumber polypeptide, pretreatment technology