为进一步提高金线莲苷的分离纯化效率，该研究以深度共熔溶剂提取的金线莲苷粗品为基础，比较了大孔树脂法和硅胶柱层析法分离金线莲苷的效果，并优化了硅胶柱层析法分离金线莲苷的条件。实验结果表明，DM130型大孔树脂对金线莲苷的吸附量仅为24.69 mg/g，解吸率为17.89%，且无法选择性的分离金线莲苷，而硅胶柱层析法对金线莲苷的分离效果较好；硅胶柱层析法分离金线莲苷的最佳条件为：洗脱溶剂为乙酸乙酯:乙醇:乙酸=6:4:0.2（V/V/V），上样量为0.60 g，洗脱速率为1.25 mL/min，在此条件下，金线莲苷的回收率可达79.29%；硅胶柱层析分离组分经半制备型高效液相色谱纯化所得的金线莲苷纯度大于99.00%。该研究建立了一种简单高效的金线莲苷分离纯化流程，有利于金线莲产业的发展，为后续的研究工作提供理论支持。

In order to improve separation and purification efficiency of kinsenoside from Anoectochilus roxburghii, in this study, the deep eutectic solvent crude extract of kinsenoside was prepared for the further separation, and the effect of macroporous resin chromatography and silica gel column chromatography on the separation of kinsenoside was compared. In addition, the conditions for the separation of kinsenoside by silica gel column chromatography were optimized. The results showed that the adsorption capacity and desorption rate of kinsenoside on DM130 macroporous resin were 24.69 mg/g and 17.89% respectively, and macroporous resin could not selectively separate kinsenoside while silica gel column chromatography had a good effect on the separation of kinsenoside. The optimal conditions for the separation of kinsenoside by silica gel column chromatography were measured as elution solvent of ethyl acetate/ethanol/acetic acid (6:4:0.2, V/V/V), loading volume of 0.60 g and elution rate of 1.25 mL/min. The kinsenoside recovery rate of 79.29% was obtained under these conditions. Then the elute fraction could be purified to >99.00% purity by semi-preparative high performance liquid chromatography. In this work, a simple and efficient process was established to separate and purify kinsenoside from the deep eutectic solvent crude extract, which is beneficial to the development of Anoectochilus roxburghii industry and provides theoretical support for subsequent research work.

广东省科技计划项目（2019A0103008）