Abstract: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.
