大孔吸附树脂-硅胶柱层析法部分纯化藜麦β-蜕皮激素

Partial Purification ofβ-ecdysteroid from Quinoa by Macroporous Adsorption Resins and Silica Gel Column Chromatography

本研究采用大孔吸附树脂-硅胶柱层析法分离纯化藜麦β-蜕皮激素。通过静态-动态吸附解吸实验确定大孔吸附树脂最佳纯化工艺:大孔吸附树脂为D101型,上样质量浓度为25 mg/mL、上样流速2.0 mL/min、洗脱剂乙醇体积分数30%和洗脱流速2.0 mL/min,得到纯度为12.12%的β-蜕皮激素。通过薄层色谱与硅胶柱层析实验,确定最佳洗脱条件为:洗脱剂V(三氯甲烷)∶V(甲醇)=4∶1。采用高效液相色谱法(HPLC)测定纯度,藜麦β-蜕皮激素的纯度提高至40.78%。藜麦β-蜕皮激素具有一定的抗氧化活性,其粗提物和纯化后产物总抗氧化能力分别为(0.44±0.01)mol/g和(6.46±0.46)mol/g;DPPH自由基清除能力的IC50值分别为(1.61±0.02)mg/mL和(0.16±0.01)mg/mL,表明部分纯化后其抗氧化活性提高。

The technology of purificationβ-ecdysteroid from quinoa was investigated by macroporous adsorption resin and silica gel column chromatography.The optimum separation conditions,studied by static-dynamic adsorption and desorption experiment,were as follows:macroporous adsorption resin of D101,sample concentration of 25 mg/mL,adsorption flow rate of 2.0 mL/min,eluent of 30%ethanol,desorption flow rate of 2.0 mL/min,the purity ofβ-ecdysteroid was 12.12%.Based on the thin-layer chromatography and silica gel column chromatography,it was found that the crude extracts could be purified on the conditions of eluent trichloromethane/methanolthe=4∶1(V/V).The purity of quinoaβ-ecdysteroid was 40.78%determined by high performance liquid chromatography(HPLC).Quinoaβ-ecdysteroid had antioxidant capacity,and the total antioxidant capacity of its crude extract and purification product were(0.44±0.01)mol/g and(6.46±0.46)mol/g;the IC50 values of DPPH free radical scavenging ability were(1.61±0.02)mg/mL and(0.16±0.01)mg/mL,indicating that the antioxidant capacity of quinoaβ-ecdysteroid was improved by purification.