目的 通过制备键合β-环糊精琼脂凝胶微球（AG-β-CD）作为分离介质，对大豆异黄酮粗品中大豆苷元进行高效分离纯化。方法 利用琼脂为原料，通过乳化、交联并键合功能基团β-环糊精（β-CD）制备AG-β-CD；将其作为一种分离介质，通过考察流动相、洗脱体积流量和微球负载量3个方面，确定AG-β-CD分离纯化大豆苷元的工艺并进行验证，质谱和核磁共振鉴定其结构；通过与其他2种黄酮类物质表没食子儿茶素没食子酸酯（EGCG）和葛根素在C₁₈反相柱色谱保留行为比较，结合autoDOCK4.0进行分子模拟，以及流动相乙腈体积分数变化对3种黄酮类物质在AG-β-CD固定相上的保留时间的变化曲线证明其色谱机制。结果 大豆异黄酮粗品中主要成分为大豆苷元（57.14%），AG-β-CD的负载量为1.33 mg/mL（大豆异黄酮粗品总量/柱体积），体积流量为2 BV/h时，通过20%乙醇洗脱2 BV、40%乙醇洗脱1.33 BV、70%乙醇洗脱6～7 BV，得到质量分数≥95%（平均质量分数96.98%）的大豆苷元，收率为97.86%。色谱机制实验表明AG-β-CD具有亲水和反相双重保留与分离作用。结论 AG-β-CD能够高效地分离纯化大豆苷元。

Objective Daidzein was efficiently purified by agar gel microspheres bonded β-cyclodextrin (AG-β-CD). Methods Using agar as raw material, after emulsification, crosslinking, and bonding β-CD as functional group, AG-β-CD was synthesized for the purification of daidzein, and the purification process was determined and proved with mobile phase, flow rate, and loading capacity of microspheres. The structure of daidzein was identified by MS and NMR, AG-β-CD was chromatographically evaluated with daidzein, EGCG, and puerarin as the following tripartition such as difference of retention behavior on C₁₈ reversed phase column chromatography, molecular simulation by autoDOCK4.0, and retention time curves on AG-β-CD with different contents of acetonitrile. Results The main component of soybean isoflavone was daidzein (57.14%). The loading quantity of AG-β-CD was 1.33 mg/mL, flow rate was 2 BV/h, eluted by 2 BV of 20% ethanol, 1.33 BV of 40% ethanol, and 6-7 BV of 70% ethanol, the content was ≥95%, purity of daidzein (96.98%) was obtained with 97.86% yield. Chromatographic mechanism research showed that AG-β-CD had hydrophilic interaction chromatography and reversed-phase chromatography. Conclusion AG-β-CD is capable of highly efficient purification of daidzein.

厦门市海洋经济创新发展区域示范项目（13CZP004SF27）