Box-Behnken设计优化金雀花中总黄酮纯化工艺研究

Study on Purification Process of Total Flavonoids from Caragana Altaica by the Box-Behnken Design

目的研究大孔吸附树脂纯化金雀花中总黄酮的工艺条件。方法以乌鲁木齐石人沟采集的金雀花为原料,以超声提取的方法得到总黄酮粗提物,对比7种大孔吸附树脂对金雀花中总黄酮的吸附-解吸效果,采用Box-Behnken设计优化总黄酮纯化工艺,以树脂的吸附率和解吸率为指标,考察进样液浓度、进样体积、进样流速、洗脱液、洗脱体积、洗脱流速等因素对纯化效果的影响。结果筛选得到AB-8型大孔吸附树脂为最适树脂,并获得最佳吸附条件和解吸条件,吸附条件:进样浓度为1.14 mg·mL-1、进样体积为2.8 BV、进样流速为2.5 mL·min-1;解吸条件:洗脱液浓度为70%、洗脱体积为2.0 BV、洗脱流速为3.0 mL·min-1。在此条件下的总黄酮百分含量从1.72%提高到了22.95%,纯度升高了13.3倍。结论本纯化工艺操作简便、可行,为金雀花中总黄酮的进一步研究提供了实验依据。

OBJECTIVE To study the process condition of purification the total flavonoids from Caragana altaica by macroporous adsorption resin. METHODS The Caragana altaica which collected from Shirengou in Urumqi were used as raw materials to obtain the crude flavonoids by ultrasonic extraction. The adsorption and desorption effects of seven macroporous resin on the total flavonoids in Caragana altaica were compared and optimized by Box-Behnken design. The flavonoid purification process, based on the adsorption rate and desorption rate of the resin, investigated the effects of factors such as the concentration of injection solution, injection volume, injection flow rate, the eluent, elution volume and the elution flow rate on the purification effect. RESULTS The AB-8 macroporous adsorption resin was selected as the optimum resin, and the optimal adsorption conditions and desorption conditions were obtained. The adsorption conditions were as followings:the injection concentration was 1.14 mg·mL-1, and the injection volume was 2.8 BV. The sample flow rate was 2.5 mL·min-1;desorption conditions:the eluent concentration was 70%, the elution volume was 2.0 BV, and the elution flow rate was 3.0 mL·min-1. The content under this condition increased from 1.72% to 22.95%, and the purity increased by 13.3 times. CONCLUSION The purification process is simple and feasible, and provides experimental basis for the further study of total flavonoids in Caragana altaica.