响应面法优化大孔树脂纯化菟丝子总黄酮工艺

目的 优化大孔树脂纯化菟丝子总黄酮工艺。方法 采用单因素实验,以总黄酮、金丝桃苷的吸附率和解析率为指标,筛选最佳大孔树脂型号。以总黄酮、金丝桃苷吸附量为指标,考察吸附时间、漏点浓度、上样液pH对纯化工艺的影响。以总黄酮、金丝桃苷洗脱率为指标,考察洗脱液流速、洗脱液体积、洗脱液浓度对纯化工艺的影响。在此基础上,以总黄酮与金丝桃苷含量的归一值为指标,利用Box-Behnken响应面法优化并确立最佳工艺。结果 菟丝子总黄酮纯化的最佳树脂型号为AB-8型,最佳工艺条件为:上样液pH 3.9、洗脱液体积19.6BV,洗脱液浓度74%,验证实验的总OD值为0.982 5,与模型预测值接近。纯化前后,总黄酮含量由18.95 mg·g^-1提高至85.68 mg·g^-1,金丝桃苷含量由4.12 mg·g^-1提高至16.85 mg·g^-1。结论 优选的纯化工艺方法稳定、可行,可用于菟丝子总黄酮的纯化。

Optimization of Purification Process of Total Flavonoids from Cuscuta chinensis by Macroporous Resin by Response Surface Methodology

OBJECTIVE To optimize the purification process of total flavonoids from Cuscuta chinensis by macroporous resin. METHODS Single factor experiment was used to select the best type of macroporous resin based on the adsorption rate and resolution rate of total flavonoids and hyperoside. The effects of adsorption time, concentration of leakage point and pH on the purification process were investigated with the adsorption amount of total flavonoids and hyperoside as indexes. The effects of eluent flow rate, eluent volume and eluent concentration on the purification process were investigated with the elution rate of total flavonoids and hyperoside as indexes. On this basis, the Box-Behnken response surface methodology was used to optimize the optimum process using the normalized value of total flavonoids and hyperoside content as the index. RESULTS The optimal resin type for the purification of total flavonoids from Cuscuta chinensis was AB-8, and the optimal process conditions were as follows:pH 3.9, eluent volume 19.6BV, eluent concentration 74%. The total OD value of validation test was 0.982 5, which was close to the predicted value. Before and after purification, the content of total flavonoids was increased from 18.95 mg·g^-1 to 85.68 mg·g^-1, and the content of hyperoside increased from 4.12 mg·g^-1 to 16.85 mg·g^-1. CONCLUSIONS The optimized purification technology is stable and feasible, and could be used for the purification of total flavonoids from Cuscuta chinensis.