Box-Behnken设计响应面法优化黑果枸杞总黄酮纯化工艺

Study on purification of total flavonoids from Lycium ruthenicum Murr.by Box-Behnken design response-surface methodology

目的应用Box-Behnken设计响应面法优化黑果枸杞总黄酮纯化工艺条件及相关参数。方法考察6种大孔吸附树脂静态、动态吸附和解吸性能,用单因素试验分别对上样浓度、上样体积、上样pH、洗脱液浓度、洗脱液体积和洗脱流速进行考察,以黑果枸杞总黄酮纯化后含量为指标,再从6个单因素中选出对解吸附试验影响最显著的3个因素进行三因素三水平Box-Behnken设计响应面实验。结果NKA-9型大孔树脂为纯化黑果枸杞总黄酮的最佳树脂,最佳纯化工艺条件为总黄酮提取液浓度为4 mg/mL,pH为4.0,按5 BV进行上样,再用3 VB的80%乙醇在2 mL/min流速下进行洗脱,黑果枸杞总黄酮含量可以从4.54%提高到43.19%。结论此工艺对黑果枸杞总黄酮的纯化效果较好,工艺简单且稳定性良好,适用于工业化生产。

Objective Application of Box-Behnken design response surface methodology to optimize the purification conditions and related parameters of total flavonoids from Lycium ruthenicum Murr.Methods Static,dynamic adsorption and desorption properties of six macroporous adsorption resins were studied.The sample concentration,volume,pH,eluent concentration,eluent volume and eluting flow rate were investigated by single factor test.Taking the purified content of total flavonoids of Lycium ruthenicum Murr.as the index,the 3 factors that had the most significant effect on the desorption test were selected from 6 single factors to carry out the three-factor and three-level Box-Behnken design response surface experiment.Results NKA-9 macroporous resin was the best resin for the purification of total flavonoids from Lycium ruthenicum Murr..The optimum purification conditions were as follows:the concentration of total flavonoids was 4 mg/mL,pH was 4.0,the sample was added at 5 BV,and eluted with 80%ethanol of 3 VB at the flow rate of 2 mL/min,the content of total flavonoids of Lycium ruthenicum Murr.could be increased from 4.54%to 43.19%.Conclusion This process has good purification effect on total flavonoids of Lycium ruthenicum Murr..It’s simple and stable,and also suitable for industrial production.