目的 联用熵权法、Plackett-Burman设计(PBD)与Box-Behnken设计(BBD),多目标筛选甘草黄酮类成分的纯化工艺。方法 在构建甘草HPLC指纹图谱的基础上,以6种黄酮类成分甘草素葡萄糖芹糖苷、甘草苷、芹糖异甘草苷、异甘草素葡萄糖芹菜苷、异甘草苷和新异甘草苷的回收率为检测指标,筛选12种大孔树脂中最佳型号。熵权法赋予6种黄酮类成分的回收率客观权重,计算综合指标作为评价标准,运用PBD筛选显著性影响因素,结合BBD优选大孔树脂富集甘草黄酮的最佳工艺参数。结果 ADS-7型大孔树脂对甘草中6种黄酮类成分的富集选择性最高。最佳工艺参数为上样pH值4.5,上样质量浓度0.20 g/mL,上样量1.0 g/g,上样体积流量0.6 mL/min,6.7 BV的83%乙醇以1.0 mL/min体积流量洗脱。纯化后6种黄酮类成分的回收率为86%~97%,模型综合评价预测值为93.32%,实验所得综合评价为93.05%,相对偏差1.17%。结论 联用熵权法、PBD与BBD筛选甘草黄酮类成分纯化工艺是科学可行的,可为实现中药有效成分分离纯化的多目标寻优提供借鉴。

Objective To multi-objectively optimize the purification process parameters of licorice flavonoids using entropy-weight method, Plackett-Burman design(PBD), and Box-Behnken design(BBD). Methods On the basis of HPLC fingerprints of licorice, the macroporous resin type was chosen using the recovery rates of six components(liquiritin apioside, liquiritigenin, isoliquiritin apioside, licuraside, isoliquiritin, and neoisoiiquiritin) as detection indexes. Weights of the recovery rates of six components were determined by entropy-weight method, in order to obtain the comprehensive index. The significantly influencing factors were firstly evaluated by PBD, then purification conditions were optimized by BBD. Results ADS-7 type resin showed a high selectivity for six components. The optimized purification technology was as follows:pH value was 4.5, sample concentration was 0.20 g/mL, ratio of sample to resin was 1.0 g/g, flow rate was 0.6 mL/min, elution dosage was 6.7 BV, ethanol concentration was 83% of eluting agent, and elution rate was 1.0 mL/min. Under the conditions, the recovery rates of six components were 86%-97%, the theoretical and actual comprehensive indexes were 93.32% and 93.05%, respectively, with a relative error of 1.17%. Conclusion Entropy-weight method combined with PBD and BBD-RSM used to optimize the purification process for the licorice flavonoids in this study is scientific and feasible, providing a new reference to realize the multi-objective optimization of the purification technology for active constituents in Chinese materia medica.

国家自然科学基金资助项目(81573868);浙江省自然科学基金资助项目(LR13H270001,LZ14H270001);浙江省卫生高层次创新人才培养工程项目