Box-Behnken响应面法优化黑果枸杞花色苷纯化工艺及其成分分析

Optimization of purification process of anthocyanins in Lycium ruthenicum Murray by Box-Behnken response surface method and analysis of its components

目的:优化黑果枸杞花色苷的大孔树脂纯化工艺,并分析其主要化学成分。方法:采用Box-Behnken响应面法考察树脂径高比、上样液浓度、上样液pH值、上样流速、洗脱乙醇体积、洗脱乙醇浓度、洗脱乙醇pH值、洗脱流速等因素水平,并进行验证试验;同时采用UPLC-LTQ Orbitrap MS对所得纯化物进行成分分析。结果:最佳纯化工艺为径高比1∶15,上样液浓度32g/L,上样液pH=2.7,上样流速1mL/min;洗脱剂为92%乙醇,洗脱剂pH=3.6,洗脱流速1mL/min,洗脱剂体积3.31BV,在此条件下,黑果枸杞花色苷的产率为8.01%。初步鉴定出纯化物中含有21种花色苷及花青素类成分,主要包括以飞燕草素、矮牵牛素和锦葵素为母核的花色苷,其中2种花色苷在黑果枸杞中首次报道。结论:采用Box-Behnken响应面法优化所得的黑果枸杞花色苷纯化工艺简单可行,且纯化效果良好,适用于黑果枸杞花色苷的纯化。

Objective:To optimize the macroporous resin purification process of anthocyanins in Lycium ruthenicum Murray and analyze its main chemical components.Methods:The Box-Behnken response surface method were used to investigate the resin aspect ratio,sample concentration,pH of the sample,loading flow rate,eluent volume,ethanol concentration,pH of the ethanol and the elution flow rate were tested and verified.The purified material was analyzed by UPLC-LTQ Orbitrap MS.Results:The best purification process was as follows:resin aspect ratio 1∶15,sample solution concentration 32 g/L,sample solution pH=2.7,sample flow rate 1 mL/min,ethanol concentration 92%,pH of the ethanol was 3.6,the elution flow rate was 1 mL/min,and the eluent volume was 3.31 BV.Under the optimal conditions,the yield of anthocyanins of was 8.01%.It was showed that 21 kinds of anthocyanin components were identified from purified substance,2 of them were identified for the first time.Conclusion:Optimization of the purification process by Box-Behnken response surface is convenient and feasible.It is suitable for the purification of anthocyanins in Lycium ruthenicum Murray.