白芍非药用部位芍头中单萜苷类有效部位的制备工艺研究

Study on preparation of monoterphene glycosides from the non-medicinal partrhizome of Paeonia lactiflora

目的优化白芍Paeonia lactiflora非药用部位芍头中单萜苷类有效部位的提取及纯化工艺。方法以芍药苷、芍药内酯苷等单萜苷类成分以及非单萜苷类成分苯甲酸、没食子酸的含量为指标,采用归一化处理得到总评归一值(overall desirability,OD);以提取时间、料液比、乙醇体积分数作为乙醇提取的关键工艺参数,运用星点设计-效应面法建立OD与工艺参数之间的数学模型,推算芍头回流提取的最佳工艺,并进行验证。在单因素试验确定大孔树脂型号的基础上,以上样体积流量、上样液质量浓度、柱径高比、洗脱体积流量作为纯化的关键工艺参数,采用可考察交互作用的正交设计实验优化提取液纯化工艺,并进行验证。结果芍头回流提取的优选工艺为加12倍量50%乙醇提取2次,每次80 min;3次工艺验证的OD为2.61、2.49、2.57,RSD为2.5%。以D-101型大孔吸附树脂纯化,0.25 g/mL药液上样,上样体积流量为0.5 mL/min,径高比为1∶4,最大上样量为3 BV,2 BV去离子水除杂,4 BV 50%乙醇以1 mL/min体积流量洗脱。3次纯化工艺验证综合评分分别为0.7333、0.7024、0.7297,RSD为2.3%;单萜苷总转移率为74.17%,纯化物中单萜苷质量分数为59.85%。结论得到的提取和纯化工艺高效且稳定,可除去芍头中大部分非单萜苷成分,获得单萜苷类有效部位。

Objective To optimize the extraction and purification process of active parts of monoterpene glycosides from the nonmedicinal part-rhizome of Paeonia lactiflora.Methods The contents of multiple components were used as indexes,including monoterpenoid glycosides,such as paeoniflorin and albiflorin,and non-monoterphene glycosides components,such as benzoic acid and gallic acid.The normalization method was used to obtain the overall desirability(OD).The extraction time,solid-to-liquid ratio and ethanol volume fraction,as the key process parameters,were optimized through central composite design-response surface methodology,and the results were verified.Based on the single factor tests used to determine the type of macroporous resin,the loading volume flow rate,loading solution mass concentration,column diameter to height ratio,and elution volume flow rate were the key process parameters for purification.Orthogonal design experiments that can investigate the interactions were used to optimize the extraction liquid purification process and the results were verified.Results The optimal reflux extraction process of rhizome of P.lactiflora was to extract twice with 12 times of 50%ethanol,80 minutes eachtime.The OD values of three validation experiment were 2.61,2.49,2.57,and the RSD was 2.5%.D-101 macroporous adsorption resin was selected for the purification,0.25 g/mL drug solution was loaded,the sample volume flow rate was 0.5 mL/min,the diameter-to-height ratio was 1:4,the maximum sample load was 3 BV,2 BV deionized water was used for impurity removal,and 4 BV 50%ethanol was eluted at the volume flow rate of 1 mL/min.The comprehensive scores of the three purification process verifications were 0.7333,0.7024,0.7297,the RSD was 2.3%,and the total transfer rate of monoterpene glycosides was 74.17%,and the mass fraction of monoterpene glycosides in the purified product was 59.85%.Conclusion The optimum extraction process is efficient and steady,which can remove most of non-monoterpene glycosides in the rhizome of P.lactiflora,and obtain the active fraction of the monoterpene glycosides in the rhizome of P.lactiflora.