半枝莲中总生物碱的大孔树脂纯化工艺优化研究

Studies on Optimization of Purification Process of Total Alkaloids from Scutellaria barbata by Macroporous Resin

目的探讨大孔吸附树脂纯化半枝莲中总生物碱的最优条件。方法以盐酸小檗碱为对照品,采用紫外分光光度法测定各条件下的吸光度,通过进行动态吸附/解吸试验、静态吸附/解吸试验,筛选出了最佳型号的大孔树脂,并以比泄漏量不超过10%为指标,确定上样终点;设计L9(3^4)正交试验,以比泄漏量、吸附率和解吸率为指标,考察了上样液浓度、上样液pH、上样体积流量、径高比四个上样参数对大孔树脂纯化半枝莲中总生物碱的影响;以总生物碱含量为指标,对洗脱液种类、洗脱液浓度、洗脱剂量、洗脱剂流速四个洗脱剂参数进行考察,记录实验结果并对进行相关的极差分析。同时对所得总生物碱进行了初步抗肿瘤活性实验。结果本实验根据动态吸附/解吸试验及静态吸附/解吸试验,确定最佳型号的大孔树脂是AB-8型号的大孔树脂;最优条件是上样液浓度为0.010g·L^-1,上样液pH为9,体积流量为0.5BV/h,径高比为1∶4,洗脱液为体积份数65%~90%甲醇、洗脱液量6BV、洗脱流速2BV/h。结论本研究筛选出了大孔吸附树脂纯化半枝莲中总生物碱的最优纯化条件,工艺简单,纯化效果好,也为半枝莲中总生物碱的进一步开发利用提供了科学的实验依据。

OBJECTIVE To screen the optimum conditions for purification of total alkaloids from Scutellaria barbata by macroporous adsorption resin.METHODS The berberine hydrochloride was used as the reference substance.The absorbance of each condition was measured by ultraviolet spectrophotometry.Through dynamic adsorption/desorption test and static adsorption/desorption test,the optimum type of macroporous resin was selected,and the end point of sampling was determined with no more than 10% the specific leakage.The orthogonal test of L9(3^4) was designed,with the specific leakage,adsorption and desorption rate as the index.The effects of four parameters on the purification of total alkaloids from S.barbata by macroporous resin were investigated,including the concentration of sample solution,the pH of sample solution,the volume flow rate of sample solution and the ratio of diameter to height.Taking the content of total alkaloids as the index,the four parameters of eluent,including the type of eluent,the concentration of eluent,the dosage of eluent and the flow rate of eluent,were investigated,and the experimental results were recorded and the related range analysis was carried out.At the same time,the preliminary anti-tumor activity of the total alkaloids was tested.RESULTS According to dynamic adsorption/desorption test and static adsorption/desorption test,the best type of macroporous resin was AB-8 macroporous resin.The optimal conditions were as follows:the concentration of sample solution was 0.010 g·L^-1,the pH of sample solution was 9,the volume flow rate was 0.5 BV/h,the ratio of diameter to height was 1∶4,the eluent was 65%~90% methanol,the eluent volume was 6 BV,and the eluent flow rate was 2 BV/h.CONCLUSION The optimum conditions for purification of total alkaloids from S.barbata by macroporous adsorption resin were screened.The process is simple and the purification effect is good.It also provides a scientific experimental basis for further deveLopment and utilization of total alkaloids in S.barbata.