土圞儿总生物碱的提取和纯化工艺研究

Study on Extraction and Purification of the Total Alkaloids from Apios Fortunei Maxim.

目的确定土圞儿中总生物碱最佳的提取、纯化条件,为其开发和利用提供科学的理论依据。方法采用L_9(3~4)正交试验,加酸水加热回流提取,考察溶剂用量、提取时间、提取次数3个因素对土圞儿中总生物碱得率的影响。结合静态和动态吸附实验,分别比较了6种不同型号的大孔树脂(D101,H-30,LSD-001,HPD450,DM130,AB-8)对土圞儿总生物碱的吸附和解吸性能,并以吸附量、洗脱量为考察指标,对大孔树脂纯化总生物碱的条件进行了筛选。以盐酸小檗碱为对照品,采用紫外分光光度法测定土圞儿总生物碱的含量。结果土圞儿中总生物碱的最佳提取工艺条件为15倍量酸水加热回流提取2次,每次1 h。HPD450型树脂具有最佳的吸附及洗脱参数,其最佳纯化条件:上样液总生物碱浓度为1.6 mg·m L^(-1),pH 8.0,先以5倍柱体积的水洗去杂质,再以4倍柱体积40%乙醇洗脱,最后用4倍柱体积90%乙醇洗脱,得总生物碱量是纯化前总生物碱量80%左右。结论该提取、纯化条件简单可行,具有良好的应用前景。

OBJECTIVE To provide a theoretical basis for development and ulitization of the total alkaloids of Apios fortunei Maxim. by optimizing the best extraction and purification process. METHODS The effects of the ratio of solid-to-liquid, extraction time and extraction times on total alkaloid yield of Apios fortunei Maxim. were investigated by L9（3^4） orthogonal test. By the static and dynamic adsorption experiments, six types of macroporous adsorption resins（D101, H-30, LSD-001, HPD450, DM130, AB-8） were evaluated to find the optimum one in purificating efficiency with measuring the adsorption and eluting ratio of the total alkaloids as indexes and used berberine hydrochloride as a comparison to determine the content of the total alkaloids of Apios fortunei Maxim.. RESULTS Ratio of solid-to-liquid was 1∶15, extracting twice and 1 h every time were optimal. The HPD450 macroporous adsorption resin provided the optimal adsorption and elution parameters through the comparison of 6 different types of macroporous resin. The best adsorption capacity was achieved with the following condition： the concentration of extract liquid was 1.6 mg·m L^-1, pH 8.0,then the resin was washed by 5 BV water and 4 BV 40% ethanol to remove impurity and alkaloids were desorbed by 4 BV 90% ethanol. The total alkaloid was 80% of it before purifiction. CONCLUSION The process is simple, feasible and has a good prospect.