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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 determine the optimum extraction and purification conditions of total alkaloids in soil, and provide a scientific theoretical basis for its development and utilization. Methods The orthogonal experiment of L_9 (3 ~ 4) was carried out, and the extract was heated with acid and heated to reflux. The effects of solvent dosage, extraction time and extraction times on the total alkaloid yield were studied. Combined with static and dynamic adsorption experiments, the adsorption and desorption properties of total alkaloids from the soil were compared with six kinds of macroporous resins (D101, H-30, LSD-001, HPD450, DM130 and AB- The conditions of total alkaloids purification by macroporous resin were screened by adsorption amount and elution amount. Taking berberine hydrochloride as reference substance, the total alkaloid content in soil was determined by UV spectrophotometry. Results The optimal extraction conditions of total alkaloids in soil were 15 times the amount of acid water heated to reflux twice, 1 h each time. HPD450 resin has the best adsorption and elution parameters. The optimal purification conditions are as follows: the total alkaloid concentration of the sample solution is 1.6 mg · mL -1, pH 8.0, and washed with 5 column volumes of water first Impurities, and then 4 times the column volume of 40% ethanol, and finally with 4 column volumes of 90% ethanol elution, the total alkaloid amount is about 80% of total alkaloids before purification. Conclusion The extraction and purification conditions are simple and feasible, and have good application prospects.