目的 优化蟾酥中脂溶性成分华蟾酥毒基、脂蟾毒配基的提取及纯化工艺。方法 以华蟾酥毒基和脂蟾毒配基总量为考察指标，以乙醇浓度、溶剂倍量、提取时间为考察因素，采用正交试验确定最佳提取工艺参数；通过单因素考察并结合Box-Behnken响应面法，以展开剂比例、柱高与直径比、吸附剂与上样量的比值为考察因素，筛选并确定最佳纯化工艺。结果 确定最佳提取工艺为加入10倍量85%的乙醇提取90 min；最佳纯化工艺为展开剂环己烷-氯仿-丙酮（4：3：3），柱高与直径比为7：1，吸附剂-上样量为5.5：1，2种蟾毒配基纯化后的总量可达66.51%。通过3批放大工艺验证，表明模型拟合度良好，采用硅胶柱色谱分离纯化蟾毒配基，具有操作简便、分离速度快、分离效果好等优点。结论 所选工艺合理、可行，为以后该产品的产业化应用提供技术参考和依据。

Objective To optimize the extraction and purification technology of liposoluble constituents from Bufonis Venenum. Methods The total content of cinobufagin and resibufogenin was taken as the index, and the ethanol potency, solvent multiplication and extraction time were taken as the investigation factors. The optimum extraction process parameters was determined by orthogonal test. Through single factor investigation experiment combined with Box-Behnken response surface method, taking the expansion agent ratio, ratio of diameter to height, adsorbent and sample quantity as the investigation factors, the optimum purification process was selected and determined. Results It was determined that the optimum extraction process for the addition was 10 times 85% ethanol for 90 minutes' extraction, the best purification process for the expansion agent cyclohexane-chloroform-acetone was 4:3:3, column height and diameter was 7:1, adsorbent and sample volume was 5.5:1. The content of two kinds of toad poison base purified was up to 66.51%. Through the verification of three batches of amplification process, it was shown that the model fit well, and the separation and purification of toad poison ligand by silicone column chromatography had the advantages of simple operation, fast separation speed, and good separation effect. Conclusion The selected process is reasonable and feasible, which provides technical reference and basis for the industrialization application of the product in the future.

国家自然科学基金资助项目（81373660）