中文摘要：

目的：优化蟾酥中脂溶性成分蟾毒灵、华蟾酥毒基及酯蟾毒配基的提取工艺，并建立了测定蟾酥提取物中上述3种成分含量的HPLC方法。方法：以蟾酥中脂溶性成分蟾毒灵、华蟾酥毒基及酯蟾毒配基含量为指标，采用L9（3^4）正交试验对蟾酥提取工艺进行了优化；建立的HPLC法，Diamonsil—C18色谱柱（4．6mm×250mm，5μm），流动相乙腈-水（55：45），流速1．0mL·min，检测波长296nm。结果：最佳提取工艺条件为A2B3D2，即75倍量95％乙醇80℃提取120min；所建立的HPLC方法蟾毒灵、华蟾酥毒基及酯蟾毒配基均得到很好的分离，且重现性、精密度、线性关系良好，最小检测限分别为14．6，9．0，11．8ng·mL^-1。结论：确定了蟾酥最佳提取条件，HPLC能准确测定蟾酥提取物中蟾毒灵、华蟾酥毒基及酯蟾毒配基的含量，方法简便，结果准确。

英文摘要：

Objective：Study on extract method of liposoluble constituent bufalin, cinobufagin and resibufogenin in Bufons Venenum by orthogonal experiment design and established HPLC method for determination the assay of three effective constituents above. Method ：An orthogonal test was adopted in this study. The content of bufalin, cinobufagin and resibufogenin was used as index for optimizing the extraction condition. In established HPLC method , the colmnn ： Diamonsil-C18 column （4.6mm×250mm,5μm）, mobile phase ： acetonitrile-water （55 ： 45 ） , the flow rate ： 1.0 mL · min ^-, the detection wavelength 296 nm. Result ： The best extraction technology was to extract for 120 min with 75 times 95% ethanol in 80 ℃. The calibration curves of bufalin, cinobufagin, resibufogenin were linear and LOD were 14.6,9.0,11.8 ng·mL^-1respectively. The repeatability and precision of the HPLC method was good. Conclusion：Confirmed the best extract method and established HPLC method to determine the assay of three constituents in Bufons Venenum extraction was simple and accurate.