目的 采用响应面分析法优化女贞子中总三萜酸的提取工艺,以均匀实验确定纯化工艺,使总三萜酸的含量达到50%以上。方法 应用HPLC-UV,建立女贞子中总三萜酸含量的测定方法,通过乙醇浓度、溶剂倍量、提取时间和提取次数等单因素考察,确定乙醇提取次数,应用响应面法优化乙醇浓度、溶剂倍量和提取时间。应用醇提水沉除去水溶性杂质;石油醚除去叶绿素、油脂等脂溶性杂质;乙醇沉淀得到女贞子总三萜酸;设计均匀实验分别确定石油醚的用量、纯化次数和时间,乙醇的浓度和用量。结果 女贞子总三萜酸以齐墩果酸计,提取工艺为体积分数80%的乙醇、10倍量、提取2次,每次0.5 h。纯化工艺为12倍量石油醚,纯化1次,每次2 h;10倍量体积分数60%的乙醇沉淀,纯化1次。结论 用响应面分析法和均匀实验设计法来优化女贞子三萜酸的提取纯化工艺,工艺简单,可产业化。

OBJECTIVE To optimize the extraction and purification process of total triterpenoid acid in Ligustri lucidi fructus using response surface method and uniform test. Finally, the content of total triterpenoid acid reached more than 50%.METHODS The content determination of total triterpenoid acid in Ligustri lucidi fructus was established by HPLC.Use ethanol as extraction solvent and through the single factor investigation about the ethanol concentration, solvent amount, extraction time and extraction times,we determined the number of ethanol extraction.In addition to extraction times, the other three factors were further optimized by response surface method.Then purified by the alcohol extraction-water precipitation method to remove the water-soluble ingredients. Petroleum ether was used to remove the chlorophyll, grease and other small polarity ingredients; using ethanol to remove the alcohol soluble components whose polarity is bigger than fructus ligustri lucidi terpene acids. Design the uniform experiment respectively to determine the amount of petroleum ether, frequency and time, also ethanol concentration and dosage. RESULTS The content of oleanolic acid and ursolic acid is larger than other compounds in total triterpenoid acid in Ligustri lucidi fructus, and they are isomers.When the results are in the equivalent of oleanolic acid, the optimum extraction process was determined as 80%, 10 times the amount of ethanol, extracting for 2 times, each time 0.5 h.The optimal purification process is 12 times the amount of petroleum ether, purified once, 2 h every time.Then 10 times amount of 60% ethanol to purify for once. CONCLUSION Optimizing the extraction and purification process of total triterpenoid acid in Ligustri lucidi fructus by using response surface method and uniform experiment is simple, feasible and worth promoting.