朱登辉 张靖柯 李孟 石静亚 刘娟娟 魏俊俊 郑晓珂 冯卫生

摘 要 目的：优化山茱萸叶总三萜提取工艺。方法：在山茱萸叶充分溶胀的基础上，采用加热回流法提取其总三萜。在单因素试验中考察乙醇体积分数、液料比、提取时间和提取次数对山茱萸叶总三萜含量的影响;以齐墩果酸为对照品，采用紫外-可见分光光度法测定总三萜含量。在单因素试验的基础上，固定提取次数为3次，以总三萜含量为响应值，以乙醇体积分数、液料比、提取时间为响应因素，采用Box-Behnken设计-响应面法优化山茱萸叶总三萜的提取工艺并进行验证。结果：山茱萸叶总三萜的最佳提取工艺为乙醇体积分数73%、液料比38 ∶ 1（mL/g）、提取时间60 min。3次验证试验结果显示，山茱萸叶总三萜的含量分别为6.92%、6.91%、6.84%，平均含量为6.89%（RSD=0.63%），与预测值（7.28%）的相对误差为5.36%。结论：优化的提取工艺稳定、可靠，可用于山茱萸叶总三萜的提取。

关键词 山茱萸叶;总三萜;Box-Behnken设计;响应面法;提取工艺

ABSTRACT   OBJECTIVE： To optimize the extraction technology for total triterpenes from the leaves of Cornus officinalis. METHODS： Based on the full swelling of the leaves of C. officinalis， total triterpenes was extracted with heating reflux method. The effects of ethanol concentration， liquid-solid ratio， extraction time and extraction times on the contents of total triterpenes from the leaves of C. officinalis were investigated by single factor test. Using oleanolic acid as control， the contents of total triterpenes were detected by UV spectrometry. On the basis of single factor test， fixing the times of extraction as 3 times， taking the contents of  total triterpenes as response value， using ethanol volume fraction， solid-liquid ratio and extraction time as factors， Box-Behnken design-response methodology was used to optimize the extraction technology of total triterpenes from the leaves of C. officinalis， and the optimized extraction technology was validated. RESULTS： The optimal extraction technology of total triterpenes from the leaves of C. officinalis were as follows as ethanol concentration of 73%， liquid-to-material ratio of 38 ∶ 1 （mL/g）， extraction time of 60 min. Results of 3 validation tests showed that the contents of total triterpenes from the leaves of C. officinalis were 6.92%， 6.91%， 6.84%; the average content was 6.89% （RSD=0.63%）， relative error of which with the predicted value （7.28%） was 5.36%. CONCLUSIONS： The optimized technology is stable and reliable， and can be used for the extraction of total triterpenes from leaves of C. officinalis.

KEYWORDS   Leaves of Cornus officinalis;Total triterpenes;Box-Behnken design; Response surface methodology; Extraction process

山茱萸為山茱萸科植物山茱萸Cornus officinalis Sieb. et Zucc.的干燥成熟果肉，具有补益肝肾、收涩固脱的功效，可用于治疗眩晕耳鸣、腰膝酸痛等症[1]。该药主要分布于我国河南、浙江、湖南等地[2]。近年来，研究人员发现，山茱萸叶也有药用价值，有文献报道山茱萸叶具有细胞毒活性和抗氧化活性[3-5]。但是作为非传统药用部位的山茱萸叶在山茱萸果实采收过程中多作为废弃物处理，造成了药用资源的浪费。

本课题组在前期研究中发现，山茱萸叶中含有大量三萜类化合物，而三萜类化合物已知的生物活性有保肝、抗菌、降血糖、抗氧化、抗炎、调节免疫、利尿、抗病毒等[6-13]。……