betway必威登陆网址 (betway.com )学报››2021,Vol. 42››Issue (10): 739-743.DOI:10.3969/j.issn.2097-0005.2021.10.004

• 基础研究 •上一篇下一篇

益母草二萜类化合物的提取工艺及其抑菌活性

曹付威, 刘福林, 任莹莹, 陈文笙, 蒋美玲, 马敬傲, 范凯丽, 郭越生, 李欣然, 高艳霞()

  1. betway必威登陆网址 (betway.com )生命科学学院,山东 泰安 271016
  • 出版日期:2021-10-25发布日期:2021-11-16
  • 通讯作者:高艳霞
  • 作者简介:曹付威,本科在读。
  • 基金资助:
    泰安市科技发展计划(2018NS0087);山东省大学生创新创业训练计划(S201910439074);国家级大学生创新创业训练计划(S202010439015)

Eextraction technology of diterpenoids fromLeonurusjaponicasand its antimicrobial activity

Fuwei Cao, Fulin Liu, Yingying Ren, Wensheng CHen, Meiling Jiang, Jingao Ma, Kaili Fan, Yuesheng Guo, Xinran Li, Yanxia Gao()

  1. College of Life Sciences,Shandong First Medical University & Shandong Academy of Medical Sciences,Taian 201016,China
  • Online:2021-10-25Published:2021-11-16
  • Contact:Yanxia Gao

摘要: 目的

利用正交试验法优化益母草二萜提取工艺,并探究其抑菌活性。

方法

充分溶涨益母草后,采用加热回流法提取制备益母草二萜。以穿心莲内酯为对照品,采用紫外分光光度法测定益母草中总二萜的含量。首先对乙醇浓度、料液比、提取时间和提取次数进行单因素试验,考察其对二萜含量的影响;根据单因素试验结果,固定提取次数为3次,进行乙醇浓度、料液比、提取时间的L9(33)正交试验,优化益母草二萜提取工艺并验证。再用乙酸乙酯对浸提物进行萃取,使用不同比例的石油醚∶丙酮溶剂进行硅胶柱层析,初步得到益母草二萜粗提物。采用琼脂扩散法对其进行金黄色葡萄球菌和大肠杆菌抑菌实验研究。

结果

益母草总二萜的最佳提取工艺为:乙醇90%、料液比1 g∶30 mL、提取时间2 h。经验证试验可知,该工艺的二萜含量平均为3.38%。硅胶柱层析洗脱剂石油醚∶丙酮的最佳比例为7∶3。该粗提物能明显抑制金黄色葡萄球菌的生长,对大肠杆菌则基本没有影响。

结论

优化的提取工艺稳定高效益,能用于益母草二萜的提取,后续的抑菌试验说明益母草二萜具有抑菌活性,为进一步开发利用益母草奠定基础。

关键词:益母草,二萜类化合物,提取工艺,抑菌

Abstract:

Objective

To study the optimal extraction technology and antimicrobial activity of diterpenoids fromLeonurusjaponicas.

Methods

After the full swelling ofL.japonicas,total diterpenoids was extracted using heating reflux method. The content of total diterpenoids inL. japonicaswas determined by ultraviolet spectrophotometry with andrographolide as control. Firstly, the effects of ethanol concentration, solid-liquid ratio, extraction time and extraction times on the content of diterpenoids were investigated by single factor tests. According to the single factor test results, fixing the extraction times as 3 times, the L9(33) orthogonal test of ethanol concentration, solid-liquid ratio and extraction time was conducted to optimize the extraction technology of diterpenoids fromL. japonicas,and the optimized extraction technology was verified. Then the extract was further leached by ethyl acetate. Moreover, different proportions of petroleum ether and acetone were investigated using silica gel column chromatography and crude extract of diterpenoids was initially obtained. On this basis, the effect of these crude diterpenoids on antimicrobial activity ofStaphylococcus aureusandEscherichia coliwere studied using disk agar diffusion method.

Results

The optimal extraction conditions of total diterpenoids fromL. japonicaswere as follows∶ ethanol concentration of 90%, solid-liquid ratio of 1 g∶30 mL and extraction time of 2 h. The average content of diterpenoids from this process was 3.38%. The optimum ratio of petroleum ether to acetone for silica gel column eluent was 7∶3. In addition, the crude extract could significantly inhibit the growth ofS. aureus, but had little effect onE. coli.

Conclusion

The optimized extraction process is stable and efficient.L. Japonicascontains active substances with antibacterial effect. This lays a foundation for further isolation and identification of active components.

Key words:diterpenoids,extract,antimicrobial activity