不同提取方式对连翘、荆芥、薄荷挥发油成分及抗菌活性的影响

Effects of different extraction methods on composition and antibacterial activity of volatile oil from Forsythiae Fructus, Schinzonepetae Herba, and Menthae Haplocalycis Herba

目的对连翘Forsythiae Fructus、荆芥Schizonepetae Herba、薄荷Menthae Haplocalycis Herba及其配伍挥发油进行化学成分及抑菌活性分析,探讨挥发油不同提取方法与其成分及抑菌活性的关系。方法采用气相色谱-质谱联用技术(GC-MS),对连翘、荆芥、薄荷挥发油成分及其配伍后成分的变化进行比较分析,并采用纸片琼脂扩散法、微量稀释法分别测定抑菌圈直径及最低抑菌浓度(MIC),评价单味连翘、荆芥、薄荷挥发油和配伍提取挥发油及单提混合后的挥发油对常见的4种致病菌的抑菌活性。结果连翘、荆芥与薄荷混合提取后,所得到的挥发油主要成分及含量均发生了变化。连翘-荆芥混合提取的挥发油中,缺失了连翘挥发油中含有的7种成分和荆芥挥发油中的7种成分,而新增了8种成分。连翘-薄荷混合提取后,缺失了连翘挥发油中的6种成分和薄荷挥发油中的8种成分,并新增8种成分。荆芥-薄荷混合提取后,缺失了荆芥挥发油中的4种成分和薄荷挥发油中的7种成分,新增了7种成分。连翘-荆芥-薄荷混合提取后,缺失了连翘挥发油中的6种成分、荆芥挥发油中的4种成分、薄荷挥发油中的2种成分,连翘和薄荷共有成分缺失1种,荆芥和薄荷共有成分缺失2种,新增了9种成分。与单提混合挥发油相比,3种药材混合提取组中胡薄荷酮的相对含量明显下降。抑菌实验显示,不同提取方式所得挥发油的抑菌效果不同,单味挥发油及单提混合挥发油对大肠埃希菌、金黄色葡萄球菌、绿脓杆菌、白色念珠菌的抑菌效果均优于配伍提取挥发油组。结论挥发油是解表中药的重要药效成分,中药挥发油采用不同的提取方式对挥发油的得率、成分及药效有一定的影响,最终影响其疗效,应对此加以关注。

Objective To compare the chemical composition and antibacterial activity of the volatile oils from Forsythiae Fructus（FF）, Schizonepetae Herba（SH）, Menthae Haplocalycis Herba（MH）, and their compatibility, and to explore the influence of different extraction methods on components and antibacterial activity of volatile oil. Methods GC-MS was employed to analyze the composition of the volatile oil from FF, SH, and MH extracted separately, and their mixture extracted, as well as physical mixing volatile oil of FF, SH, and MH extracted separately. The diameter of the inhibition zone and the minimum inhibitory concentration（MIC） were measured using the paper agar diffusion method and micro dilution method. The antibacterial activities of volatile oil on four common pathogens of respiratory tract were evaluated. Results When FF, SH, and MH were mixed and extracted, the main obtained components and contents of volatile oil changed. Compared to separately extracted volatile oil, eight new components emerged, seven components from FF and seven components from SH disappeared in FF and SH mixture extracted volatile oil. In FF and MH mixture extracted volatile oil, compared with separately extracted, six components from FF and eight components from MH disappeared, and eight new components emerged. As to SH and MH mixture extracted volatile oil, four components from SH and seven components from MH disappeared, and seven new components emerged. In the three herbs mixture extracted volatile oil, six components from FF, four components from SH, and two components from MH disappeared, one commom component of FF and MH, two common components of SH and MH disappeared, however, nine new components emerged. The content of pulegone significantly decreased in the three herbs mixture extracted volatile oil compared with that in separately extracted. The antibacterial experiments showed that the antibacterial effects of the volatile oils obtained by different extraction methods were different. The antibacterial effects of volatile oil extracted separately and their physical mixing volatile oil on Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans were better than that from mixture extracted. Conclusion Volatile oil is an important effective component of Chinese patent medicines with relieving exterior syndrome function. Different extraction methods have a certain influence on the yield and composition of volatile oil, which ultimately affects the efficacy of drugs. Therefore, we should pay more attention to extraction method and process of volatile oil in these prescriptions.