艾叶挥发油化学成分分析和抗真菌活性的研究

摘要

目的 探讨艾叶挥发油化学成分及其单体化合物对常见食品致腐败真菌的抗菌作用.方法 采用水蒸气蒸馏法提取艾叶挥发油,经气相色谱-质谱(GC/MS)法分析,通过薄层色谱(TLC)法和硅胶柱层析法从艾叶挥发油中分离纯化出单体化合物,并用TLC法和GC/MS法定性鉴别,核磁共振(NMR)技术对其分子结构进行确认.采用菌丝生长速率法计算艾叶挥发油及其分离的单体化合物对疫霉、黑曲霉、粉红聚端孢、青霉、链格孢的抑菌率,二倍梯度稀释法测定其最低抑菌浓度(MIC).结果 艾叶挥发油共检测出30种化合物,其中桉树脑和4-松油烯醇的含量分别为22.71％和7.34％.TLC法和GC/MS法分析样品和对照品,其Rf值和保留时间基本相同,经NMR技术确定桉树脑和4-松油烯醇的分子结构.艾叶挥发油对5种真菌(疫霉、黑曲霉、粉红聚端孢、青霉、链格孢)的抑菌率为15.2％～74.5％,桉树脑对5种真菌的抑菌率为25.6％～69.4％,4-松油烯醇对5种真菌抑菌率为18.2％～72.9％.艾叶挥发油对疫霉、黑曲霉、粉红聚端孢、青霉的MIC为50 μL/mL,对链格孢的MIC为100 μL/mL,桉树脑和4-松油烯醇对5种真菌的MIC分别为50、40 μL/mL.结论 首次从艾叶挥发油中分离纯化出桉树脑和4-松油烯醇.艾叶挥发油及其分离的这2种单体化合物对5种真菌均有抗真菌活性,抗菌活性随浓度的增加而增强.%Objective To study the violate compounds and antifungal activity of volatile oil extracted from Artemisia argyi Levl.et Vant.Methods Steam distillation was used to extract volatile oil from Artemisia argyi that was subjected to gas chromatography/mass(GC/MS)analysis,the monomer compounds were separated using thin-layer chromatography(TLC)combined with silica gel column chromatography,TCL method and GC/MS qualitative was employed to identify,the molecular structure was confirmed by nuclear magnetic resonance(NMR).The inhibition rate of volatile oil its monomers on Phytophthoracapsici,Aspergillus niger,Trichotheciumroseum,Bgbfngnvb.nhg and Alternaria were determined by mycelial growth rate method.The minimum inhibitory concentration(MIC)was determined by two fold gradient dilution method.Results 30 compounds were detected from the essential oil,including eucalyptol and terpinen-4-ol with their relative content 22.71％and 7.34％,respectively.After being separated,,the molecular structure of eucalyptol and terpinen-4-ol are validated by NMR.The range of antifungal rate of essential oils is 15.2％～74.5％,range of eucalyptol and terpinen-4-ol was 25.6％～69.4％and 18.2％～72.9％,respectively.The MIC of essential oil against Artemisia argyi was 100 μL/mL,the others were 50 μL/mL.The MIC of eucalyptol against the 5 kinds of fungi was 50 μL/mL and the MIC of terpinen-4-ol against the 5 kinds of fungi was 40 μL/mL.Conclusion Eucalyptus and terpinen-4-ol were isolated for the first time from the essential oil of Artemisia argyi L,and were found to be antifungal activity against the 5 kinds of fungi available,and the antifungal activity increased with the increasing of its concentration.