油松叶低极性成分的GC-MS法分析及其抗真菌活性研究

Analysis of low polarity chemical constituents of Pinus tabulaeformis Carr.leaves by GC-MS method and study on their antifungal activity

目的:研究油松叶低极性部位的化学成分及其抗真菌活性。方法:油松叶经80%乙醇热提取后用石油醚萃取得到低极性部位,采用GC-MS法初步分析化学成分,所测结果与标准谱库进行对比分析,用峰面积归一法计算各个成分的相对含量。采用棋盘式微量稀释法测定低极性部位联合氟康唑对耐氟康唑白念珠菌的80%最小抑菌浓度(MIC80)。结果:从低极性部位中检测并鉴定出30个化学成分,相对含量最高的是15-羟基脱氢枞酸甲酯(11.73%),其次是脱氢枞酸甲酯(7.67%);油松叶的石油醚提取物、80%乙醇提取物及低极性部位联合氟康唑对耐药白念珠菌的MIC_(80)分别为>250μg/ml、1.95μg/ml和3.91μg/ml。结论:油松叶80%乙醇提取物及低极性部位都具有协同氟康唑抗耐药白念珠菌活性。油松叶低极性部位的协同作用可能与其含有的倍半萜和二萜类化合物有关。

Objective:To investigate the main chemical constituents and antifungal activity of the low polarity extract from Pinus tabulaeformis Carr.leaves.Methods:Low polarity extract was obtained by petroleum ether following extraction of Pinus tabulaeformis Carr.leaves by 80%ethanol.Then,the chemical constituents of the extract were preliminarily analyzed by GC-MS method and the obtained results were compared and analyzed with the standard mass spectral data.The contents of various components were calculated using the area normalization method.The 80%minimal inhibitory concentrations(MIC80)of fluconazole-resistant Candida albicans strains by low polarity extract in combination with fluconazole were determined by checkerboard microdilution assay.Results:Thirty components were detected and identified from the low polarity extract,in which the highest relative content was 15-hydroxydehydroabietic acid methyl ester(11.73%),followed by methyl dehydroabietate(7.67%).The MIC80 data of the petroleum ether extract,the 80%ethanol extract and the low polarity extract from the Pinus tabulaeformis Carr.leaves in combination with fluconazole against fluconazole-resistant Candida albicans were respectively>250μg/ml,1.95μg/ml and 3.91μg/ml.Conclusion:In combination with fluconazole,the 80%ethanol extract of Pinus tabulaeformis Carr.leaves and its low polarity extract all had synergistic activity on fluconazole-resistant Candida albicans.The synergistic effect of the low polarity extract of Pinus tabulaeformis Carr.leaves might be related to the sesquiterpenes and diterpenoids of the extract.