Abstract: Objective　To investigate the activity of Acorus tatarinowii extracts against dust mites, and to isolate and characterize active ingredient of A. tatarinowii extracts. Methods　The essential oil components were extracted from A. tatarinowii rhizome powder by rotary evaporation with methanol as solvents, followed by petroleum ether extraction and rotary evaporation. The essential oil was mixed with Tween⁃80 at a ratio of 1∶1 and diluted into concentrations of 1.000 00%, 0.500 00%, 0.250 00%, 0.125 00%, 0.062 50% and 0.031 25%, while diluted Tween⁃80 served as controls. A. tatarinowii essential oil at each concentration (200 μL) was transferred evenly to filter papers containing 100 adult mites, with each test repeated in triplicate, and controls were assigned for each concentration. Following treatment at 25 ℃ and 75% relative humidity for 24 h, the mean corrected mortality of mites was calculated. The essential oil components were separated by silica gel column chromatography, and the essential oil was prepared in the positive column of medium pressure; and then, each component was collected. Silica gel column chromatography was run with the mobile phase that consisted of petroleum ether solution containing 10% ethyl acetate and pure ethyl acetate, detection wavelength of 254 nm, positive silica gel column as the chromatography column, and room temperature as the column temperature. Each component of the purified A. tatarinowii essential oil was diluted into 1.000 00% for acaricidal tests. The components with less than 100% acaricidal activity were discarded, and the remaining components were diluted into 50% of the previous⁃round tests for subsequent acaricidal tests. The components with acaricidal activity were subjected to high⁃performance liquid chromatography, liquid chromatography⁃mass spectrometry and pulsed⁃Fourier transform nuclear magnetic resonance spectroscopy. The structure of active monomer compounds was determined by standard spectral library retrieval and literature review. Results　A. tatarinowii essential oil at concentrations of 1.000 00%, 0.500 00%, 0.250 00% and 0.125 00% killed all dust mites, and the corrected mortality was all 100%. Exposure to A. tatarinowii extracts at an effective concentration of 0.062 5% for 24 hours resulted in 94.33% mortality of dust mites. Six components (A to F) were separated using gel column chromatography, and components D and E both showed a 100% acaricidal activity against dust mites at a concentration of 0.500 00%. In addition, Component D was identified as isoeugenol methyl ether, and Component E as β⁃asarinol. Conclusion　The extract of A. tatarinowii essential oil has acaricidal activity, and the isoeugenol methyl ether shows a remarkable acaricidal activity against dust mites.

Key words: Dust mite, Acorus tatarinowii, Acaricidal activity, Isoeugenol methyl ether

摘要： 目的　分析石菖蒲提取物对粉尘螨的杀灭活性，并对其有效成分进行分离鉴定。方法 以甲醇为溶剂，采用旋蒸法从石菖蒲根茎磨粉中获得冷浸提取物，再以石油醚进行萃取，再次旋蒸获得石菖蒲精油成分。将石菖蒲精油与吐温⁃80以1∶1比例混匀后用蒸馏水进行倍比稀释至1.000 00%、0.500 00%、0.250 00%、0.125 00%、0.062 50%和0.031 25%等6个浓度，同时将吐温⁃80用蒸馏水以同样比例稀释作为对照组。取200 μL各浓度石菖蒲精油分别均匀滴加于有100只成螨的滤纸上，每一浓度重复实验3次，各浓度石菖蒲精油均设置1个对照组，于25 ℃，75%相对湿度培养箱内处理24 h后，计算螨虫校正死亡率平均值并进行统计分析。采用硅胶柱层析法分离精油成分，石菖蒲精油经正向柱中压制备，收集其中各个组分。运行条件：流动相为含10%乙酸乙酯的石油醚溶液和纯乙酸乙酯，检测波长为254 nm，色谱柱使用正向硅胶柱，柱温为室温。将分离纯化得到的石菖蒲精油各组份均稀释至1.000 00%浓度进行杀螨活性测定，舍弃杀螨活性低于100%的组分后，将保留组分稀释至上轮50%浓度后再进行多轮杀螨活性测定。将有杀螨效果的组分采用高效液相色谱、液相色谱⁃质谱联用仪和超导脉冲傅里叶变换核磁共振波谱仪进行质谱和核磁分析，通过计算机标准谱图库检索并与文献资料比较，确定活性单体化合物结构。结果 1.000 00%、0.500 00%、0.250 00%和0.125 00%浓度石菖蒲精油可将实验粉尘螨全部杀灭，校正死亡率均为100%。石菖蒲提取物有效浓度0.062 5%时对粉尘螨的24 h致死率可达94.33%。采用硅胶柱层析法对其进行分离纯化，分离出A ~ F等6个组分，对分离出的不同成分进行杀螨活性对比，发现D、E组分杀螨效果良好，在0.500 00%浓度下均达到100%的杀螨率。质谱和核磁分析结果显示，D组分为异丁香酚甲醚、E组分为β⁃细辛脑。结论 石菖蒲精油提取物具有杀螨活性，其中异丁香酚甲醚对粉尘螨有显著杀灭活性。

关键词: 粉尘螨, 石菖蒲, 杀螨活性, 异丁香酚甲醚