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丁香酚的分离及其杀多子小瓜虫活性研究
引用本文:姚嘉赟,徐洋,袁雪梅,蔺凌云,尹文林,潘晓艺,郝贵杰,沈锦玉.丁香酚的分离及其杀多子小瓜虫活性研究[J].水产科学,2016(6):669-674.
作者姓名:姚嘉赟  徐洋  袁雪梅  蔺凌云  尹文林  潘晓艺  郝贵杰  沈锦玉
作者单位:浙江省淡水水产研究所,农业部淡水渔业健康养殖重点实验室,浙江省鱼类健康与营养重点实验室,浙江湖州,313001
基金项目:国家自然科学基金青年科学基金资助项目(31302211);浙江省公益技术应用研究项目(2014C32055).
摘    要:利用不同极性大小的溶剂石油醚、乙酸乙酯、氯仿、甲醇回流提取丁香制备粗提物,进行杀灭多子小瓜虫幼虫和包囊的药效活性追踪,确定丁香杀虫的活性部位为石油醚提取部分,进而利用硅胶柱层析和中压制备液相等分离技术结合活性追逐试验追踪分离丁香石油醚提取物(对多子小瓜虫幼虫和包囊的100%致死质量浓度分别为50mg/L和80mg/L),最终分离获得一种杀虫活性物质,经质谱、核磁氢谱、核磁碳谱等多种波谱分析技术确定该活性物质为丁香酚。体外杀虫结果表明,丁香酚对多子小瓜虫具有较强的杀灭作用,其对幼虫15min、1h、2h、3h和4h的半数致死质量浓度分别为6.21(5.85~6.65)mg/L、3.50(3.24~3.84)mg/L、2.90(2.62~3.22)mg/L、2.33(2.10~2.50)mg/L和2.15(1.98~2.32)mg/L,对幼虫4h的100%杀灭的质量浓度为3.5mg/L,对包囊6h的100%杀灭质量浓度为3.5mg/L。结果表明,丁香酚是一种具有较好开发前景的杀多子小瓜虫药物。

关 键 词:丁香酚  多子小瓜虫  活性追踪  杀虫活性

Isolation of Active Compounds from Eugenia caryophy llata against Ichthyophthirius multi filiis in Vitro
Abstract:Ichthyophthirius multif iliis is a holotrichous protozoan that invades the gills and skin surfaces of fish and can cause morbidity and high mortality in most species of freshwater fish worldwide .The pres‐ent study was undertaken to investigate the antiparasitic activity of crude extracts and pure compounds from Eugenia caryophyllata .The dried E .caryophyllata was extracted successively in a separating fun‐nel with petroleum ether ,ethyl acetate ,chloroform and methanol ,in w hich only the petroleum ether ex‐tract showed promising activity (100% effectiveness against I .multif iliis theronts and encysted tomonts at the concentration of 50 .0 mg/L and 80 .0 mg/L respectively ) and therefore ,was fractionated on silica gel column chromatography in a bioactivity‐guided isolation affording a compound showing potent activity . The structure of the compound was elucidated as clove oil by hydrogen and carbon‐13 nuclear magnetic reso‐nance spectrum and electron ionization mass spectrometry .The in vitro tests revealed that clove oil was found to be 100% effective against encysted tomonts at the concentration of 16 .0 mg/L . The results showed that clove oil has potent anti‐parasitic efficacy with the 15 min‐,1 h‐,2 h‐,3 h‐and 4 h‐LC50 (95%Confidence Intervals) of 6 .21 (5 .85—6 .65) mg/L ,3 .50 (3 .24—3 .84) mg/L ,2 .90 (2 .62—3 .22) mg/L , 2 .33 (2 .10—2 .50) mg/L and 2 .15 (1 .98—2 .32) mg/L against theronts of I .multi f iliis ,respectively . The findings provide important information for the potential application of clove oil in the therapy of seri‐ous infection caused by I .multi f iliis .
Keywords:clove oil  Ichthyophthirius multif ilis  active site  bioactivity-guided isolation
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