赤子爱胜蚓(Eisenia fetida)EROD活性测定的HPLC法的建立  

Establishment of EROD Activity Determination in Earthworm(Eisenia fetida)by HPLC

作　　者：李玲玲 宋玉芳[2] 史奕[2] LI Lingling;SONG Yufang;SHI Yi(School of Life Science,Hefei Normal University,Hefei 230009,China;Key Laboratory of Pollution Ecology and Environmental Engineering,Institute of Applied Ecology,Chinese Academy of Sciences,Shenyang 110016,China)

机构地区：[1]合肥师范学院生命科学学院,安徽合肥230009 [2]中国科学院沈阳应用生态研究所污染生态与环境工程重点实验室,辽宁沈阳110016

出　　处：《沈阳大学学报（自然科学版）》2021年第1期33-40,共8页Journal of Shenyang University：Natural Science

基　　金：国家自然科学基金面上资助项目(21377139);安徽省自然科学基金资助项目(2008085QC103);合肥师范学院高层次人才科研启动基金资助项目(2020rcjj49).

摘　　要：以赤子爱胜蚓(Eisenia fetida)为供试动物,通过微粒体提取、孵育和HPLC测定等系列条件的摸索,建立了测定其微粒体7-乙氧基-3-异吩(噁)唑酮-脱乙基酶(EROD)活性的高效液相色谱(HPLC)法.结果表明,以赤子爱胜蚓整体为材料,采用差速离心法获取微粒体进行体外孵育,反应液采用C18反相柱分离,在激发波长530 nm,发射波长582 nm条件下能够成功测定其EROD活性.组间和组内回收率在93%~118%之间,变异系数均小于15%,方法稳定.在氰戊菊酯暴露验证试验中发现氰戊菊酯暴露7 d后,赤子爱胜蚓EROD活性被显著诱导,且呈现显著的量效关系,表明蚯蚓EROD活性可以作为生物标志物指示土壤拟除虫菊酯暴露的风险状况.HPLC method was established for the determination of 7-ethoxyresorufin-o-deethylase(EROD)activity of Eisenia fetida by a series of experiments which include microsomes extraction,incubation and High Performance Liquid Chromatography(HPLC)measurement.The results showed that the microsomes of whole worms were obtained by ultracentrifugation for in vitro incubation,and the reaction solution was separated by C18 reversed phase column.The EROD activity could be determined at the excitation wavelength of 530 nm and emission wavelength of 582 nm.The recovery rates of intra-day and inter-day were between 93%and 118%,and the coefficient of variations were all<15%.After exposure of fenvelerate for 7 days,EROD activity in earthworms was significantly induced in a dose-response relationship,indicating that EROD activity in earthworms could be served as one potential sensitive biomarker to diagnose the toxicity of pyrethroids.

关 键 词：CYP1A1 蚯蚓 高效液相色谱 氰戊菊酯 生物标志物 

分 类 号：X171.5[环境科学与工程—环境科学]