摘要
将异丙甲草胺与3-巯基丙酸反应合成出带羧基的异丙甲草胺半抗原MMPA;用活泼酯法将半抗原与载体牛血清白蛋白(BSA)和卵清白蛋白(OVA)分别偶联,制备免疫抗原MMPA-BSA和包被抗原MMPA-OVA,紫外光谱鉴定后用MMPA-BSA免疫新西兰大白兔,成功制备出抗异丙甲草胺多克隆抗体,并建立异丙甲草胺的间接竞争酶联免疫吸附分析(icELISA)法。该方法半抑制浓度(IC50)为34.3ng/mL,检测限(IC10)为6.3ng/mL,检测范围(IC20～IC80)为12.3～99.2ng/mL。该抗体对其他结构类似物无明显交叉反应。将该方法应用于环境水样中异丙甲草胺检测,回收率在89.5%～107.9%之间,相对标准偏差9.2%～14.5%。所建立的异丙甲草胺免疫分析方法特异性强、准确度高、灵敏度高,可用于环境水样中异丙甲草胺的快速检测。
Using one step reaction,metolachlor was modified with 3-marcapropanoic acid（3-MPA） to obtain the hapten 3-（2-（（2-ethyl-6-methylphenyl）（1-methoxypropan-2-yl）amino）-2-oxoethylthio）propanoic acid（MMPA）,which was then coupled to bovine serum albumin（BSA） as immunogen（MMPA-BSA） and ovalbumin（OVA） as coating antigen（MMPAOVA） using active ester method,respectively.New Zealand rabbits were immunized with MMPA-BSA.Based on the obtained polyclonal antibody,an indirect competitive ELSIA（icELISA） was developed successfully for the detection of metolachlor and demonstrated excellent performance.The 50% inhibition concentration（IC50） was 34.3 ng/mL and the detectable range（IC20–IC80） was 12.3–99.2 ng/mL,while the limit of detection（LOD,IC10） was 6.3 ng/mL.The antibody showed low cross-reactivity（10.9%） towards S-（-）-metolachlor.However,no significant cross-reactivity towards other structurally related compounds was found.The recoveries of spiked samples were in the range of 89.5%–107.9% with RSD ranging from 9.2% to 14.5%.Our results indicated that the proposed icELISA method is highly specific,sensitive,accurate and suitable for the rapid detection of herbicide metolachlor in water samples.
出处
《食品科学》
EI
CAS
CSCD
北大核心
2013年第10期126-129,共4页
Food Science
基金
公益性行业(农业)科研专项(201003008-08)
国家自然科学基金面上项目(30700663
21105030)
广东省科技计划项目(2010A032000001-4)
福建省科技计划重点项目(2012Y0003)
关键词
异丙甲草胺
酶联免疫吸附分析
抗体
metolachlor
enzyme-linked immuno sorbent assay（ELISA）
antibody
