食品科学 ›› 2013, Vol. 34 ›› Issue (10): 126-129.doi: 10.7506/spkx1002-6630-201310027

• 分析检测 • 上一篇    下一篇

酶联免疫吸附法检测除草剂异丙甲草胺

张炽坚,王 强,唐秋实,刘英菊,孙远明,唐允健,雷红涛   

  1. 1.农业部农产品贮藏保鲜质量安全风险评估实验室,广东省食品质量安全重点实验室,广东 广州 510642;
    2.华南农业大学理学院,生物材料研究所,广东 广州 510642
  • 收稿日期:2012-04-06 修回日期:2013-04-27 出版日期:2013-05-25 发布日期:2013-05-07
  • 通讯作者: 雷红涛 E-mail:immunoassay@126.com
  • 基金资助:

    高等学校博士点基金优先发展领域项目;广州市科技计划应用基础专项重点项目;国家重点基础研究发展计划(973)项目;中国科学院“百人计划”项目

Determination of of the Herbicide Metolachlor by Enzyme-Linked Immunosorbent Assay

ZHANG Chi-jian,WANG Qiang,TANG Qiu-shi,LIU Ying-ju,SUN Yuan-ming,TANG Yun-jian,LEI Hong-tao   

  1. 1. Key Laboratory of Risk Assessment of Agricultural Products in Storage, Ministry of Agriculture, Guangdong Provincial Key Laboratory of Food Quality and Safety, Guangzhou 510642, China;2. Institute of Biomaterials, College of Sciences, South China Agricultural University, Guangzhou 510642, China
  • Received:2012-04-06 Revised:2013-04-27 Online:2013-05-25 Published:2013-05-07
  • Contact: Hongtao Lei E-mail:immunoassay@126.com

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。

关键词: 异丙甲草胺, 酶联免疫吸附分析, 抗体

Abstract:

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.

Key words: metolachlor, enzyme-linked immuno sorbent assay (ELISA), antibody

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