用于检测环境样品中五氯硝基苯残留的酶联免疫吸附测定法的开发。

Development of an enzyme-linked immunosorbent assay for the detection of pentachloronitrobenzene residues in environmental samples.

作者信息

Xu Ting, Shao Xiao L, Li Qing X, Keum Young S, Jing Hong Y, Sheng Wei, Li Ji

机构信息

College of Resource and Environment Sciences, China Agricultural University, Beijing 100094, China.

出版信息

J Agric Food Chem. 2007 May 16;55(10):3764-70. doi: 10.1021/jf063287d. Epub 2007 Apr 25.

DOI:10.1021/jf063287d

PMID:17455949

Abstract

A competitive enzyme-linked immunosorbent assay (ELISA) for pentachloronitrobenzene (PCNB), a fungicide and chemical intermediate, was developed using a polyclonal antiserum produced against a hapten-protein conjugate of pentachlorophenoxypropionic acid-bovine serum albumin (BSA). An indirect competitive ELISA of PCNB showed an IC50 of 37 ng/mL and a limit of detection (LOD) of 7 ng/mL. The ELISA can tolerate up to 10% (v/v) methanol, 5% (v/v) acetonitrile, or 5% (v/v) acetone without significant fluctuation of Amax and IC50. The assay sensitivity showed little change in a range of pH from 6 to 8 and concentrations of 0.05-0.2 M NaCl in the assay buffer. Very low cross-reactivities were observed for some structurally related compounds except for hexachlorobenzene (12%). The average recoveries of PCNB from fortified well water, river water, and soil samples were in ranges of 88-94, 80-91, and 70-81%, respectively. The correlations between the gas chromatographic and ELISA results were excellent (r 2 >or= 0.97, slopes from 0.86 to 1.10) for those fortified samples. The ELISA is a good alternative tool for monitoring PCNB residues in environmental samples.

摘要

利用针对五氯苯氧基丙酸 - 牛血清白蛋白（BSA）半抗原 - 蛋白质偶联物产生的多克隆抗血清，开发了一种用于检测杀菌剂和化学中间体五氯硝基苯（PCNB）的竞争性酶联免疫吸附测定（ELISA）方法。PCNB的间接竞争性ELISA显示IC50为37 ng/mL，检测限（LOD）为7 ng/mL。该ELISA在含有高达10%（v/v）甲醇、5%（v/v）乙腈或5%（v/v）丙酮的情况下仍能耐受，且最大吸光度（Amax）和IC50无显著波动。在pH值为6至8以及测定缓冲液中NaCl浓度为0.05 - 0.2 M的范围内，该测定方法的灵敏度变化很小。除六氯苯（12%）外，对一些结构相关化合物的交叉反应率非常低。从加标井水、河水和土壤样品中回收PCNB的平均回收率分别在88 - 94%、80 - 91%和70 - 81%的范围内。对于那些加标样品，气相色谱法和ELISA结果之间的相关性非常好（r²≥0.97，斜率为0.86至1.10）。该ELISA是监测环境样品中PCNB残留的良好替代工具。