Synthesis of three haptens for the class-specific immunoassay of O,O-dimethyl organophosphorus pesticides and effect of hapten heterology on immunoassay sensitivity
Synthesis of three haptens for the class-specific immunoassay of O,O-dimethyl organophosphorus pesticides and effect of hapten heterology on immunoassay sensitivity
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DOI：
10.1016/j.aca.2008.03.050
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发表时间：
2008-05-19
影响因子：
6.2
通讯作者：
Fan, Ming Tao
中科院分区：
文献类型：
--
作者：
Liang, Ying;Liu, Xian Jin;Fan, Ming Tao
开发了一种针对 O,O-二甲基有机磷农药（包括马拉硫磷、乐果、酚酯、亚胺硫磷、甲硫磷、杀螟硫磷、甲基对硫磷和倍硫磷）的通用且广泛的类别特异性酶联免疫吸附测定法。合成了具有不同间隔臂的三种半抗原。半抗原与牛血清白蛋白（BSA）缀合作为免疫原，并与卵清蛋白（OVA）缀合作为包被抗原。用免疫原对兔子进行免疫，产生六种多克隆抗血清，并使用竞争性间接酶联免疫吸附测定针对每种包被抗原进行筛选，以选择合适的抗血清。还研究了半抗原异源性对免疫测定灵敏度的影响。对马拉硫磷最具选择性的抗体-抗原组合被进一步优化，并测试了对共溶剂、pH 和离子强度变化的耐受性。在最佳条件下，估计的 IC50 值为马拉硫磷 30.1 μg L-1、乐果 28.9 μg L-1、酚妥特 88.3 μg L-1、亚胺硫磷 159.7 μg L-1、甲硫磷 191.7 μg L-1、杀螟硫磷 324.0 μg L-1、483.9甲基对硫磷为μg L-1，倍硫磷为788.9μg L-1。强化大白菜样品中马拉硫磷、乐果、酚酞、硫磷胺和甲硫磷的回收率为 77.1% 至 104.7%。该测定可用于 O,O-二甲基有机磷农药多残留测定的监测研究。 (C) 2008 年由 Elsevier B.V. 出版
A general and broad class-specific enzyme-linked immunosorbent assay was developed for the O,O-dimethyl organophosphorus pesticides, including malathion, dimethoate, phenthoate, phosmet, methidathion, fenitrothion, methyl parathion and fenthion. Three haptens with different spacer-arms were synthesized. The haptens were conjugated to bovine serum albumin (BSA) for immunogens and to ovalbumin (OVA) for coating antigens. Rabbits were immunized with the immunogens and six polyclonal antisera were produced and screened against each of the coating antigens using competitive indirect enzyme-linked immunosorbent assay for selecting the proper antiserum. The effect of hapten heterology on immunoassay sensitivity was also studied. The antibody-antigen combination with the most selectivity for malathion was further optimized and tested for tolerance to cosolvent, pH and ionic strength changes. The IC50 values, under optimum conditions, were estimated to be 30.1 mu g L-1 for malathion, 28.9 mu g L-1 for dimethoate, 88.3 mu g L-1 for phenthoate, 159.7 mu g L-1 for phosmet, 191.7 mu g L-1 for methidathion, 324.0 mu g L-1 for fenitrothion, 483.9 mu g L-1 for methyl parathion, and 788.9 mu g L-1 for fenthion. Recoveries of malathion, dimethoate, phenthoate, phosmet and methidathion from fortified Chinese cabbage samples ranged between 77.1% and 104.7%. This assay can be used in monitoring studies for the multi-residue determination of O,O-dimethyl organophosphorus pesticides. (C) 2008 Published by Elsevier B.V.