Abad A, Moreno M J, Pelegrí R, Martínez M I, Sáez A, Gamón M, Montoya A
Laboratorio Integrado de Bioingeniería, Universidad Politécnica de Valencia, Spain.
J Chromatogr A. 1999 Feb 12;833(1):3-12. doi: 10.1016/s0021-9673(98)00856-5.
Carbaryl, carbofuran and methiocarb are three of the most important N-methylcarbamate pesticides. In the present work, the application of laboratory-developed monoclonal antibody-based enzyme-linked immunosorbent assays (ELISAs) to the determination of these compounds in fruits and vegetables is described. Cucumbers and strawberries were spiked with the three carbamates at 10, 50 and 200 ppb. After extraction and clean-up, samples were analyzed by immunoassay and by HPLC with post-column derivatization and fluorescence detection (US Environmental Protection Agency Method 531.1). Results obtained by ELISA correlated well with those obtained by HPLC, both in terms of accuracy and precision. Recoveries were in the 60-90% range by ELISA and in the 50-90% range by HPLC, depending on the particular combination of commodity, pesticide, and fortification level under consideration. ELISAs were also applied to the analysis of non-purified sample extracts with excellent results: recoveries close to 100% were obtained, while maintaining similar precision values. This approach avoids the use of solid-phase extraction columns, saves time, and considerably increases the sample throughput. Results clearly indicate that the developed immunoassays may be suitable for the quantitative and reliable determination of carbaryl, carbofuran and methiocarb in fruits and vegetables even without including clean-up steps. These considerations make these ELISAs very useful analytical tools for monitoring and regulatory programs, without the need of complex and expensive instrumentation.
西维因、克百威和灭虫威是三种最重要的氨基甲酸酯类农药。在本研究中，描述了实验室开发的基于单克隆抗体的酶联免疫吸附测定法（ELISA）在水果和蔬菜中这些化合物测定中的应用。黄瓜和草莓分别添加了浓度为10、50和200 ppb的这三种氨基甲酸酯。经过提取和净化后，样品通过免疫测定以及采用柱后衍生化和荧光检测的高效液相色谱法（美国环境保护局方法531.1）进行分析。ELISA获得的结果在准确性和精密度方面与HPLC获得的结果相关性良好。ELISA的回收率在60 - 90%范围内，HPLC的回收率在50 - 90%范围内，具体取决于所考虑的商品、农药和强化水平的特定组合。ELISA还应用于非纯化样品提取物的分析，结果极佳：回收率接近100%，同时保持相似的精密度值。这种方法避免了使用固相萃取柱，节省了时间，并显著提高了样品通量。结果清楚地表明，即使不包括净化步骤，所开发的免疫测定法也可能适用于水果和蔬菜中西维因、克百威和灭虫威的定量和可靠测定。这些因素使得这些ELISA成为监测和监管计划非常有用的分析工具，无需复杂且昂贵的仪器。
