Ein kompetitiver Immunoassay zur Bestimmung des Herbizids Fluazifop in Trink‐ und Grundwasser

F. Poppe, K. Rubach
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引用次数: 0

Abstract

A competitive solid-phase enzyme immunoassay using rabbit polyclonal antibodies was developed for the detection of the herbicide fluazifop [(RS)-2-[4-(5-trifluoro-methyl-2-pyridyloxy)phenoxy]propionic acid] in drinking water and groundwater. Present regulatory limits for drinking water in Germany were taken as the critical level. The carrier protein was bovine serum albumin; horseradish peroxidase was used as marker enzyme with 3,3',5,5'-tetramethylbenzidine as substrate. A high concentration of high-affinity antibodies in the serum, optimization of test conditions (antibody and enzyme tracer concentration, incubation time etc.), and very low cross reactivities to substances of similar structures led to a highly sensitive and specific ELISA with a detection limit below 0.1 μg/L for fluazifop as free acid. On testing the suitability of the assay's use as a screening test with one hundred drinking-water samples, the three samples which had been spiked in the laboratory were recognized as positive with respect to their fluazifop content. Confirmation by gas chromatography-mass spectrometry showed the test results of two other samples to be false positive. False negative results did not appear. The concentration was in the detection limit region of 0.1 μg/L.
一个竞争的Immunoassay确定Herbizids Fluazifop在喝‐和地下水
建立了兔多克隆抗体竞争固相酶免疫分析法检测饮用水和地下水中除草剂氟唑吡肟[(RS)-2-[4-(5-三氟甲基-2-吡啶氧基)苯氧基]丙酸。目前德国饮用水的监管限值被视为临界水平。载体蛋白为牛血清白蛋白;以辣根过氧化物酶为标记酶,以3,3',5,5'-四甲基联苯胺为底物。血清中高亲和抗体的浓度较高,实验条件(抗体和酶示踪剂浓度、孵育时间等)的优化,以及与相似结构物质的交叉反应性极低,使得ELISA对游离酸氟唑吡酯的检测限在0.1 μg/L以下,具有较高的灵敏度和特异性。在对100个饮用水样品进行筛选试验的适用性测试时,在实验室加标的三个样品的氟唑吡酯含量被认为是阳性的。经气相色谱-质谱联用证实,另外两份样品的检测结果为假阳性。没有出现假阴性结果。浓度在0.1 μg/L的检出限范围内。
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