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应用作者所在实验室自制的、对溴氰菊酯具有特异性亲和力的多克隆抗体和酶标半抗原,建立了包被抗体-酶标半抗原直接竞争酶联免疫吸附分析法(DC-ELISA)。小白菜样品经超声提取、浓缩和10%甲醇定容后采用DC-ELISA法测定,并采用高效液相色谱法进行验证。结果表明:在抗体包被浓度为2.0 mg/L、辣根过氧化物酶(HRP)标记溴氰菊酯半抗原稀释2.0×105倍、包被介质为p H 7.2 PB、反应介质为含0.125 mol/L Na Cl p H 6.8 PB的优化条件下,DCELISA法检测溴氰菊酯的线性范围为0.10~10 mg/L,溴氰菊酯对抗体-酶标半抗原反应的抑制中浓度(IC50)为1.68 mg/L,相对标准偏差(RSD,n=5)为2.11%,IC10值为0.16 mg/L。在5、1和0.2 mg/kg 3个添加水平下,DC-ELISA法测定小白菜中溴氰菊酯的平均回收率分别为97%、106%和95%,RSD(n=5)分别为8.5%、8.1%和8.1%。高效液相色谱法同步测定小白菜中添加0.2 mg/kg溴氰菊酯的回收率为107%。本研究建立的DC-ELISA法可用于小白菜中溴氰菊酯含量的快速测定。
A direct competitive enzyme-linked immunosorbent assay (DC-ELISA) was developed for the detection of polyclonal antibody and enzyme-labeled hapten, which was made by our laboratory in our laboratory and had specific affinity for deltamethrin. . The cabbage samples were extracted by ultrasound, concentrated and quantified by 10% methanol, then determined by DC-ELISA and verified by high performance liquid chromatography. The results showed that the HRP labeled deltamethrin hapten diluted 2.0 × 105 times and the coating medium was p H 7.2 PB at the concentration of 2.0 mg / L, the reaction medium was 0.125 The linear range of deltamethrin detected by DCELISA was 0.10-10 mg / L under the optimal conditions of mol / L Na Cl p H 6.8 PB. The inhibitory concentration of deltamethrin on the antibody-hapten response (IC50 ) Was 1.68 mg / L with relative standard deviation (RSD, n = 5) of 2.11% and IC10 of 0.16 mg / L. The average recoveries of deltamethrin in Chinese cabbage by DC-ELISA were 97%, 106% and 95% at 3, 5, 1 and 0.2 mg / kg levels, respectively. The RSDs 8.5%, 8.1% and 8.1%. The simultaneous determination of cabbage by 0.2 mg / kg deltamethrin by HPLC was 107%. The DC-ELISA method established in this study can be used for the rapid determination of deltamethrin content in Chinese cabbage.