孙宁浩, 周引怡, 王翠翠, 刘曙照. 直接竞争酶联免疫吸附分析法测定小白菜中溴氰菊酯的残留量[J]. 农药学学报, 2016, 18(2): 219-226. DOI: 10.16801/j.issn.1008-7303.2016.0029
    引用本文: 孙宁浩, 周引怡, 王翠翠, 刘曙照. 直接竞争酶联免疫吸附分析法测定小白菜中溴氰菊酯的残留量[J]. 农药学学报, 2016, 18(2): 219-226. DOI: 10.16801/j.issn.1008-7303.2016.0029
    SUN Ninghao, ZHOU Yinyi, WANG Cuicui, LIU Shuzhao. Determination of deltamethrin residue in Chinese cabbage by direct competitive enzyme-linked immunosorbent assay[J]. Chinese Journal of Pesticide Science, 2016, 18(2): 219-226. DOI: 10.16801/j.issn.1008-7303.2016.0029
    Citation: SUN Ninghao, ZHOU Yinyi, WANG Cuicui, LIU Shuzhao. Determination of deltamethrin residue in Chinese cabbage by direct competitive enzyme-linked immunosorbent assay[J]. Chinese Journal of Pesticide Science, 2016, 18(2): 219-226. DOI: 10.16801/j.issn.1008-7303.2016.0029

    直接竞争酶联免疫吸附分析法测定小白菜中溴氰菊酯的残留量

    Determination of deltamethrin residue in Chinese cabbage by direct competitive enzyme-linked immunosorbent assay

    • 摘要: 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pH 7.2 PB、反应介质为含0.125 mol/L NaCl pH 6.8 PB的优化条件下,DC-ELISA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。

       

      Abstract: The deltamethrin polyclonal antibody and the horseradish peroxidase labeled hapten was employed to establish the direct competitive enzyme-linked immunosorbent assay(DC-ELISA) for the determination of deltamethrin residue in Chinese cabbage. After ultrasonic extraction, concentration and dissolution with 10% methanol, the samples were determined by DC-ELISA and verified by HPLC. The results showed that the optimized concentration for antibody immobilization was 2.0 mg/L, and for HRP-hapten was 2×105 times diluted. The optimized coating medium was pH 7.2 PB and the optimized reaction medium was pH 6.8 PB containing 0.125 mol/L NaCl. Under the optimized conditions, the linear concentration range for the deltamethrin determination method using DC-ELISA was from 0.10 mg/L to 10 mg/L. The half maximal inhibitory concentration(IC50) of deltamethrin to antibody HRP-hapten reaction was 1.68 mg/L with relative standard deviation(RSD) of 2.11%(n=5). And the IC10 was 0.16 mg/L. The average recoveries of deltamethrin from Chinese cabbage determined by DC-ELISA were 97%, 106% and 95% with RSDs of 8.5%, 8.1% and 8.1%(n=5) at the spiked levels of 5 mg/kg, 1 mg/kg and 0.2 mg/kg respectively. The recovery of deltamethrin from Chinese cabbage determined by HPLC was 107% at the spiked level of 0.2 mg/kg. The DC-ELISA could be used for the rapid determination of deltamethrin residue in Chinese cabbage.

       

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