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。
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 p H 7.2 PB and the optimized reaction medium was p H 6.8 PB containing 0.125 mol/L Na Cl. 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.
出处
《农药学学报》
CAS
CSCD
北大核心
2016年第2期219-226,共8页
Chinese Journal of Pesticide Science
基金
国家自然科学基金项目(21247002)
国家高技术研究发展计划("863"计划)项目(2009AA03Z331)
关键词
直接竞争酶联免疫吸附分析
高效液相色谱
小白菜
溴氰菊酯
残留
direct competitive enzyme-linked immunosorbent assay（DC-ELISA）
high performance liquid chromatography
Chinese cabbage
deltamethrin
residue
