农产品中三唑磷农药残留化学发光酶免疫分析方法研究

Development of a Chemiluminescent Enzyme Immunoassay for Detection of Triazophos in Agricultural Products

运用棋盘法确定直接竞争CLEIA法抗体和酶标半抗原的最适工作浓度，以4-（咪唑-1-基）苯酚（4-IMP）作为增强剂，运用L2，（5^6）正交设计实验优化增敏液配方，建立了农产品中三唑磷农药残留的化学发光酶免疫分析方法。优化的最适工作条件：抗体包被浓度为2．0μg／mL，酶标半抗原浓度为0．006μg／mL；增敏液最佳配方为：6．4％DMF，0．01mmol／LpH9．0的Tris—HCl缓冲液，0．6mmol／L鲁米诺，2mmol／LH202，1mmol／L4-IMP。优化后方法灵敏度为0．489ng／mL，线性范围为0．16—20．00ng／mL，相关系数为0．9964。该方法能实现对苹果、节瓜、大米、柑橘、荔枝及甘蓝等样品中三唑磷残留的快速、灵敏检测，加标回收率为82．8％-118．4％，与GC—MS法的相关性（r2）为0．957。结果表明，该方法可用于农产品中三唑磷农药的快速筛查。

A chemiluminescent enzyme immunoassay（CLEIA） method for the detection of triazophos pesticide residue in agricultural products was developed. Checkerboard method was used to get the working concentrations of the immunoassay. Based on the experiment, the concentration of antibody was selected as 2. 0 μg/mL, and the concentration of enzyme labeled hapten was chosen as 0. 006 μg/mL. Chemiluminescent enhanced solution （CES） was optimized by L25 （56 ） orthogonal experiment and the optimized conditions for the CES were as follows ： percentage of organic solvent （DMF） in the CES： 6. 4% , buffer： 0. 01 mmoL/L Tris - HCl （pH 9.0）, concentration of luminol： 0. 6 mmol/L, concentration of H2O2： 2 mmol/L, and concentration of4-IMP： 1 mmol/L. The sensitivity for detection was 0. 489 ng/mL at a practical working concentration range of 0. 16 -20. 00 ng/mL. The recoveries of triazophos in apple, zucchini, flee, orange, litchi and cabbage samples were in the range of 82. 8% - 118.4% . The confirmation experiment showed that the analytical result of CLE- IA method was in agreement with that of the GC - MS method. The developed CLEIA method could be employed in the screening of triazophos residue in apple, zucchini, rice and other samples.