Key Laboratory for Agro-Products Quality and Safety, Inst. of Quality Standards, Testing Technology for Agro-Products, Chinese Academy of Agricultural Sciences, Beijing, 100081, China.
J Food Sci. 2012 May;77(5):T99-T104. doi: 10.1111/j.1750-3841.2012.02659.x. Epub 2012 Apr 10.
A direct competitive chemiluminescent enzyme immunoassay (CLEIA) for triazophos was developed, which was based on the anti-THHe IgG monoclonal antibody and a heterogeneous enzyme tracer (THHu-HRP). Several components of chemiluminescent enhanced solution (CES) were optimized. The results showed that 1 mM of p-iodo-phenol, 0.625 mM of luminol, and 4 mM of H(2)O(2) had the best performance. Based on the study of CES, the influence of several factors (assay buffer, blocking substance, and solvent) on the immunoassay was investigated. The sensitivity for detection, IC(50) value was 0.87 ng/mL at a practical working concentration range between 0.04 ng/mL and 5 ng/mL and the limit of detection for triazophos was 0.063 ng/mL. The average recovery of triazophos added to lettuce, carrot, apple, water, and soil were 78.71%, 67.52%, 118.3%, 117.2%, and 122.0%, respectively. Finally, comparison between the methods of CLEIA and high-performance liquid chromatography-tandem mass spectrum (HPLC-MS/MS) was performed. The results obtained from CLEIA were in agreement with those of HPLC-MS/MS.
建立了一种测定三唑磷的竞争化学发光酶免疫分析方法（CLEIA），该方法基于抗-THHe IgG 单克隆抗体和异质酶示踪剂（THHu-HRP）。优化了化学发光增强液（CES）的几个组成部分。结果表明，1 mM 的对碘苯酚、0.625 mM 的鲁米诺和 4 mM 的 H2O2 具有最佳性能。基于 CES 的研究，考察了几种因素（测定缓冲液、封闭物质和溶剂）对免疫分析的影响。在实际工作浓度范围内（0.04ng/mL 至 5ng/mL），检测的灵敏度为 0.87ng/mL，IC50 值为 0.87ng/mL，检测限为 0.063ng/mL。三唑磷在生菜、胡萝卜、苹果、水和土壤中的平均回收率分别为 78.71%、67.52%、118.3%、117.2%和 122.0%。最后，对 CLEIA 和高效液相色谱-串联质谱（HPLC-MS/MS）方法进行了比较。CLEIA 得到的结果与 HPLC-MS/MS 的结果一致。
