为了快速准确检测食品中的日落黄，本研究通过羰基二咪唑法，将日落黄与载体蛋白偶联，并使用紫外光谱进行鉴定，结果显示日落黄检测抗原和免疫抗原成功合成；通过免疫小鼠制备了效价高达1:64000以上的抗日落黄单克隆抗体，研制了日落黄ELISA检测试剂盒。试剂盒性能检测分析结果显示，试剂盒标准曲线的线性检测范围为62.50~1000 ng/mL，检测限为1.50 ng/mL，检测灵敏度IC50值为6.50 ng/mL；试剂盒中，用于检测的单克隆抗体对日落黄的交叉反应率为100%，对诱惑红有弱交叉反应，反应率为2%，与其他2种色素添加剂未见交叉反应；检测市售果汁、水果罐头和果冻类样品的批内、批间回收率分别为89.18%~101.13%、87.69%~97.90%，且批内、批间试验的相对标准偏差均小于10%，说明试剂盒具有良好的灵敏度和准确性，且操作方便快捷。本研究应用免疫学方法成功地制备了日落黄酶联免疫试剂盒，并检测了水果类制品中添加的日落黄，为大批量测定食品中日落黄含量提供了高效准确的实验方法。

In order to detect quickly and accurately sunset yellow in food, the sunset yellow was coupled with a carrier protein by the carbonyl diimidazole method and ultraviolet spectroscopy was used for identification of sunset yellow in this study. The results showed that the antigen and immune antigen for sunset yellow detection were successfully synthesized. A monoclonal antibody with a titer of 1:64000 was prepared by immunizing mice and an ELISA kit was developed for detecting sunset yellow. The results of the kit’s detection and analysis performance showed that the linear detection range of the kit’s standard curve was from 62.50~1000 ng/mL with the detection limit being 1.50 ng/mL and the detection sensitivity IC50 value as 6.50 ng/mL; the cross-reaction rate of the monoclonal antibody towards sunset yellow in this kit was 100%. A weak cross-reaction towards allure red (reaction rate: 2%) was found, with no cross-reactions towards other two color additives were detected. The intra-assay recoveries and inter-assay recoveries of the commercially available samples such as fruit juices, canned fruits and jellies were in the ranges of 89.18%~101.13% and 87.69%~97.90%, respectively, with the relative standard deviations of the intra-assay and inter-assay tests less than 10%. These results demonstrated that the ELISA kit was sensitive, accurate, convenient and fast. In this study, an immunological method was used to prepare successfully an ELISA kit for sunset yellow detection, and the sunset yellow added in fruit products was detected, which provides an efficient and accurate experimental method for large-volume detection of the sunset yellow content in food in large quantities.