Preparation and Application of ELISA Kitfor Detection of Sunset Yellow
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Abstract:
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.
