ELISA adsorption analysis method for detecting monoclonal antibodies of sudan red NO.1 in foodstuff

Abstract

The present invention discloses a monoclonal antibody enzyme-linked immunosorbent analysis method (ELISA) which is used for determining the content of the Sudan dye 1. The method is characterized in that the modified compound which is used for synthesizing the Sudan dye 1 is coupled with protein to prepare immunogen and coating antigen. The anti-Sudan dye 1 monoclonal antibody is prepared with the hybridoma technology which comprises immunization, fusion, screening and other steps. The antibody is used for immunoassay of the Sudan dye, and the concentration range of the standard curve is 0.1 to 100 ng / mL; the concentration of the IC50 is 1.1 to 2.0ng / mL; the rate of the cross reaction between the antibody and the Sudan dye 2 to 4 is 0.95 to 33.9 percent; the antibody can hardly perform the cross reaction with the other 6 kinds of addable pigments of the food; therefore, the established ELISA not only has high sensitivity but also has high specificity. The 6 kinds of food sold in the market are extracted by methanol after being labeled; the extracted solution can be determined by the ELISA after being appropriately diluted; the recovery of standard addition of the Sudan dye 1 is 88.2 to 110.5 percent; and the relative standard deviation is 1.5 to 17.4 percent. The HPLC and the ELISA are used for analyzing the labeled sample, and the two methods have good relativity (r is equal to 0.9840, and n is equal to 9).

Description

1. Technical field [0001] The invention relates to a monoclonal antibody enzyme-linked immunosorbent assay method (ELISA) for determining the content of Sudan Red No. 1 in food, belonging to the research field of food safety supervision or food analysis. 2. Background technology [0002] Sudan red belongs to a class of artificially synthesized azo compounds. It is a red oil-soluble dye, which is mainly used in petroleum, engine oil and some other industrial solvents. Sudan red dyes include four substances, Sudan Red No. 1-4, all of which contain azo and naphthalene compound structures, which have obvious toxic effects on the liver and kidney organs of the human body, and are carcinogenic. Because Sudan red is cheap and not easy to fade, it is used by some unscrupulous merchants for food dyeing, which poses a great threat to human health. Our country and many other countries prohibit its use in food production. At the beginning of 2007, the AQSIQ announced the product quali...

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Problems solved by technology

The inventor has applied for a Chinese patent, 200710049361.5, on the enzyme-linked immunosorbent assay method for determining the content of Sudan Red No. 1 in food based on polyclonal antibodies, but the polyclonal antibodies have some shortcomings, such as inconsistent with the immunoaffinity of the test substance The total amount of antibodies is limited, and it is impossible to continuously provide antibodies with the same performance, etc.

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