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Enzyme-linked immuno sorbent assay (ELISA) method for detecting melamine content in food

A technology of melamine content, which is applied in the research field of food safety supervision or food analysis, can solve the problem of insufficient detection sensitivity, and achieve the effect of simple sample processing, low test cost and large test volume

Inactive Publication Date: 2013-01-16
SUZHOU UNIV
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  • Application Information

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Problems solved by technology

However, the detection sensitivity of the reported methods is not high enough to meet the needs of melamine detection in real life.

Method used

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  • Enzyme-linked immuno sorbent assay (ELISA) method for detecting melamine content in food
  • Enzyme-linked immuno sorbent assay (ELISA) method for detecting melamine content in food
  • Enzyme-linked immuno sorbent assay (ELISA) method for detecting melamine content in food

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Embodiment

[0075] 1. Preparation of melamine modification

[0076] Melamine is a small molecular compound without immunogenicity, and cannot directly immunize animals to produce antibodies. The molecular structure of melamine must be chemically modified reasonably and effectively so that it has active groups, so that it can be cross-linked with the carrier protein to obtain immunity Original and coated antigens. The present invention has prepared three kinds of melamine modifications, i.e. Hapten A, Hapten B and Hapten C:

[0077] 1) Synthesis of melamine hapten A (Hapten A)

[0078] Weigh 3~5mmol of 2-chloro-4,6-diamino-1,3,5-triazine (CAAT) into the reaction bottle, dissolve with anhydrous methanol, then add 5~10mmol of p-aminobenzoic acid to dissolve in 10~ 15mmol potassium hydroxide in anhydrous methanol. The reaction was refluxed for 5-12 hours until the CAAT reaction was complete. The reaction mixture was filtered and washed three times each with absolute ethanol and distilled ...

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Abstract

The invention discloses an enzyme-linked immuno sorbent assay (ELISA) method for detecting the melamine content in a food. The method is characterized in that three different melamine hapten derivatives are synthesized and are linked with a carrier protein to synthesize three immunogens and three envelope antigens; and a monoclonal antibody is prepared by using a hybridoma monoclonal antibody preparation technology. Different amounts of melamine are added into four samples, namely pure milk, milk powder, chicken feeds and pig feeds, and are directly measured by ELISA; the labeling recycling rate is 72.6 to 133.3 percent; the relative standard deflection is 0.8 to 18.9 percent; when the samples are labeled, measured results are detected and compared by using high performance liquid chromatography (HPLC) and ELISA; a regression curve is Y which is equal to 1.5589x-1.4004; a linear relevant coefficient is 0.9902; and a result shows that the two methods are relatively high in relevance.

Description

technical field [0001] The invention relates to an enzyme-linked immunosorbent assay method (ELISA) for detecting melamine content in food, which belongs to the research field of food safety supervision or food analysis. Background technique [0002] Melamine, the chemical name is 2,4,6-triamino-1,3,5-triazine, also known as melamine, cyanuramide, tripolyamide, etc., is an important nitrogen heterocyclic organic chemical raw material , is widely used. The main use of melamine is as a raw material for the production of melamine formaldehyde resin (MF), which is widely used in wood, plastic, paper, textile, leather and other industries; melamine can also be used as a flame retardant, water reducing agent, formaldehyde cleaner, etc. Since the melamine molecule contains a large amount of nitrogen, the conventional "Kjeldahl method" cannot rule out the interference of such "pseudoprotein nitrogen" when measuring the protein content in feed or food. This non-food chemical raw ma...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/531
Inventor 邓安平曹碧云杨红宋娟
Owner SUZHOU UNIV
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