Recent cases of adulteration with melamine have led to the need for rapid and reliable screening methods. The discovery of melamine in pet food, animal feed, milk and protein sources including wheat gluten, rice protein concentrate, and corn gluten created an urgent need for rapid methods for detecting melamine in food. Melamine (2,4,6-triamino-1,3,5-triazine) is used in the production of plastics, in finishers for paper, in fertilizer, as a flame retardant, and in the manufacture of wrinkle-free textiles.
The recently released Melamine Plate kit (Abraxis, Warminster, Pa.) displayed a limit of detection of 9 ng/ml for melamine in phosphate-buffered saline (PBS) and approximately 1 µg/ml for melamine added to dog food. Using the Melamine ELISA, both extraction protocols yielded identical results with a dog food sample adulterated with melamine. The recovery of melamine spiked into gravy from dog food using UD was app. 75%. In conclusion, the ELISA for melamine proved to be a useful alternative to more cumbersome methods.Assay procedure
The Melamine plate kit is a competitive enzyme-labeled immunoassay. Melamine is extracted from a sample by blending or shaking with extraction solution. The Melamine HRP conjugate, sample extract and calibrators are pipetted into the test wells coated with Melamine antibody to initiate the reaction. During the 30 minute incubation period, Melamine from the sample and Melamine HRP conjugate compete for binding to Melamine antibody. Following this 30 minute incubation, the contents of the well are removed and the wells are washed to remove any unbound Melamine, and Melamine HRP conjugate. After washing with the diluted wash solution, a clear substrate is then added to the wells and any bound enzyme conjugate causes the conversion to a blue color. Following a 20 minute incubation, the reaction is stopped and the amount of color in each well is read. The color of the unknown samples is compared to the color of the calibrators and the Melamine concentration of the samples is derived.
The Melamine assay has an estimated minimum detectable concentration in buffer of 10 ppb. Read more
The Melamine ELISA kit has been evaluated along with other commercial enzyme-linked immunosorbent assay (ELISA) test kits for the detection of triazines. Please contact firstname.lastname@example.org
for the article “Detection of Melamine Using Commercial Enzyme-Linked Immunosorbent Assay Technology
” by Garber, E. A. E. Journal of Food Protection, Vol.71, No. 3, 2008, Pages 590-594
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