The Glyphosate AOAC ELISA Plate Test is a direct competitive ELISA based kit on the recognition of Glyphosate by polyclonal antibodies. The sample to be tested is derivatized and then added to microtiter wells coated with goat anti-rabbit antibodies. A rabbit anti-Glyphosate antibody solution is added to the wells with the derivatized samples and allowed to incubate for 30 minutes. The Glyphosate enzyme conjugate is then added and a competitive reaction occurs between the Glyphosate, which may be present in the sample, and the enzyme labeled Glyphosate for the binding sites of the rabbit anti-Glyphosate antibodies bound by the goat anti-rabbit antibodies immobilized on the microtiter plate. The reaction is allowed to continue for 60 minutes. After a washing step and addition of the substrate solution, a color signal is generated. The intensity of the blue color is inversely proportional to the concentration of Glyphosate present in the sample. The color reaction is stopped after a specified time and the color is evaluated using an ELISA reader. The concentrations of the samples are determined by interpolation using the standard curve constructed with each run.
The AOAC ELISA method showed good correlation to the LC-MS/MS reference method performed by the independent laboratories involved in approving the test method under AOAC Performance Tested™ License Number 072104.
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