Enzyme Immunoassay for the rapid quantitative determination of Zearalenone in cereals and beer/gyle.
Background: Zearalenone in addition to fumonisin, deoxynivalenol and other trichothecenes belongs to the fusarium toxins. These toxins are already produced on the field in consequence of a contact of the cereals by fusarium species. These moulds infect grain and other types of food like peanuts and beans already during their growth. When a considerable amount of zearalenone contaminated feed is taken up by cows, it can also be detected in their milk. Even in beer it could be found. Zearalenone shows a strong estrogen-like activity. Thus zearalenone can cause an enlargement of the uterus, diminution of the ovarian glands and even infertility. Zearalenone is one of the main contaminants of farm products, which can be taken up by humans and animals.
In the European Union the limits are 20 – 400 ppb for food products. Thus a monitoring of food and feed with respect to the concentration of zearalenone is obligatory.
The Zearalenone ELISA represents a highly sensitive detection system and is particulary capable of the rapid quantification of zearalenone contaminations in cereals and beer.
Description: The Zearalenone quantitative test is based on the principle of the enzyme-linked immunosorbent assay. An antibody binding protein is coated on the surface of a microtiter plate. Zearalenone containing samples or standards, a zearalenone-peroxidase conjugate and an antibody directed against zearalenone are given into the wells of the microtiter plate. The conjugate competes with the zearalenone of samples/standards for the limited number of antibody sites. Simultaneously the anti-zearalenone antibody is bound to the antibody-binding protein coated on the microtiter plate. After 10 minutes incubation at room temperature the wells are washed with diluted washing solution to remove unbound material. A substrate solution is added and incubated for 10 minutes, resulting in the development of a blue colour. The colour development is inhibited by the addition of a stop solution, and the colour turns yellow. The yellow colour is measured photometrically at 450 nm. The concentration of zearalenone is indirectly proportional to the colour intensity of the test sample.
- The kit contains reagents for 96 determinations;
- Microtiter plate consisting of 12 strips with 8 breakable wells;
- Zearalenone Standards: 0, 10, 25, 75, 200, 500 ppb;
- ELISA reader at 450 nm;
- Standard range: 10 - 500 ppb;
- Sensitivity analytical: 5 ppb.