Enzyme immunoassay for the quantitative determination of Pecan nut in food.
Background: The pecan tree (Carya illinoinensis) belongs to the family of Juglandiaceae. The amount of protein in pecan nut is about 9%. Some of these proteins are known for being allergenic, like the 2S albumin Car I 1 or the 11S legumin Car I 4 . Compared to other nuts the allergenic potential of pecan nuts is still slightly characterized. Pecan nut allergies are relatively seldom, but can be very distinct in particular cases. For pecan nut allergic persons hidden pecan nut allergens in food are a critical problem. Already very low amounts of pecan nut can cause allergic reactions, which may lead to anaphylactic shock in severe cases. Because of this, pecan nut-allergic persons should strictly avoid the consumption of pecan nut containing food. Cross-contamination, mostly in consequence of the production process, is often noticed. This explains why in many cases the existence of pecan nut residues in food cannot be excluded. For this reason sensitive detection systems for pecan nut residues in foodstuff are required. Pecan nut ELISA represents a highly sensitive detection system for pecan nut and is particularly capable of the quantification of residues in cookies, cereals, ice-cream, chocolate and sausage.
Description: The Pecan nut quantitative test is based on the principle of the enzyme linked immunosorbent assay. An antibody directed against pecan nut proteins is bound on the surface of a microtiter plate. Pecan nut containing samples or standards are given into the wells of the microtiter plate. After 20 minutes incubation at room temperature the wells are washed with diluted washing solution to remove unbound material. A peroxidase conjugated second antibody directed against pecan nut proteins is given into the wells and after 20 minutes of incubation the plate is washed again. A substrate solution is added and incubated for 20 minutes, resulting in the development of a blue colour. The colour development is terminated by the addition of a stop solution, and the colour turns yellow. The yellow colour is measured photometrically at 450 nm. The concentration of pecan nut is directly 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;
- Pecan nut Standards: 0, 2, 6, 20, 60 ppm;
- ELISA reader at 450 nm;
- Standard range: 2 - 60 ppm;
- Sensitivity analytical: 0.2 ppm.