Enzyme immunoassay for the quantitative determination of Chloramphenicol in food and urine.
Background: Due to its outstanding antibacterial properties, chloramphenicol is an often used antibiotic in the production of milk, meat and eggs. In humans it leads to haematotoxic side effects, like the chloramphenicol induced aplastic anemia. This has caused low limits in Germany, e.g. 1 µg/kg for milk, meat and eggs.
Till now the chloramphenicol concentration was determined by radioimmunoassay or by gas chromatography. However, compared with conventional methods, enzyme immunoassays show some essential advantages. There is no need to work with radioactive material, the required assay time is shorter and the sensitivity is better than with chromatographic methods.
The Chloramphenicol test provides a rapid, sensitive and reliable assay for the determination of chloramphenicol in food and urine. 40 samples can be assayed in duplicate within 60 minutes.
Description: The Chloramphenicol 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. Chloramphenicol containing samples or standards and an antibody directed against chloramphenicol are given into the wells of the microtiter plate. The chloramphenicol contained in samples or standards will bind to the antibody which reacts with the binding protein coated onto the microtiter plate. After 30 minutes incubation at room temperature a chloramphenicol-peroxidase conjugate is added into the wells without a preceding washing step to saturate free antibody binding sites. After additional 15 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 15 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 chloramphenicol 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;
- Chloramphenicol Standards (0, 0.05, 0.1, 0.5, 1, 5 ng/mL);
- ELISA reader at 450 nm;
- Standard range: 0.05 - 5 ng/mL;
- Sensitivity analytical: 0.03 ng/mL.