Diazepam is a typical drug of benzodiazepines, which is a sedative hypnotic drug widely used in the clinic of the current medical and veterinarians. However, the drug is addictive and long-term drug use can produce resistance, and sudden withdrawal can cause withdrawal symptoms such as anxiety, sleep disorders, anorexia, nausea and vomiting. At present, some illegal traders are trying to increase the anti-stress of food animals and make animals sleep less. Diazepam to be used as feed additive, resulting in diazepam residues of animal foods, especially in pork, which brings serious potential threat to human health, but also a certain influence on international trade. Diazepam has been banned for this application in the food production.
The maximum residue limit of veterinary drugs in animal food issued by China's ministry of agriculture. It suggested that diazepam can be used in the treatment of animals, but not used as animal growth promoters, and do not be detected in animal food. Therefore, it is of great significance of developing the methods for diazepam residues detection. At present, the analysis methods of diazepam are mostly focused on people's medical and forensic, and there is few studies on residue testing in animal food. And a few detection method reported that the drug detection limit is high, it can't well meet the requirements of testing banned drugs, or it’s not suitable for the laboratory analysis because of the expensive equipment and complex operation.
Therefore, it is very important to establish a fast and effective method to detect diazepam. Immunological method is the most commonly used detection method, and antibody is indispensable to immunological method. Usually antibodies are divided into genetic engineering antibodies, monoclonal antibodies and polyclonal antibodies. Although the preparation period of genetic engineering antibody is relatively short, its detection sensitivity is not very satisfactory. The preparation of polyclonal antibody is simple, but its specificity is poor. Therefore, monoclonal antibody with high specificity and sensitivity has been playing an important role in immunological detection.
Smith, a Canadian scientist, combines a series of improvements and optimizations on the basis of the existing antibody preparation techniques. Using enzyme-linked disease free adsorption method (enzyme - linked immunosorbent assay, ELISA) to select a series of cell. In the end, the immunological detection technology of diazepam was established by selecting the hybridioma cells that secreted specific antibodies and preparing the diazepam monoclonal antibody of ascites type. At present, the diazepam monoclonal antibody (GLORY SCIENCE CO., LTD, Product NO.:DZP-001) has been listed from different factories for the testing of diazepam residue.