Vomitoxin, also known as deoxynivalenol (DON), the chemical name is 3α, 7α, 15-trihydroxyfusarium-9-en-8-one, belonging to the trichothecenes family Compounds, mainly produced by Fusarium graminearum, Fusarium oxysporum, Fusarium moniliformes, Fusarium mycoides, Fusarium pink, Fusarium nivalis and other Fusarium species. In addition, strains of the genus Cephalosporium, Trichoderma, and Trichoderma can produce the toxin. There are more than 150 kinds of trichothecenes toxins, which are a class of powerful immunosuppressants. The typical symptom caused by them is decreased feed intake. Therefore, these toxins are also called feed antifeedant toxins. DON is one of the most important toxins, mainly from the genus Fusarium, especially Fusarium graminearum and Fusarium flavus. Because it can cause vomiting in pigs, it is also known as DON.
DON is classified as a Class 3 carcinogen. They have high cytotoxic and immunosuppressive properties, and therefore pose a threat to human and animal health, especially with significant effects on immune function. DON is widely present in the world, mainly contaminating wheat, barley, corn and other cereal crops, and also contaminating food products. When people ingest DON-contaminated food, it will cause anorexia, vomiting, diarrhea, fever, unsteady standing, and unresponsiveness. Other symptoms of acute poisoning, and in severe cases, damage the hematopoietic system and cause death. Because the proportion of grains and grains in the traditional Chinese diet is much higher than that of the Western diet, the harm of DON is more prominent. The content of DON in grains and feeds has strict limits. The limit standard of DON in grains in China is 1.0 mg/kg.
In the liquid chromatography for the detection of vomit toxin in China, the immunoaffinity column of emetic toxin (deoxynivalenol) is often used. The immunoaffinity column can selectively adsorb deoxynivalenol in the sample solution, so as to play a very specific purification role of deoxynivalenol.
Using the antigen antibody reaction, the antibody connection within the cylinder, the sample after extraction, filtration, slowly through deoxidizing snow sickle bacteria leaven immune affinity chromatography column, endotoxin in immune affinity column with antibodies, immune affinity column after washing to remove without being combined with other irrelevant material, reoccupy methanol elution, and then used to detect. After the purification column, it can be directly used for the determination of the content of liquid phase deoxygenated fusarium niuforniol, which can improve the accuracy of the detection method and achieve the purpose of rapid determination.
Reference standard “GB 5009.111-2016 National Food Safety Standard Determination of deoxynivalenol and its acetylated derivatives in food”, Welch vomit toxin (deoxynivalenol) DON immunoaffinity column completed in line with the standard requirements. The flour samples were purified by DON immunoaffinity column using Welch vomit toxin (deoxynivalenol) and then detected.
Temperature return: after the immunoaffinity column is removed from the low temperature, it is restored to room temperature and the liquid in the column is released;
Sample loading: until the purification liquid is all sample loading, discard;
Rinsing: 5mL phosphate buffer, 5mL water, discard, drain the column;
Elution: add 2mL methanol to elution and drain the column;
Concentration: The eluent was placed in a 50℃ water bath with nitrogen blown to dry, and the volume was constant with 20% methanol water to 1mL. The eluent was filtered by a 0.22μm filter membrane, and then measured on the machine.
The chromatographic conditions
Column: Welch Ultisil® XB-C18 4.6×150mm, 5μm;
Mobile phase: water: methanol (80:20);
Flow rate: 1.0mL/min;
Injection volume: 20μL;
Recovery results are shown below
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