Su B ₁ And Citrinin The Gold Standard Research Paper Preparation Techniques Simultaneously Detect Aflatoxin

Aflatoxin B1(AFB1) and Citrinin (CIT) belong to fungi toxin, Which is a kind of secondary metabolites, produced by aspergillus and penicillium genus of fungi in the process of growth. AFB1and CIT usually through contaminated grain and animal foods moved into the food chain, which are great harmful to human’s liver and kidney, meanwhile constitute a potential threat to human health and national security. Therefore, it is very important to develop a rapid detection mothed about AFB1and CIT at the same time. The main contents and conclusions are as follows:1. An aflatoxin B1artificial antigen (AFB1-BSA) was synthesized by N-hydroxy-succinimide method and a citrinin artificial antigen (CIT-BSA) was synthesized by active ester method. Then combined with ultraviolet spectrophotometry and high performance liquid chromatography (HPLC)-mass spectrometry, the results showed the aflatoxin B1artificial antigen and citrinin artificial antigen was synthesized successfully, And the coupling ration of AFB1-BSA and CIT-BSA were5.13:1and7.2:1respectively.2. Colloidal gold particles were prepared by citrate-natrium method. The optimal pH for the colloidal gold labeled was when it is added20uL/ml0.1mol/L K2CO3solution, and the optimal antibody protein was of60ug/ml. Then the anti-AFB1and anti-CIT were marked by colloidal gold solution, respectively. AFB1and CIT immune probes were prepared, and was identified through the spot gold immune infiltration experiment, the result suggested that gold probe had an immune activity.3. The AFB1artificial antigen and CIT artificial antigen was used as envelope antigen, which was immobilized on the nitrocellulose membrane detection of lines T1and T2respectively. And the goat anti-mouse second antibody was coated on nitrocellulose membrane as the control line (C line). Then assembled colloidal gold immuno-chromatographic strip allowed to detect AFB1and CIT meanwhile. To explore the optimal experimental conditions and to determine the parameters of preparation of strip, such as the dilution formula and the dilution degree of the gold marked antibody, the optimum concentration of the coated antigen of the line T1and T2, and the second antibody concentration of the C line, were determined.4. The indicators of test strip were optimized after test strip has been assembled. Series of concentrations standard solution of AFB1and CIT was tested, the strip of the detection limit is5ng/ml to AFB1and50ng/ml to CIT. There is no cross reaction to AFT B2, M1, zearalenone and fumonisin. The test strip can be saved more than three months at4℃5. The test strip can simultaneously rapid detection, and this technique is possessed of high sensitivity, strong specificity, and the operation is simple and convenient, so the method can be used in the field of rapid detection. This research laid a foundation for the simultaneously rapid detection of a variety of toxins of test strip.