Preparation Of Furazolidone Metabolite Colloidal Gold Test Strip

Because of the low price and easily obtained, Nitrofuran drugs, which belong to a class ofsynthetic antibiotics, are widely used in aquaculture in order to prevent the disease. The half-lifeperiod of Nitrofuran drugs is very short, it can be metabolized inside animals, the metabolitescan steadily exist. The nitrofuran metabolite in aquatic products seriously threat to the health ofpeople, so the nitrofuran drugs are worldwide banned from use as fishery drugs.At present, LC-MS/MS is a commonly used method for detection of nitrofurans metabolites,but this method need expensive equipment, complex pre-treatment, high cost and long analysistime. However, Gold Immunochromatographic Assay has advantages of time saving, low cost,high efficiency, and the repeatability of test results are good, and it is suitable for on-site testingmass screening and rapid detection. The purpose of the research was to develop the colloidalgold immunochromatographic assay of furazolidone metabolite, and preliminary build a rapiddetection method for furazolidone metabolite, and provide the basis for the development ofcommercial strip.The research optimized the method of preparation of colloidal gold, choosing three kinds ofreducing agent to prepare the colloidal gold, and contrasting the performance parameters ofcolloidal gold, then choosing the best reductive methods. The research choose the sodium citrateto prepare the colloidal gold by comparing, and the particle size of colloidal gold is about18nm.Using the colloidal gold labeled monoclonal antibodies of AOZ to get jinbiao probe, thenCPAOZ-BSA and the sheep anti-mouse immunoglobulins were coated onto a nitrocellulosemembrane. Nitrocellulose membrane, gold conjugate pad, sample pad and absorbent pad wereassembled onto a sheet and cut into individual strips. The optimum pH of colloidal gold labeledantibodies was9.0, the quantity of colloidal gold labeled antibodies was7.8μg/mL, theconcentration of CPAOZ-BSA was1.0mg/mL as the test line, the concentration of sheepanti-mouse IgG was1.0mg/mL, the sensitivity of strips was80ng/mL, the test could be finishedwithin10minutes.The research tested the specificity of the strip, the result showed that there were not crossreactions with SEM, CPSEM, AHD, CPAHD.In a word, the strip is used to detect AOZ, it shows that the strip is time saving, sensitive,easy to be justified and suitable for the on-site detection. These key techniques will help us research and develop a great strip to detect AOZ on-site more quickly.