Immunoaffinity stir bar for adsorbing chloramphenicol, and preparation method and application thereof
A technology of chloramphenicol and immunophile, which is applied in the field of food safety testing, can solve the problems of low specificity, non-commercialization, environmental pollution, etc., and achieve the effects of strong specificity, lower analysis cost and saving organic solvents
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Embodiment 1
[0042] 1. Preparation of chloramphenicol mouse monoclonal antibody
[0043] The CAP-BSA immune antigen was synthesized by the mixed acid anhydride method, and 20 mg of chloramphenicol (Chloramphenicol, CAP) was dissolved in 1 mL of N,N-dimethylformamide (N,N-Dimethylformamide, DMF) and 1 mL of 1,4 - Dioxane (1,4-dioxane) mixed solution, then add 5 mL of isobutyl chloroformate (Isobutyl chloroformate), 4 ℃ low-speed stirring for 30 min, dropwise added to the BSA solution (50 mg of BSA dissolved In 1 mL of 0.02 mol / L PBS), stirred at 4 °C for 4 h at low speed, and the PBS solution (0.01 mol / L) was stirred and dialyzed at 4 °C for 5 days.
[0044] The coated antigen CAP-OVA was synthesized by the diazonium method. Take 80 mg of CAP, dissolve it in 1.5 mL of absolute ethanol, add 12 mL of hydrochloric acid (1 mol / L), add 65 mg of zinc powder, and heat in a water bath at 70 °C for 50 min. Add NaNO dropwise at 4°C and pH 1-2 2 (0.5 mol / L), and stir in the dark until the starch pot...
Embodiment 3
[0062] Immunoaffinity stir bar was used for the extraction of chloramphenicol in water. The blank water sample was passed through a 0.45 μm water filter membrane to remove impurities, and 5 mL of the filtered water sample was accurately weighed, placed in a 10 mL centrifuge tube, an appropriate amount of chloramphenicol standard solution was added, vortexed, and the immunophile Put it and the glass rod into the water sample after removing the mixing, and shake it on the shaker for 30 min. Take the oscillated glass rod out of the centrifuge tube, wash it with ultrapure water, put it into a 2 mL centrifuge tube filled with 700 μL of methanol, shake it on a shaker for 20 min to elute, dry it with nitrogen, and use the mobile phase Reconstitution, filtration with 0.22 μm microporous membrane, separation and detection by high performance liquid chromatography-mass spectrometry. The results showed that chloramphenicol and succinic chloramphenicol could be detected, indicating that...
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