Testing kit for enrofloxacin and testing method thereof
A technology of enrofloxacin and reagent kits, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of limited application, expensive, cumbersome sample processing, etc., and achieve the effects of high sensitivity, simple structure, and convenient use
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Embodiment 1
[0012] Example 1: Preparation of kit and detection of food of animal origin
[0013] Preparation of immune antigen:
[0014] The artificial immune antigen of the present invention combines enrofloxacin (ENR) with a macromolecule carrier protein by a mixed anhydride method, and uses synthetic ENR-BSA as an immune antigen to immunize animals.
[0015] The carrier protein is bovine serum albumin (BSA). Weigh 30mg~300mg bovine serum albumin, dissolve in 6mL~10mL of 50% dioxane aqueous solution, adjust the pH value to 9~10, and make it A liquid; weigh 10mg~40mg ENR, dissolve in 2mL dioxane Set it at 4°C for 10 minutes, add 20 μL of tri-n-butylamine and 10 μL of isobutyl chloroformate in sequence, stir at 4°C for 30 minutes, and use it as liquid B. Add liquid A dropwise to liquid B at 4°C, and stir overnight. The reaction solution was dialyzed against distilled water for 72 hours, during which the distilled water was changed 5 times. Or purified by Sephadex G50 column and eluted...
Embodiment 2
[0056] Implementation example 2 preparation kit
[0057] ENR-BSA antigen preparation:
[0058] Weigh 12mg ENR, 40mg N-hydroxysuccinamide (NHS), 35mg carbodiimide (EDC) and dissolve in N,N′-dimethylformamide (DMF), stir at room temperature for 24h to prepare A solution; Dissolve 50mg of BSA in 3mL of 0.01mol / mL pH7.4 PBS to prepare solution B. Add solution A dropwise to solution B, stir while adding, react at room temperature for 3 hours, dialyze the reaction solution in distilled water for 72 hours, and change distilled water 5 times during the period. Or purified by Sephadex G50 column and eluted with 0.05mol / L pH8.0 carbonate buffer solution. Collect the dialysis purified liquid or the column eluate of the first peak, aliquot and store at -20°C for immunogen use.
[0059] Preparation of polyclonal enrofloxacin antibody: same as in Example 1, omitted.
[0060] Eu 3+ -Preparation of goat anti-rabbit antibody: same as Example 1, omitted.
[0061] Preparation of solid-phas...
Embodiment 3
[0068] The reagent provided by the kit is the same as that in Example 1, and is used to detect honey samples.
[0069] Reagents that should be prepared by the laboratory:
[0070] (1) 0.02mol / L PB buffer solution: weigh Na 2 HPO 4 12H 2 O 5.16g, NaH 2 PO 4 0.87g into the volumetric flask, add water to dissolve and make the volume to 1L;
[0071] (2) dichloromethane solution;
[0072] (3) 0.02mol / L PB buffer solution: weigh Na 2 HPO 4 12H 2 O 5.16g, NaH 2 PO 4 0.87g into the volumetric flask, add water to dissolve and make the volume to 1L;
[0073] (4) Phosphate buffer working solution: pH7.2, take 1.45g disodium hydrogen phosphate (containing 12 crystal water), 0.1g potassium dihydrogen phosphate (anhydrous), 8.0g sodium chloride, dissolve in water and set Dissolve to 1000mL.
[0074] Matters needing attention before determination: same as Example 1.
[0075] The specific detection steps are as follows:
[0076] Accurately weigh 1.0g±0.01g of honey sample into ...
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