Preparation method of zeranol complete antigen
A technology of zearalanol and complete antigen, applied in chemical instruments and methods, animal/human protein, serum albumin, etc., can solve the problems of ZER antibody preparation and detection reports, low detection sensitivity, etc., and achieve strong practicability , High antigen titer, simple operation
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Embodiment 1
[0019] A preparation method of zearalanol complete antigen, comprising the following steps:
[0020] (1) Weigh 2 mg of ZER and dissolve it in 3 mL of acetone, add excess potassium carbonate 10 mg and 1 microliter of ethyl 4-bromobutyrate, at 60 ° C
[0021] Under reflux for 24h;
[0022] (2) After the completion of the reaction, stand at room temperature in the dark for 2 days, remove the precipitate by suction filtration, dry the filtrate by rotary evaporation, use 2 mL of Φ=20% NaOH-methanol solution for alkaline hydrolysis at 80 degrees Celsius for 1 hour, and dry the solution by rotary evaporation;
[0023] (3) Dissolve in 2 mL of double-distilled water, adjust the pH to 3.5 with 2 mol / L sulfuric acid, then repeatedly extract with 2-3 mL of ethyl acetate for 5 times, discard the water phase, filter once with saturated saline, and add anhydrous sulfuric acid Sodium drying, after suction filtration, the solution was rotary evaporated to dryness, and the yellow oil collected...
Embodiment 2
[0031] A preparation method of zearalanol complete antigen, comprising the following steps:
[0032] (1) Weigh 2 mg of ZER and dissolve it in 3 mL of acetone, add excess potassium carbonate 10 mg and 1 microliter of ethyl 4-bromobutyrate, at 60 ° C
[0033] Under reflux for 24h;
[0034] (2) After the completion of the reaction, stand at room temperature in the dark for 2 days, remove the precipitate by suction filtration, dry the filtrate by rotary evaporation, use 2 mL of Φ=20% NaOH-methanol solution for alkaline hydrolysis at 80 degrees Celsius for 1 hour, and dry the solution by rotary evaporation;
[0035] (3) Dissolve in 2 mL of double-distilled water, adjust the pH to 3.5 with 2 mol / L sulfuric acid, then repeatedly extract with 2-3 mL of ethyl acetate for 5 times, discard the water phase, filter once with saturated saline, and add anhydrous sulfuric acid Sodium drying, after suction filtration, the solution was rotary evaporated to dryness, and the yellow oil collected...
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