Indoxacarb hapten, antigen, antibody, detection device as well as preparation and application of indoxacarb hapten, antigen and antibody
A detection device, indoxacarb technology, applied in the field of antigen, indoxacarb hapten, detection device, antibody, can solve the problems that cannot meet the on-site supervision and law enforcement of the regulatory department, it is difficult to achieve rapid detection, the process is complicated, etc., to achieve detection Low cost, easy operation and high reaction yield
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Embodiment 1
[0027] In this example, the technical indoxacarb is selected from 95% of the technical indoxacarb supplied by Jingbo Agrochemical Technology Co., Ltd., the sodium borohydride is selected from the analytically pure sodium borohydride supplied by Sinopharm Chemical Reagent Co., Ltd., and the ethyl acetate is selected Analytical pure ethyl acetate supplied by Sinopharm Chemical Reagent Co., Ltd., anhydrous sodium sulfate 99% anhydrous sodium sulfate supplied by Sinopharm Chemical Reagent Co., Ltd., and 99.5% anhydrous pyridine supplied by Sinopharm Chemical Reagent Co., Ltd. Pyridine and succinic anhydride were selected from the analytically pure succinic anhydride supplied by Sinopharm Chemical Reagent Co., Ltd., and N,N-dimethylformamide was selected from 99.8% N,N-dimethylformamide supplied by Sinopharm Chemical Reagent Co., Ltd. Tri-n-butylamine selects analytically pure tri-n-butylamine supplied by Sinopharm Group Chemical Reagent Co., Ltd., and isobutyl chloroformate selects...
Embodiment 2
[0028] Example 2 Synthesis and identification of indoxacarb hapten
[0029] The preparation method of indoxacarb hapten comprises the following steps:
[0030] S1. Take a single-neck bottle, weigh 1.0g of indoxacarb original drug into the single-neck bottle, dissolve with 20ml of anhydrous methanol, add 0.21g of sodium borohydride in batches under ice bath conditions, stir at room temperature, and monitor the reaction by TLC Completely, after evaporating the solvent, add 100ml of distilled water, add ethyl acetate for extraction and liquid separation to obtain an organic phase, after drying the organic phase with anhydrous sodium sulfate, purify by column chromatography to obtain 0.8g of an intermediate whose structural formula is as follows: Formula (II) shows:
[0031] (II);
[0032] S2. Take 400 mg of the intermediate, dissolve it with anhydrous pyridine, add 0.2 g of succinic anhydride, heat up to 80 ° C and react overnight, evaporate the solvent after the reaction, ad...
Embodiment 3
[0033] Example 3 Synthesis of Indoxacarb immunization antigen
[0034] The indoxacarb immunization antigen was prepared by the active ester method, 70 mg of indoxacarb hapten was weighed, dissolved in 3 ml of DMF, 36 mg of NHS and 41 mg of EDC.HCl were added, and the reaction was carried out at room temperature for 6 h to prepare an activation solution; 64 mg of BSA was weighed to dissolve In 3.2mL 0.1M pH9.0 boric acid buffer, add 1ml DMF, 0.9mL above activation solution, react at room temperature for 4 hours and then dialyze with PBS (0.01 mol / L, pH=7.4 phosphate buffer solution), change every 4 hours After dialysis, 4000 rpm / min was centrifuged for 5 min, and the supernatant was taken to obtain the indoxacarb hapten-BSA conjugate, which was the antigen for indoxacarb immunization. save.
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