An electrochemical immunosensor for detecting pesticide triazolium tin and its preparation method
An immunosensor, the technology of triazolyl tin, is applied in the field of electrochemical immunosensors for detecting large sterically hindered pesticide triazolyl tin and its preparation, and achieves the effects of not interfering with antigen-antibody reactions, being convenient to carry, and being firmly labeled
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Embodiment 1
[0048] Electrolyte: 0.1mol / L KCl+5mmol / L K 4 [Fe(CN) 6 ]+5mmol / L K 3 [Fe(CN) 6 Example 1: Preparation method of electrochemical immunosensor for detecting pesticide triazolium tin
[0049] 1. Synthesis of triazolidine artificial antigen
[0050] (1) Synthesis of triazotin hapten: Weigh 193g~195g tricyclic tin and dissolve it with 24~26mL methanol, then slowly add dropwise to 58g~59g and dissolve it in 48~52mL 80℃~85℃ hot water The 5-mercapto-3-amino-1,2,4-triazole (5-mercapto-3-amino-1,2,4-triazole is insoluble in cold water and methanol, and is Solubility in water is very high, so the reaction selects high-temperature reaction), and after stirring for 1-2 hours, a large amount of white precipitates are formed. After the white precipitates are filtered, they are washed with boiling water for 3-4 times, and after drying, they are recrystallized with 95% ethanol to obtain three Azotin hapten ACT-H1; the reaction equation is as follows:
[0051]
[0052] (2) Synthesis of...
Embodiment 2
[0059] Embodiment 2: Utilize the method for detecting triazotin by electrochemical immunosensor
[0060] 1. Preparation of triazolin monoclonal antibody: immunize 6-8 week-old SPF female Balb / c mice with triazolin antigen ACT-H1-BSA, and the animal immunization process is as follows: take 100 μL of aliquoted triazolin The antigen and the equal volume of Freund’s complete adjuvant are emulsified into an emulsion by mutual push of a syringe, emulsified to drops, and can remain intact and not dispersed when dropped into water, and float on the water surface in a drop shape, that is, the emulsification is complete; use the back subcutaneous, abdominal cavity and foot pads Multi-point injection of 0.2mL-0.3mL per mouse (containing antigen 100μg-150μg), 3 weeks later, the same dose of artificial antigen mixed with an equal volume of Freund's incomplete adjuvant to form an emulsion, and the same dose and method were injected for booster immunization; then, every Booster immunization ...
Embodiment 3
[0066] Example 3: Optimization of the reaction conditions of the triazolidine electrochemical immunosensor
[0067] In immunosensors, optimizing the conditions during the immunization process is very important for the performance of the sensor, the dilution ratio of the antigen buffer used for immobilization, the dilution ratio of the antibody buffer that binds to the antigen, the amount of antibody that binds to the hapten, The time and temperature of the immune reaction have a great influence on the accuracy and sensitivity of the sensor. This experiment focuses on the concentration of naphthol solution, nano-Co 3 o 4 / Pan solution dilution ratio, antigen-antibody dilution ratio, pH value of potassium ferricyanide buffer dilution were optimized, the same experiment in the optimization process set three parallel experiments, the data take the average value of three experiments, the experimental results are as follows Figure 5 As shown, this experiment determined triazolium...
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