Preparation method and application of environmental estrogen electrochemical immunosensor
An environmental estrogen and immunosensor technology is applied in the field of preparation of environmental estrogen electrochemical immunosensors, which can solve the problems of cumbersome detection process, high cost, complicated instruments and the like, achieves less reagent consumption, simple method, and maintains biological activity. Effect
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Embodiment 1
[0045] (1) Weigh 0.5mg graphene and add it to a concentration of 1.0mg·mL -1 Chitosan solution, ultrasonic 30min to disperse uniformly, prepared 0.5mg·mL uniformly dispersed -1 Graphene-chitosan solution (GS-Chit);
[0046] (2) Weigh 15mg of SBA-15 and add it to 3mL with a concentration of 10mg·mL -1 Chlorauric acid (HAuCl 4 ), diluted with water to 40mL, stirred for 20h under sealed conditions, and 6000r·min -1 Centrifuge at a high speed for 15 minutes, and discard the supernatant. Then add 1mmol·L dropwise -1 sodium borohydride until the color of the solution no longer changes, stirred for 10 minutes, washed by centrifugation, and dried in vacuum at room temperature to obtain powdered AuSBA-15;
[0047] (3) Take the powdered AuSBA-15 in step (2) and dilute it with water to 3.5 mg·mL -1 AuSBA-15 solution, take 500 μL of glutaraldehyde with a mass fraction of 2.5%, and 750 μL with a concentration of 3.5 mg mL -1 AuSBA-15 solution and 1000 μL concentration of 1.0 mg·mL ...
Embodiment 2
[0055] (1) Weigh 1.0mg graphene and add it to a concentration of 1.5mg·mL -1 In the chitosan solution, ultrasonic 30min to disperse evenly, to prepare uniformly dispersed 1.0mg mL -1 Graphene-chitosan solution (GS-Chit);
[0056] (2) Weigh 30mg of SBA-15 and add it to 3mL with a concentration of 10mg·mL -1 Chlorauric acid (HAuCl 4 ), diluted with water to 40mL, stirred for 20h under sealed conditions, and then added 1mmol·L dropwise -1 Sodium borohydride until the solution turns brown and the color does not change, stir for 10 minutes, centrifuge and wash, and dry in vacuum at room temperature to obtain powdered AuSBA-15;
[0057] (3) Take the powdered AuSBA-15 in step (2) and dilute it with water to 3.5 mg·mL -1 For the AuSBA-15 solution, take 500 μL of glutaraldehyde with a mass fraction of 2.5%, and the concentration of 1000 μL is 3.5 mg·mL -1 AuSBA-15 solution and 1500 μL concentration of 1.0 mg·mL -1 Ab in a 4mL centrifuge tube, shake at 4°C for 20h to incubate the ...
Embodiment 3
[0065] (1) Weigh 1.5mg graphene and add it to a concentration of 3.0mg·mL -1 Chitosan solution, ultrasonic 30min to disperse evenly, prepared uniformly dispersed 1.5mg·mL -1 Graphene-chitosan solution (GS-Chit);
[0066] (2) Weigh 45mg of SBA-15 and add it to 3mL with a concentration of 10mg·mL -1 Chlorauric acid (HAuCl 4 ), diluted with water to 40mL, stirred for 20h under sealed conditions, and then added 1mmol·L dropwise -1 Sodium borohydride until the solution turns brown and the color does not change, stir for 10 minutes, centrifuge and wash, and dry in vacuum at room temperature to obtain powdered AuSBA-15;
[0067] (3) Take the powdered AuSBA-15 in step (2) and dilute it with water to 3.5 mg·mL -1 AuSBA-15 solution, respectively take 500 μL of glutaraldehyde with a mass fraction of 2.5%, and 1250 μL with a concentration of 3.5 mg mL -1 1000 μL of AuSBA-15 solution and a concentration of 1.0 mg·mL -1 Ab in a 4mL centrifuge tube, shake at 4°C for 30h to incubate the...
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