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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 high cost, complicated instruments, cumbersome detection process, etc., and achieves simple method, less reagent consumption, and maintenance of biological activity. Effect

Active Publication Date: 2012-10-24
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] One of the technical tasks of the present invention is to provide a highly sensitive and specific environmental estrogen electrochemical immunosensor for overcoming the deficiencies in the existing technology for detecting environmental estrogen, such as complex instruments, high cost, and cumbersome detection process. Preparation method, its preparation technology is mature and reliable;

Method used

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  • Preparation method and application of environmental estrogen electrochemical immunosensor
  • Preparation method and application of environmental estrogen electrochemical immunosensor
  • Preparation method and application of environmental estrogen electrochemical immunosensor

Examples

Experimental program
Comparison scheme
Effect test

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 AuSBA-15 solution and 1000 μL concentration of 1.0 mg·mL -1 Ab in a 4mL centrifuge tube, shake at 4°C for 30h to incubate the anti...

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Abstract

The invention relates to a preparation method and application of an environmental estrogen electrochemical immunosensor, belonging to the technical field of electrochemical detection. The sensitivity of the sensor is obviously improved by adopting the characteristics that the conductivity of chitosan dispersed graphene is high, the stability is high, the specific surface area of a gold hybrid mesoporous silicon dioxide nano composite material is large, the biocompatibility is good, the catalytic efficiency is high and the like. The immunosensor is constructed by adopting the method of layer-by-layer self-assembly of the chitosan dispersed graphene, cross-linking agent, composite solution of nano materials and antibodies, and bovine serum albumin. In combination with specificities of antigens and antibodies, by virtue of an electrochemical workstation instrument, by recording current change before and after corresponding antigens modify the sensor, the environmental estrogens in water and food can be directly detected. The environmental estrogen electrochemical immunosensor prepared by adopting the preparation method has the advantages that the sensitivity is high, the specificity is good, the operation is easy to conduct and the detection limit is low, and the sensitive, rapid and accurate detection of various environmental estrogens in actual samples can be realized.

Description

technical field [0001] The invention relates to a preparation method and application of an environmental estrogen electrochemical immunosensor. Specifically, it is a label-free electrochemical immunosensor prepared based on a gold hybrid mesoporous silicon dioxide nanocomposite material, which is used for measuring environmental estrogen in water and food, and belongs to the technical field of electrochemical detection. Background technique [0002] Environmental estrogens (EEs) belong to environmental endocrine disruptors, which are a type of environmental toxins that will produce effects similar to estrogen after entering the human body. These toxins can interfere with our endocrine system, causing endocrine disorders, which can lead to a series of health problems. Environmental estrogens come from many sources and are ubiquitous in everyday life. Common ones include plastic-packaged food, shampoo, shower gel and other daily chemical products, fish, shrimp, shellfish and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327G01N27/36
Inventor 魏琴杜斌黎荣霞吴丹马洪敏李贺李玉阳张勇孙蒙闫良国
Owner UNIV OF JINAN
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