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Electrochemical immunosensor for detecting estrol and bisphenol A, preparation method and application thereof

An immunosensor and electrochemical technology, which is applied in the field of food safety detection and analytical chemistry, can solve the problems of cumbersome operation process, high cost of use and maintenance, difficulty in meeting the needs of on-site rapid detection, etc., and achieve the effect of reducing energy consumption

Active Publication Date: 2019-06-21
NANJING NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these analytical methods require large-scale analytical instruments, the operation process is cumbersome, the use and maintenance costs are high, and it is difficult to meet the increasing needs of on-site rapid detection. Therefore, it is necessary to study and establish more efficient, fast, sensitive, and low-cost analytical methods

Method used

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  • Electrochemical immunosensor for detecting estrol and bisphenol A, preparation method and application thereof
  • Electrochemical immunosensor for detecting estrol and bisphenol A, preparation method and application thereof
  • Electrochemical immunosensor for detecting estrol and bisphenol A, preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] The synthesis of embodiment 1N-(hexestroloxy group) acetyl cysteamine

[0039] 1) Synthesis of α-bromo N-acetyl cysteamine

[0040] Dissolve N-acetylcysteamine (119mg, 1mmol) in 10ml THF, cool to 0°C, add N-bromosuccinimide (NBS, 195mg, 1.1mmol) in batches, and continue to react at 0°C for 2 hours After the solvent was removed by rotary evaporation, the product was purified by a neutral alumina column to obtain 135 mg of the product α-bromo-N-acetylcysteamine (68% yield).

[0041] 2) Synthesis of N-(Hestestroloxy)acetylcysteine

[0042] Dissolve 99mg (0.5mmol) of α-bromo-N-acetylcysteamine in 1ml of acetone to obtain liquid A; dissolve 122mg (0.5mmol) of hexestrol in 5ml of dioxane to obtain liquid B, and slowly Add solution B dropwise, then slowly add 0.5ml of sodium carbonate solution (1mol / L) dropwise, and stir at 70°C for 12h. Suction filtration, the product obtained after the filtrate was rotary evaporated to remove the solvent was dissolved in 15ml of boiling c...

Embodiment 2

[0045]The detection of embodiment 2 to diethylstilbestrol standard sample

[0046] 1) Preparation of electrochemical immunosensor:

[0047] A glassy carbon electrode with a diameter of 3 mm was sequentially filled with Al with a diameter of 0.3 μm and 0.05 μm 2 o 3 Grinding with polishing powder, followed by ultrasonic cleaning with absolute ethanol-distilled water and distilled water for 2 minutes, and then rinsed with distilled water. Place the above electrode in 1% chloroauric acid solution for constant potential electrochemical deposition (voltage is -0.2V, deposition time 30s), rinse with deionized water and then immerse in 1mmol / L N-(hexestrol Oxygen) acetylcysteamine solution, placed in a refrigerator at 4°C for 24 hours to react, and then the electrochemical immunosensor modified by nano-gold / N-(hexestradioloxy)acetylcysteamine can be obtained.

[0048] 2) Detection of diethylstilbestrol standard sample:

[0049] Immerse the electrochemical immunosensor described i...

Embodiment 3

[0050] Embodiment 3 is to the detection of hexestrol standard sample

[0051] Prepare the electrochemical immunosensor modified by nano-gold / N-(hexestroloxy) acetylcysteamine in exactly the same way as in Example 2, step 1), and immerse it in a total volume of 50 μL containing a concentration of 0.5 mg / L of estrol monoclonal antibody and a series of standard solutions of different concentrations of estrol, then use the same method as in step 2) of Example 2 to perform EIS scanning and data processing. EIS curve as image 3 As shown, the concentration of the graph from top to bottom is 0ng / mL, 0.5ng / mL, 1ng / mL, 5ng / mL, 10ng / mL, 20ng / mL, 100ng / mL, 200ng / mL, 500ng / mL, 1000ng / mL and 2000ng / mL. The resulting change in charge transfer resistance ΔR ct It has a linear relationship with the logarithmic value lgC of estrol concentration when the concentration of estrol is between 0.5 and 2000ng / mL, and the working curve is as follows Figure 4 As shown, the linear correlation coef...

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Abstract

The invention discloses an electrochemical immunosensor for detecting diethylstibestrol and bisphenol A and a preparation method and application thereof. The sensor comprises a substrate glassy carbon electrode, wherein gold nanoparticles are deposited on the surface of the substrate glassy carbon electrode, and N-(hexestrol oxyl) acetyl cysteamine is modified on the surfaces of the gold nanoparticles through a self-assembly method. The N-(hexestrol oxyl) acetyl cysteamine is modified on the surface of the sensor by adopting a chemical method, so that the electrochemical immunosensor can be regenerated through simple treatment after use, and can be repeatedly utilized, and consumption of the material is reduced. The detection limit of the sensor for detecting the diethylstibestrol and the bisphenol A is 0.25ng / mL; the detection limit of diethylstilbestrol is 0.15ng / mL; the detection limit of the bisphenol A is 0.20ng / mL; the linear range of the bisphenol A is 0.5-1,000ng / mL; the linear ranges of the others are 0.5-2,000ng / mL; and the electrochemical immunosensor has a relatively low detection limit and a relatively wide linear range.

Description

technical field [0001] The invention belongs to the technical field of food safety detection and analytical chemistry, and relates to an electrochemical immunosensor for detecting estrol and bisphenol A, a preparation method and application thereof. Background technique [0002] Diethylstilbestrol (DES), hexestrol (HEX) and dienestrol (DIEN) (see formula 1 for structure) are a class of artificially synthesized drugs with estrogen effects, because they can promote animal growth. , improve feed conversion rate and reduce fat synthesis, etc. It has been approved as a growth-promoting agent and widely used in animal husbandry. The abuse of such drugs will eventually endanger human health through the food chain, causing various serious consequences, such as abnormal development, tumors, fetal deformities, etc. [0003] Bisphenol A (bisphenol A, BPA) (see formula 1 for structure) is widely used in the production of various polymer materials such as polycarbonate and epoxy resin, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/327
CPCG01N27/327
Inventor 赵波王雅娟邵科峰徐红平朱永强
Owner NANJING NORMAL UNIVERSITY