Electrochemical immunosensor for detecting diethylstilbestrol, preparation method and application thereof

An immunosensor, diethylstilbestrol technology, applied in the direction of material electrochemical variables, biological testing, material inspection products, etc., can solve the problems of poor specificity, achieve high sensitivity, rapid on-site detection, and wide application range

Inactive Publication Date: 2011-11-30
NANJING NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The modification process of the working electrode of the invention is simple and can be reused, the test has good reproducibility and high sensitivity, and the detection limit can reach 1ng / g, but the specificity of the method is not good

Method used

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  • Electrochemical immunosensor for detecting diethylstilbestrol, preparation method and application thereof
  • Electrochemical immunosensor for detecting diethylstilbestrol, preparation method and application thereof
  • Electrochemical immunosensor for detecting diethylstilbestrol, preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1 Immunosensor preparation

[0052] An electrochemical immunosensor for detecting diethylstilbestrol is obtained by modifying a graphene-diethylstilbestrol-chitosan complex after depositing nano gold on the surface of a glassy carbon electrode.

[0053] Its preparation method is as follows:

[0054] 1) Preparation of graphite oxide: 50 mL concentrated H 2 SO 4 +10 g K 2 S 2 o 8 +10gP 2 o 5 Mixed, 12 g of graphite powder was added to the above mixed solution, and reacted for 6 h. Add 2 L of water and leave overnight. 460 mL concentrated H 2 SO 4 (refrigerator freezing treatment), the oxidized graphite powder was added to the concentrated H 2 SO 4 Stir in. Control the temperature below 10 ℃, slowly add 60 g KMnO 4 , the mixture was reacted at 35 °C for 2 h, 920 mL of deionized water was slowly added, kept below 50 °C, and stirred for 2 h. Add 2.8 L of water and 50 mL of 30 % H 2 o 2 , stirred for one day. Rinse with 5 L of 10% HCl and centrifuge....

Embodiment 2

[0062] Example 2 Modified electrode cyclic voltammetry scan

[0063] Cyclic voltammetry studies were carried out on different modified electrodes, and the results are as follows: Figure 4 . Bare electrode (curve a) at 2mM K 3 [Fe(CN) 6 ] The cyclic voltammetry curves in solution show a pair of distinct Fe(CN) 6 3- / 4- Redox peak; when the electrode is modified with graphene / diethylstilbestrol / chitosan (curve b), the peak current increases significantly, and graphene shows excellent electrochemical activity, which enhances the electron transfer; when the electrode is modified After nano-gold / graphene / diethylstilbestrol / chitosan (curve c), the peak current increases even more, and the composite material of nano-gold and graphene has more superior electrochemical performance; After the modified electrode was incubated in the incubation solution containing diethylstilbestrol antibody (curve d), the peak current decreased. This was because the diethylstilbestrol antibody react...

Embodiment 3

[0065] Example 3 Detection condition optimization

[0066] The effect of different detection conditions, such as the time of immune response and antibody concentration, was determined by the DPV method.

[0067] 1. Immune response time

[0068] The immunosensor prepared in Example 1 was incubated with the incubation solution containing the same antibody concentration for 0, 10, 20, 40, 60 and 70 minutes, washed with PBS, and incubated at 2 mM K 3 [Fe(CN) 6 ] solution in a differential pulse voltammetry (DPV) scan, the results are as follows Figure 5 . The DPV peak current of the nanogold / graphene / diethylstilbestrol / chitosan modified electrode (immunosensor) decreased rapidly within 60 minutes as the incubation time increased, and rose slightly at 70 minutes ( Figure 5 ). The current response no longer decreases when the incubation time is increased. Therefore, 60 min was chosen as the optimal incubation time for the immune response.

[0069]2. Effect of antibody conce...

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Abstract

The invention relates to a non-labeled electrochemical immune sensor for detecting diethylstilbestrol and a preparation method and application of the sensor. The immune sensor comprises a substrate electrode; and after nanometer gold is deposited on the surface of the substrate electrode, a graphene-diethylstilbestrol-chitosan complex is modified, and non-specific active loci are sealed by bovineserum albumin. The preparation method comprises the following step of: after depositing the nanometer gold on the surface of the substrate electrode, enveloping graphene and the diethylstilbestrol byusing chitosan and fixing to the surface of the electrode. In the immune sensor, on the basis of a competitive mode of immune reaction, the immune reaction is monitored by taking K3Fe(CN)6 as a probeand by differential pulse voltammetry, so the immune sensor can be used for detecting the diethylstilbestrol. The non-labeled electrochemical immune sensor has the characteristics of high sensitivityand specificity, quick on-site detection, low cost and the like; and a detection method is simple and wide in application range, the detection limit can reach 0.1 ng / ml, and the linear range is between 0.5 and 1,500 ng / ml.

Description

technical field [0001] The invention belongs to the technical field of food safety detection and analytical chemistry, and relates to a detection device and method for diethylstilbestrol, in particular to an electrochemical immunosensor for measuring artificially synthesized estrogen (Diethylstilbestrol, DES) in food and its A preparation method and a method for measuring diethylstilbestrol by using the sensor. Background technique [0002] Diethylstilbestrol is a synthetic non-steroidal estrogen, which was artificially synthesized in London in 1938. It can produce the same pharmacological and therapeutic effects as natural estradiol, and is mainly used for functions caused by hypoestrogenism and hormone imbalance Sexual bleeding, amenorrhea, and can also be used before stillbirth induction to increase the sensitivity of the myometrium to oxytocin. Diethylstilbestrol also has the functions of promoting protein metabolism and synthesis, increasing animal daily gain and reduc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/48G01N27/30G01N33/53
Inventor 赵波陈昌云王传现邵科峰颜妍张芹陈园园
Owner NANJING NORMAL UNIVERSITY
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