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Preparation method and detection method of nanocomposite material, sdm electrochemical aptamer sensor

A nanocomposite material and sensor technology, applied in the field of electrochemical detection, can solve the problems of inability to meet the actual needs of trace SDM, low specificity and sensitivity of detection results, cumbersome and time-consuming operations, etc., achieve high specificity, improve detection sensitivity, The effect of enhancing the electrochemical response signal

Inactive Publication Date: 2019-06-25
CHONGQING MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these methods have high requirements for experimental conditions and technical personnel, cumbersome and time-consuming operations, expensive instruments and equipment, and high detection costs; in addition, the specificity and sensitivity of the detection results are often low, and usually cannot meet the actual needs of trace SDM detection

Method used

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  • Preparation method and detection method of nanocomposite material, sdm electrochemical aptamer sensor
  • Preparation method and detection method of nanocomposite material, sdm electrochemical aptamer sensor
  • Preparation method and detection method of nanocomposite material, sdm electrochemical aptamer sensor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 Preparation of P-rGO-C 60 -Pt@Au nanocomposite

[0047] Follow the steps below:

[0048] (1) P-rGO-C 60 Preparation of nanocomposites:

[0049] 1) Add 5mL toluene to 5mg C 60 powder, and then use ultrasonic treatment to obtain a deep purple solution, then add 5mL ultrapure water, stand to separate layers, and then continue ultrasonic treatment for 10-12h until the C of the toluene phase 60 Transfer to the water phase, wait until the toluene is completely evaporated, and finally obtain a brownish yellow solution, and finally filter it with a 0.45 μm filter to obtain pure C 60 aqueous solution.

[0050] 2) ultrasonically disperse 5 mg of graphene oxide (rGO) in 5 mL of ultrapure water, add 5 mL of C prepared in step 1) 60 aqueous solution, and then 40 μL of PDDA (20%) was added dropwise. After stirring for 30 minutes, add 100 μL of hydrazine hydrate (80%) dropwise and react at 90°C for 4 hours; finally centrifuge at 10,000 r / min for 5 minutes to wash exce...

Embodiment 2

[0055] Example 2 Preparation of an electrochemical DNA aptamer sensor for SDM detection

[0056] Follow the steps below: (The construction principle is as follows figure 1 shown)

[0057] 1) Treat the DNA single strand of the thiol-labeled SDM-binding aptamer (SBA) with 10 mM TCEP solution at room temperature for 1 hour, and set aside.

[0058] 2) Use 0.3μm and 0.05μm Al respectively 2 o 3 The glassy carbon electrode (GCE) was polished into a mirror surface, and then the electrodes were ultrasonically treated in the order of ultrapure water, absolute ethanol, and ultrapure water for 5 minutes each, and dried at room temperature for later use.

[0059] 3) Place the electrode obtained in step 2) in 0.5M H 2 SO 4 Electrochemical cleaning was carried out in the medium, and the potential sweep was 0.2-1.6V until a stable cyclic voltammogram was obtained, and then rinsed with ultrapure water again and dried in air.

[0060] 4) 8 μL of the P-rGO-C prepared in Example 1 60 The ...

Embodiment 3

[0064] Example 3 Detection of SDM by electrochemical DNA aptamer sensor

[0065] Using the electrochemical DNA aptamer sensor constructed in Example 2 to detect SDM, follow the steps below:

[0066] 1. Draw the working curve

[0067] 1) The modified electrodes from Step 3) to Step 7) of Example 2 were respectively placed in 0.1M Na 2 HPO 4 , 0.1M KH 2 PO 4 , 10M KCl and 2mM MgCl 2 Characterized in 0.1M PBS, measured its electrochemical response signal, the results are as follows figure 2 Shown: (a) bare GCE; (b) dropwise addition of P-rGO-C 60 ; (c) Incubation of GOD; (d) dropwise addition of P-rGO-C 60 -Pt@Au; (e) combined with SBA; (f) blocked by GOD.

[0068] 2) Add 20 μL of the target substance sulfadimethoxine at different concentrations dropwise to the electrode of the aptamer sensor prepared in Example 2, and measure the current changes thereof respectively.

[0069] 3) According to the linear relationship between the obtained current change value and the loga...

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Abstract

The invention discloses a P-rGO-C60 nanocomposite material and a P-rGO-C60-Pt@Au nanocomposite material, and also discloses an electrochemical DNA aptamer sensor for SDM detection, wherein the electrochemical DNA aptamer sensor is prepared by the following method: 1) treating an SDM aptamer by using a TCEP solution for standby application; 2) polishing a glassy carbon electrode into a mirror surface, processing the electrode, and drying for standby application; 3) electrochemically cleaning the electrode, swashing with water, and drying; 4) dropwise adding 6-10 [mu]L of P-rGO-C60 solution intothe surface of the glassy carbon electrode, drying, dropwise adding a GOD solution to the electrode; 5) dropwise adding the P-rGO-C60-Pt@Au solution to the electrode surface, and drying at room temperature; 6) taking an SDM binding aptamer, dropwise adding on the electrode, and incubating at room temperature; and 7) immersing the electrode in the GOD solution, and thus obtaining the product. A method for detecting the SDM by using the sensor is also disclosed.

Description

technical field [0001] The invention relates to the technical field of electrochemical detection, in particular to a nanocomposite material, a preparation method and a detection method of an SDM electrochemical aptamer sensor. Background technique [0002] Sulfadimethoxine (SDM) is a sulfonamide drug that can effectively act on most Gram-positive and negative bacteria. One of the commonly used sulfa drugs. However, the irrational use of SDM leads to drug residues in animal feces or tissues; on the one hand, the manure containing residual drugs is used as fertilizer for agricultural cultivation to pollute the soil and affect the growth of microorganisms in the soil; Foods of animal origin with drug residues can also endanger their health. In order to protect the environment and the health of consumers, the governments of many countries, such as the European Commission and China, have adopted 100ng / mL as the maximum residue limit of SDM in food of animal origin. [0003] Tr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/48G01N27/30
CPCG01N27/30G01N27/308G01N27/48
Inventor 白丽娟母昭德游欢
Owner CHONGQING MEDICAL UNIVERSITY
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