Method for detecting 1,8-diaminonaphthalene based on electrochemical DNA biosensor

A biosensor, diaminonaphthalene technology, applied in the field of electrochemical detection and chemical sensors, can solve the problems of DNA membrane uniformity, poor stability, and less detection of aromatic amine pollutants, etc. The method is simple, easy to implement, and resistant to environmental factors The effect of strong interference performance and low cost

Inactive Publication Date: 2012-09-26
BEIJING NORMAL UNIVERSITY
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  • Abstract
  • Description
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Problems solved by technology

[0005] At present, the application of electrochemical DNA biosensors to the detection of aromatic amine pollutants is still relatively small, and all of them use DNA sequences of natural organisms.
Although these DNA biosensors have high sensitivity to aromatic amine pollutants, their disadvantages are the poor uniformity and stability of the DNA membrane modified to the electrode surface by adsorption.
So far, the application of artificially synthesized disulfide-modified DNA sequences to prepare gold electrode sensors for the detection of aromatic amines has not been reported. On the surface of the gold electrode, the DNA membrane structure is highly ordered and stable, and the DNA base sequence can be designed and adjusted according to the experiment, so as to prepare a specific and highly sensitive DNA biosensor, which is perfecting the existing detection technology in our country. It is of great significance and good application value

Method used

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  • Method for detecting 1,8-diaminonaphthalene based on electrochemical DNA biosensor
  • Method for detecting 1,8-diaminonaphthalene based on electrochemical DNA biosensor
  • Method for detecting 1,8-diaminonaphthalene based on electrochemical DNA biosensor

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Embodiment 1

[0031] 1) Put the gold electrode on the buckskin polishing cloth with 0.05μm α-Al 2 o 3 The powder is polished to a mirror surface. After polishing, the surface dirt is first washed away, and then ultrasonically cleaned in ethanol and deionized water, each time for 2 to 3 minutes, and repeated three times; then in 1mol / L H 2 SO 4 The solution was scanned by cyclic voltammetry until it was stable, and finally rinsed with deionized water and dried with nitrogen to obtain a gold electrode with a clean surface;

[0032] 2) High ionic strength Tris-NaClO with pH=7.4 4 Buffer solution to dissolve solid hairpin DNA, in which Tris concentration is 20mM, NaClO 4 The concentration is 300mM, mixed with a vortex shaker, and left at room temperature for 12 hours, under which conditions the hairpin DNA structure is formed;

[0033] 3) Soak the activated gold electrode in the pre-prepared 5 μM hairpin DNA solution, and let it stand at room temperature for 4-5 days to obtain the hairpin D...

Embodiment 2

[0035] With hairpin DNA modified electrode of the present invention as working electrode, with Ag / AgCl (saturated KCl solution) electrode as reference electrode, platinum electrode as counter electrode to construct three-electrode system, electrolyte solution is 20mmol / L Tris-NaClO of pH=7.4 4 2mM K prepared in buffer solution 3 [Fe(CN) 6 ] / K 4 [Fe(CN) 6 ], the concentration of 1,8-diaminonaphthalene solution preparation is respectively: 0.1nmol / L, 0.3nmol / L, 0.6nmol / L, 3nmol / L, 6nmol / L, 30nmol / L, 60nmol / L, 120nmol / L, 300nmol / L and 600nmol / L. The PARSTAT2273 electrochemical comprehensive test system was used for electrochemical impedance measurement. The experiment shows that the hairpin DNA modified electrode has excellent performance, fast response time, high sensitivity, and the detection linear range of 1,8-diaminonaphthalene: 0.6~120nmol / L.

[0036] Table 1 Electrochemical impedance change (ΔR CT )

[0037]

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Abstract

Disclosed is a method for detecting 1,8-diaminonaphthalene based on an electrochemical DNA biosensor. The invention belongs to the technical field of electrochemical detection and chemical sensors for detecting 1,8-diaminonaphthalene based on a gold electrode modified by hairpin DNA, and relates to preparation of the gold electrode modified by hairpin DNA which has an electrochemical response to the 1,8-diaminonaphthalene and detection of the 1,8-diaminonaphthalene by adopting the electrode. Specifically, the preparation of the gold electrode comprises using hairpin DNA modified by dithio as a raw material, and self-assembling on the gold electrode surface through the action of gold-sulfur bonds to prepare the electrode modified by the hairpin DNA. Changes in impedance of a DNA film are caused by interaction between the electrode modified by the hairpin DNA and the 1,8-diaminonaphthalene, and detection of 1,8-diaminonaphthalene can be carried out according to responded changes in the impedance. A method for preparing the DNA biosensor of the invention has the advantages of mild conditions, simplicity and convenience, good stability, high sensitivity and simple operation.

Description

technical field [0001] The invention belongs to the technical field of electrochemical detection and chemical sensors, and relates to the preparation of an electrochemical DNA biosensor based on hairpin DNA and the use of the prepared hairpin DNA biosensor for detecting 1,8-diaminonaphthalene. Background technique [0002] Aromatic amines are an important class of environmental pollutants and have been listed as one of the priority environmental pollutants for monitoring. Aromatic amines are widely used in industry. As important chemical raw materials and fine chemical intermediates, they are often used in printing and dyeing, pharmaceuticals, food, medicine, explosives, pesticides, cosmetics, plastics, rubber, textiles, papermaking, ceramic glazing and In the production process of paint and other processes, especially in the printing and dyeing industry, wastewater often contains high concentrations of aromatic amine intermediates, and some dyes eventually release aromatic ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/26G01N27/327
Inventor 刘新会梁刚李晓宏巩文雯陶莉付文君
Owner BEIJING NORMAL UNIVERSITY
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