Aptamer sensor for mercury ion detection, as well as preparation method and application of aptamer sensor

An aptamer sensor, mercury ion technology, applied in the field of food safety detection and analytical chemistry, can solve the problems of large-scale equipment, high cost, cumbersome and time-consuming processing process, etc., and achieve the effect of strong anti-interference ability and strong specificity

Inactive Publication Date: 2017-06-13
南京百圃环境科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although these methods have high accuracy, they often require large-scale instruments and equipment, and the cost is relatively high. The processing process is cumbersome and time-consuming, which cannot meet the needs of high efficiency and low cost in actual monitoring.

Method used

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  • Aptamer sensor for mercury ion detection, as well as preparation method and application of aptamer sensor
  • Aptamer sensor for mercury ion detection, as well as preparation method and application of aptamer sensor
  • Aptamer sensor for mercury ion detection, as well as preparation method and application of aptamer sensor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] The preparation of embodiment 1 nanometer gold-aptamer conjugate

[0029] 1) Place 10ml of prepared nano-gold solution in a 10ml centrifuge tube, discard the supernatant after centrifuging at 12000 rpm for 15 minutes, and dissolve the lower supernatant in 10ml of phosphate buffer solution (pH = 7.4), and put it in a refrigerator at 4°C for later use;

[0030] 2) Dissolve the aptamer in TE buffer solution (0.1mmol / L), add 2μl of TCEP solution (10mmol / L) to it, and react at room temperature for 2 hours;

[0031] 3) Add thiol-activated aptamer solution to 1ml of the nano-gold solution described in step 1), react at room temperature for 16 hours, and slowly add NaCl solution to it several times until the final concentration of NaCl is 0.1mol / L , care should be taken to prevent gold nanocoagulation during the addition process;

[0032] 4) React the solution described in step 3) at room temperature for 48 hours, centrifuge at 12000 rpm for 30 minutes, discard the supernatan...

Embodiment 2

[0033] Embodiment 2 is to the detection of mercury ion standard sample

[0034] 1) Preparation of aptasensor:

[0035] A glassy carbon electrode with a diameter of 3 mm was polished on a polished flannelette with 0.3 μm alumina powder, rinsed with distilled water, and then ultrasonicated in absolute ethanol-distilled water (V / V=1 / 1) and distilled water for 30 s. Insert the above electrode into 3mmol / L chloroauric acid solution for constant potential electrochemical deposition (voltage is -0.2V, deposition time 60s), rinse with deionized water and then immerse in the hexanedithiol solution with a concentration of 1mmol / L in 37°C for 1 hour. Rinse the above electrodes with deionized water, drop-coat 20 μl of the nano-gold-aptamer solution described in Example 1 on the surface, and incubate in a refrigerator at 4°C for 24 hours to obtain nano-gold / hexanedisulfide Alcohol / nano gold / aptamer modified aptasensor;

[0036] 2) Detection of mercury ion standard sample

[0037] Immer...

Embodiment 3

[0038] The mensuration of adding standard mercury ion in lake water of embodiment 3

[0039] Measure 2.0ml of lake water sample. The lake water is taken from the inner lake of South Normal University. Add different volumes of mercury ion standard solutions to prepare a series of samples to be tested containing different concentrations of mercury ions with a total volume of 10.0ml. Immerse the aptamer sensor described in step 1) of Example 2 into the above-mentioned sample to be tested, incubate at 37°C for 30 minutes, rinse with Tris-HCl buffer and place in K containing 2mmol / L 3 [Fe(CN) 6 ] in the KCl solution (0.1mol / L) to carry out electrochemical impedance spectroscopy (EIS) scanning, with the charge transfer resistance change value Δ R obtained in the same method as step 2) in Example 2 ct and ΔR ct -lgC working curve, calculate the concentration C of mercury ions, and the detection recovery results are shown in Table 1.

[0040] Table 1 shows the recovery rate of the ...

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Abstract

The invention discloses an aptamer sensor for mercury ion detection, as well as a preparation method and application of the aptamer sensor. The sensor comprises a substrate glassy carbon electrode, wherein nano-gold particles are deposited on the surface of the substrate glassy carbon electrode; the surface of nano-gold particles is modified with ethanthiol; and a nano-gold-aptamer conjugate is modified to the nano-gold particles on the surface of the electrode through the self-assembly effect of Au-S bond. The aptamer sensor has a relatively strong amplification effect by adopting dual-layer nano-gold modification, has ultrahigh sensitivity for detecting mercury ions reaching 1.1pg/mL, and has a linear range of 5-2000pg/mL; meanwhile, the aptamer has strong interference resistance due to strong specificity, and does not interfere with Ag<+>, Ba<2+>, Cd<2+>, K<+>, Na<+>, Pb<2+>, Cu<2+> and other ions with 200 times of concentration.

Description

technical field [0001] The invention belongs to the technical field of food safety detection and analytical chemistry, and relates to an aptamer sensor for mercury ion detection, a preparation method and application thereof. Background technique [0002] Mercury is a highly toxic global environmental pollutant that seriously endangers the environment and human health. Mercury pollution mostly comes from solid waste incineration and fossil fuel combustion, which can exist in the atmosphere for a long time and have seriously polluted some water sources and soil. Moreover, inorganic mercury can be converted into more toxic methylmercury under the action of some microorganisms in the marine environment, and accumulate in all links of the food chain, and finally enter the human body through the food chain, and will be deposited in the brain, liver and other organs. Produce chronic poisoning, damage kidney, brain, stomach and intestinal tract, and even cause death. Therefore, th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327
CPCG01N27/3278
Inventor 赵波徐红平邵科峰魏梦
Owner 南京百圃环境科技有限公司
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