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Electrochemical transducer for mercury ion detection and manufacturing method and detection method thereof

A detection method and electrochemical technology, applied in the field of biological analysis, can solve the problems of narrow detection range and insufficient sensitivity, and achieve the effects of high specificity, high detection sensitivity and length reduction.

Active Publication Date: 2014-07-02
CAPITAL NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The sensitivity and selectivity of these sensors have been greatly improved, but most of these methods require additional reagents during detection, which is not a simple method for one-step detection, and the sensitivity is not high enough or the detection range is narrow

Method used

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  • Electrochemical transducer for mercury ion detection and manufacturing method and detection method thereof
  • Electrochemical transducer for mercury ion detection and manufacturing method and detection method thereof
  • Electrochemical transducer for mercury ion detection and manufacturing method and detection method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0045] Example 1: Preparation of Hg using Probe A and Probe B 2+ Detection of electrochemical sensors (based on Method A).

[0046] 1 μM Probe A (full thymine (T) oligonucleotide chain with terminal sulfhydryl modification) was reduced in 100 μM tris[2-carboxyethyl]phosphine (TCEP), 10 mM Tris, 1.0 M NaCl, pH=8.0 at room temperature 1h, soak unmodified and clean gold electrodes in it, and self-assemble overnight at room temperature. The assembled electrode was rinsed twice with 10mM Tris, 1.0M NaCl, pH = 8.0, then placed in an aqueous solution containing 1mM mercaptohexanol (MCH) to block for 30 minutes, and then washed with 10mM Tris, 1.0M NaCl, pH = 8.0 Rinse three times, dry and set aside; Dilute the ferrocene-labeled probe B (full adenine (A) oligonucleotide complementary strand) to 50nM with hybridization solution 1 / 15M PB, 0.3M NaCl, pH=7.4 , soak the gold electrode assembled with all thymine (T) oligonucleotide chains in it, react at 25°C for 3h, and then wash three t...

Embodiment 2

[0047] Example 2: Preparation of Hg using Probe C and Probe D 2+ Detection of electrochemical sensors (based on method B).

[0048] 1 μM Probe C (full thymine (T) oligonucleotide chain with terminal sulfhydryl modification) was reduced in 100 μM tris[2-carboxyethyl]phosphine (TCEP), 10 mM Tris, 1.0 M NaCl, pH=8.0 at room temperature 1h, soak unmodified and clean gold electrodes in it, and self-assemble overnight at room temperature. The assembled electrode was rinsed twice with 10mM Tris, 1.0M NaCl, pH = 8.0, then placed in an aqueous solution containing 1mM mercaptohexanol (MCH) to block for 30 minutes, and then washed with 10mM Tris, 1.0M NaCl, pH = 8.0 Rinse three times and dry it for later use; Dissolve the probe D (full thymine (T) oligonucleotide chain complementary to all adenine (A) oligonucleotide complementary chain) labeled ferrocene to 50 mM Dithiothreitol (DTT), 2% triethylamine (TEA) aqueous solution, room temperature, reaction 20min. Use NAP-5column to pass t...

Embodiment 3

[0049] Embodiment 3: Utilize the electrochemical sensor prepared by probe A and B to investigate Hg 2+ reaction time kinetics.

[0050] The electrochemical sensors prepared with probes A and B were soaked to 1 nM Hg 2+ Soak in the buffer solution (1 / 15M PB, 0.3M NaCl, pH=7.4) at room temperature, take out the electrode every reaction for 5 minutes, wash with 1 / 15M PB, 0.1M NaCl, pH=7.4 solution for 3 times, and use a multi-channel constant Potentiometer (VMP3) to do SWV scan and analyze the results ( figure 2 ).

[0051] The results showed that Hg 2+ Inducing the release of probe B is a relatively rapid process, and 1nM Hg 2+ The reaction was basically completed within 20 min. At 25min, the SWV peak current no longer decreases.

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Abstract

The invention relates to a mercury ion electrochemical transducer based on oligonucleotide chains and a manufacturing method and a detection method thereof and belongs to the technical field of bioanalysis. The method for manufacturing the sensor comprises the following steps of: fixing a full thymine (T) oligonucleotide chain subjected to chemical modification to a gold electrode; and soaking the gold electrode fixed with the full thymine (T) oligonucleotide chain into hybridization solution of full adenine (A) oligonucleotide complementary chain. The Hg2<+> can be specifically covalently bound with T bases on two adjacent full thymine (T) oligonucleotide chains, a stable intermolecular T-Hg2<+>-T structure is formed, and release of the full adenine (A) oligonucleotide complementary chain which is hybridized with the full thymine (T) oligonucleotide chain is induced. The complementary chain is released through mercury ions, so that the electrochemical signal is reduced, and the electrochemical detection of the mercury ions is realized. The detection sensitivity is greatly improved, and the electrochemical transducer has high ion selectivity.

Description

technical field [0001] The invention relates to the production and application method of an oligonucleotide chain-based mercury ion electrochemical sensor, which belongs to the technical field of biological analysis. Background technique [0002] Mercury is a highly toxic global environmental pollutant, especially its high mobility, persistence, methylation, bioaccumulation, and food chain amplification. The health of animals, plants and humans is also a great threat. The annual emission of mercury in the world is about 15,000 tons, mainly from mercury mines, metallurgy, chlor-alkali industry, electrical industry and combustion of fossil fuels. Mercury exists in various forms in the environment, water-soluble divalent mercury ions (Hg 2+ ) is one of the most common and stable forms of mercury pollution. [0003] How to effectively measure the content of mercury ions in the environment is still a challenge for the majority of analysts. At present, the traditional mercury ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/416
Inventor 娄新徽赵滔
Owner CAPITAL NORMAL UNIVERSITY
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