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Enzyme based electrochemical biosensing method applied to detection of trace mercury ions

A biosensing and mercury ion technology, applied in the field of environmental analysis, achieves the effects of high sensitivity, high selectivity, and simple operation

Active Publication Date: 2014-12-10
DALIAN UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are few studies on the detection of mercury ions by enzyme-based electrochemical biosensors, and the enzymes used to construct electrochemical biosensors are mainly concentrated on streptavidin-catalase (HRP) and urease, and these two The minimum detection limit of the enzyme-based electrochemical biosensor for mercury ions is much higher than that of the other two (based on nanomaterials or DNA structural transformation) electrochemical biosensors

Method used

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  • Enzyme based electrochemical biosensing method applied to detection of trace mercury ions
  • Enzyme based electrochemical biosensing method applied to detection of trace mercury ions
  • Enzyme based electrochemical biosensing method applied to detection of trace mercury ions

Examples

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Effect test

Embodiment 1

[0029] Embodiment 1: An enzyme-based electrochemical biosensing method for trace mercury ion detection, such as figure 1 shown, including the following steps:

[0030] (1) Grind the gold electrode with a diameter of 2 mm in the suspension of alumina polishing powder with a particle size of 1 μm, 0.3 μm and 0.05 μm for 5-10 minutes successively, and clean it ultrasonically with ethanol and high-purity water for 5 minutes after each grinding, After the electrode is dry, immerse it in the mixed solution of newly prepared concentrated sulfuric acid and 30% hydrogen peroxide (volume ratio 3:1) for 30 minutes, then wash it with high-purity water, and immerse it in 0.5M H 2 SO 4 solution, carry out cyclic voltammetry scanning until the scanning curve is stable, and finally wash with high-purity water and use N 2 blow dry.

[0031] (2) Add 10 μL of probe DNA (pDNA) incubation buffer containing 1 μM to the surface of the cleaned gold electrode, incubate it at 4°C for 24 hours, and for...

Embodiment 2

[0036] Embodiment 2: a kind of enzyme-based electrochemical biosensing method for trace mercury ion detection, such as figure 1 shown, including the following steps:

[0037] (1) Grind the gold electrode with a diameter of 2 mm in the suspension of alumina polishing powder with a particle size of 1 μm, 0.3 μm and 0.05 μm for 5-10 minutes successively, and clean it ultrasonically with ethanol and high-purity water for 5 minutes after each grinding, After the electrode is dry, immerse it in the mixed solution of newly prepared concentrated sulfuric acid and 30% hydrogen peroxide (volume ratio 3:1) for 30 minutes, then wash it with high-purity water, and immerse it in 0.5M H 2 SO 4 solution, carry out cyclic voltammetry scanning until the scanning curve is stable, and finally wash with high-purity water and use N 2 blow dry.

[0038] (2) Add 10 μL of probe DNA (pDNA) incubation buffer containing 1 μM dropwise onto the cleaned gold electrode surface, incubate it at 4 °C for 24 ...

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Abstract

The invention discloses an enzyme based electrochemical biosensing method applied to detection of trace mercury ions, belonging to the field of environment analysis. By virtue of specific actions of thymine and mercury ions and circle amplification function of nucleotide excision enzyme III, the trace mercury ions can be detected; probe DNA hybridizes with target DNA by capturing mercury ions to form double-chain DNA of a special structure; consequently, pDNA is digested by ExoIII, while tDNA is free and continues to hybridize with other pDNAs, and the process is repeated; the pDNA remained after digestion is in direct proportion to [Ru(NH3)6]<3+> electrical signal strength, so that mercury ions can be detected. The enzyme based electrochemical biosensing method has the characteristics of high sensitivity, high selectivity and low cost; the linear detection range of mercury ions is 0.01-500nM; the detection limit is 1pM.

Description

technical field [0001] The invention belongs to the field of environmental analysis and relates to an enzyme-based electrochemical biosensing method for detecting trace mercury ions. Background technique [0002] As a non-essential element, mercury ions can cause permanent damage to human tissues and internal organs [Bontidean I, Mortari A, Leth S, Brown N L, Karlson U, Larsen M M, Vangronsveld J, Corbisier P, Csoregi E. Environ . Pollut., 2004, 131(2), 255-262]. Once mercury ions in sewage enter the human body through the food chain, it is difficult for the human body's own metabolic system to excrete it from the body [Yu C J, Cheng T L, Tseng W L. Biosens. Bioelectron.2009,25(1):204-210] . As far as the toxicity of mercury ions is concerned, heritable toxicity can occur even if the human body is exposed to low-concentration mercury sources for a long time. Therefore, it is of great significance to develop new and simple detection methods for the efficient detection of t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327G01N27/26
Inventor 赵慧敏甘小荣全燮
Owner DALIAN UNIV OF TECH
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