Detection kit and detection method of mercury ion

A technology for detection kits and detection methods, which is applied in the field of mercury ion detection, can solve problems such as limitations and low ion concentration tolerance, and achieve the effects of low cost, improved catalytic activity, and high stability

Inactive Publication Date: 2014-11-19
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the Au/Hg 2+ Nano-alloys have low tolerance to the ion concentration in the detection sample, therefore, in Au/Hg 2+ When nanomimetic enzymes are applied to actual s

Method used

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  • Detection kit and detection method of mercury ion
  • Detection kit and detection method of mercury ion
  • Detection kit and detection method of mercury ion

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] (1) Take 80 μL of gold nanoparticles with an average particle diameter of 15 nm prepared by the reduction method of diluted sodium citrate (concentration of 6×10 -10 M), 10 μL of 2 μM ssDNA solution and 10 μL of sodium chloride solution (1.0 M) are mixed thoroughly, and incubated for 30 min at room temperature. Then centrifugation to remove the free oligonucleotide, while retaining the complex of the oligonucleotide and the gold nanoparticles;

[0035] (2) Place 10 μL of the mixed solution in step (1) into the microtiter plate, add 44 μL 1.1 mM TMB solution, 28 μL 5 M H 2 O 2 Solution, 40 μL Hg of different concentrations (0-1000nM) 2+ Solution, 78 μL of sodium citrate citrate buffer solution (0.04 M, pH 4.5), shake and mix thoroughly. Measure the absorbance at 650 nm when the mixed solution reacts for 10 min. Hg obtained by this method 2+ The detection standard curve such as image 3 As shown, the detection sensitivity can reach 3.0 nM, and the detection linear range is ...

Embodiment 2

[0040] (1) Take 80 μL of diluted gold nanoparticles prepared by sodium citrate reduction method with an average of 30 nm (concentration of 4×10 -10 M), mix 10 μL of 3 μM ssDNA solution and 10 μL of sodium chloride solution (1.0 M) thoroughly, and incubate for 30 min at room temperature. Then centrifugation to remove the free oligonucleotide, while retaining the complex of the oligonucleotide and the gold nanoparticles;

[0041] (2) Put 10 μL of the mixed solution in step (1) into the microtiter plate, add 44 μL 1.5 mM TMB solution, 28 μL 5.5 M H 2 O 2 Solution, 40 μL Hg of different concentrations (0-1000nM) 2+ Solution, 78 μL of sodium citrate citrate buffer solution (0.04 M, pH 3.5), shake and mix thoroughly. Measure the absorbance at 650 nm when the mixed solution reacts for 10 min. Hg obtained according to this example 2+ According to the test results, a standard curve similar to that of Example 1 can also be obtained.

[0042] (3) Hg in tap water 2+ The detection: the tap wat...

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Abstract

The invention discloses a detection kit and a detection method of mercury ions. The kit includes: gold nanoparticles which are used for combining with Hg<2+> to form an horseradish peroxide mimetic enzyme; a single-chain nucleic acid which is used for combining with the gold nanoparticles to enhance a catalytic activity of the horseradish peroxide mimetic enzyme; a characteristic substrate of the horseradish peroxide mimetic enzyme; and auxiliary reagents including a sodium citrate buffer solution which is required when an oxidized characteristic substrate is catalyzed by the horseradish peroxide mimetic enzyme. The detection method is carried out mainly on the basis of the detection kit. By means of the kit and the method, high-sensitivity detection of Hg<2+> can be achieved with a linear range of the detection of the Hg<2+> being 10-1000 nM and a sensitivity being 3.0 nM. The kit and the method are simple, convenient and quick, are low in cost, are high in stability and are suitable for being used in detection of the Hg<2+> in samples of environment, food and the like.

Description

technical field [0001] The invention particularly relates to a method for improving the stability of nano-gold-mercury ion mimic enzyme by using single-stranded nucleic acid and applying it to mercury ion detection, which belongs to the field of nanotechnology. Background technique [0002] Enzymes are a class of biocatalysts, proteins with catalytic functions. Chemical reactions can be catalyzed in a suitable environment, but due to the inherent characteristics of enzymes, their applications are also limited, such as proteases are easy to denature and hydrolyze, and high costs and strict conditions of use also limit their applications. Therefore, it is particularly important to develop a mimetic enzyme with similar catalytic activity. Mimic enzymes are a class of artificially synthesized catalysts that have no protein structure but have similar catalytic performance to natural enzymes. Since 2007, Yan Xiyun's research group first discovered magnetic ferric iron tetroxide ...

Claims

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Application Information

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IPC IPC(8): G01N21/31
Inventor 彭池方泮秋立刘春丽刘丽强
Owner JIANGNAN UNIV
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