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Method for quantitative detection of mercury ions through gold label silver stain and kit thereof

A technology of gold standard silver staining and quantitative detection, which is applied in the preparation of test samples, material analysis by observing the influence of chemical indicators, and analysis by making materials undergo chemical reactions, etc., to achieve high sensitivity, low price, Ease of pervasive effect

Inactive Publication Date: 2012-10-31
NINGBO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are no relevant research reports on the application of gold-labeled silver staining technology to the quantitative detection of mercury ions at home and abroad.

Method used

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  • Method for quantitative detection of mercury ions through gold label silver stain and kit thereof
  • Method for quantitative detection of mercury ions through gold label silver stain and kit thereof

Examples

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Effect test

Embodiment example 1

[0038] A method for the quantitative detection of mercury ions by gold-labeled silver staining of the present invention specifically comprises the following steps:

[0039] (1) Preparation of silanized glass slides

[0040] Place the glass slide in piranha washing solution, heat it at 90-100°C for 20-40 min, take it out, wash it three times with distilled water, and place it under N 2 After drying in the atmosphere, immerse in 1-3wt% 3-aminopropyltriethoxysilane (APTES) solution, silanize at room temperature for 2-6h, then wash with ethanol, vacuum at 110-130°C After drying for 1-3 hours, a silanized glass slide is obtained, and the piranha washing solution is obtained by mixing 98% concentrated sulfuric acid and 30% hydrogen peroxide at a volume ratio of 7:3;

[0041] (2) Preparation of specific DNA-nanogold probe solution

[0042] Mix 30-40 ul of 100 uM DNA with the sequence HS-5'-TTTTTTTTTT-3' and 0.5-1.5 mL of nano-gold solution with a concentration of 20-25 nM, shake we...

Embodiment example 2

[0055] The present invention is a kit for the quantitative detection of mercury ions by gold-labeled silver staining, comprising silanized glass slides, specific DNA-nano-gold probes and silver staining reagents, and the specific DNA-nano-gold probes are combined with nano-gold Specific DNA, the sequence of the specific DNA is as follows: HS-5'-TTTTTTTTTT-3'.

[0056] In this specific example, the diameter of the gold nanoparticle is 15-30 nm, and the structure of the specific DNA-nanogold probe is AuNPs-HS-5'-TTTTTTTTTT-3'. The silver staining reagent is composed of component A and component B, and the component A is AgNO containing 0.25 g / mL 3 Solution, component B is 0.0425 g / mL of hydroquinone, 0.064 g / mL of citric acid, and 0.059 g / mL of trisodium citrate mixed in an equal volume ratio. Before use, component A and component B Mix at a volume ratio of 3:100 to obtain the silver staining reagent.

[0057] In this specific example, the process of silanizing the glass slide...

Embodiment example 3

[0059] The establishment of the quantitative relationship between the gold-labeled silver-stained signal gray value and the concentration of mercury ions of the present invention specifically comprises the following steps:

[0060] (1) Silanization of glass slides

[0061] The slides were placed in piranha washing solution, heated at 90°C for 40 min, removed, washed three times with distilled water, and placed in N 2 After drying in the atmosphere, immerse in 2% 3-aminopropyltriethoxysilane (APTES) solution for silanization treatment at room temperature for 6 h, then wash with ethanol and then vacuum dry at 110 °C for 3 h. The above piranha washing solution is 98% concentrated sulfuric acid and 30% hydrogen peroxide are obtained by mixing in a volume ratio of 7: 3;

[0062] (2) Preparation of specific DNA-nanogold probe solution

[0063] Mix 34 μL of 100 μM DNA with the sequence HS-5'-TTTTTTTTTT-3' and 1 mL of 22.4 nM nano-gold solution, shake well, incubate at room temperat...

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Abstract

The invention discloses a method for the quantitative detection of mercury ions through gold label silver stain and a kit thereof. The method is characterized by comprising the following steps of: heating, washing and drying a glass slide, immersing the glass slide in an APTES (3-Aminopropyltriethoxysilane) solution, performing silanization treatment on the glass slide at room temperature, then cleaning the glass slide with ethanol, and drying in vacuum at 110-130 DEG C for 1-3h to obtain the silanization glass slide; mixing DNA and a nanogold solution, vibrating to reach a uniform state, incubating at room temperature, and adding water for dilution to obtain a specific DNA-nanogold probe solution; mixing the specific DNA-nanogold probe solution with a sample solution, dripping the mixture on the silanization glass slide, drying a sample point at room temperature completely, dripping a silver staining reagent, standing in a black box, then, immediately rinsing the glass slide with distilled water and drying; and finally, performing quantitative analysis on gold label silver stain signals, and calculating to obtain the accurate concentration of the mercury ions in the sample solution to be detected. The method has the advantages of high sensitivity, high selectivity, availability for quantitative analysis of mercury ion concentration, simplicity for operation and availability for field detection.

Description

technical field [0001] The invention relates to a method for detecting mercury ions, in particular to a method for quantitatively detecting mercury ions by gold-labeled silver staining and a kit thereof. Background technique [0002] Mercury (Hg), the only liquid metal at room temperature, is one of the heavy metals. In recent years, the mining, smelting, processing and commercial manufacturing activities of heavy metal mercury have been increasing, and a large amount of mercury has entered the atmosphere, water and soil for retention, accumulation and migration. Mercury is highly toxic and remains in the organism and the environment for a long time, which seriously threatens human health. Therefore, many countries and regions such as the United States, the European Union, Canada, and my country have strictly limited the mercury content in food, daily necessities, and the environment. Mercury Pollution detection and monitoring has become a major concern. Mercury in nature i...

Claims

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Application Information

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IPC IPC(8): G01N21/78G01N1/30
Inventor 王泽波郭智勇郝婷婷马青青杜书平
Owner NINGBO UNIV
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