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Portable copper ion concentration detection method based on click chemistry

A portable detection and click chemistry technology, applied in the field of analytical chemistry, can solve the problems of inability to realize on-site detection, inconvenient to carry, and expensive, and achieve the effects of good activity and stability, easy control, and low cost.

Active Publication Date: 2013-06-19
FUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, a common disadvantage of these methods is that large-scale instruments and equipment are required, which are not only expensive, complicated to operate, but also inconvenient to carry, and cannot realize on-site detection

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0028] 1. Synthesis of DNA compounds modified with alkynyl and sucrose invertase. References (Hermanson, G. T. Bioconjugate Techniques; Elsevier: London, 2008). Specific steps are as follows:

[0029] A. Mix 30 μM sulfhydryl-modified DNA with 60 μM TCEP in PBS buffer solution and incubate at room temperature for 1 hour;

[0030] B. Mix 11.30 mg / mL sucrose invertase and 1 mg Sulfo-SMCC in a buffer solution (0.1 M PBS, PH=7.3, 0.1 M NaCl) for 5 minutes, shake on a shaker at room temperature for 1 hour, and then centrifuge the mixture Separate and take the supernatant;

[0031] C. Mix the substances prepared in A and B, and react at room temperature for 48 hours.

[0032] 2. The preparation of magnetic beads modified with sucrose invertase and the detection of copper ions, the specific steps are as follows:

[0033](1) Dissolve the magnetic beads modified with streptavidin in a buffer solution consisting of 0.1M NaCl, 0.1M PBS with pH=7.3, and 0.05% Tween-20, add 0.3 μM DNA m...

Embodiment 2

[0038] The detection of copper ion concentration in human serum, the specific steps are as follows:

[0039] Add 0.6 mM sodium ascorbate to 0.3 μM mixture of the two modified DNAs and dilute 10 5 Double human serum sample, incubated at room temperature for (1.5±0.05) hours, then separated the mixture with a magnet, washed the magnetic beads with 50 μL buffer solution three times and added 10 μL buffer solution, then added 20 μL 1M sucrose solution, reacted at 40 °C for 15 minutes, and transferred Take 2.5 μL to measure with a blood glucose meter and record the data. According to the linear equation Y=0.86807+1.66028lgX of the copper ion concentration obtained in Example 1 and the blood glucose meter reading, the copper ion concentration in the four groups of human serum samples is calculated to be 19.23 pM, 16.74 pM, 16.74 pM, 16.74 pM, 16.74 pM.

Embodiment 3

[0041] In order to detect the specificity of the present invention, the copper ions used in step 2 of Example 1 are replaced with other interfering substances, which are respectively potassium ions, sodium ions, magnesium ions, silver ions, calcium ions, nickel ions and iron ions, and copper ions The concentration of ions and other interfering species was 1 nM. Such as figure 2 (A), from left to right are the blood glucose meter signals shown in different interfering substances, from left to right are copper ions (6.5 mM), potassium ions (1.1 mM), sodium ions (0.9 mM), Magnesium ions (1.4 mM), silver ions (1.8 mM), calcium ions (2.3 mM), nickel ions (1.4 mM) and iron ions (1.7 mM), the corresponding color changes of the color windows of blood glucose test strips are as follows: figure 2 As shown in (B), from 0 to 7 are copper ions, potassium ions, 6.5 sodium ions, magnesium ions, silver ions, calcium ions, nickel ions and iron ions, except for 0 which is obviously green, th...

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Abstract

The invention provides a portable copper ion concentration detection method based on click chemistry and belongs to the field of analytical chemistry. Under the condition that sodium ascorbate reduces cupric into cuprous as a catalyst, a 1,3-dipole cycloaddition reaction is carried out between NDA modified with an azide group and DNA modified with an alkynyl group, so that DNA modified with cane sugar invertase is fixed on magnetic beads; The magnetic beads are separated from a solution, the cleaned magnetic beads are dissolved into a cane sugar solution to carry out enzymatic hydrolysis, and then a reaction solution after enzymatic hydrolysis is measured by adopting a blood glucose meter, so that copper ion concentration can be detected. The portable copper ion concentration detection method provided by the invention takes the blood glucose meter as a platform and can portably and quickly detect the copper ion concentration on site; and the portable copper ion concentration detection method can be applied to copper ion detection in a human body serum sample, and the portable copper ion concentration detection method has the advantages of easy operation, low cost, high sensitivity and good specificity.

Description

technical field [0001] The invention relates to a click chemistry-based portable method for detecting copper ion concentration, which belongs to the field of analytical chemistry. Background technique [0002] Copper is an indispensable trace element in animals (E.Merian, Metals and their compounds in the environment, VCH, Weinheim, 1991, p.893.). In 1945, Braude proposed to add 10 times the amount of copper required by animal physiology to feed. It has a growth-promoting effect and can significantly improve the growth performance of animals, so copper has become one of the necessary additives for animal feed. In addition, copper also has antibacterial and antimicrobial effects. Increasing copper in the feed can enhance the immune function of the animal body (Zhou Mingrong, Wang Jinfa, Wang Zongyuan, Shao Keming, Copper, zinc, manganese, selenium on the growth performance and blood of broilers Research on the influence of some physiological and biochemical indicators in 199...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/78C12N15/11
Inventor 林振宇周津陈国南郭隆华邱彬
Owner FUZHOU UNIV
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