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Kanamycin electrochemical detection method based on azido alkyne cycloaddition and electrochemical regulation of atom transfer radical polymerization

A kanamycin and atom transfer technology, applied in the field of life science analysis, can solve the problems of expensive instruments, poor specificity, and complicated detection procedures, and achieve the effects of improved sensitivity, high stability and reproducibility

Inactive Publication Date: 2021-02-09
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Among them, the microbial detection method is simple to operate, economical and practical, but has poor specificity and can only detect biologically active substances, which has certain limitations.
Physical and chemical detection methods mainly include high performance liquid chromatography, liquid chromatography-mass spectrometry, capillary electrophoresis, etc. The methods have high sensitivity, but the detection procedures are complicated and the instruments are expensive

Method used

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  • Kanamycin electrochemical detection method based on azido alkyne cycloaddition and electrochemical regulation of atom transfer radical polymerization
  • Kanamycin electrochemical detection method based on azido alkyne cycloaddition and electrochemical regulation of atom transfer radical polymerization
  • Kanamycin electrochemical detection method based on azido alkyne cycloaddition and electrochemical regulation of atom transfer radical polymerization

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Embodiment 1

[0024] Embodiment 1. Feasibility verification

[0025] Firstly, a detection system is constructed on the electrode surface, such as figure 1 shown. Immediately afterwards, the feasibility verification of the detection system is carried out, and the experimental steps are the same as the content of the invention (1)-(6). Such as figure 2 As shown, firstly, without kanamycin (Kana), the hairpin DNA / MCH / PBIB / FMMA modified electrode (curve b) has a weaker current intensity. On the contrary, after adding Kana, the hairpin DNA / MCH / Kana / PBIB / FMMA modified electrode (curve a) can observe an obvious current peak at 0.3 V, which is because a large amount of FMMA was introduced to the gold electrode through eATRP, resulting in a very large current signal. Through comparative experiments, the feasibility of the electrochemical detection method of kanamycin based on azide-alkyne cycloaddition and electrochemical regulation of atom transfer radical polymerization was fully proved.

Embodiment 2

[0026] Embodiment 2. Sensitivity experiment

[0027] The experimental procedure is the same as the content of the invention (1)-(6), respectively detecting different concentrations of kanamycin (100fg / mL, 1pg / mL, 10pg / mL, 100pg / mL, 1ng / mL, 10ng / mL, 100ng / mL) to study the detection performance of the inventive method for kanamycin. Such as image 3 As shown, the current intensity increases with the concentration of kanamycin, because the higher the concentration of kanamycin, the farther away from the electrode and the exposed N 3 The higher the number, the more CuAAC is carried out and the more initiator PBIB is introduced, which in turn leads to more FMMA electrochemical polymer monomers and stronger current intensity. The logarithmic value of the maximum peak intensity of the current and the concentration of kanamycin presents a good linear relationship in the range of 100fg / mL to 100ng / mL, and the linear equation is I=0.7716+0.4280logC(R 2 =0.9989), the detection limit w...

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Abstract

The invention discloses a kanamycin electrochemical detection method based on azido alkyne cycloaddition and electrochemical regulation of atom transfer radical polymerization, and belongs to the technical field of life science analysis. According to the detection principles, a kanamycin hairpin-like aptamer modified with an N3 group is fixed on the surface of a gold electrode, N3 is far away fromthe surface of the electrode due to change of aptamer conformation when kanamycin appears, azido alkyne cycloaddition reaction with PBIB is carried out under the catalysis of Cu<+>, and then a largeamount of FMMA is connected and polymerized at the tail end of the unfolded hairpin aptamer, and finally, it is determined that the current intensity is positively correlated with the concentration ofthe kanamycin by square wave voltammetry, a standard curve between electrochemical signals and the kanamycin concentration is drawn, so that a linear equation is obtained; the electrochemical signalof a to-be-detected sample is measured, and sensitive detection of the kanamycin can be realized. The method provided by the invention has high selectivity on kanamycin, is simple, quick and sensitive, has good practical value and is convenient to popularize.

Description

technical field [0001] The invention relates to an electrochemical detection method for kanamycin based on azide-alkyne cycloaddition and electrochemically regulated atom transfer radical polymerization, which belongs to the field of life science analysis technology. Background technique [0002] Kanamycin is a basic aminoglycoside antibiotic extracted from the culture solution of Streptomyces, Micromonas or semi-synthetically extracted from natural products. Kanamycin is soluble in water and has a strong effect on Gram-negative bacilli. It is cheap and can be used as an important anti-infection drug in clinic. At present, kanamycin is also one of the veterinary drugs commonly used in my country's agriculture, animal husbandry and aquaculture. Because kanamycin in food of animal origin can accumulate in the human body, it will cause a certain degree of harm to the human body. For example, kanamycin can affect the cochlear nerve, which can induce ototoxicity and cause heari...

Claims

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Application Information

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IPC IPC(8): G01N27/327G01N27/48
CPCG01N27/3277G01N27/48
Inventor 许媛媛李意鲍芳
Owner NANJING AGRICULTURAL UNIVERSITY
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