Electrochemical aptamer electrode for kanamycin detection and preparation method of electrochemical aptamer electrode

A kanamycin and electrochemical technology is applied in the field of detection sensors to achieve the effects of fast online detection, sensitive detection and simple operation

Inactive Publication Date: 2014-12-24
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In order to solve the shortcomings of high detection cost, complicated instrument operation and professional operators in the existing detection methods in the above prior art, a nucleic acid-based aptamer was designed Electrochemical aptamer electrode with novel sandwich configuration for kanamycin detection

Method used

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  • Electrochemical aptamer electrode for kanamycin detection and preparation method of electrochemical aptamer electrode
  • Electrochemical aptamer electrode for kanamycin detection and preparation method of electrochemical aptamer electrode
  • Electrochemical aptamer electrode for kanamycin detection and preparation method of electrochemical aptamer electrode

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preparation example Construction

[0040] (1) Preparation of graphene oxide (GO):

[0041] Graphene oxide (GO) was prepared according to classical methods. Briefly, 1.0 g of graphite powder was added to 0.5 g of sodium nitrate (NaNO 3 ) and 3.0 g potassium permanganate (KMnO 4 ) with a mass fraction of 98% in 50 mL of concentrated sulfuric acid (H 2 SO 4 )middle. When the temperature is lower than 20 °C, react for 1 h, then raise the temperature to 35 °C, and continue to stir for 30 min. Then slowly add about 100 mL of ionized water, and after stirring continuously for 20 min, add 50 mL of H 2 o 2(30%), to reduce the residual oxidant, the solution gradually turned bright yellow; filtered while hot, washed with HCl solution and deionized water until no sulfate was detected in the filtrate. The precipitate was dried in a thermostat at 60 °C to obtain GO.

[0042] (2) Synthesis of graphene-polyaniline (GR-PANI) composite

[0043] 2.75 mL of aniline (99%) dissolved in 100 mL of 1 mol L -1 The concentratio...

Embodiment 1

[0049] A method for preparing a sandwich type electrochemical sensor of the present invention, comprising the following steps:

[0050] a. The glassy carbon electrode is first polished in 0.3 and 0.05??m alumina slurry until it becomes a mirror surface, and then rinsed with secondary water;

[0051] b. Add 10 μL of the prepared GR-PANI dropwise on the surface of the electrode and dry it at room temperature overnight; the modified electrode is immersed in the prepared PAMAM-Au solution and kept for 3 h;

[0052] c. After washing several times with secondary water, dissolve 5 μmol L in 10 μL -1 Add the PBS solution of Kanamycin Antibody (KAb) dropwise on the surface of the electrode, keep it at room temperature for 90 min, rinse the electrode with secondary water and PBS to remove unbound KAb;

[0053] d, 10 μL BSA (0.1%) and poly(A) 10 Strand (single-stranded DNA with 10 adenine bases, 5 μmol L -1 ) is used to block the unbound sites on the electrode surface, that is, a sand...

Embodiment 2

[0061] Dissolve 5 μmol L in 10 μL -1 Add the PBS solution of Kanamycin antibody (KAb) dropwise on the surface of the electrode, change the incubation time (30, 60, 90, 120, 150, 180 min) at room temperature, and rinse the electrode with secondary water and PBS to remove unbound KAb, making different electrodes, used to select the best incubation time of KAb, the others are the same as in Example 1.

[0062]

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Abstract

The invention relates to the technical field of detection sensors and particularly relates to an electrochemical aptamer electrode for kanamycin detection. Graphene-polyaniline compound, polyamide-amine dendritic high molecule-gold nano compound, kanamycin anti-body are modified on a glass carbon electrode in sequence from bottom to up; the surface of the electrode is sealed with BSA (Bovine Serum Albumin) and single chain DNA (Deoxyribose Nucleic Acid) containing 10 adenine nucleotide. The preparation method is simple, stable in performance and good in electrode repeating performance and is suitable for practical applications of kanamycin detection in food safety and bio-sensor industrialization; the prepared electrode is low in technical cost and is suitable for the requirement of low cost in industrialization; according to a sandwich type electrochemical sensing system fixed by using the glass carbon electrode as a fixing carrier and based on a DNA aptamer, rapid and online kanamycin detection in food is realized and the detection limit is 4.6x10<-6>mu gmL<-1>.

Description

technical field [0001] The invention relates to the technical field of detection sensors, in particular to an electrochemical aptamer electrode for kanamycin detection, and also to a preparation method of the electrochemical aptamer electrode. Background technique [0002] Kanamycin (Ka) is an aminoglycoside antibiotic produced by the fermentation of Streptomyces sp. and is widely used to treat mistranslation-induced infections and indirectly inhibit translocation during protein synthesis. Like other aminoglycosides, Ka can accumulate in animals and transfer to the food chain, which may cause potential hazards to human health, such as hearing loss, nephrotoxicity and drug allergic reactions. EU organizations have established maximum residues of Ka in edible tissues and milk. In this regard, it is crucial to establish a sensitive and highly selective method for the detection of Ka in clinical diagnosis and in food safety analysis. The currently reported methods for detect...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/30
Inventor 黄加栋徐伟王玉刘素崔敏李杰王虹智郭玉娜许颖崔杰邱婷婷
Owner UNIV OF JINAN
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