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CN-121978184-A - Preparation method and application of gold nanoparticle-laser-induced graphene composite material modified electrode with molecular imprinted polymer and aptamer double recognition

CN121978184ACN 121978184 ACN121978184 ACN 121978184ACN-121978184-A

Abstract

The invention discloses a preparation method and application of a gold nanoparticle-laser-induced graphene composite material modified electrode for molecular imprinting polymer and aptamer double recognition, and belongs to the technical field of flexible electrochemical sensors and kanamycin detection. According to the invention, the gold nanoparticle-laser-induced graphene composite material is used for modifying the electrode as a matrix material, the kanamycin aptamer is fixed through gold-sulfur bonds by utilizing the gold nanoparticle on the surface of the electrode, and the molecularly imprinted polymer layer is further modified, so that an electrochemical sensing electrode with a molecularly imprinted polymer and aptamer dual identification interface is constructed, and the electrochemical sensing electrode is used as a high-performance sensitive element of a sensor, and shows good analysis performance in kanamycin detection of an actual fish sample. The modified electrode prepared by the invention can sensitively detect KANA within the concentration range of 10.0 pmol/L-1.0 mmol/L, the detection limit is as low as 3.3pmol/L, and the selectivity and the stability are good.

Inventors

  • SUN WEI
  • HUANG JITAN
  • GENG JIAQI
  • Lin Faman
  • Lv Yanxin
  • LIU TAO
  • HUANG YITIAN

Assignees

  • 海南师范大学

Dates

Publication Date
20260505
Application Date
20260206

Claims (10)

  1. 1. The preparation method of the gold nanoparticle-laser-induced graphene composite material modified electrode with the double recognition of the molecularly imprinted polymer and the aptamer is characterized by comprising the following steps of: (1) Preparing a gold nanoparticle-laser-induced graphene composite material modified electrode; (2) Mixing an aptamer solution and a kanamycin solution, incubating, and then applying the mixture to the surface of the gold nanoparticle-laser-induced graphene composite material modified electrode to obtain a kanamycin aptamer-gold nanoparticle-laser-induced graphene electrode; (3) And placing the kanamycin aptamer-gold nanoparticle-laser-induced graphene electrode in an aniline solution for electropolymerization, and eluting to obtain the gold nanoparticle-laser-induced graphene composite material modified electrode with double recognition of the molecularly imprinted polymer and the aptamer.
  2. 2. The preparation method of the gold nanoparticle-laser-induced graphene composite material modified electrode according to claim 1 is characterized in that in the step (1), the gold nanoparticle-laser-induced graphene composite material modified electrode is prepared by uniformly mixing chloroauric acid solution and liquid polyimide, coating the mixture on the surface of a substrate, and curing the mixture to obtain a flexible precursor film; The concentration of the chloroauric acid solution is 10-90 mmol/L, and the dosage ratio of the chloroauric acid solution to polyimide is 200 mu L to 4g; the curing temperature is 90-110 ℃, and the curing time is 20-40 min; The laser wavelength of the laser induction treatment is 450nm, the engraving depth of the laser induction treatment is 20-40%, and the laser power of the laser induction treatment is 1.65-2.48W; the temperature of the heat curing treatment is 120-140 ℃, and the time of the heat curing treatment is 20-40 min.
  3. 3. The preparation method of claim 1, wherein in the step (2), the concentration of the aptamer solution is 10-20 μmol/L, the concentration of the kanamycin solution is 10-20 μmol/L, and the volume ratio of the aptamer solution to the kanamycin solution is 4:1, 3:2, 1:1, 2:3 or 1:4.
  4. 4. The preparation method according to claim 1, wherein in the step (2), the incubation temperature is 37 ℃, the incubation volume is 50-150 μl, and the incubation time is 10-30 min.
  5. 5. The preparation method of the aniline compound according to claim 1, wherein in the step (3), the concentration of the aniline solution is 1-5 mmol/L, the aniline solution is phosphate buffer solution containing aniline, and the pH value of the phosphate buffer solution is 6.0-8.0.
  6. 6. The preparation method according to claim 1, wherein in the step (3), the electropolymerization mode is cyclic voltammetry, the number of turns of the electropolymerization is 5-30, the scanning rate of the electropolymerization is 0.05-0.25V/s, and the voltage window of the electropolymerization is-0.2-0.6V.
  7. 7. The preparation method of claim 1, wherein in the step (3), the eluting solution is a mixed solution of methanol and acetic acid, the volume ratio of the methanol to the acetic acid is 9:1, and the eluting time is 10-30 min.
  8. 8. A gold nanoparticle-laser-induced graphene composite modified electrode with double recognition of a molecularly imprinted polymer and an aptamer prepared by the preparation method according to any one of claims 1-7.
  9. 9. An electrochemical sensor, characterized by comprising the gold nanoparticle-laser-induced graphene composite modified electrode with double recognition of a molecularly imprinted polymer and an aptamer according to claim 8.
  10. 10. Use of the molecularly imprinted polymer of claim 8 and aptamer dual-recognition gold nanoparticle-laser-induced graphene composite modified electrode or the electrochemical sensor of claim 9 for detecting kanamycin.

Description

Preparation method and application of gold nanoparticle-laser-induced graphene composite material modified electrode with molecular imprinted polymer and aptamer double recognition Technical Field The invention belongs to the technical field of flexible electrochemical sensors and kanamycin detection, and particularly relates to a preparation method and application of a gold nanoparticle-laser-induced graphene composite material modified electrode with double recognition of a molecularly imprinted polymer and an aptamer. Background Kanamycin (KANAMYCIN, KANA) is widely used as an important representative of aminoglycoside antibiotics for the treatment of various infectious diseases caused by gram-positive and gram-negative bacteria due to its broad-spectrum antibacterial activity. The antibiotics can inhibit protein synthesis by irreversibly binding to bacterial ribosomal 30S subunit, thereby exerting bactericidal effect. However, excessive use or abuse of KANA in the fields of livestock breeding and aquaculture is susceptible to its residues and accumulation in animal tissues, dairy products, and food products such as poultry eggs. After long-term intake of foods of animal origin containing KANA residues, the drugs can accumulate in the body, thereby causing ototoxicity (such as hearing loss, vestibular function injury) and nephrotoxicity (such as tubular injury, renal function decline), which form a potential threat to public health. Therefore, the establishment of a quick, sensitive and reliable KANA residue detection method has important significance for guaranteeing food safety and human health. Currently, methods commonly used for KANA residue analysis mainly include enzyme-linked immunosorbent assay (ELISA), liquid chromatography-mass spectrometry (LC-MS), high Performance Liquid Chromatography (HPLC), surface Plasmon Resonance (SPR), and the like. Although ELISA method has better specificity, is easy to be interfered by matrix and has cross reaction, while LC-MS and HPLC methods have higher sensitivity and accuracy, the ELISA method depends on expensive instruments, has complex pretreatment, takes longer time, has high professional requirements on operators, and the SPR technology is limited by equipment cost, signal stability and other problems. In general, the traditional methods have the defects of large reagent consumption, complicated operation flow, high detection cost, difficulty in realizing on-site rapid detection and the like. In recent years, electrochemical sensing technology has received wide attention in the field of antibiotic residue detection because of its remarkable advantages of simple equipment, rapid response, low cost, easy miniaturization, and realization of real-time detection. The method is based on the change of signals such as current, potential or impedance caused by the electrochemical behavior of the target object on the surface of the electrode, so that the qualitative or quantitative analysis of the target object is realized. By designing a high-selectivity identification interface, the identification capability and the detection sensitivity of the electrochemical sensor to KANA can be further improved. Therefore, developing a novel KANA detection method based on electrochemical sensing is not only helpful for making up the defects of the traditional analysis technology, but also provides a technical path with prospect for realizing on-site and rapid screening of antibiotic residues in food. Disclosure of Invention In order to solve the technical problems, the invention provides a preparation method and application of a gold nanoparticle-laser-induced graphene composite material modified electrode with double recognition of a molecularly imprinted polymer and an aptamer. In order to achieve the above purpose, the present invention provides the following technical solutions: the invention provides a preparation method of a gold nanoparticle (AuNPs) -laser-induced graphene (LIG) composite material modified electrode with double recognition of Molecularly Imprinted Polymers (MIPs) and an aptamer (Apt), which comprises the following steps: (1) Preparing a gold nanoparticle-laser-induced graphene composite material modified electrode; (2) Mixing an aptamer solution and a kanamycin solution, incubating, and then applying the mixture to the surface of the gold nanoparticle-laser-induced graphene composite material modified electrode to obtain a kanamycin aptamer-gold nanoparticle-laser-induced graphene electrode; (3) And placing the kanamycin aptamer-gold nanoparticle-laser-induced graphene electrode in an aniline solution for electropolymerization, and eluting to obtain the gold nanoparticle-laser-induced graphene composite material modified electrode with double recognition of the molecularly imprinted polymer and the aptamer. Further, in the step (1), the preparation of the gold nanoparticle-laser-induced graphene composite material modified electrode comprises the steps