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Method for detecting kanamycin sulfate by double-signal enhanced surface enhanced Raman spectroscopy

A surface-enhanced Raman and spectral detection technology, which is applied in Raman scattering, measuring devices, and material analysis through optical means, can solve problems such as complex detection procedures, high detection limits, and low detection limit sensitivities

Active Publication Date: 2021-04-27
SHANDONG AGRICULTURAL UNIVERSITY
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Problems solved by technology

Among them, high-performance liquid chromatography and chromatography-mass spectrometry are used to detect the content of kanamycin sulfate in food accurately and reliably, but kanamycin sulfate lacks a chromophore, and pre-column derivatization is required when using fluorescence or ultraviolet detectors for detection or post-column derivatization, so the detection procedure is more complicated and requires higher requirements for experimenters; although the microbiological method is simple to operate, economical and practical, but the detection limit is high and can only detect biologically active substances; ELISA has high specificity , high sensitivity, but the preparation of antibodies is difficult and the cost is high; capillary electrophoresis technology is applied to the detection and analysis of kanamycin sulfate fast and requires a small amount of sample, but the repeatability is poor and the detection limit is high, and field amplification is required , solid phase extraction and other techniques
Therefore, there are still challenges in the detection of kanamycin sulfate in food in terms of low detection limit, high sensitivity, fast and reliable

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  • Method for detecting kanamycin sulfate by double-signal enhanced surface enhanced Raman spectroscopy
  • Method for detecting kanamycin sulfate by double-signal enhanced surface enhanced Raman spectroscopy
  • Method for detecting kanamycin sulfate by double-signal enhanced surface enhanced Raman spectroscopy

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preparation example Construction

[0053] In one embodiment of the present invention, a method for preparing a size-controllable gold nanorod as a Raman substrate is provided, comprising the steps of:

[0054] Mix 5mL 0.2mol / L CTAB solution with 5mL 0.0005mol / L HAuCl 4 The solution is mixed. Add 0.6 mL of freshly prepared 0.01 mol / L NaBH to the stirred solution 4 solution, namely the seed solution.

[0055] 4.5mL of 2.5×10 -4 mol / L HAuCl 4 4.5 mL of 0.1 mol / L CTAB solution was mixed with 0.05 mL of 0.1 mol / L ascorbic acid solution to obtain a growth solution.

[0056] Take 1 mL of the seed solution and add it to 9.05 mL of the growth solution, and react for 4 hours to obtain liquid A. Take 1 mL from solution A and add it to 9.05 mL of newly prepared growth solution, and react for 4 hours to obtain solution B. Take 1 mL from solution B and add it to 9.05 mL of newly prepared growth solution, and react for 12 hours to obtain single crystal gold nanorods.

[0057] 3.64g CTAB and 66mg HAuCl 4 ·3H 2 Dissolv...

Embodiment 1

[0067] 1. Preparation of size-controllable gold nanoparticles:

[0068] (1) Preparation of single crystal gold nanoparticles:

[0069] Mix 5mL 0.2mol / L CTAB solution with 5mL 0.0005mol / L HAuCl 4 The solution is mixed. Add 0.6 mL of freshly prepared 0.01 mol / L NaBH to the stirred solution 4 solution, namely the seed solution.

[0070] 4.5mL of 2.5×10 -4 mol / L HAuCl 4 4.5 mL of 0.1 mol / L CTAB solution was mixed with 0.05 mL of 0.1 mol / L ascorbic acid solution to obtain a growth solution.

[0071]At 28°C, add 1 mL of seed solution to the growth solution and react for 4 hours (solution A); take 1 mL from solution A and add new growth solution, and react for 4 hours (solution B); take 1 mL of solution B and add new In the growth solution, after 12 hours of reaction, a single crystal gold nanorod is obtained.

[0072] (2) Dissolution of single crystal gold nanorods: 3.64g CTAB and 66mg HAuCl 4 ·3H 2 O was dissolved in 100 mL of deionized water, 1.5 mL of the above solution ...

Embodiment 2

[0081] Using the surface-enhanced Raman spectroscopy analysis method based on the nucleic acid aptamer-controlled metal-doped carbon quantum dot catalytic switch and the size-controllable gold nanoparticle double-enhanced signal established in Example 1 to analyze the kanamycin sulfate in the milk sample detection, as follows:

[0082] (1) Accurately take 1 g of milk sample into a 50 mL centrifuge tube, add 30 mL of 5% phosphoric acid solution by volume, shake and extract for 10 min, and add 3 mL of 50% by volume trichloroacetic acid solution. After vortex mixing, centrifuge at 944×g for 10 min. This process was repeated twice and the supernatants were pooled. Concentrate the components to be tested under reduced pressure, redissolve in water to 1 mL, and filter through a 0.22 μm filter membrane to obtain the solution to be tested.

[0083] (2) Add 200 μL of 0.03 mol / L nucleic acid aptamer into a 5 mL test tube, add 50 μL of the solution to be tested and 70 μL of metal zinc-...

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Abstract

The invention discloses a method for detecting kanamycin sulfate by double-signal enhanced surface enhanced Raman spectroscopy, and belongs to the technical field of food safety detection. In the invention, gold nanorod with a controllable surface plasma resonance peak is used as a Raman substrate, and a cascade oxidation-reduction reaction catalyzed by metal-doped carbon quantum dots and a switching structure controlled by a nucleic acid aptamer are combined to establish a kanamycin sulfate detection system with double enhanced Raman signals, so that the sensitivity of detecting kanamycin sulfate is further improved. The method is low in detection limit, good in specificity, simple, convenient and rapid, and suitable for ultra-sensitive detection of kanamycin sulfate in food.

Description

technical field [0001] The invention relates to the technical field of food safety detection, in particular to a method for detecting kanamycin sulfate by surface-enhanced Raman spectroscopy with double signal enhancement. Background technique [0002] With the improvement of people's quality of life, the problem of antibiotic drug residues has become one of the focus issues that consumers pay attention to. Kanamycin sulfate is one of the high-efficiency aminoglycoside antibiotics widely used in the treatment of animal infectious diseases in the food production and manufacturing industry. Its structural formula is as follows: [0003] [0004] Kanamycin sulfate is composed of amino sugar molecules and amino cyclic alcohols linked by ether bonds. It mainly kills bacteria quickly by interfering with the synthesis of proteins in bacteria. Kanamycin sulfate remaining in foods such as meat, eggs, milk, etc. will accumulate in the human body. Long-term intake of foods with exc...

Claims

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Application Information

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IPC IPC(8): G01N21/65
CPCG01N21/658
Inventor 徐志祥王汐沫陈晨徐龙华孔非凡
Owner SHANDONG AGRICULTURAL UNIVERSITY
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