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CN-122016746-A - Fluorescence detection method for detecting histamine, hydrogel and preparation method and application thereof

CN122016746ACN 122016746 ACN122016746 ACN 122016746ACN-122016746-A

Abstract

The invention provides a fluorescence detection method for detecting histamine, a hydrogel and a preparation method and application thereof, belongs to the technical field of biogenic amine detection, and particularly provides a fluorescence detection method for detecting histamine, which is mainly used for mixing copper nanocluster solution, carbon dot solution and histamine solution with different concentrations, then fixing the volume, detecting fluorescence emission spectrums of various systems under 365 nm excitation wavelength, and making a standard curve by using the ratio F 437 /F 605 of fluorescence intensity at the wavelengths of 437 nm and 605 nm as an ordinate and using the concentration of histamine in various systems as an abscissa for detecting the concentration of histamine in a sample to be detected.

Inventors

  • YANG CHUNLEI
  • ZHANG HONGWEI
  • XU GUIJU
  • HOU CHENGHAO

Assignees

  • 山东省农业科学院

Dates

Publication Date
20260512
Application Date
20260203

Claims (12)

  1. 1. A fluorescence detection method for detecting histamine, comprising the steps of: Mixing the copper nanocluster solution, the carbon dot solution and the histamine solution with different concentrations, fixing the volume to the same volume, reacting and standing to obtain mixed solutions of all systems, detecting fluorescence emission spectra of all systems under 365 nm excitation wavelength, and using the ratio F 437 /F 605 of fluorescence intensity of 437 nm and 605 nm emission wavelength as an ordinate and the concentration of histamine in all systems as an abscissa to manufacture a standard curve for detecting the concentration of histamine in a sample to be detected.
  2. 2. The detection method as claimed in claim 1, wherein in the fluorescence detection method, deionized water is used for fixing the volume to the same volume, and the mixed solution of each system is obtained by standing for more than 90min in room temperature reaction; The solvent of the copper nanocluster solution is ethanol; the final concentration of the copper nanoclusters added into each system is 180-200 mug/mL; When a standard curve is prepared, the final concentration of histamine is 40-120 mu M; the volume of the carbon dot solution added into each system is the same; The volume ratio of the carbon dot solution added into each system to the volume after volume fixing is 1 (15-20); The method comprises the steps of mixing a copper nano-cluster solution, a carbon dot solution and a sample solution to be detected by using the same method as that for manufacturing a standard curve, using deionized water to fix the volume to the same volume, obtaining mixed solutions of each system after reaction and standing, detecting and recording fluorescence emission spectra under 365 nm excitation wavelength, using the ratio F 437 /F 605 of fluorescence intensities at 437 nm and 605 nm emission wavelengths, and calculating the content of histamine in the sample to be detected by using the prepared standard curve.
  3. 3. The method of claim 1, wherein the fluorescence detection method comprises the steps of: Mixing 100 mu L of copper nanocluster solution, 3 mg/mL of carbon dot solution and histamine solution with different concentrations, respectively using deionized water to fix the volume to 1.5 mL, standing for 90 and min to obtain mixed solutions of the systems, detecting fluorescence emission spectra of the systems at 365 nm excitation wavelength, using the ratio F 437 /F 605 of fluorescence intensity of 437 nm and 605 nm emission wavelength as an ordinate, and using the concentration of histamine in the systems as an abscissa to prepare a standard curve for detecting the concentration of histamine in a sample to be detected.
  4. 4. The method of claim 1, wherein the linear range of the method is 40-120. Mu.M and the detection limit is 0.37. Mu.M.
  5. 5. The hydrogel is characterized by comprising copper nanoclusters, a carbon dot solution and polyvinyl alcohol.
  6. 6. The method for preparing the hydrogel according to claim 5, comprising the steps of: uniformly mixing the copper nanoclusters, the carbon dot solution and the polyvinyl alcohol solution to obtain a mixture, and repeatedly freezing and thawing to prepare the hydrogel.
  7. 7. The method of manufacturing as claimed in claim 6, comprising the steps of: 1 (9-10) mg/mL is stirred for 30-40min at 85-90 ℃ to obtain a mixture, the mixture is frozen at-18 to-20 ℃, melted at room temperature and the freezing and thawing process is repeated for more than three times to obtain hydrogel; Or the hydrogel is prepared into different shapes by a mold for use.
  8. 8. Use of the detection method according to any one of claims 1 to 4, the hydrogel according to claim 5 or the hydrogel prepared by the method according to any one of claims 6 to 7 in any one of the following: ① Detecting histamine in a sample; ② The freshness of the samples was evaluated.
  9. 9. The use of claim 8, wherein the sample is an aquatic product.
  10. 10. A method for detecting histamine in a sample using a hydrogel, comprising the steps of: Placing the hydrogel according to claim 5 or the hydrogel prepared by the method according to any one of claims 6 to 7 in contact with or immersing the hydrogel in a sample to be tested, and determining the change of histamine in the sample to be tested according to the color change by irradiating the hydrogel with ultraviolet light.
  11. 11. The method of claim 10, wherein the ultraviolet light is at a wavelength of 365 nm; In the method, the color of the hydrogel is gradually changed from yellow to orange as the histamine content in the sample to be tested is increased.
  12. 12. The detection method according to any one of claims 1 to 4, the hydrogel according to claim 5, or the preparation method according to any one of claims 6 to 7, characterized in that the preparation method of copper nanoclusters comprises the steps of: mixing copper nitrate, 3, 5-bis (trifluoromethyl) thiophenol and dichloromethane according to the mass volume ratio of (0.36-0.50) 1 (120-140) g/mL/mL, stirring at room temperature for more than 8 h, centrifugally collecting precipitate, washing with ethanol for more than 3 times, and drying at the temperature of below 100 ℃ to obtain copper nanoclusters; The preparation method of the carbon dot solution comprises the following steps: Mixing and dissolving glutathione, sodium citrate dihydrate and deionized water according to the mass volume ratio of (0.10-0.12) (1.00-1.03) (27-30) g/g/mL, reacting at 180-200 ℃ for 4-5 h, cooling to room temperature, collecting the product, dialyzing and purifying the product in deionized water, and keeping the dialysis cut-off molecular weight of 500Da and the dialysis cut-off molecular weight of more than 24 h to obtain the carbon dot solution.

Description

Fluorescence detection method for detecting histamine, hydrogel and preparation method and application thereof Technical Field The invention belongs to the technical field of biogenic amine detection, and particularly relates to a fluorescence detection method for detecting histamine, a hydrogel, a preparation method and application thereof. Background Food safety is an important problem worldwide. During processing, storage and transportation, the seafood may be contaminated with internal and external microorganisms, resulting in spoilage and release of biogenic amines. Histamine is one of the most common biogenic amines, which is produced by decarboxylation of histidine by endogenous decarboxylase. Intake of foods containing excessive histamine is associated with food poisoning, and may cause adverse reactions such as headache, skin problems, arterial distension, hypotension, etc., and even pose a threat to human life. The histamine content in food reflects the metabolic activity of microorganisms and can be used as a key index for evaluating food quality and safety. Therefore, it is important to develop a simple and efficient histamine determination strategy. Currently, various analytical methods have been developed for histamine detection, including gas chromatography, mass spectrometry, high performance liquid chromatography, colorimetry, electrochemistry, and immunoassays, among others. However, some of the above methods, such as chromatography, often require complex instrumentation and the sample preparation process is time consuming. Fluorescent sensors are favored for their high sensitivity, low cost detection, rapid response speed, and simple operation. In recent years, many metal-based fluorescent nanomaterials, such as semiconductor quantum dots, metal nanoclusters, and the like, have been used as specific units of various fluorescent sensors. However, these materials often encounter challenges including poor performance in solid state lighting applications, photobleaching, the need for toxic precursors, complex manufacturing processes, and high costs, which have prevented their practical use in detection. Therefore, it is necessary to develop a simple and sensitive fluorescence and visual rapid detection method. Disclosure of Invention Aiming at the defects of the prior art, the invention provides a fluorescence detection method for detecting histamine, a hydrogel, a preparation method and application thereof. The technical scheme of the invention is as follows: a fluorescence detection method for detecting histamine, comprising the steps of: Mixing the copper nanocluster solution, the carbon dot solution and the histamine solution with different concentrations, fixing the volume to the same volume, reacting and standing to obtain mixed solutions of all systems, detecting fluorescence emission spectra of all systems under 365 nm excitation wavelength, and using the ratio F 437/F605 of fluorescence intensity of 437 nm and 605 nm emission wavelength as an ordinate and the concentration of histamine in all systems as an abscissa to manufacture a standard curve for detecting the concentration of histamine in a sample to be detected. According to the preferred fluorescence detection method, deionized water is used for fixing the volume to the same volume, and the mixture solution of each system is obtained after reaction and standing for more than 90 min at room temperature; The solvent of the copper nanocluster solution is ethanol; the final concentration of the copper nanoclusters added into each system is 180-200 mug/mL; When a standard curve is prepared, the final concentration of histamine is 40-120 mu M; the volume of the carbon dot solution added into each system is the same; The volume ratio of the carbon dot solution added into each system to the volume after volume fixing is 1 (15-20); The method comprises the steps of mixing a copper nano-cluster solution, a carbon dot solution and a sample solution to be detected by using the same method as that for manufacturing a standard curve, using deionized water to fix the volume to the same volume, obtaining mixed solutions of each system after reaction and standing, detecting and recording fluorescence emission spectra under 365 nm excitation wavelength, using the ratio F 437/F605 of fluorescence intensities at 437 nm and 605 nm emission wavelengths, and calculating the content of histamine in the sample to be detected by using the prepared standard curve. According to a preferred embodiment of the present invention, the fluorescence detection method comprises the steps of: Mixing 100 mu L of copper nanocluster solution, 3 mg/mL of carbon dot solution and histamine solution with different concentrations, respectively using deionized water to fix the volume to 1.5 mL, standing for 90 and min to obtain mixed solutions of the systems, detecting fluorescence emission spectra of the systems at 365 nm excitation wavelength, using the ratio F 437/F605