CN113640258A - Thin film type fluorescence sensor and preparation method and application thereof - Google Patents
Thin film type fluorescence sensor and preparation method and application thereof Download PDFInfo
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- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
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- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/02—Use of particular materials as binders, particle coatings or suspension media therefor
- C09K11/025—Use of particular materials as binders, particle coatings or suspension media therefor non-luminescent particle coatings or suspension media
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
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- C09K11/08—Luminescent, e.g. electroluminescent, chemiluminescent materials containing inorganic luminescent materials
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- G—PHYSICS
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- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
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- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
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Abstract
The invention provides a thin film type fluorescence sensor and a preparation method and application thereof. The film type fluorescence sensor provided by the invention judges the freshness of fish meat through the fluorescence intensity change in the storage period, realizes rapid nondestructive detection, and has high sensitivity and accuracy.
Description
Technical Field
The invention relates to the field of food detection, in particular to a thin film type fluorescence sensor and a preparation method and application thereof.
Background
The aquatic products are extremely easy to decay and deteriorate in the storage and transportation process due to the conditions of high nutrition, high water activity, attachment of a large amount of microorganisms, nearly neutral pH value and the like. Spoilage of aquatic products not only increases sales costs, but also increases the risk of food-borne diseases for consumers. Freshness detection is one of the important means for preventing aquatic product spoilage. The amino acid and protein in the aquatic product are decomposed under the action of enzyme and bacteria to generate volatile alkaline nitrogen-containing substances called volatile basic nitrogen (TVB-N), and the content of the TVB-N is closely related to the content of enzyme and microorganism in the aquatic product, so that the TVB-N is used as an important index for measuring the freshness of animal food. However, the traditional laboratory chemical detection method needs to use alkali liquor to absorb volatile amine-containing substances and use standard acid solution to perform titration, so that the operation is complex, and time and labor are wasted. Compared with the traditional methods, the freshness nondestructive detection method, such as a near infrared spectrum technology, a machine vision technology, a gas-sensitive sensing technology and the like, has certain advantages, but still has the problems of complex data processing, incapability of realizing online detection, low accuracy, poor stability and the like, and can not completely realize ideal rapid nondestructive detection. Therefore, it is urgently needed to develop a quick freshness detection method which is simple, convenient, easy, low in cost, safe and efficient.
Carbon Dots (CDs) are widely used in the fields of cell imaging, visible light catalysis and the like due to the advantages of fluorescence characteristics, low biological toxicity, easiness in synthesis and the like. Zhao Li et al synthesized for Cu by solvolysis of citric acid and histidine2+The measured high-selectivity nitrogen-doped carbon quantum dot (N-CQDs) fluorescence sensor realizes the detection of Cu in environmental water samples and fruit juice samples2+And (6) detecting. Li Yingping and the like adopt a hydrothermal synthesis method, Thiamine Nitrate (TN) is used as a single raw material to prepare N, S-CDs with the fluorescence maximum excitation wavelength of 368nm, and the determination of lemon yellow in two energy beverages and soda beverages is realized based on a fluorescence quenching principle. CDs have been applied as fluorescent sensors for food safety detection. Although there are many reports on nondestructive freshness detection, there are few studies on nondestructive freshness detection using CDs as a fluorescence sensor. The invention discloses a visible light/fluorescence double-signal indicator (bromothymol blue-carbon quantum dot indicator), which generates a change of visible light color and fluorescence color by reacting with volatile components generated by meat decay, and monitoring the freshness of meat products can be realized by utilizing the change of the visible light color and the fluorescence color. The indicator film is based on bromothymol blue-carbon quantum dotsThe fluorescence quenching principle is adopted, and volatile components generated by meat decay recover the fluorescence of the carbon quantum dots. However, the color change of bromothymol blue is greatly influenced by carbon dioxide, which limits the application of the indicating film in modified atmosphere packaging, and the bromothymol blue has irritation and is difficult to avoid contacting meat products when used in food packaging, so whether the bromothymol blue is compounded with food safety grade, whether the toxic effect of the bromothymol blue is enriched in the meat products or not is clearly researched. At present, the problem that freshness is difficult to rapidly detect in a nondestructive mode in the fish storage and transportation process is urgently needed to be solved in the field, and therefore a more effective technical means is provided for quality monitoring and tracking of fish.
Disclosure of Invention
The invention provides a thin film type fluorescence sensor and a preparation method and application thereof. The thin-film fluorescence sensor provided by the invention has the advantages that the fluorescence intensity changes in the storage period, the freshness of fish meat is judged, the rapid nondestructive detection is realized, the TVB-N content of the fish meat is quantitatively detected, and the thin-film fluorescence sensor has high sensitivity and accuracy.
The invention provides a film type fluorescence sensor which is prepared from raw materials including N and S co-doped carbon quantum dots, a matrix and a blending modifier, wherein the matrix is gelatin, and the blending modifier is nafion solution. According to the invention, the (N, S-CDs-nafion/gelatin) film type fluorescence sensor better reflects the freshness of the fish meat through the fitting relation between the fluorescence intensity and the fish meat TV B-N, can quantitatively detect the TVB-N content of the fish meat, has higher sensitivity and accuracy, and provides a more effective technical means for monitoring and tracking the quality of the fish meat. Particularly, the N, S co-doped carbon quantum dots (N, S-CDs) are prepared by taking citric acid or fruit acid as a carbon source, methionine as a nitrogen source and a sulfur source, gelatin is taken as a matrix, nafion is taken as a blending modifier, the (N, S-CDs-nafion/gelatin) film type fluorescence sensor is manufactured, under the action of nafion, the N, S-CDs are better dispersed in the gelatin matrix, and the sensitivity of the N, S-CDs-nafion/gelatin film type fluorescence sensor is improved.
According to the film type fluorescence sensor provided by the invention, in the raw material of the N, S co-doped carbon quantum dot, a carbon source is fruit acid, the fruit acid is preferably citric acid or malic acid, and a nitrogen source and a sulfur source are methionine; preferably, the molar ratio of the fruit acid to the methionine is 1-4: 1-2, preferably 2: 1. According to the invention, by adopting the specific carbon source, the nitrogen source and the sulfur source and doping the nitrogen element and the sulfur element into the CDs in one step under the dosage ratio, the method has the advantages of low cost and simplicity and convenience in operation, and the N, S-CDs have abundant functional groups on the surface, so that the selectivity and the sensitivity to ammonia are improved.
According to the thin film type fluorescence sensor provided by the invention, the mass-to-volume ratio of the N, S co-doped carbon quantum dots, the gelatin and the nafion solution is 2-2000 mg: 1-5 g: 1-10 ml, and the concentration of the nafion solution is 20-40%. In the invention, by adopting the gelatin and nafion solution, particularly under the dosage proportion, the dispersion degree of N, S-CDs in the gelatin matrix can be improved, thereby improving the fluorescence intensity of the film type fluorescence sensor and the response characteristic to ammonia.
The invention also provides a preparation method of the thin film type fluorescence sensor, which comprises the following steps:
1) mixing the gelatin aqueous solution with the nafion solution to obtain a blended solution;
2) mixing the N, S co-doped carbon quantum dots with the blending solution, dissolving, and performing ultrasonic degassing to obtain a mixed solution of N, S-CDs;
3) preparing a film from the mixed solution, and then cutting the film. The (N, S co-doped carbon quantum dot) film type fluorescence sensor synthesized by the invention is used as a sensor for detecting freshness, is used in storage and transportation environments of fish, and can achieve rapid nondestructive detection on the quality of the fish.
According to the preparation method of the film-type fluorescence sensor provided by the invention, in the step 1), the volume ratio of the gelatin aqueous solution to the nafion solution is 100 ml: 1-10 ml; preferably, the mass volume ratio of gelatin to water in the gelatin aqueous solution is 1-5 g: 100ml, the concentration of the nafion solution is 20-40%, preferably 30%.
According to the preparation method of the thin film type fluorescence sensor provided by the invention, in the step 2), the mass-to-volume ratio of the N, S co-doped carbon quantum dots to the blending solution is 2-2000 mg: 101-110 ml; and/or in the step 3), the thickness of the film is 0.1-3.0 mm, and the cutting size is 1 multiplied by 2 cm.
According to the preparation method of the thin film type fluorescence sensor provided by the invention, the preparation steps of the N, S co-doped carbon quantum dots comprise: and mixing the aqueous solution of the fruit acid and the aqueous solution of the methionine for reaction, and then cooling, filtering and drying to obtain the N, S-CDs.
According to the preparation method of the thin film type fluorescence sensor, in the step of preparing the N, S co-doped carbon quantum dots, the reaction temperature is 130-180 ℃, preferably 150 ℃, and the reaction time is 18-30 hours, preferably 24 hours; and/or the drying temperature is 50-70 ℃, preferably 60 ℃, and the drying time is 10-15 hours, preferably 12 hours. In the invention, by adopting the reaction method and conditions for preparing the N, S co-doped carbon quantum dots, particularly by controlling the reaction temperature and the reaction time, the synthesized N, S-CDs have better fluorescence characteristics and water solubility under the preferable condition range.
The invention also provides application of the film type fluorescence sensor or the film type fluorescence sensor prepared by the preparation method in fish freshness detection.
The application of the film-type fluorescence sensor in fish freshness detection provided by the invention comprises the following steps: placing a fish sample and the thin film type fluorescent sensor in a test container; acquiring fluorescence emission spectrum intensity of the thin film type fluorescence sensor; the freshness of the fish sample is judged by the change of the fluorescence intensity of the film type fluorescence sensor.
According to the application of the film-type fluorescent sensor in fish freshness detection, the calibration of the film-type fluorescent probe comprises the following steps:
1. fish samples were purchased from fresh salmon with regular quality assurance.
2. Cutting 30-50 g of salmon, respectively putting the salmon and a film type fluorescence sensor (N, S-CDs-nafion/gelatin film) into a sealed container, and storing in a refrigerator at 10 ℃;
3. measuring the intensity change delta I of fluorescence emission spectra every 8 hours;
the preferred operation is as follows: 1) ultraviolet-visible spectrum analysis: performing ultraviolet spectrum analysis on a sample by adopting an ultraviolet-visible spectrophotometer under the conditions that the scanning wavelength is 200-600 nm, the sampling interval is 2nm and the photometric mode is Abs, wherein the wavelength at the strongest absorption peak is the optimal excitation wavelength lambdex of N, S-CDs; 2) measurement of fluorescence emission spectral intensity Δ I: recording a fluorescence emission spectrum of the film sample under the condition that the wavelength of the excitation light is lambda ex by using a fluorescence spectrophotometer, wherein the fluorescence intensity reaches a peak value Imax at the optimal emission spectrum lambda em; recording the fluorescence intensity Imax at λ em;
4. fitting the peak intensity Imax of the fluorescence emission spectrum of the thin-film fluorescence sensor with the TVB-N of the salmon to obtain a calibration curve of the fluorescence (emission spectrum) intensity I and freshness of the thin-film fluorescence sensor.
In the invention, the fluorescence intensity of the N, S-CDs-nafion/gelatin film type fluorescence sensor changes along with the increase of the storage time of the salmon, and the shelf life end point of the aquatic product is determined according to the detected TVB-N and the fish freshness maximum limit value specified by the national standard (according to the specification of GB 2733 + 2015 safety standard for fresh and frozen aquatic products, the TVB-N value of the marine fish is not more than 30mg/100 g). The seawater fish TVB-N is more than or equal to 30mg/100g for deterioration, and the peak value of fluorescence emission light of the N, S-CDs-nafion/gelatin film is lower along with the increase of storage time; the TVB-N of salmon fillets was measured at regular intervals and the change in fluorescence intensity of N, S-CDs-nafion/gelatin films was recorded. And finally, judging the freshness of the meat food according to the change of the fluorescence emission light intensity of the N, S-CDs-nafion/gelatin film type sensor, and realizing rapid nondestructive detection.
The invention has the beneficial effects that:
1) the N and S co-doped carbon quantum dots are prepared to be used as a fluorescence sensor, the fluorescence sensor generates fluorescence intensity change through reaction with volatile components generated by fish decay, and the freshness of fish products is monitored by utilizing the change of the fluorescence intensity; compared with the existing visible light indicator, the N, S-CDs has stable fluorescence performance, is sensitive to the change of a fluorescence indicating signal of a target detection object, has low detection limit, and can realize the quantitative detection of fish freshness through the fitting with TVB-N.
2) The N, S-CDs-nafion/gelatin film prepared by the invention is synthesized by nafion and gelatin, the brittleness of the gelatin film is greatly reduced through blending modification of nafion solution to the gelatin, meanwhile, nafion provides a large number of cation channels, the absorption of the composite film to putrefactive gas is improved, and the interference of anions to detection results is reduced; meanwhile, the nafion solution greatly improves the dispersion degree of the N, S-CDs in the gelatin, so that the N, S-CDs-nafion/gelatin film has higher sensitivity response to volatile gas generated by fish decay.
3) The method for detecting fish freshness based on the fluorescent sensor can effectively detect the current TVB-N value of fish according to the fluorescence intensity color change of the film probe in the fish storage and transportation process, accurately evaluate the fish freshness and realize the rapid nondestructive detection of the fish freshness; expensive detection instruments are not needed, the detection cost is low, and the operation is simple and convenient; the thin film type fluorescence sensor is nontoxic, low in cost, high in stability and beneficial to commercial application.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings needed to be used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art that other drawings can be obtained according to these drawings without creative efforts.
FIG. 1 is an X-ray diffraction (XRD) pattern of N, S-CDs according to example of the present invention;
FIG. 2 is a graph showing an ultraviolet-visible absorption spectrum, a fluorescence excitation spectrum and an emission spectrum of N, S-CDs according to example N, S-CDs of the present invention, wherein (1) is a photograph of N, S-CDs under sunlight, and (2) is a photograph of N, S-CDs under an ultraviolet lamp (365 nm);
FIG. 3 is a schematic diagram showing the change of fluorescence intensity of N, S-CDs-nafion/gelatin films of different ammonia concentrations under the conditions of exciting light at 350nm and emitting light at 445nm in the embodiment of the present invention, wherein the ammonia concentration: a.0; b.2; c 10; d.20; e.30; f.50; g.65.0; h.80, the unit is mmol/L;
FIG. 4 is a calibration graph of ammonia concentration and fluorescence intensity according to an embodiment of the present invention;
FIG. 5 is a graph showing the anti-interference curve of the N, S-CDs-nafion/gelatin film of example N, S-CDs-nafion/gelatin film of the present invention;
FIG. 6 is a graph showing the stability results of N, S-CDs-nafion/gelatin film of example of the present invention, wherein (a) is a graph showing the change of fluorescence intensity with time under UV lamp irradiation (365 nm); graph (b) is a graph showing the change in fluorescence intensity with time in sunlight;
FIG. 7 is a graph showing the reproducibility of the N, S-CDs-nafion/gelatin film of example of the present invention;
FIG. 8 is a graph showing the change in fluorescence intensity and TVB-N content of salmon when the N, S-CDs-nafion/gelatin film of example of the present invention and salmon are stored at 10 ℃;
FIG. 9 is a graph showing the calibration of the change in fluorescence intensity with the TVB-N content of salmon in the example of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and the following embodiments are used for illustrating the present invention and are not intended to limit the scope of the present invention. The examples do not show the specific techniques or conditions, according to the technical or conditions described in the literature in the field, or according to the product specifications.
In the present invention, the instruments and the like used are conventional products which are purchased from regular vendors, not indicated by manufacturers. The process is conventional unless otherwise specified, and the starting materials are commercially available from the open literature. The examples do not show the specific techniques or conditions, according to the technical or conditions described in the literature in the field, or according to the product specifications.
In the embodiment of the present invention, expressions such as N, S-CDs-nafion/gelatin film, N, S-CDs-nafion/gelatin type fluorescence sensor, and the like refer to the film type fluorescence sensor.
Example 1
This example provides the preparation of N, S-CDs, comprising the steps of: respectively preparing 200ml of 0.2mol/L malic acid aqueous solution and 200ml of 0.1mol/L methionine aqueous solution; mixing the malic acid aqueous solution and the methionine aqueous solution according to the volume ratio of 1: 1, reacting for 24 hours in an oil bath at 150 ℃, cooling the reaction liquid to room temperature, and filtering to obtain a supernatant; and (3) drying the supernatant in a vacuum drying oven at 60 ℃ for 12h to obtain a yellow product, namely N, S-CDs, and storing the yellow product in a dark place. As shown in FIG. 1, the peaks of the synthesized N, S-CDs at 20.9 ℃ and 27 ℃ are respectively characterized by 0.21nm of the graphitic carbon (100) plane and 0.26nm of the graphitic carbon (020) plane, and the synthesized N, S-CD is proved to have good crystallinity.
Example 2
This example provides an N, S-CDs-nafion/gelatin thin film type fluorescence sensor, which is prepared by the following specific steps: dissolving 1g of gelatin in 100ml of ultrapure water, adding 1ml of a commercially available 30% nafion solution, and fully stirring and dissolving to obtain a nafion modified gelatin blending solution; adding 20mg of N, S-CDs prepared in example 1 into the blending solution, fully stirring and dissolving, and then carrying out ultrasonic degassing; pouring the mixed solution of the N, S-CDs into a glass slide, and manufacturing an N, S-CDs-nafion/gelatin film by using a spin coater, wherein the thickness of the film is 1-1.5 mm; the finished film was cut to 1X 2cm for subsequent freshness fluorescence detection applications.
Example 3
The N, S-CDs-nafion/gelatin thin film type fluorescence sensor prepared in example 2 was used for NH3The detection comprises the following specific processes:
1. placing a 1 x 2 cm-sized N, S-CDs-nafion/gelatin film in an air chamber, introducing mixed gas of ammonia gas and air into the air chamber for 1min, and detecting the change of the fluorescence intensity of the film, wherein the concentration of the ammonia gas is respectively 0, 2, 10, 20, 30, 50, 65, 80, unit: mmol/L, the specific operation process is as follows:
(1) ultraviolet-visible spectrum analysis: an HACH ultraviolet-visible spectrophotometer is adopted to carry out ultraviolet spectrum analysis on a film sample under the conditions that the scanning wavelength is 200-600 nm, the sampling interval is 2nm, and the photometric mode is Abs, the wavelength at the strongest absorption peak is the optimal excitation wavelength of N, S-CDs, as shown in figure 2, the optimal excitation wavelength of the N, S-CDs synthesized by the method is 350 nm.
(2) Measurement of fluorescence emission spectral intensity I: an Edinburgh FS5 type fluorescence spectrophotometer is adopted, under the conditions that the width of an excitation slit is 5.0nm, the width of an emission slit is 5.0nm, and the optimal excitation wavelength (Ex) of a film sample is 350nm, the optimal emission wavelength (Em) is 445 nm. The excitation wavelength was 350nm, and the fluorescence emission spectra of the film samples at different concentrations of ammonia gas and the peak fluorescence intensity Imax at 445nm of the emission spectrum were recorded, as shown in fig. 3.
2. The peak intensity I of the fluorescence emission spectrum of the N, S-CDs-nafion/gelatin film was fitted to the ammonia gas concentration, and as shown in fig. 4, the N, S-CDs-nafion/gelatin film of this example had a good linear response to ammonia gas. Fig. 5 shows that the N, S-CDs-nafion/gelatin thin film type fluorescence sensor has high selectivity to volatile amine gases and good anti-interference performance. FIG. 6 shows that the N, S-CDs-nafion/gelatin film type fluorescence sensor has good photobleaching resistance and stability. FIG. 7 shows that the N, S-CDs-nafion/gelatin film type fluorescence sensor has better repeatability.
Example 4
The N, S-CDs-nafion/gelatin film prepared in example 2 is used for rapid nondestructive detection of fish freshness, and the specific process is as follows:
(1) fish samples were purchased from fresh salmon with regular quality assurance. In the embodiment of the invention, the TVB-N detection of fish meat: measuring according to a micro-diffusion method in GB 5009.228-2016 (national standard for measuring volatile basic nitrogen in food safety) of food;
(2) cutting 30g of each salmon sample, putting the cut salmon sample and the N, S-CDs-nafion/gelatin film into a plastic culture dish, sealing the salmon sample and the N, S-CDs-nafion/gelatin film with a preservative film, and storing the salmon sample in a high-precision low-temperature incubator at 10 ℃;
(3) the fluorescence emission spectrum intensity Imax of the N, S-CDs-nafion/gelatin thin film in the film storage process is measured every 8 hours, and the specific operation is as follows: the fluorescence emission spectrum of the sample was recorded using an Edinburgh FS5 type fluorescence spectrophotometer at an excitation slit width of 5.0nm, an emission slit width of 5.0nm and an excitation wavelength λ ex 350nm, and the fluorescence intensity peak Imax at an emission spectrum λ em 445nm was shown in FIGS. 8 and 9. And judging the freshness of the salmon through the change of the fluorescence intensity. When the fluorescence intensity decreased by about 20%, the TVB-N content of salmon exceeded 30mg/100g, indicating salmon spoilage.
Although the invention has been described in detail hereinabove with respect to a general description and specific embodiments thereof, it will be apparent to those skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (10)
1. The thin film type fluorescence sensor is characterized by being prepared from raw materials including N and S co-doped carbon quantum dots, a matrix and a blending modifier, wherein the matrix is gelatin, and the blending modifier is nafion solution.
2. The thin film type fluorescence sensor of claim 1, wherein in the raw material of the N, S co-doped carbon quantum dots, a carbon source is fruit acid, the fruit acid is preferably citric acid or malic acid, and a nitrogen source and a sulfur source are methionine; preferably, the molar ratio of the fruit acid to the methionine is 1-4: 1-2, preferably 2: 1.
3. The thin film type fluorescence sensor according to claim 1 or 2, wherein the mass-to-volume ratio of the N, S co-doped carbon quantum dots, the gelatin and the nafion solution is 2-2000 mg: 1-5 g: 1-10 ml, and the concentration of the nafion solution is 20-40%.
4. The method for producing a thin film type fluorescence sensor according to any one of claims 1 to 3, comprising:
1) mixing the gelatin aqueous solution with the nafion solution to obtain a blended solution;
2) mixing the N, S co-doped carbon quantum dots with the blending solution, dissolving, and performing ultrasonic degassing to obtain a mixed solution of N, S-CDs;
3) preparing a film from the mixed solution, and then cutting the film.
5. The method for manufacturing a thin film type fluorescence sensor according to claim 4, wherein the volume ratio of the gelatin aqueous solution to the nafion solution in step 1) is 100 ml: 1-10 ml; preferably, the mass volume ratio of gelatin to water in the gelatin aqueous solution is 1-5 g: 100ml, the concentration of the nafion solution is 20-40%, preferably 30%.
6. The preparation method of the thin film type fluorescence sensor according to claim 4 or 5, wherein in the step 2), the mass-to-volume ratio of the N, S co-doped carbon quantum dots to the blending solution is 2-2000 mg: 101-110 ml; and/or in the step 3), the thickness of the film is 0.1-3.0 mm, and the cutting size is 1 multiplied by 2 cm.
7. The preparation method of the thin film type fluorescence sensor according to any one of claims 4 to 6, wherein the preparation step of the N, S co-doped carbon quantum dot comprises the following steps: and mixing the aqueous solution of the fruit acid and the aqueous solution of the methionine for reaction, and then cooling, filtering and drying to obtain the N, S-CDs.
8. The preparation method of the thin film type fluorescence sensor according to claim 7, wherein in the step of preparing the N, S co-doped carbon quantum dot, the reaction temperature is 130-180 ℃, preferably 150 ℃, and the reaction time is 18-30 hours, preferably 24 hours; and/or the drying temperature is 50-70 ℃, preferably 60 ℃, and the drying time is 10-15 hours, preferably 12 hours.
9. Use of the film-type fluorescence sensor according to any one of claims 1 to 3 or the film-type fluorescence sensor prepared by the method according to any one of claims 4 to 8 for detecting freshness of fish meat.
10. Use according to claim 9, characterized in that it comprises the following steps: placing a fish sample and the thin film type fluorescent sensor in a test container; acquiring fluorescence emission spectrum intensity of the thin film type fluorescence sensor; the freshness of the fish sample is judged by the change of the fluorescence intensity of the film type fluorescence sensor.
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