Surface-enhanced Raman probes based on dual signals and their applications

Abstract

This invention relates to the field of food safety detection technology, specifically to a surface-enhanced Raman probe based on dual signals and its application. The dual-signal surface-enhanced Raman probe is silver nanoparticles modified with chloroauric acid and potassium ferrocyanide. The preparation method includes: incubating potassium ferrocyanide solution and chloroauric acid solution for 1-60 min; reducing silver nitrate with a reducing agent to obtain a silver nanoparticle solution, centrifuging, removing the supernatant, resuspending in water to the original volume to obtain a silver nanoparticle resuspension; adding solution 1 to the silver nanoparticle resuspension and incubating for 1-60 min; centrifuging at 8000-12000 rpm for 8-15 min, removing the supernatant, washing, drying, and sealing for storage. This invention provides the application of a dual-signal surface-enhanced Raman probe in compound detection. This invention improves the detection sensitivity of target analytes, lowers the detection limit of analytical methods, and expands the linear range of analytical detection.

Description

Technical Field

[0001] This invention belongs to the field of food safety testing technology, and specifically relates to a surface-enhanced Raman probe based on dual signals and its application. Background Technology

[0002] Adenine, a purine-based organic compound with the molecular formula C5H5N5, is an important component of deoxyribonucleic acid (DNA) and an essential base for both DNA and RNA. Adenine plays a vital role in life processes such as energy transfer, neurotransmitter release, cell signaling, and prevention of cardiac arrhythmias. Detecting changes in adenine levels in body fluids can predict the extent of DNA damage and indicate the development of certain diseases, such as cancer and AIDS. Therefore, sensitive analysis and detection of adenine levels have significant clinical implications.

[0003] Ceftriaxone sodium is a third-generation cephalosporin antibiotic with the molecular formula C. 18 H 16 N8Na2O7S3·3H2O. Currently, ceftriaxone sodium is one of the most widely used cephalosporin antibiotics, widely applied in the clinical treatment of infectious diseases due to its inhibitory or bactericidal activity against pathogenic microorganisms. However, the toxic side effects caused by overuse and the environmental problems resulting from its abuse have also attracted widespread attention. Achieving a simple and rapid detection method for ceftriaxone sodium is a fundamental prerequisite for evaluating the efficacy and toxic side effects of antibiotics.

[0004] Malachite green, listed as a prohibited drug and other compound in food animals, is a triphenylane organic compound once used in aquaculture for sterilization and insecticidal purposes. However, its metabolism in humans and animals produces carcinogenic, teratogenic, and mutagenic substances, posing a significant threat to human health. my country stipulates that malachite green and its metabolite, colorless malachite green, must not be detected in pollution-free aquatic products. Despite this, due to its low price and good sterilization effect, the illegal addition of malachite green to aquaculture continues. Therefore, the analysis and detection of malachite green is of great significance to human health.

[0005] Currently, the main methods for detecting adenine, ceftriaxone sodium, and malachite green include chromatography, fluorescence analysis, and capillary electrophoresis. However, these methods rely heavily on large instruments, require sophisticated pretreatment, involve complex procedures, necessitate professional operation, and are expensive. They are also ill-suited for the rapid detection needs of many applications, limiting their widespread adoption. Surface-enhanced Raman spectroscopy (SERS) has emerged as a trend in rapid detection technologies, offering advantages such as speed, high sensitivity, non-destructive testing, and low cost. However, the variability between Raman substrates makes the non-reproducibility of SERS a major obstacle for SERS sensing platforms. Therefore, the development of dual-signal SERS analysis methods holds promise for solving the current problem of SERS non-reproducibility. Summary of the Invention

[0006] To address the aforementioned problems, this invention provides a surface-enhanced Raman probe based on dual signals and its applications.

[0007] The primary objective of this invention is to provide a surface-enhanced Raman probe based on dual signals, wherein the probe is silver nanoparticles modified with chloroauric acid and potassium ferrocyanide.

[0008] Preferably, the molar ratio of chloroauric acid, potassium ferrocyanide, and silver nanoparticles is (0.1-10):(0.1-10):(3×10⁻⁶). -7 ).

[0009] The second objective of this invention is to provide a method for preparing a surface-enhanced Raman probe based on dual signals, comprising the following steps: S1. Take potassium ferrocyanide solution and chloroauric acid solution, incubate for 1-60 min to obtain solution 1; S2. Reduce silver nitrate with a reducing agent to obtain a silver nanoparticle solution. Centrifuge at 8000~12000 rpm for 8~15 min, remove the supernatant, and resuspend in water to the original volume to obtain a silver nanoparticle resuspension. S3. Add solution 1 prepared in step S1 to the silver nanoparticle resuspension prepared in step S2, and incubate for 1-60 min; centrifuge at 8000-12000 rpm for 8-15 min, remove the supernatant, wash, dry, and seal to obtain a surface-enhanced Raman probe based on dual signals. Preferably, the molar ratio of potassium ferrocyanide to chloroauric acid in step S1 is 0.1-10:0.1-10, and the volume ratio is 1-100:1-100.

[0010] Preferably, the volume ratio of the silver nanoparticle resuspension to solution 1 is 1-5:1-4.

[0011] Preferably, the reducing agent is hydroxylamine hydrochloride, sodium citrate, or sodium borohydride.

[0012] A third objective of this invention is to provide an application of a dual-signal surface-enhanced Raman probe in the detection of compounds that do not contain cyano groups and do not react with cyano groups.

[0013] Preferably, the compound is adenine, ceftriaxone sodium, and / or malachite green.

[0014] Preferably, the compound is malachite green.

[0015] The fourth objective of this invention is to provide a surface-enhanced Raman spectroscopy detection method based on dual signals, comprising the following steps: S11. Take the sample to be tested, perform pretreatment, centrifuge, and collect the sample supernatant; S12. Add the surface-enhanced Raman probe based on dual signals to pure water to prepare a resuspension, and incubate it with the sample supernatant for 1-60 min to obtain the solution to be detected. S13. Perform Raman spectroscopy detection on the solution to be tested.

[0016] Preferably, the sample to be tested includes at least one of adenine, ceftriaxone sodium, and malachite green.

[0017] Preferably, in step S12, the volume ratio of the resuspension to the sample supernatant is 1-5:1-3.

[0018] Beneficial effects of this invention: (1) The cyano signal intensity of silver nanoparticles modified with chloroauric acid and potassium ferrocyanide is significantly improved; (2) It can meet the requirement of increasing the Raman signal intensity of the modified Raman substrate, improve the sensitivity of the target analyte detection, reduce the detection limit of the analytical method, and expand the linear range of the analytical detection; (3) It can analyze and detect electrically neutral adenine, negatively charged ceftriaxone sodium, and positively charged malachite green in the sample to be tested. Attached Figure Description

[0019] Figure 1 This is a schematic diagram illustrating the technical principle of the present invention.

[0020] Figure 2 These are transmission electron microscope (TEM) images of the Raman probe of this invention before and after etching with silver nanoparticles.

[0021] Figure 3 These are Raman spectra of Raman probes prepared from different components of this invention; component a consists of silver nanoparticles, chloroauric acid, and potassium ferrocyanide; component b consists of silver nanoparticles and potassium ferrocyanide; component c consists of silver nanoparticles, chloroauric acid, and potassium ferrocyanide; and component d consists of silver nanoparticles and potassium ferrocyanide.

[0022] Figure 4These are Raman spectra of adenine at different concentrations according to the present invention; Solid is the Raman spectrum of adenine; Blank is the Raman spectrum of blank.

[0023] Figure 5 This is the adenine standard curve diagram of the present invention.

[0024] Figure 6 These are Raman spectra of ceftriaxone sodium at different concentrations according to the present invention; Solid is the Raman spectrum of ceftriaxone sodium; Blank is the blank Raman spectrum.

[0025] Figure 7 This is the standard curve diagram of ceftriaxone sodium of the present invention.

[0026] Figure 8 These are Raman spectra of malachite green at different concentrations according to the present invention; Solid is the Raman spectrum of malachite green; Blank is the Raman spectrum of blank.

[0027] Figure 9 This is the malachite green standard curve diagram of the present invention. Detailed Implementation

[0028] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to the accompanying drawings and specific embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and do not constitute a limitation thereof.

[0029] A surface-enhanced Raman probe based on dual signals is silver nanoparticles modified with chloroauric acid and potassium ferrocyanide; preferably, the molar ratio of chloroauric acid, potassium ferrocyanide, and silver nanoparticles is (0.1-10):(0.1-10):(3×10⁻⁶). -7 ); The probe is prepared by the following method: S1. Take potassium ferrocyanide solution and chloroauric acid solution, incubate for 1-60 min to obtain solution 1; S2. Reduce silver nitrate with a reducing agent to obtain a silver nanoparticle solution. Centrifuge at 8000~12000 rpm for 8~15 min, remove the supernatant, and resuspend in water to the original volume to obtain a silver nanoparticle resuspension. S3. Add the solution 1 prepared in step S1 to the silver nanoparticle resuspension prepared in step S2 and incubate for 1-60 min; centrifuge at 8000-12000 rpm for 8-15 min, remove the supernatant, wash, dry and seal to obtain the surface-enhanced Raman probe based on dual signals. Preferably, in step S1, the molar ratio of potassium ferrocyanide to chloroauric acid is 0.1-10:0.1-10, and the volume ratio is 1-100:1-100. Preferably, the molar ratio of potassium ferrocyanide to chloroauric acid is 1:2 and the volume ratio is 3:1; Preferably, the molar ratio of potassium ferrocyanide to chloroauric acid is 3:2 and the volume ratio is 5:2; Preferably, the volume ratio of the silver nanoparticle resuspension to solution 1 is 1-5:1-4; Preferably, the volume ratio of the silver nanoparticle resuspension to solution 1 is 5:4; Preferably, the reducing agent is hydroxylamine hydrochloride, sodium citrate, or sodium borohydride.

[0030] A surface-enhanced Raman spectroscopy detection method based on dual signals includes the following steps: S11. Take the sample to be tested, perform pretreatment, centrifuge, and collect the sample supernatant; S12. Add the surface-enhanced Raman probe based on dual signals to pure water to prepare a resuspension, and incubate it with the sample supernatant for 1-60 min to obtain the solution to be detected. S13. Perform Raman spectroscopy detection on the solution to be tested; Preferably, in step S12, the volume ratio of the resuspension to the sample supernatant is 1-5:1-3; Preferably, the sample to be tested includes at least one of adenine, ceftriaxone sodium, and malachite green; Preferably, the sample to be tested is fish meat.

[0031] like Figure 1 The technical principle of the analytical method in this invention is illustrated in the schematic diagram. First, a solution of silver nanoparticles (AgNPs) is synthesized, centrifuged, resuspended, and stored at 4°C for later use; this is the silver nanoparticle resuspension. Second, potassium ferrocyanide (K4Fe(CN)6) and chloroauric acid (HAuCl4) are incubated for a certain period of time (resulting in solution 1), which is then added to a certain amount of the silver nanoparticle resuspension for further incubation to obtain solution 2. Next, solution 2 is centrifuged, resuspended, and incubated with the target analyte to obtain solution 3. Finally, solution 3 is subjected to Raman spectroscopy, and the concentration of the target analyte is determined based on the ratio of its characteristic peak signal intensities. The surface-enhanced Raman probe and its detection method of the present invention can be widely used in food safety testing to detect compounds in food or medicine. However, some substances cannot be detected. The range of undetectable targets includes: substances that easily oxidize silver nanoparticles, such as strong oxidants like potassium permanganate; organic or inorganic substances containing cyano groups, such as cyanomethyl cyanate and other cyano compounds; substances that easily replace the cyano groups on the surface of silver nanoparticles; and substances that easily react with cyano groups, such as sodium azide and other azide compounds.

[0032] Example 1: Fabrication of a surface-enhanced Raman probe based on dual signals The specific steps for fabricating a surface-enhanced Raman probe based on dual signals are as follows: (1) Incubation of potassium ferrocyanide and chloroauric acid: Take 300 μL of 1 mM potassium ferrocyanide solution and incubate with 100 μL of 2 mM chloroauric acid solution for 5 min to obtain solution 1; (2) Synthesis and surface modification of silver nanoparticles: Silver nitrate was reduced by hydroxylamine hydrochloride to obtain a silver nanoparticle solution; 1000 μL of silver nanoparticle solution was placed in a centrifuge tube and centrifuged at 10000 rpm for 10 min. The supernatant was removed and water was added to resuspend the solution to the original volume to obtain a silver nanoparticle resuspension; then, the solution 1 prepared in step (1) was added to 500 μL of silver nanoparticle resuspension and incubated for 20 min. After incubation, the solution was centrifuged at 10000 rpm for 10 min. The supernatant was removed, washed, dried and sealed for storage to obtain a surface-enhanced Raman probe based on dual signals.

[0033] Figure 2 The transmission electron microscope (TEM) image of the prepared Raman probe is shown. This TEM image illustrates that after etching, silver nanoparticles, chloroauric acid, and potassium ferrocyanide react to produce morphological changes, indirectly indicating that cyano groups were successfully modified on the surface of silver nanoparticles.

[0034] Example 2: Fabrication of a surface-enhanced Raman probe based on dual signals The specific steps for fabricating a surface-enhanced Raman probe based on dual signals are as follows: (1) Incubation of potassium ferrocyanide and chloroauric acid: Take 200 μL of 3 mM potassium ferrocyanide solution and incubate with 80 μL of 2 mM chloroauric acid solution for 15 min to obtain solution 1; (2) Synthesis and surface modification of silver nanoparticles: silver nitrate was reduced by sodium citrate reducing agent to obtain silver nanoparticle solution; 1000 μL of silver nanoparticle solution was placed in a centrifuge tube and centrifuged at 10000 rpm for 10 min. The supernatant was removed and water was added to resuspend the solution to the original volume to obtain silver nanoparticle resuspension; then, the solution 1 prepared in step (1) was added to 400 μL of silver nanoparticle resuspension and incubated for 30 min. After incubation, the solution was centrifuged at 10000 rpm for 10 min. The supernatant was removed, washed, dried and sealed for storage to obtain surface-enhanced Raman probe based on dual signals.

[0035] Example 3: Raman signal intensity testing based on a surface-enhanced Raman probe with dual signals The performance of the surface-enhanced Raman probe based on dual signals prepared in this invention was tested by comparison. The experiment was divided into 4 groups (a, b, c, d), with group a being the experimental group and groups b, c, and d being the control group, as detailed below: (1) Incubation of potassium ferrocyanide and chloroauric acid: Take 300 μL of 1 mM potassium ferrocyanide solution and incubate with 100 μL of 2 mM chloroauric acid solution for 5 min to obtain solution 1; (2) Incubation of potassium ferricyanide and chloroauric acid: Take 300 μL of 1 mM potassium ferricyanide solution and incubate with 100 μL of 2 mM chloroauric acid solution for 5 min, and then obtain solution 2; (3) Synthesis and surface modification of silver nanoparticles: Silver nitrate was reduced using hydroxylamine hydrochloride as a reducing agent to obtain a silver nanoparticle solution; 1000 μL of the silver nanoparticle solution was placed in a centrifuge tube and centrifuged at 10000 rpm for 10 min. The supernatant was removed, and the solution was resuspended in water to the original volume to obtain a silver nanoparticle resuspension; next, solution 1 prepared in step (1) was added to 400 μL of the silver nanoparticle resuspension and incubated for 30 min. After incubation, the solution was centrifuged at 10000 rpm for 10 min. The supernatant was removed, and the solution was resuspended in pure water to the original volume to obtain solution a; then, 300 μL of the solution was added to the resuspension. 1 mM potassium ferrocyanide solution was added to 400 μL of silver nanoparticle resuspension and incubated for 30 min. After incubation, the solution was centrifuged at 10000 rpm for 10 min. After removing the supernatant, pure water was added and the solution was resuspended to the original volume to obtain solution b. Then, solution 2 prepared in step (2) was added to 400 μL of silver nanoparticle resuspension and incubated for 30 min. After incubation, the solution was centrifuged at 10000 rpm for 10 min. After removing the supernatant, pure water was added and the solution was resuspended to the original volume to obtain solution c. Subsequently, 300 μL of 1 mM potassium ferrocyanide solution was added to 400 μL of silver nanoparticle resuspension and incubated for 30 min. After incubation, the solution was centrifuged at 10000 rpm for 10 min. After removing the supernatant, pure water was added and the solution was resuspended to the original volume to obtain solution d. (4) Incubate solutions a, b, c, and d from (3) with 300 μL of 2 mM 4-mercaptobenzoic acid for 20 min, respectively; then, use a Raman spectroscopy instrument with a laser wavelength of 785 nm, a laser power of 10%, and a range of 200-3000 cm⁻¹. -1 The Raman spectra of each solution were measured with an integration time of 2 seconds and a cumulative count of 3. The spectra are shown below. Figure 3 As shown, group a is the Raman probe prepared in this invention, composed of silver nanoparticles, chloroauric acid, and potassium ferrocyanide; group b uses silver nanoparticles as the Raman probe and utilizes the cyano group in potassium ferrocyanide as the Raman signal tag, composed of silver nanoparticles and potassium ferrocyanide; group c replaces the potassium ferrocyanide in group a with potassium ferrocyanide during the preparation process, composed of silver nanoparticles, chloroauric acid, and potassium ferrocyanide; group d is composed of silver nanoparticles and potassium ferrocyanide. 4-MBA, or 4-mercaptobenzoic acid, was used as a model molecule to verify the signal intensity of the system.

[0036] Figure 3Raman spectra of Raman substrates prepared with different components are shown. The figures show that group a significantly enhances the Raman signal intensity of the cyano group of silver nanoparticles, unlike the combination of cyano group Raman signal intensities in groups b, c, and d. Firstly, comparing the Raman spectra of groups a and b demonstrates that chloroauric acid significantly enhances the cyano group signal of silver nanoparticles. Secondly, comparing the Raman spectra of components a and c demonstrates that potassium ferrocyanide significantly enhances the cyano group signal of silver nanoparticles. Thirdly, comparing the Raman spectra of components b and d demonstrates that the cyano group signal generated by potassium ferrocyanide and silver nanoparticles is stronger than that generated by potassium ferrocyanide. Fourthly, comparing the Raman spectra of components c and d demonstrates that chloroauric acid slightly enhances the cyano group signal of silver nanoparticles. In summary, the surface-enhanced Raman probe based on dual signals prepared in this invention achieves the strongest signal and best effect.

[0037] Example 4: Construction of a dual-signal surface-enhanced Raman spectroscopy analysis method I. Adenine Analysis Method Preparation of adenine standard solution: Accurately weigh 0.0014 g of adenine standard and dissolve it in 1000 μL of pure water to prepare a 10 mM adenine standard solution; then take different volumes of the 10 mM adenine standard solution and dilute it with pure water to prepare 0.1, 0.5, 1.0, 5.0, 8.0, 10.0, and 20.0 μM solutions, each with a volume of 1000 μL.

[0038] The surface-enhanced Raman probe prepared in Example 1 or Example 2 was added to pure water to prepare a resuspension, which was then mixed with the prepared adenine standard solution and incubated at room temperature for 1-60 min. Then, a Raman spectroscopy instrument with a laser wavelength of 785 nm and a laser power of 10% was used, with a range of 200-3000 cm⁻¹. -1 The Raman spectra of each solution were measured with an integration time of 2 seconds and a cumulative count of 3. The spectra are shown below. Figure 4 As shown; a standard curve was obtained with the concentration of adenine standard solution as the x-axis and the ratio of the cyano group to the adenine Raman spectral signal intensity as the y-axis. Figure 5 The results from the standard curve show that the linear range is 0.1-20 μM, the limit of detection (LOD) is 0.074 μM, and the signal-to-noise ratio is 3.

[0039] II. Analysis of Ceftriaxone Sodium Preparation of ceftriaxone sodium standard solution: Accurately weigh 0.0060 g of ceftriaxone sodium standard and dissolve it in 1000 μL of pure water to prepare a 10 mM ceftriaxone sodium standard solution; then take different volumes of the 10 mM ceftriaxone sodium standard solution and dilute it with pure water to prepare 1000 μL of each of the following solutions: 0.1, 0.5, 1.0, 5.0, 8.0, 10.0, and 20.0 μM.

[0040] The surface-enhanced Raman probe prepared in Example 1 or Example 2 was added to pure water to prepare a resuspension, which was then mixed with the prepared ceftriaxone sodium standard solution and incubated at room temperature for 1-60 min. Then, a Raman spectroscopy instrument with a laser wavelength of 785 nm and a laser power of 10% was used, with a range of 200-3000 cm⁻¹. -1 The Raman spectra of each solution were measured with an integration time of 2 seconds and a cumulative count of 3. The spectra are shown below. Figure 6 As shown; a standard curve was obtained with the concentration of ceftriaxone sodium standard solution as the abscissa and the ratio of the cyano group to the Raman spectral signal intensity of ceftriaxone sodium as the ordinate. Figure 7 The results from the standard curve show that the linear range is 0.1-10 μM, the limit of detection (LOD) is 0.050 μM, and the signal-to-noise ratio is 3.

[0041] III. Malachite Green Analysis Preparation of malachite green standard solution: Accurately weigh 0.0036 g of malachite green standard and dissolve it in 1000 μL of pure water to prepare a 10 mM malachite green standard solution; then take different volumes of the 10 mM malachite green standard solution and dilute them with pure water to prepare 1000 μL of each of the following solutions: 0.1, 0.5, 1.0, 5.0, 8.0, 10.0, and 20.0 μM.

[0042] The surface-enhanced Raman probe prepared in Example 1 or Example 2 was added to pure water to prepare a resuspension, which was then mixed with the prepared malachite green standard solution and incubated at room temperature for 1-60 min. Then, a Raman spectroscopy instrument with a laser wavelength of 785 nm and a laser power of 10% was used, with a range of 200-3000 cm⁻¹. -1 The Raman spectra of each solution were measured with an integration time of 2 seconds and a cumulative count of 3. The spectra are shown below. Figure 8 As shown; a standard curve was obtained with the concentration of malachite green standard solution as the abscissa and the division relationship between the cyano group and the Raman spectral signal intensity of malachite green as the ordinate. Figure 9 The results from the standard curve show that the linear range is 0.1-2.0 μM, the limit of detection (LOD) is 0.032 μM, and the signal-to-noise ratio is 3.

[0043] IV. Application of Test Samples Take a solution or meat sample, pretreat it, centrifuge it, and collect the supernatant; incubate the surface-enhanced Raman probe prepared in this invention with the sample supernatant for 1-60 min to obtain the test solution; measure the Raman spectrum of the test solution to determine the substances contained in the sample and their content; when the sample may contain at least two of adenine, ceftriaxone sodium and malachite green, in addition to Raman spectroscopy analysis, principal component analysis (PCA) or linear regression analysis should also be performed to finally determine the substances contained in the sample.

[0044] Although embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are exemplary and should not be construed as limiting the present invention. Those skilled in the art can make changes, modifications, substitutions, and variations to the above embodiments within the scope of the present invention.

[0045] The specific embodiments of the present invention described above do not constitute a limitation on the scope of protection of the present invention. Any other corresponding changes and modifications made in accordance with the technical concept of the present invention should be included within the scope of protection of the claims of the present invention.

Claims

1. A surface-enhanced Raman probe based on dual signals, characterized in that: The probe is silver nanoparticles modified with chloroauric acid and potassium ferrocyanide; the molar ratio of chloroauric acid, potassium ferrocyanide, and silver nanoparticles is (0.1-10):(0.1-10):(3×10⁻⁶). -7 The probe is used to detect compounds that do not contain cyano groups and cannot react with cyano groups. The preparation method includes the following steps: S1. Take potassium ferrocyanide solution and chloroauric acid solution, incubate for 1-60 min to obtain solution 1; S2. Reduce silver nitrate with a reducing agent to obtain a silver nanoparticle solution. Centrifuge at 8000~12000 rpm for 8~15 min, remove the supernatant, and resuspend in water to the original volume to obtain a silver nanoparticle resuspension. S3. Add the solution 1 prepared in step S1 to the silver nanoparticle resuspension prepared in step S2 and incubate for 1-60 min; centrifuge at 8000-12000 rpm for 8-15 min, remove the supernatant, wash, dry and seal to obtain the surface-enhanced Raman probe based on dual signals.

2. The surface-enhanced Raman probe based on dual signals according to claim 1, characterized in that: In step S1, the molar ratio of potassium ferrocyanide to chloroauric acid is (0.1-10):(0.1-10), and the volume ratio is (1-100):(1-100).

3. The surface-enhanced Raman probe based on dual signals according to claim 2, characterized in that: The volume ratio of the silver nanoparticle resuspension to solution 1 is (1-5):(1-4).

4. The surface-enhanced Raman probe based on dual signals according to claim 3, characterized in that: The reducing agent is hydroxylamine hydrochloride, sodium citrate, or sodium borohydride.

5. The application of the surface-enhanced Raman probe based on dual signals as described in claim 1 in the detection of compounds, characterized in that: The compounds are adenine, ceftriaxone sodium, and / or malachite green.

6. The application according to claim 5, characterized in that: The compound is malachite green.

7. A surface-enhanced Raman spectroscopy detection method based on dual signals, using the surface-enhanced Raman probe based on dual signals as described in claim 1, characterized in that, Includes the following steps: S11. Take the sample to be tested, perform pretreatment and centrifugation, and collect the sample supernatant; S12. Add the surface-enhanced Raman probe based on dual signals to pure water to prepare a resuspension, and incubate it with the sample supernatant for 1-60 min to obtain the solution to be detected. S13. Perform Raman spectroscopy detection on the solution to be tested.

8. The surface-enhanced Raman spectroscopy detection method based on dual signals according to claim 7, characterized in that: The sample to be tested includes at least one of adenine, ceftriaxone sodium, and malachite green.

9. The surface-enhanced Raman spectroscopy detection method based on dual signals according to claim 8, characterized in that: In step S12, the volume ratio of the resuspension to the sample supernatant is (1-5):(1-3).

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