Method for building enantiomer multi-mode sensing platform based on plasma nano-enzyme

Abstract

The invention discloses an enantiomer multi-mode sensing platform building method based on plasma nano enzyme, and relates to the technical field of sensing. The multi-mode sensing technology is adopted, multiple signal outputs are integrated, the detection precision and reliability are remarkably improved, the sensor can provide multi-dimensional detection information, self-verification is achieved, analysis is more accurate and credible, and particularly, when extremely similar chiral molecules are recognized, the detection precision and reliability are greatly improved. The multi-mode sensing technology not only overcomes the limitation of a single technology, but also greatly expands the recognition range, improves the analysis precision and applicability of the enantiomer in a complex environment, shows important practical application value, combines three modes of chemiluminescence, temperature measurement and smartphone-assisted RGB, is used for recognition of the chiral molecular enantiomer, and has good application prospects. The core innovation point of the platform is that through multi-dimensional signal output of multi-mode sensing, the limitation of a traditional single-mode sensing technology is overcome, and the range and reliability of chiral molecule enantiomer recognition are remarkably improved.

Description

Technical Field

[0001] The present invention relates to the field of sensing technology, and specifically to a method for constructing an enantiomer multimodal sensing platform based on plasma nanozymes. Background Art

[0002] Currently, most traditional chiral molecule detection technologies rely on single-mode sensors, such as optical activity, chromatography, and mass spectrometry. However, due to the extreme structural similarity of chiral molecular enantiomers, single-mode detection methods are easily affected by experimental conditions and instrumentation, resulting in unstable measurement results, poor sensitivity, and large errors. This is particularly problematic for molecular recognition and high-precision detection in complex environments.

[0003] See the following prior art:

[0004] Chinese patent publication number CN107402203B, titled "A method for rapid chiral identification of tartrate enantiomers by colorimetric and ultraviolet spectroscopy," relates to the preparation and application of thiourea detection reagents.

[0005] The Chinese patent publication number is CN112834379B, the patent name is a cysteine chiral recognition sensor, which belongs to the field of chiral recognition technology;

[0006] Chinese patent publication number CN113237934B, titled "A method for preparing chiral silver sulfide quantum dots / few-layer carbon nitride composites for electrochemiluminescence chiral recognition," belongs to the field of nanomaterial preparation and molecular recognition technology;

[0007] The above technology has the following technical problems:

[0008] Insufficient sensitivity and reliability: Traditional single-mode sensing technology often finds it difficult to achieve high sensitivity and high reliability when detecting chiral molecular enantiomers with extremely similar structures due to the limitations of a single mode. It is easily affected by environmental interference, resulting in inaccurate detection results.

[0009] Limited scope of application: Single-mode sensing technology is difficult to apply to the identification of multiple chiral molecules, especially for chiral molecule analysis tasks in complex environments, and cannot meet diverse detection needs.

[0010] Lack of portability: Most existing methods for detecting chiral molecular enantiomers require complex equipment and operating procedures, making it difficult to achieve portable detection, which limits their promotion in practical applications.

[0011] Therefore, a new solution to the above problems needs to be proposed. Summary of the Invention

[0012] The purpose of the present invention is to provide a method for constructing an enantiomeric multimodal sensing platform based on plasmon nanozymes to solve the design problems raised in the background technology.

[0013] To achieve the above objectives, the present invention provides the following technical solution: a method for constructing an enantiomer multimodal sensing platform based on plasmon nanozymes, comprising at least the following steps:

[0014] S1: Preparation of plasmonic nanozymes. By element doping, cobalt is loaded onto gold nanoparticles to synthesize AuCoNPs. As plasmonic nanozymes and sensing materials, AuCoNPs have unique optical and catalytic properties.

[0015] S2: Construction of a multimodal sensing platform based on AuCoNPs solution, integrating chemiluminescence, temperature measurement, and smartphone-assisted RGB modes. The multimodal sensing platform uses Glu as a model analyte and can distinguish between L-Glu and D-Glu, achieving highly sensitive and specific detection of Glu.

[0016] Furthermore, the preparation of AuCoNPs in S1 is a citric acid reduction method, and the specific process includes at least the following steps:

[0017] A solution to be treated was prepared by mixing 100 μL of 1% chloroauric acid, 125 μL of 50 mmol / L cobalt chloride solution, and 9800 μL of water;

[0018] The solution to be treated was added to a round-bottom flask equipped with a condenser, heated to boiling in an oil bath and refluxed continuously for 5 min;

[0019] Then, 0.25 mL of a 2% sodium citrate solution was quickly injected into the boiled solution to be treated, causing the solution to turn red, thereby obtaining an AuCoNPs solution.

[0020] When the AuCoNPs solution is required to remain in liquid form, store it directly in a brown glass bottle away from light.

[0021] Furthermore, the chemiluminescence is a CL mode, and the CL mode distinguishes L / D-Glu according to different chemiluminescence signal intensities;

[0022] The identification of Glu enantiomers under the CL mode comprises at least the following steps:

[0023] 50 μL of L / D-Glu solution with different concentrations was injected into the wells of a 96-well hydrogel microplate;

[0024] Then, 50 μL of 5-fold diluted AuCoNPs solution was immediately added and incubated at 37°C for 0.5 h;

[0025] Remove the supernatant and rinse the wells with ultrapure water;

[0026] Then 50 μL of the molar concentration of 5.0×10 -7 Luminol and 50 μL of 0.05 M hydrogen peroxide were injected into the microwell to collect CL signals.

[0027] Furthermore, the preparation of the 96-well hydrogel microplate comprises at least the following steps: adding 50 μl of a 1 wt% chitosan solution into the wells of the 96-well microplate and drying at 50° C. to form a film as a binder for the PVA / SA hydrogel;

[0028] Then, 50 μL of a mixed solution containing 0.5% SA and 0.25 wt % PVA was added to the wells and frozen at −20 °C;

[0029] After 1 hour, 100 μL of a 0.05 wt % calcium chloride solution was added, and after cross-linking at room temperature for 1 hour, the excess liquid was removed and the microplate was washed with ultrapure water.

[0030] Furthermore, the temperature measurement is the TP mode, which systematically records the temperature signal and analyzes the reaction conditions at different concentrations to distinguish the different reactivities of L / D-Glu from the rate and amplitude of temperature increase;

[0031] The identification of Glu enantiomers under the TP mode comprises at least the following steps:

[0032] Fill a transparent test tube with 100 μL of L / D-Glu solution of different concentrations;

[0033] Then, 100 μL of 5-fold diluted AuCoNPs solution was added;

[0034] The reaction solution was irradiated with NIR-660 nm laser for 3 min, and a handheld digital thermometer was inserted into the liquid surface to collect the temperature signal of the solution.

[0035] Furthermore, the smartphone-assisted RGB, or RGB mode, identifies the difference between L / D-Glu solution and AuCoNPs by monitoring the color change produced by the reaction between the two. That is, by collecting and analyzing RGB values, especially grayscale values, the reaction differences between L-Glu and D-Glu under the same experimental conditions can be accurately distinguished. Ultimately, the data obtained through the mobile phone video mode and color recognition application, combined with the grayscale value calculated by the weighted average method of difference, can provide a reliable basis for the identification of L-Glu and D-Glu.

[0036] The identification of Glu enantiomers under the RGB mode comprises at least the following steps:

[0037] 50 μL of L / D-Glu solution of different concentrations was loaded into a 96-well white microplate;

[0038] Then, 50 μL of 5-fold diluted AuCoNPs solution was added;

[0039] The reaction was carried out at a constant temperature of 37°C for 5 min and then transferred to a dark box with 5W fluorescent lamps on both sides;

[0040] The color signal in the 96-well microplate was collected using the mobile phone video mode;

[0041] Use a color recognition application to read the R, G, and B values in each well;

[0042] The grayscale value is calculated using the differential weighted average method.

[0043] Compared with the prior art, the present invention has the following beneficial effects:

[0044] 1. Improved detection accuracy and reliability: By integrating three signal modes—chemiluminescence (CL), temperature change (TP), and RGB color analysis—this technology effectively overcomes the limitations of single-mode sensors in identifying chiral molecular enantiomers. The synergistic effect of these three modes provides more comprehensive information for analysis, reduces experimental error, and enhances the reliability and accuracy of results.

[0045] 2. Broaden the recognition range of chiral molecular enantiomers: The multimodal sensing platform can not only identify L-type and D-type enantiomers, but also maintain high recognition accuracy in complex environments, broadening the application range of enantiomers. Especially when the enantiomer structures are very similar, the output of multiple modes can effectively enhance the discrimination.

[0046] 3. Strong anti-interference ability: When faced with complex analytical environments and experimental conditions, the use of multimodal sensing technology significantly improves anti-interference ability, reduces instability caused by environmental factors, experimental errors, and equipment differences, and thus enhances the reliability of enantiomer identification. BRIEF DESCRIPTION OF THE DRAWINGS

[0047] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings required for describing the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work.

[0048] Figure 1 A multi-mode schematic diagram of the present invention as a whole;

[0049] Figure 2TEM image of AuCoNPs of the present invention;

[0050] Figure 3 The XPS graphs of AuNPs and AuCoNPs of the present invention are shown;

[0051] Figure 4 Schematic diagram of the CL signal of AuCoNPs of the present invention;

[0052] Figure 5 Schematic diagram of the photothermal conversion effect of AuCoNPs of the present invention;

[0053] Figure 6 Schematic diagram of the color conversion effect of AuCoNPs of the present invention. DETAILED DESCRIPTION

[0054] The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments.

[0055] Example 1:

[0056] See also Figure 1 A method for constructing an enantiomeric multimodal sensing platform based on plasmon nanozymes comprises at least the following steps:

[0057] S1: Preparation of plasmonic nanozymes. By element doping, cobalt is loaded onto gold nanoparticles to synthesize AuCoNPs. As plasmonic nanozymes and sensing materials, AuCoNPs have unique optical and catalytic properties.

[0058] The preparation of AuCoNPs in S1 is a citric acid reduction method, which includes at least the following steps:

[0059] A solution to be treated was prepared by mixing 100 μL of 1% chloroauric acid, 125 μL of 50 mmol / L cobalt chloride solution, and 9800 μL of water;

[0060] The solution to be treated was added to a round-bottom flask equipped with a condenser, heated to boiling in an oil bath and refluxed continuously for 5 min;

[0061] Then, 0.25 mL of a 2% sodium citrate solution was quickly injected into the boiled solution to be treated, causing the solution to turn red, thereby obtaining an AuCoNPs solution.

[0062] When the AuCoNPs solution is required to remain in liquid form, store it directly in a brown glass bottle away from light.

[0063] To demonstrate the unique optical and catalytic properties of AuCoNPs, we conducted the following operations:

[0064] The morphology and dispersion of AuCoNPs were observed by TEM. AuCoNPs showed well-dispersed spherical structures with a uniform particle size of approximately 15-20 nm ( Figure 2 A and B). After adding L-Glu, AuCoNPs showed more obvious aggregation ( Figure 2 C), while after adding D-Glu, AuCoNPs showed weak aggregation ( Figure 2 D) The differential aggregation behavior between L-Glu and D-Glu treated samples may be due to their stereospecific interactions with AuCoNPs. The stronger interaction between L-Glu and AuCoNPs leads to more pronounced aggregation of AuCoNPs.

[0065] XPS was used to characterize the elemental composition and chemical state of AuNPs and AuCoNPs. Figure 3 As shown, the binding energy of Co2p in AuCoNPs is located at 781.3 eV and 796.7 eV, corresponding to Co 2+ The characteristic peaks of Co2p3 / 2 and Co 2p1 / 2. The binding energies at 786.3eV and 803.5eV correspond to the oscillator satellite peaks. XPS results show that cobalt is mainly in the form of Co 2+ exists in the form of and is dispersed on the surface of AuCoNPs.

[0066] In order to subsequently build the CL model, the following proofs are proposed: AuCoNPs promote chemiluminescence (CL) signals;

[0067] Figure 4 It was shown that a 5-fold dilution of AuCoNPs solution could promote the CL signal of the luminol-hydrogen peroxide (Luminol-H2O2) system by about 304 times. EPR showed that three types of reactive oxygen species (ROS) were involved in the AuCoNPs-catalyzed Luminol-H2O2 reaction.

[0068] In order to subsequently build the TP model, the following proofs are proposed: photothermal conversion of AuCoNPs;

[0069] In order to evaluate the photothermal conversion effect of AuCoNPs, 200 μL of 5-fold diluted AuCoNPs solution was heated at a power density of 2.4 W / cm 2 The temperature was monitored by a pen-type digital thermometer. Figure 5As shown in A, no significant temperature increase was observed after 10 minutes of continuous irradiation. This is because the UV absorption peak of AuCoNPs is located at 520 nm, which prevents them from absorbing the energy of NIR-660 nm laser ( Figure 5 B). However, after the addition of Glu enantiomers to AuCoNPs, the UV absorption peak of AuCoNPs red-shifted from 520 nm to around 660-670 nm ( Figure 5 D), which enables them to absorb a large amount of NIR-660nm laser energy and convert it into heat. Figure 5 As shown in Figure C, the temperature increased significantly after continuous irradiation of the AuCoNPs solution containing Glu enantiomers for 10 minutes.

[0070] In order to subsequently build the RGB model, the following proofs are proposed: color conversion of AuCoNPs;

[0071] like Figure 6 As shown in A, the synthesized AuCoNPs exhibit a bright red color. Due to the localized surface plasmon resonance (LSPR) effect of AuCoNPs, the L / D-Glu enantiomers promote their aggregation, resulting in changes in the absorption, scattering spectra, and color in the visible light range ( Figure 6 B). AuCoNPs containing L-Glu appear purple, and AuCoNPs containing D-Glu appear pink.

[0072] S2: A multimodal sensing platform was constructed based on AuCoNPs solution, integrating chemiluminescence, temperature measurement, and smartphone-assisted RGB modes. Glu was used as a model analyte in the multimodal sensing platform, which was able to distinguish between L-Glu and D-Glu, achieving highly sensitive and specific detection of Glu.

[0073] The CL mode distinguishes L / D-Glu based on different chemiluminescence signal intensities;

[0074] The identification of Glu enantiomers in CL mode includes at least the following steps:

[0075] 50 μL of L / D-Glu solution with different concentrations was injected into the wells of a 96-well hydrogel microplate;

[0076] Then, 50 μL of 5-fold diluted AuCoNPs solution was immediately added and incubated at 37°C for 0.5 h;

[0077] Remove the supernatant and rinse the wells with ultrapure water;

[0078] Then 50 μL of the molar concentration of 5.0×10 -7 Luminol and 50 μL of 0.05 M hydrogen peroxide were injected into the microwell to collect CL signals.

[0079] The preparation in a 96-well hydrogel microplate includes at least the following steps: adding 50 μL of a 1 wt % chitosan solution into the wells of a 96-well microplate and drying at 50° C. to form a thin film as a binder for the PVA / SA hydrogel;

[0080] Then, 50 μL of a mixed solution containing 0.5% SA and 0.25 wt % PVA was added to the wells and frozen at −20 °C;

[0081] After 1 hour, 100 μL of a 0.05 wt % calcium chloride solution was added, and after cross-linking at room temperature for 1 hour, the excess liquid was removed and the microplate was washed with ultrapure water.

[0082] The TP mode systematically records the temperature signal and analyzes the reaction conditions at different concentrations, distinguishing the different reactivities of L / D-Glu based on the rate and amplitude of temperature increase.

[0083] The identification of Glu enantiomers in TP mode includes at least the following steps:

[0084] Fill a transparent test tube with 100 μL of L / D-Glu solution of different concentrations;

[0085] Then, 100 μL of 5-fold diluted AuCoNPs solution was added;

[0086] The reaction solution was irradiated with NIR-660 nm laser for 3 min, and a handheld digital thermometer was inserted into the liquid surface to collect the temperature signal of the solution.

[0087] The smartphone-assisted RGB mode monitors the color change produced by the reaction between L / D-Glu solution and AuCoNPs to identify the difference between the two. This is to accurately distinguish the reaction differences between L-Glu and D-Glu under the same experimental conditions by collecting and analyzing RGB values, especially grayscale values. Ultimately, the data obtained through the mobile phone video mode and color recognition application, combined with the grayscale value calculated by the weighted average method of difference, can provide a reliable basis for the identification of L-Glu and D-Glu.

[0088] The identification of Glu enantiomers in RGB mode includes at least the following steps:

[0089] 50 μL of L / D-Glu solution of different concentrations was loaded into a 96-well white microplate;

[0090] Then, 50 μL of 5-fold diluted AuCoNPs solution was added;

[0091] The reaction was carried out at a constant temperature of 37°C for 5 min and then transferred to a dark box with 5W fluorescent lamps on both sides;

[0092] The color signal in the 96-well microplate was collected using the mobile phone video mode;

[0093] Use a color recognition application to read the R, G, and B values in each well;

[0094] The grayscale value is calculated using the differential weighted average method.

[0095] Using Glu as a model analyte, the mechanism for identifying Glu enantiomers in chemiluminescence mode is also related to PVA / SA hydrogels. SA is the primary raw material for hydrogel formation and provides abundant chiral sites. PVA enhances the hydrogel's mechanical properties, allowing SA to form a stable hydrogel with only a small amount of calcium ions, thus avoiding the effects of high-concentration ion solutions on the stability of AuCoNPs. SA has a significantly higher affinity for Na₃Citt than the Glu enantiomer, indicating that Na₃Citt is the primary binding site for SA in the AuCoNPs / Glu enantiomer mixture. When AuCoNPs / L-Glu and AuCoNPs / D-Glu mixed solutions are added to the SA / PVA hydrogel, the strong affinity of L-Glu for AuCoNPs occupies a large number of Na₃Cit sites on the surface, preventing SA from binding and anchoring in the PVA / SA hydrogel. In contrast, the weaker affinity of D-Glu for AuCoNPs leaves Na3Cit sites abundant, allowing it to fully bind to SA and be immobilized in the SA / PVA hydrogel. Therefore, we speculate that the different chemiluminescence signals are generated due to the difference in the amount of AuCoNPs immobilized in the hydrogel after the addition of Glu enantiomers. Due to the local surface plasmon resonance of AuCoNPs, the addition of Glu enantiomers causes different degrees of aggregation and color changes. Under the NIR-660nm laser, AuCoNPs convert light into heat, allowing the identification of Glu enantiomers through TP signals. RGB imaging further allows for rapid and clear differentiation of Glu enantiomers.

[0096] Table 1 summarizes the analytical performance, LOD, and enantioselectivity of the three modes for glutamate enantiomers. Overall, CL mode offers the highest sensitivity, TP mode provides rapid and direct detection, and RGB mode enables low-cost and portable detection using portable devices. The combination of these three modes enables multidimensional identification and quantitative analysis of Glu enantiomers, with self-verification capabilities, improving the accuracy of Glu enantiomer identification.

[0097] Table 1

[0098]

[0099] In summary:

[0100] Most existing technologies are single-mode sensing. Due to the extreme similarity of enantiomers, a single signal mode is easily affected by experimental conditions and equipment, resulting in instability in detection results and increasing the possibility of errors. In contrast, multimodal sensing technology significantly improves the accuracy and reliability of detection by integrating multiple signal outputs. This sensor can provide multi-dimensional detection information, achieve self-verification, and make the analysis more accurate and reliable. Especially when identifying extremely similar chiral molecules, multimodal sensing technology not only overcomes the limitations of a single technology, but also greatly expands the recognition range, improves the analytical accuracy and applicability of enantiomers in complex environments, and demonstrates important practical application value. The present invention combines three modes: chemiluminescence, temperature measurement, and smartphone-assisted RGB, for the identification of chiral molecule enantiomers. The core innovation of this platform is that the multi-dimensional signal output of multimodal sensing overcomes the limitations of traditional single-mode sensing technology and significantly improves the range and reliability of chiral molecule enantiomer identification.

[0101] Example 2:

[0102] Based on the above embodiments, this embodiment proposes a portable test strip device based on AuCoNPs to improve portability and rapid detection effects.

[0103] When AuCoNPs are needed, the obtained AuCoNPs solution is freeze-dried to obtain AuCoNPs.

[0104] Prepare sample pads and test strips;

[0105] The AuCoNPs liquid is coated on the conjugate pad and then dried, which allows the AuCoNPs to react with the amino acid enantiomers in the sample under the conjugate pad.

[0106] SA hydrogel is coated on the detection line of the test strip. SA hydrogel can interact with the AuCoNPs-amino acid enantiomer complex to form a detection signal. The role of the SA hydrogel is to capture and fix those that have already formed the AuCoNPs-amino acid enantiomer complex, thereby forming a visual signal.

[0107] Carry out overall packaging to avoid exposure to light and moisture.

[0108] The application of the portable test strip device comprises at least the following steps:

[0109] When the amino acid enantiomer solution is added to the sample pad, the amino acid enantiomer preferentially interacts with the AuCoNPs;

[0110] The AuCoNPs and amino acid enantiomer complex solution flows toward the absorbent pad by capillary action.

[0111] They are then captured by the detection lines of the SA hydrogel to identify the enantiomers.

[0112] By integrating the technology proposed in Example 1 into a portable test strip device, it has better operational convenience and rapid detection capabilities. Through optimized design, the time it takes for the sample liquid to flow through the sample pad to the detection line is about 3 minutes, which meets the requirements of rapid detection. It also controls the reaction time between AuCoNPs and the analyte, greatly improving the efficiency and accuracy of on-site detection.

[0113] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above and that the invention can be embodied in other specific forms without departing from the spirit or essential characteristics of the invention. Therefore, the embodiments should be considered in all respects as illustrative and non-restrictive, and the scope of the invention is defined by the appended claims, not the foregoing description, and all variations within the meaning and range of equivalents of the claims are intended to be included therein. Any reference sign in a claim should not be construed as limiting the claim to which it relates.

Claims

1. A method for constructing an enantiomeric multimodal sensing platform based on plasmon nanozymes, characterized by: At least the following steps are included: S1: Preparation of plasmonic nanozymes. By element doping, cobalt is loaded onto gold nanoparticles to synthesize AuCoNPs. As plasmonic nanozymes and sensing materials, AuCoNPs have unique optical and catalytic properties. S2: Construction of a multimodal sensing platform based on AuCoNPs solution, integrating chemiluminescence, temperature measurement, and smartphone-assisted RGB modes. The multimodal sensing platform uses Glu as a model analyte and can distinguish between L-Glu and D-Glu, achieving highly sensitive and specific detection of Glu.

2. The method for constructing an enantiomeric multimodal sensing platform based on plasmon nanozymes according to claim 1, characterized in that: The preparation of AuCoNPs in S1 is a citric acid reduction method, and the specific process includes at least the following steps: A solution to be treated was prepared by mixing 100 μL of 1% chloroauric acid, 125 μL of 50 mmol / L cobalt chloride solution, and 9800 μL of water; The solution to be treated was added to a round-bottom flask equipped with a condenser, heated to boiling in an oil bath and refluxed continuously for 5 min; Then, 0.25 mL of a 2% sodium citrate solution was quickly injected into the boiled solution to be treated, causing the solution to turn red, thereby obtaining an AuCoNPs solution. When the AuCoNPs solution is required to remain in liquid form, store it directly in a brown glass bottle away from light.

3. The method for constructing an enantiomeric multimodal sensing platform based on plasmon nanozymes according to claim 1, characterized in that: The chemiluminescence is a CL mode, and the CL mode distinguishes L / D-Glu according to different chemiluminescence signal intensities; The identification of Glu enantiomers under the CL mode comprises at least the following steps: 50 μL of L / D-Glu solution with different concentrations was injected into the wells of a 96-well hydrogel microplate; Then, 50 μL of 5-fold diluted AuCoNPs solution was immediately added and incubated at 37°C for 0.5 h; Remove the supernatant and rinse the wells with ultrapure water; Then 50 μL of the molar concentration of 5.0×10 -7 Luminol and 50 μL of 0.05 M hydrogen peroxide were injected into the microwell to collect CL signals.

4. The method for constructing an enantiomeric multimodal sensing platform based on plasmon nanozymes according to claim 3, characterized in that: The preparation of the 96-well hydrogel microplate comprises at least the following steps: Fifty microliters of a 1 wt% chitosan solution was added to a 96-well microplate well and dried at 50°C to form a thin film as a binder for the PVA / SA hydrogel; Then, 50 μL of a mixed solution containing 0.5% SA and 0.25 wt % PVA was added to the wells and frozen at −20 °C; After 1 hour, 100 μL of a 0.05 wt % calcium chloride solution was added, and after cross-linking at room temperature for 1 hour, the excess liquid was removed and the microplate was washed with ultrapure water.

5. The method for constructing an enantiomeric multimodal sensing platform based on plasmon nanozymes according to claim 1, characterized in that: The temperature measurement is the TP mode, which systematically records the temperature signal and analyzes the reaction conditions at different concentrations to distinguish the different reactivities of L / D-Glu from the rate and amplitude of temperature increase; The identification of Glu enantiomers under the TP mode comprises at least the following steps: Fill a transparent test tube with 100 μL of L / D-Glu solution of different concentrations; Then, 100 μL of 5-fold diluted AuCoNPs solution was added; The reaction solution was irradiated with NIR-660 nm laser for 3 min, and a handheld digital thermometer was inserted into the liquid surface to collect the temperature signal of the solution.

6. The method for constructing an enantiomeric multimodal sensing platform based on plasmon nanozymes according to claim 1, characterized in that: The smartphone-assisted RGB is an RGB mode. In this RGB mode, the difference between L / D-Glu solution and AuCoNPs is identified by monitoring the color change produced by the reaction between the two. That is, by collecting and analyzing RGB values, especially grayscale values, the reaction differences between L-Glu and D-Glu under the same experimental conditions can be accurately distinguished. Ultimately, the data obtained by the mobile phone video mode and color recognition application, combined with the grayscale value calculated by the weighted average method of difference, can provide a reliable basis for the identification of L-Glu and D-Glu. The identification of Glu enantiomers under the RGB mode comprises at least the following steps: 50 μL of L / D-Glu solution of different concentrations was loaded into a 96-well white microplate; Then, 50 μL of 5-fold diluted AuCoNPs solution was added; The reaction was carried out at a constant temperature of 37°C for 5 min and then transferred to a dark box with 5W fluorescent lamps on both sides; The color signal in the 96-well microplate was collected using the mobile phone video mode; Use a color recognition application to read the R, G, and B values in each well; The grayscale value is calculated using the differential weighted average method.

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