A hyperbranched nanocomposite noble metal particle, a preparation method thereof and an application thereof

Abstract

The present invention discloses a hyperbranched nanocomposite noble metal particle, wherein the hyperbranched nanocomposite noble metal particle is a core-shell structure in which a noble metal is coated with a hyperbranched polyglycidol-n-dodecyl-polyethylene glycol copolymer HPG-C12-PEO, and the noble metal is selected from one or a combination of at least two of palladium, platinum or gold. The present invention also provides a preparation method of the above-mentioned hyperbranched nanocomposite noble metal particle and its application in the detection of small molecule metabolites by laser desorption ionization mass spectrometry. The preparation method provided by the present invention is simple, and the application of the provided hyperbranched nanocomposite noble metal particle in the detection of small molecule metabolites improves the accuracy and sensitivity of the detection.

Description

Technical Field

[0001] The present invention relates to the technical field of small molecule metabolite detection matrices, and particularly relates to a hyperbranched nanocomposite noble metal particle, a preparation method thereof, and an application thereof. Background Art

[0002] Liquid biopsy allows the detection of target biomarkers through non-invasive techniques from biological fluids (such as blood), offering hope for the early clinical screening of various diseases. Various biomarkers such as genetic materials and proteins have been found in human blood for the diagnosis of early cancers, but their applicability is limited by suboptimal accuracy, high cost, and lack of validation in early cancer patients. In addition to genes and proteins, the detection of metabolites can directly reflect the biochemical activities of the human body and is related to the occurrence of different diseases.

[0003] Mass spectrometry (MS), especially laser desorption / ionization (LDI) MS, is a powerful analytical tool capable of simultaneously detecting and spatially mapping various metabolites. However, in metabolic analysis, due to the low molecular abundance of metabolites and high sample complexity in specimens, it is inevitably offset by enrichment and purification. Charge and heat transfer from the matrix to the analyte play a crucial role in spectroscopic techniques. In particular, noble metals are expected to improve the LDI efficiency, generating a large density of charges and carrier relaxation in surface plasmon excitation to induce local heating. However, most current matrices are in the form of single-metal nanoparticles or their bimetallic alloys, showing unsatisfactory LDI sensitivity in clinical use. Therefore, a trimetallic alloy with a synergistic effect and mesoporous morphology will solve the limitations of single / bimetallic matrices and promote the precise diagnosis of downstream metabolic analysis. However, due to the high sample complexity and low metabolite abundance in patient specimens, the traditional analysis of low molecular weight compounds in biological fluids is hindered in clinical use.

[0004] Hyperbranched polymers and their nanomaterials possess unique physicochemical properties and attractive structural features, revolutionizing many applications, including nanocatalysis, energy storage and conversion, and nanomedicine. Matrix materials with designed molecular interfaces determine the performance of LDI-MS and have received strong attention from global research groups due to size exclusion effects and chemical specific affinities. In particular, plasmonic particles (such as silver) utilize the unique properties of surface plasmon resonance, nanoscale roughness, and "hot carrier" generation, which are produced by the quantum confinement effect of electron wave functions, but suffer from instability or oxidation, limiting their LDI process. New plasmonic particles can overcome the current major obstacles and be explored as effective matrices. Summary of the Invention

[0005] The object of the present invention is to provide a hyperbranched composite noble metal nanomaterial, a preparation method thereof and an application thereof. The preparation method is simple, and the application in the detection of small molecule metabolites improves the accuracy and sensitivity of the detection.

[0006] The technical solution provided by the present invention is as follows:

[0007] A hyperbranched nano-composite noble metal particle, wherein the hyperbranched nano-composite noble metal particle is a core-shell structure in which a hyperbranched polyglycidol-n-dodecyl-polyethylene glycol copolymer HPG-C12-PEO coats a noble metal, and the noble metal is selected from one or a combination of at least two of palladium, platinum or gold.

[0008] Or it can be understood that the hyperbranched nano-composite noble metal particle provided by the present invention includes an amphiphilic hyperbranched polymer template and a composite noble metal nanoparticle with a catalytic effect stabilized by this template. The hyperbranched polyglycidol-n-dodecyl-polyethylene glycol copolymer HPG-C12-PEO has an amphiphilic structure.

[0009] Preferably, the mass content of the noble metal in the hyperbranched nano-composite noble metal particle is 4-35%.

[0010] Preferably, the noble metals are palladium, platinum and gold, and the mass fractions are 1-5%, 1-20% and 1-10% respectively, and the total mass content is 8-35%. This combination can achieve good application effects with a small amount of effective metal components.

[0011] Preferably, the particle size of the hyperbranched nano-composite noble metal particle is 5-100 nm.

[0012] More preferably, the average particle size of the hyperbranched nano-composite noble metal particle is 8-16 nm. By regulating its average particle size, the accuracy and sensitivity of the detection of the composite noble metal particle in the detection of small molecule metabolites can be further improved.

[0013] The present invention also provides a preparation method of a hyperbranched nano-composite noble metal particle, and the preparation method includes:

[0014] (1) Add trimethylolpropane and heat to dissolve, then add potassium methoxide dissolved in anhydrous methanol, and remove methanol;

[0015] (2) Add a mixture of glycidol and polyglycidol dodecyl ether, continue to react after the reaction, add glycidol and react, and add anhydrous methanol and stir to form a homogeneous solution;

[0016] (3) The solution was neutralized with a hydrogen ion exchange resin column to obtain neutral macromolecules. After rotary evaporation to remove methanol, anhydrous ether was added, and the mixture was stirred to discard the ether. Then, it was dissolved in deionized water, dialyzed, and freeze-dried to obtain a viscous liquid HPG-C12-PEO;

[0017] (4) The noble metal precursor was dissolved in an aqueous solution of HPG-C12-PEO, and a vitamin C solution was added dropwise. Under continuous stirring, the noble metal precursor was reduced to obtain a core-shell structure with noble metal coated by HPG-C12-PEO.

[0018] In this invention, through anionic polymerization, the three-membered ring structure of glycidol was used as the branching condition. Through a one-pot synthesis method, under the condition of continuous slow feeding, a hyperbranched polymer template HPG-C12-PEO with an amphiphilic structure was synthesized. Then, through in-situ coordination-reduction method, metal nanoparticles were stabilized in the hyperbranched polymer template with an amphiphilic structure, thus synthesizing hyperbranched nanocomposite noble metal particles.

[0019] Preferably, the mass ratio of trimethylolpropane, hyperbranched polyglycidol, and polyethyleneglycol glycidyl dodecyl ether is 1-10:10-50:20-200; the number-average relative molecular mass of the polyethyleneglycol glycidyl dodecyl ether used is 600-1000.

[0020] Preferably, the relative number-average molecular mass of HPG-C12-PEO is 5000-32000.

[0021] Preferably, the heating temperature in step (1) is 50-70 °C, and the temperature for the reduction of the noble metal precursor in step (4) is 30-50 °C.

[0022] In this invention, by controlling process conditions such as the feed ratio and reaction temperature, the particle size of the hyperbranched nanocomposite noble metal particles is regulated to be between 5 and 100 nm.

[0023] Taking Pt / Pd / Au@HPG-C12-PEO as an example, the specific preparation method of the hyperbranched nanocomposite noble metal particles includes:

[0024] The three-necked flask was taken out after being dried in an oven, and metered trimethylolpropane was added. After heating to 60 °C for dissolution, the air and water vapor in the flask were expelled by the way of air extraction - nitrogen passing. Then, potassium methoxide, which had been weighed and dissolved in a small amount of anhydrous methanol (2 mL), was injected with a syringe. After magnetic stirring for 30 min, methanol was removed by vacuum pumping. Under nitrogen protection, a mixture of refined glycidol and polyethyleneglycol glycidyl dodecyl ether was continuously added dropwise by injection for 24 h, and then the reaction continued for 3 hours. Subsequently, a certain amount of glycidol was continuously injected by an injection pump for 24 h, and then the reaction continued for 3 hours. After adding 50 mL of anhydrous methanol, it was stirred into a homogeneous solution. Then, it was neutralized with a hydrogen ion exchange resin column to obtain a neutral macromolecule. After most of the methanol was removed by rotary evaporation, it was poured into 250 mL of anhydrous ether, and a viscous polymer was obtained after magnetic stirring for half an hour. The ether was discarded and this was repeated twice. Then, it was dissolved in deionized water and collected in a dialysis bag with a molecular weight cut-off of 3500, and dialyzed in deionized water for 3 days, with the water being changed 3 times a day. Finally, the liquid in the dialysis bag was freeze-dried in a vacuum freeze dryer to obtain a white viscous liquid.

[0025] Potassium tetrachloropalladate, chloroauric acid and chloroplatinic acid, these precious metal precursors, were dissolved in an aqueous solution of HPG-C12-PEO and stirred for a certain time until completely homogeneous. Subsequently, a vitamin C solution was added dropwise to the reaction system, and under the condition of continuous stirring, the above-mentioned precious metal precursors were reduced at 40 °C, and these precursors were gradually reduced under high-speed stirring, and finally Pt / Pd / Au@HPG-C12-PEO nanoparticles were obtained. The product was separated by ultracentrifugation, and finally a nanoparticle product was obtained.

[0026] The present invention also provides an application of the hyperbranched nanocomposite precious metal particles in the detection of small molecule metabolites by laser desorption ionization mass spectrometry (LDI-MS).

[0027] The sample treatment method for carrying out laser desorption ionization mass spectrometry LDI-MS detection is as follows: sample preparation is carried out by the method of covering the surface of an inorganic matrix. The method for sample preparation by covering the surface of an inorganic matrix includes: before LDI-MS detection, the hyperbranched nanocomposite precious metal particles are dispersed in water as a matrix solution with a concentration of 0.5 - 10 mg / mL; 0.01 - 1 μL of the analyte solution is dropped on a steel plate and allowed to dry naturally in air, and then 0.01 - 1 μL of the matrix solution is dropped to cover the upper layer of the analyte solution and wait for evaporation and drying.

[0028] The analyte solution is selected from serum, urine, cerebrospinal fluid, bile or tissue fluid, or tissues or excreta containing metabolite components.

[0029] The alloy particles of precious metals selected in the present invention can effectively improve the LDI efficiency, generating a large density of charges and carrier relaxation in surface plasmon excitation to induce local heating. Utilizing the unique properties of surface plasmon resonance, nanoscale roughness, and "hot carrier" generation, which are produced by the quantum confinement effect of the electron wave function, the form of molecular cleavage becomes more diverse and directional.

[0030] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0031] (1) The hyperbranched composite precious metal nanomaterials in the present invention can be used as an auxiliary matrix for LDI-MS detection, enabling rapid detection (as fast as only a few seconds) with a very small amount of body fluid (as low as 1 μL).

[0032] (2) The hyperbranched composite precious metal nanomaterials in the present invention can be controlled within 5 - 100 nm, effectively overcoming the "sweet spot effect" and high background signal interference problems in the detection of small molecule metabolites (relative molecular mass below 600 Da) by LDI-MS, improving the abundance of characteristic peaks of small molecule metabolites, and making it more convenient for testers to obtain their fingerprint spectra.

[0033] (3) The hyperbranched nano-composite precious metal particles provided by the present invention, as an inorganic matrix for laser desorption ionization mass spectrometry, can effectively improve the LDI efficiency of such composite precious metal nanoparticles, generating a large density of charges and carrier relaxation in surface plasmon excitation to induce local heating. Utilizing the unique properties of surface plasmon resonance, nanoscale roughness, and "hot carrier" generation, which are produced by the quantum confinement effect of the electron wave function, the form of molecular cleavage becomes more diverse and directional, effectively improving the detection effect of small molecule metabolites.

[0034] In summary, the present invention designs a functional nano-reactor based on the unique advantages of hyperbranched polymers. The hyperbranched polymer HPG-C12-PEO that can carry nano-precious metal particles is synthesized by a one-step reaction method. The synthesized hyperbranched polymer is used as a template for nano-precious metals, and further the synthesized hyperbranched nano-alloy particles are used as a matrix to carry out LDI-MS detection of various body fluids. This invention will contribute to the progress of constructing tools for personalized diagnosis of early diseases. Description of the Drawings

[0035] Figure 1 1H NMR analysis chart of HPG-C12-PEO prepared in Example 1;

[0036] Figure 2 13C NMR analysis chart of HPG-C12-PEO prepared in Example 1;

[0037] Figure 3Infrared spectrum of HPG-C12-PEO prepared in Example 1;

[0038] Figure 4 Transmission electron microscopy image of hyperbranched composite noble metal nanoparticles prepared in Example 2;

[0039] Figure 5 Particle size distribution diagram of dynamic light scattering test of hyperbranched nanoalloy particles prepared in Examples 2, 3, and 4;

[0040] Figure 6 XRD pattern of hyperbranched nanoalloy particles prepared in Examples 2, 3, and 4;

[0041] Figure 7 Enhanced effect of hyperbranched nanocomposite noble metal nanoparticles prepared in Example 2 on the detection of small molecule metabolites in mouse serum by Maldi-MS. Detailed implementation mode

[0042] The present invention will be further described below in conjunction with specific embodiments.

[0043] Example 1: Preparation of HPG-C12-PEO

[0044] After the three-necked flask was dried in an oven and taken out, the measured trimethylolpropane was added. After heating to 60 °C and dissolving, the air and water vapor in the flask were removed by the method of pumping air - passing nitrogen. Then, potassium methoxide weighed and dissolved in a small amount of anhydrous methanol (2 mL) was injected with a syringe. After magnetic stirring for 30 min, methanol was removed by vacuum. Under nitrogen protection, a mixture of refined glycidol and polyethylene glycol glycidyl dodecyl ether was continuously added dropwise with a syringe for 24 h, and then the reaction was continued for 3 hours. Subsequently, a certain amount of glycidol was continuously injected with a syringe pump for 24 hours, and then the reaction was continued for 3 hours. After adding 50 mL of anhydrous methanol, it was stirred into a homogeneous solution. Then, it was neutralized with a hydrogen ion exchange resin column to obtain a neutral macromolecule. After most of the methanol was removed by rotary evaporation, it was poured into 250 mL of anhydrous ether. After magnetic stirring for half an hour, a viscous polymer was obtained. The ether was discarded and repeated 2 times. Then, it was dissolved in deionized water and collected in a dialysis bag with a molecular weight cut-off of 3500, and dialyzed in deionized water for 3 days, changing the water 3 times a day. Finally, the liquid in the dialysis bag was freeze-dried in a vacuum freeze dryer to obtain a white viscous liquid.

[0045] In this example, the dosages of trimethylolpropane, hyperbranched polyglycidol, and polyethylene glycol glycidyl dodecyl ether are 0.3 mol, 25 mL, and 10 mL respectively. The number-average relative molecular mass of the polyethylene glycol glycidyl dodecyl ether used is 400, and the relative number-average molecular mass of the prepared HPG-C12-PEO is 8,600.

[0046] Example 2: Preparation of Hyperbranched Composite Noble Metal Nanomaterials Pt / Pd / Au@HPG-C12-PEO

[0047] Potassium tetrachloropalladate, chloroauric acid and chloroplatinic acid were dissolved in an aqueous solution of HPG-C12-PEO and stirred for 2 h until completely homogeneous. Subsequently, a vitamin C solution was added dropwise to the reaction system to reduce the above noble metal precursors. The noble metal precursors were gradually reduced under high-speed stirring, and finally, nanoparticles of Pt / Pd / Au@HPG-C12-PEO were obtained. The product was separated by ultracentrifugation, and finally, a nanoparticle product was obtained.

[0048] In this example, the dosages of potassium tetrachloropalladate, chloroplatinic acid and chloroauric acid were 2 mg, 10 mg and 4 mg respectively. The mass content of noble metals in the prepared hyperbranched composite noble metal nanoparticles was 8.6 wt%, and the mass fractions of palladium, platinum and gold were 1.5 wt%, 5.4 wt% and 1.7 wt% respectively. The average particle size of the prepared hyperbranched composite noble metal nanoparticles was 15.7 nm.

[0049] Example 3: Preparation of Hyperbranched Composite Noble Metal Nanomaterials Pd / Pt@HPG-C12-PEO

[0050] Potassium tetrachloropalladate and chloroplatinic acid were dissolved in an aqueous solution of HPG-C12-PEO and stirred for 2 h until completely homogeneous. Subsequently, a vitamin C solution was added dropwise to the reaction system to reduce the above noble metal precursors. The noble metal precursors were gradually reduced under high-speed stirring, and finally, nanoparticles of Pt / Pd / Au@HPG-C12-PEO were obtained. The product was separated by ultracentrifugation, and finally, a nanoparticle product was obtained.

[0051] In this example, the dosages of potassium tetrachloropalladate and chloroplatinic acid were 2 mg and 10 mg respectively. The mass content of noble metals in the prepared hyperbranched composite noble metal nanoparticles was 6.5 wt%, and the mass fractions of palladium and platinum were 2.8 wt% and 3.7 wt% respectively. The average particle size of the prepared hyperbranched composite noble metal nanoparticles was 13.6 nm.

[0052] Example 4: Preparation of Hyperbranched Composite Noble Metal Nanomaterials Au / Pt@HPG-C12-PEO

[0053] Potassium chloroaurate and chloroplatinic acid were dissolved in an aqueous solution of HPG-C12-PEO and stirred for 2 h until completely homogeneous. Subsequently, a vitamin C solution was added dropwise to the reaction system to reduce the above noble metal precursors. The noble metal precursors were gradually reduced under high-speed stirring, and finally, nanoparticles of Au / Pt@HPG-C12-PEO were obtained. The product was separated by ultracentrifugation, and finally, a nanoparticle product was obtained.

[0054] In this example, the dosages of potassium chloroaurate and chloroplatinic acid were 4 mg and 10 mg respectively. The mass content of noble metals in the prepared hyperbranched composite noble metal nanoparticles was 7.4 wt%, and the mass fractions of gold and platinum were 3.2 wt% and 4.2 wt% respectively. The average particle size of the prepared hyperbranched composite noble metal nanoparticles was 9.4 nm.

[0055] Example 5: Preparation of hyperbranched composite noble metal nanomaterial Pd / Au@HPG-C12-PEO

[0056] Potassium chloroaurate and potassium tetrachloropalladate were dissolved in an aqueous solution of HPG-C12-PEO and stirred for 2 h until completely homogeneous. Subsequently, a vitamin C solution was added dropwise to the reaction system to reduce the above-mentioned noble metal precursors. The noble metal precursors were gradually reduced under high-speed stirring, and finally nanoparticles of Pd / Au@HPG-C12-PEO were obtained. The product was separated by ultracentrifugation, and finally a nanoparticle product was obtained.

[0057] In this example, the dosages of potassium chloroaurate and potassium tetrachloropalladate were 4 mg and 2 mg respectively. The mass content of noble metals in the prepared hyperbranched composite noble metal nanoparticles was 4.5 wt%, and the mass fractions of gold and palladium were 2.8 wt% and 1.7 wt% respectively. The average particle size of the prepared hyperbranched composite noble metal nanoparticles was 8.6 nm.

[0058] Characterization test 1: 1H NMR analysis

[0059] The 1H NMR spectrum of HPG-C12-PEO prepared in Example 1 was as Figure 1 shown. Figure 1 The peaks at 0.82 and 1.29 ppm in it were the peaks of saturated hydrocarbons on n-dodecyl, and their integrals conformed to the numerical ratio of n-dodecyl. The continuous stacked peaks at 3-4 ppm in the middle were the peaks of polyglycidol and PEO in the core and outer layer. The hydrogen on the oxygen-carrying carbon was in this range. Due to the complex polymer structure, it showed a wide stacked peak. The spectrum proved the successful preparation of HPG-C12-PEO.

[0060] Characterization test 2: 13C NMR analysis

[0061] As Figure 2 shown, in 13 13C NMR, the three peaks before 40 ppm belonged to the peaks of three types of carbon on n-dodecyl. The slightly wider peak between 60-85 ppm belonged to the peak of the polymer structure. Since PEO, like HPG, had a polyether backbone structure, there was a large overlap in 13 13C NMR and it appeared as a continuous peak in the spectrum.

[0062] Characterization Test 3: Infrared Spectroscopy Detection

[0063] The infrared spectrum of HPG-C12-PEO prepared in Example 1 is given ( Figure 3 ). As shown in the spectrum, there are typical hydroxyl peaks at 3400 cm-1, peaks of long-chain alkyl groups near 2890 cm -1 , and peaks of stretching vibrations of multiple ether bonds in typical PEO at 1200 cm -1 , all indicating the presence of the above functional groups in HPG-C12-PEO.

[0064] Characterization Test 4: Transmission Electron Microscopy Analysis

[0065] Figure 4 is the transmission electron microscope image of hyperbranched composite noble metal nanoparticles.

[0066] Characterization Test 5: Transmission Electron Microscopy Analysis

[0067] Figure 5 is the particle size distribution diagram of dynamic light scattering test of hyperbranched composite noble metal nanoparticles prepared in Examples 2-4.

[0068] Characterization Test 6: X-ray Diffraction Analysis

[0069] An X-ray diffractometer was used to analyze the crystal structures of these three kinds of nanoparticles. The results are as Figure 6 shown. Since the properties of these three metals are relatively similar in the periodic table and their diffraction peak angles are relatively close in the XRD spectrum, the corresponding crystal planes can still be found according to their fingerprint spectra. The details are as Figure 6 shown. The signal peaks at angles below 40 in the spectrum belong to the peaks of hyperbranched polymers. Among them, the sharp 1-2 peaks should be caused by the crystallization diffraction brought by a certain degree of linear structure existing in the hyperbranched structure. At the same time, in the crystal structure of hyperbranched composite noble metal nanoparticles, the three metals form alloys with each other, so the diffraction peaks of the three metals in their XRD spectra have obvious enhancement and broadening.

[0070] Performance Test 1: Test on the Enhancement Effect of Hyperbranched Composite Noble Metal Nanoparticles on the Maldi-MS Detection of Serum Small Molecule Metabolites

[0071] The performance of the palladium, platinum, and gold composite nanoparticles in the hyperbranched composite noble metal nanoparticles prepared in Example 2 in LDI MS as a substrate was studied by the number of metabolic signals. In a typical mass spectrum from a single representative normal mouse in the m / z range of 100 - 800, among which, within the molecular weight range of 200 - 400 of typical small molecule metabolites, 110 - 250 strong metabolic signals were received on Pd / Pt / Au@HPG-C12-PEO using 1 μL of natural serum within 1 minute. The results are as Figure 7 shown.

[0072] In summary, the hyperbranched nano-composite noble metal particles provided by the present invention improve the accuracy and sensitivity of LDI-MS for the detection of small molecule metabolites, thereby realizing the rapid detection of small molecule metabolite fingerprint maps in various body fluids by the LDI-MS method. While enhancing the detection effect, the amount of body fluid specimens used is reduced; the preparation method provided by the present invention is simple, and nano-materials with high sensitivity and high responsiveness can be obtained.

Claims

1. Application of hyperbranched nanocomposite noble metal particles in detecting small molecule metabolites by laser desorption ionization mass spectrometry, characterized in that, The sample preparation method for laser desorption ionization mass spectrometry (LDI-MS) detection is to prepare samples by covering the surface of an inorganic matrix. Specifically: before LDI-MS detection, hyperbranched nanocomposite noble metal particles are dispersed in water as a matrix solution with a concentration of 0.5 - 10 mg / mL; 0.01 - 1 μL of the analyte solution is dropped on a steel plate and allowed to dry naturally in air, and then 0.01 - 1 μL of the matrix solution is dropped to cover the upper layer of the analyte solution and wait for evaporation and drying; The hyperbranched nanocomposite noble metal particles are core-shell structures with hyperbranched polyglycidol - n-dodecyl - polyethylene glycol copolymer (HPG-C12-PEO) coating noble metals; The noble metals are palladium, platinum, and gold, with mass fractions of 1 - 5%, 1 - 20%, and 1 - 10% respectively, and the total mass content is 8 - 35%; The particle size of the hyperbranched nanocomposite noble metal particles is 5 - 100 nm.

2. The application according to claim 1, wherein The relative number-average molecular weight of the HPG-C12-PEO is 5000 - 32000.

3. The application according to claim 1, characterized in that, The preparation method of the hyperbranched nanocomposite noble metal particles includes: (1) Add trimethylolpropane and heat to dissolve, then add potassium methoxide dissolved in anhydrous methanol and remove methanol; (2) Add a mixture of glycidol and poly(ethylene glycol) glycidyl dodecyl ether, continue to react after the reaction by adding glycidol, and add anhydrous methanol and stir to form a homogeneous solution; (3) The solution is neutralized with a hydrogen ion exchange resin column to obtain a neutral macromolecule. After rotary evaporation to remove methanol, add anhydrous ether, stir and discard the ether; then dissolve with deionized water, dialyze, and freeze-dry to obtain a viscous liquid HPG-C12-PEO; (4) Dissolve the noble metal precursor in an aqueous solution of HPG-C12-PEO, dropwise add vitamin C solution, and under continuous stirring, the noble metal precursor is reduced to obtain a core-shell structure with HPG-C12-PEO coating noble metals.

4. The application according to claim 3, characterized in that The mass ratio of trimethylolpropane, hyperbranched polyglycidol, and poly(ethylene glycol) glycidyl dodecyl ether is 1 - 10:10 - 50:20 - 200; the number-average relative molecular weight of the poly(ethylene glycol) glycidyl dodecyl ether used is 600 - 1000.

5. The application according to claim 1, characterized in that, The analyte solution is selected from serum, urine, cerebrospinal fluid, bile, or tissue fluid.

Images