An hp-aZIF@tar@au sensor, a preparation method thereof and application thereof in detection of human sweat tyrosine

Abstract

The application provides an HP-aZIF@Tar@Au sensor and a preparation method and application thereof in detection of human sweat tyrosine, and belongs to the technical field of nanomaterials.The HP-aZIF@Tar@Au sensor comprises an AAO substrate and HP-aZIF@Tar@Au composite material loaded on the surface of the AAO substrate, and raw materials of the HP-aZIF@Tar@Au composite material comprise HP-ZIF-67, HAuCl4 and L-tartaric acid.The HP-aZIF@Tar@Au sensor is simple and effective in preparation, can be used for super-sensitive and specific detection of human sweat tyrosine, is stable in process, and is high in sensitivity and good in selectivity.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of sensors, and particularly relates to an HP-aZIF@Tar@Au sensor and a preparation method thereof and application thereof in detecting human sweat tyrosine. BACKGROUND

[0002] Sweat contains rich biomarkers and metabolites, which can provide valuable insights into an individual's health, athletic performance, and potential disease risks. Diseases such as albinism and melanoma can cause abnormal expression of tyrosine in sweat, so detecting the content of tyrosine in sweat has important biomedical value.

[0003] Surface-enhanced Raman scattering (SERS) is a non-destructive molecular detection technique that reflects molecular bond vibration energy and rotational energy information, and is a "fingerprint" spectrum that can specifically identify molecules. When the target detection molecules are adsorbed to some rough metal surfaces (such as gold, silver, copper, etc.), their Raman scattering intensity will increase by 10 4 ~ 10 6 times. This technology has the advantages of high sensitivity, small sample size, simple pretreatment, non-destructive sample analysis, and no interference in simultaneous detection of multiple analytes.

[0004] Metal-organic frameworks (MOFs) are versatile and customizable materials composed of metal ions and organic ligands arranged in a crystal structure. They have a wide range of applications in various scientific fields, including chemistry, biology, and materials science. One important application of MOFs is the targeted detection of small molecules present in sweat. ZIF-67 is an organic metal framework material (MOF) with multiple advantages such as high selectivity, high sensitivity, good stability, good controllability, and multifunctionality, making it a research hotspot in the field of sweat small molecule monitoring.

[0005] However, combining SERS technology with MOFs to effectively capture and detect tyrosine molecules in sweat requires the development of new integration strategies and technologies. To this end, the present application proposes an HP-aZIF@Tar@Au sensor and a preparation method thereof and application thereof in detecting human sweat tyrosine. SUMMARY

[0006] To solve the above technical problems, the present application proposes an HP-aZIF@Tar@Au sensor and a preparation method thereof and application thereof in detecting human sweat tyrosine.

[0007] To achieve the above-mentioned purposes, the present application provides the following technical solutions:

[0008] One of the technical solutions of the present application is:

[0009] The HP-aZIF@Tar@Au sensor comprises an AAO (anodic aluminum oxide oxide) substrate and a HP-aZIF@Tar@Au composite material loaded on the surface of the AAO substrate, and raw materials for preparing the HP-aZIF@Tar@Au composite material comprise HP-ZIF-67, HAuCl4 and L-tartaric acid.

[0010] Further, the HP-ZIF-67 is ZIF-67 with a porous structure, and "HP" represents hierarchical porosity, a metal salt for preparing the HP-ZIF-67 is Co(NO3)2·6H2O, and a ligand is 2-methylimidazole.

[0011] Further, the loading amount of the HP-aZIF@Tar@Au composite material on the AAO substrate is 0.075 mg / cm 2 .

[0012] The second technical solution of the present application is:

[0013] A preparation method of the HP-aZIF@Tar@Au sensor comprises the following steps:

[0014] After the HP-ZIF-67 is dispersed with a solvent, the HAuCl4 solution is mixed and stirred, the product is centrifuged and then dispersed with a solvent again, and the NaBH4 solution is mixed and stirred in an ice water bath to obtain a mixed solution, the mixed solution is centrifuged and dried to obtain the HP-ZIF-67@Au;

[0015] The HP-ZIF-67@Au is mixed and ground with L-tartaric acid, and then refluxed in toluene, cooled, centrifuged and dried to obtain the HP-aZIF@Tar@Au composite material;

[0016] After the HP-aZIF@Tar@Au composite material is dispersed with DMF, it is added dropwise to the AAO substrate treated with hydrophilic to obtain the HP-aZIF@Tar@Au sensor.

[0017] Further, the solvent is anhydrous ethanol and deionized water, and the volume ratio of the anhydrous ethanol to the deionized water is 1:2.

[0018] Further, the molar ratio of the HP-ZIF-67 to the HAuCl4 is 2:1, and the molar ratio of Au 3+ in the mixed solution to NaBH4 in the NaBH4 solution is 3:20.

[0019] Further, the concentration of the HAuCl4 solution is 0.025 M, and the concentration of the NaBH4 solution is 0.1 M.

[0020] Further, the molar ratio of the HP-ZIF-67@Au and L-tartaric acid is 1:1.2.

[0021] Further, the concentration of the HP-aZIF@Tar@Au composite material dispersed by DMF is 2 mg / mL.

[0022] Further, the HP-ZIF-67 is dispersed by solvent, and then HAuCl4 solution is added, mixed and stirred for 3 h at room temperature, and NaBH4 solution is added, mixed and stirred for 5 h in an ice water bath.

[0023] Further, the AAO substrate is hydrophilic treated by 2-amino terephthalic acid.

[0024] Further, the diameter of the AAO substrate is 390 nm, and the thickness is 40 mu m.

[0025] The third technical scheme of the present application is as follows:

[0026] The application of the HP-aZIF@Tar@Au sensor in detecting tyrosine in human sweat.

[0027] Further, the tyrosine in the human sweat attached to the surface of the HP-aZIF@Tar@Au sensor is determined by a Raman spectrometer, and the detection limit reaches 10 -8 M.

[0028] Compared with the prior art, the present application has the following advantages and technical effects:

[0029] The HP-aZIF@Tar@Au sensor provided by the present application is prepared by a physical deposition method, which is simple and efficient, and the HP-aZIF@Tar@Au sensor is mainly used for the ultra-sensitive and specific detection of tyrosine in human sweat, and the HP-aZIF@Tar@Au sensor can realize the rapid identification of tyrosine in a complex biological environment, and has excellent sensitivity and selectivity. In practical application, tyrosine and other interfering substances can be effectively distinguished, and high-precision detection can be realized. BRIEF DESCRIPTION OF DRAWINGS

[0030] The accompanying drawings, which form a part of the present application, are included to provide a further understanding of the present application, and are incorporated herein for purposes of explanation, and are not intended to limit the present application. In the drawings:

[0031] Figure 1 The SEM characterization diagram of the HP-ZIF-67@Au prepared in Example 1;

[0032] Figure 2SEM characterization diagram of the HP-aZIF@Tar@Au composite material prepared in Example 2;

[0033] Figure 3 XRD diagram of the HP-ZIF-67@Au prepared in Example 1 and the HP-aZIF@Tar@Au composite material prepared in Example 2;

[0034] Figure 4 Raman diagram of SERS detection of different concentrations of tyrosine by the HP-aZIF@Tar@Au sensor prepared in Example 3;

[0035] Figure 5 Raman diagram of SERS detection of 9 kinds of amino acids by the HP-aZIF@Tar@Au sensor prepared in Example 3;

[0036] Figure 6 Raman test result diagram of 10 -5 M tyrosine by the HP-aZIF@Tar@Au sensor prepared in Example 3, Comparative Example 3;

[0037] Figure 7 RSD diagram of 10 -5 M tyrosine 20 test points by the HP-aZIF@Tar@Au sensor prepared in Example 3, Comparative Example 4. DETAILED DESCRIPTION

[0038] The present application will now be described in detail with specific reference being made to various examples thereof, which are not to be construed as limiting the application, but rather as being illustrative thereof. The detailed description makes reference to the following figures, in which:

[0039] It is to be understood that the terms used in the present application are merely descriptive, but are not intended to limit the application. In addition, for numerical ranges in the present application, it is to be understood that each intermediate value between the upper limit and the lower limit of the range is specifically disclosed. Each smaller range within any stated value or stated range, as well as between any stated value or stated range, is also included in the present application. The upper and lower limits of these smaller ranges can be independently included or excluded from the range.

[0040] Unless otherwise indicated, all technical and scientific terms used herein have the same meaning as those of ordinary skill in the art to which the application relates. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present application, the preferred methods and materials are described. All documents mentioned herein are incorporated by reference to disclose and describe in full the methods and / or materials which are described therein. In case of conflict between the content of the specification and that of any document incorporated herein by reference, the content of the specification prevails.

[0041] Many modifications and variations of this application can be made in the light of the above teachings without departing from the spirit or scope of the application. Other implementations of the application will be apparent to those skilled in the art from consideration of the specification and practice of the application disclosed herein. The specification and examples given are exemplary only.

[0042] As used herein, the terms "comprises", "comprising", "includes", "including", "has", "having", "contains", "containing", or variations thereof, are intended to be open-ended terms that mean inclusion, but not limited to, the listed materials and methods.

[0043] The embodiment of the present application provides a HP-aZIF@Tar@Au sensor, which comprises an AAO (anodic aluminum oxide) substrate and a HP-aZIF@Tar@Au composite material loaded on the surface of the AAO substrate, and the raw material of the HP-aZIF@Tar@Au composite material comprises HP-ZIF-67, HAuCl4 and L-tartaric acid.

[0044] In the preferred embodiment of the present application, the loading amount of the HP-aZIF@Tar@Au composite material on the AAO substrate is 0.075 mg / cm2. 2 .

[0045] In the preferred embodiment of the present application, the HP-ZIF-67 is ZIF-67 with a porous structure, and "HP" represents hierarchical porosity. The metal salt for preparing the HP-ZIF-67 is Co (NO3) 2·6H2O, and the ligand is 2-methylimidazole. The HP-ZIF-67 is specifically purchased from the Maclean Limited Company.

[0046] The embodiment of the present application further provides a preparation method of the HP-aZIF@Tar@Au sensor, which comprises the following steps:

[0047] The HP-ZIF-67 is dispersed with a solvent, and then added into a HAuCl4 solution to mix and stir. The product is centrifuged, and then dispersed with a solvent again. A NaBH4 solution is added to mix and stir in an ice water bath to obtain a mixed solution. The mixed solution is centrifuged, purified and dried to obtain the HP-ZIF-67@Au.

[0048] The HP-ZIF-67@Au is mixed and ground with L-tartaric acid, then refluxed in toluene, cooled, centrifuged and dried to obtain the HP-aZIF@Tar@Au composite material;

[0049] The HP-aZIF@Tar@Au composite material is dispersed with DMF, then added dropwise to the hydrophilic treated AAO substrate to obtain the HP-aZIF@Tar@Au sensor.

[0050] In the preferred embodiment of the present application, the solvent used in the two solvent dispersion processes in the preparation of HP-ZIF-67@Au is the same, which is an ethanol aqueous solution prepared from anhydrous ethanol and deionized water, and the volume ratio of the anhydrous ethanol and deionized water is 1:2.

[0051] In the preferred embodiment of the present application, the molar ratio of HP-ZIF-67 and HAuCl4 is 2:1, and the molar ratio of Au 3+ to NaBH4 in the NaBH4 solution is 3:20.

[0052] In the preferred embodiment of the present application, the concentration of the HAuCl4 solution is 0.025M, and the concentration of the NaBH4 solution is 0.1M.

[0053] In the preferred embodiment of the present application, the molar ratio of HP-ZIF-67@Au and L-tartaric acid is 1:1.2. The mixed grinding is preferably carried out using a agate mortar, and the grinding time is 15min.

[0054] In the preferred embodiment of the present application, after centrifugation, a washing step is further included, and the product is washed with ethanol; and the drying is specifically vacuum drying at 60℃ overnight.

[0055] In the preferred embodiment of the present application, the concentration of the HP-aZIF@Tar@Au composite material after being dispersed with DMF is 2mg / mL.

[0056] In the preferred embodiment of the present application, after the HP-ZIF-67 is dispersed with a solvent, the HAuCl4 solution is added, and the mixture is stirred at room temperature for 3h, and then the NaBH4 solution is added, and the mixture is stirred in an ice water bath for 5h.

[0057] In the preferred embodiment of the present application, the AAO substrate is subjected to hydrophilic treatment with 2-amino terephthalic acid. The specific treatment steps include: immersing the AAO substrate in a 0.1M 2-amino terephthalic acid solution for 24h, rinsing the AAO substrate clean after completing the hydrophilic treatment, and air drying at room temperature.

[0058] In the preferred embodiment of the present application, the AAO substrate has a diameter of 390 nm and a thickness of 40 μm, and is purchased from Topo Precision Film Technology Co., Ltd. of Shenzhen.

[0059] In the embodiment of the present application, "room temperature" refers to "25±3℃".

[0060] The technical solutions of the present application are further described below through examples.

[0061] Example 1

[0062] Preparation of HP-ZIF-67@Au:

[0063] The porous HP-ZIF-67 (60 mg) was dispersed in 3 mL of anhydrous ethanol and deionized water solution (v / v = 1:2) and ultrasonically treated for 10 min for sufficient dispersion, and then an aqueous solution of HAuCl4 (5.4 mL, 0.025 M, i.e., the molar ratio of HP-ZIF-67 to HAuCl4 is 2:1) was added under stirring, and the reaction mixture was stirred at room temperature for 3 h, and then the product was centrifuged (6000 rpm, 5 min) and re-dispersed in 3 mL of anhydrous ethanol and deionized water solution (v / v = 1:2) under ultrasonic treatment, and then placed in an ice water bath after stirring for 5 min, and then an aqueous solution of NaBH4 (9 mL, 0.1 M) was added to the mixture under the condition of ice water bath, and the reaction mixture was stirred in the ice water bath for 5 h, and then the reaction was completed, and the product was centrifuged and purified using anhydrous ethanol in three repeated centrifugation cycles (6000 rpm, 5 min), and dried at 60℃ for 12 h to obtain HP-ZIF-67@Au.

[0064] The SEM characterization graph of HP-ZIF-67@Au prepared in Example 1 is shown in Figure 1 As shown in the figure, the Au nanoparticles are uniformly distributed on the HP-ZIF-67, and the porous structure and complete crystal structure of the HP-ZIF-67 are retained.

[0065] Example 2

[0066] Preparation of HP-aZIF@Tar@Au composite material:

[0067] The HP-ZIF-67@Au (1 mmol, 0.28 g) prepared in Example 1 and L-tartaric acid (1.2 mmol, 0.18 g) were ground in a marble mortar for 15 min, and then refluxed in toluene for 6 h, and then the reaction mixture was cooled, centrifuged, and the product was washed with ethanol and dried under vacuum at 60℃ overnight to obtain the HP-aZIF@Tar@Au composite material.

[0068] The SEM characterization graph of the HP-aZIF@Tar@Au composite material prepared in Example 2 is shown in Figure 2 It can be seen from the figure that after the HP-ZIF-67@Au is doped with tartaric acid molecules, the HP-aZIF@Tar@Au still retains a hierarchical pore structure, and has undergone amorphous transformation.

[0069] Example 3

[0070] Preparation of the HP-aZIF@Tar@Au sensor:

[0071] The AAO substrate is hydrophilically treated with 2-amino terephthalic acid, and the specific process is as follows:

[0072] The AAO substrate is immersed in a 0.1M 2-amino terephthalic acid solution for 24h, and after the hydrophilic treatment is completed, it is washed clean with distilled water and air-dried at room temperature for standby use.

[0073] The HP-aZIF@Tar@Au composite material prepared in Example 2 is dispersed in DMF to obtain a mixture with a concentration of 2mg / mL, and by dropping 50μL of the mixture liquid onto the AAO substrate (pore size 390nm, thickness 40μm) hydrophilically treated with 2-amino terephthalic acid, the loading amount of the HP-aZIF@Tar@Au composite material on the AAO substrate is 0.075mg / cm 2 , to obtain the HP-aZIF@Tar@Au sensor.

[0074] The XRD patterns of the HP-ZIF-67@Au prepared in Example 1, the HP-aZIF@Tar@Au composite material prepared in Example 2, and the raw material HP-ZIF-67 of Example 1, as well as the XRD standard cards of Au and ZIF-67 are shown in Figure 3 It can be seen that the HP-ZIF-67 in the HP-aZIF@Tar@Au composite material is converted into amorphous MOF.

[0075] The HP-aZIF@Tar@Au sensor prepared in Example 3 is used to detect the tyrosine (Try) in the human sweat attached to the surface by a Raman spectrometer, and the SERS detection of tyrosine with different concentrations by the HP-aZIF@Tar@Au sensor prepared in Example 3 is shown in Figure 4 It can be seen that the detection limit reaches 10 -8 M.

[0076] The HP-aZIF@Tar@Au sensor prepared in Example 3 is used to detect 12 kinds of amino acids by SERS, and the Raman spectra are shown in Figure 5, proving that the HP-aZIF@Tar@Au sensor has a specific detection function for tyrosine. The present invention achieves the specific recognition of Tyr in complex sweat through the difference in adsorption energy between different amino acids and tartaric acid.

[0077] Comparative Example 1

[0078] The method provided by the patent application number 201610619910.7 has a detection limit of 3.0×10 -5 mol / L.

[0079] Comparative Example 2

[0080] The SERS detection limit (LOD) of the microneedle SERS sensor for in situ detection of tyrosinase in patent application number 202311113388.1 is 0.01 U / m.

[0081] Comparative Example 3

[0082] The preparation of HP-ZIF-67@Au was the same as in Example 1;

[0083] The HP-aZIF@Tar@Au composite material was prepared as in Example 2, except that HP-ZIF-67@Au (1 mmol, 0.28 g) and L-tartaric acid (0.6 mmol, 0.09 g) were ground in an agate mortar;

[0084] The preparation method of the HP-aZIF@Tar@Au sensor is the same as that in Example 3.

[0085] Comparative Example 4

[0086] The preparation of HP-ZIF-67@Au was the same as in Example 1;

[0087] The preparation of HP-aZIF@Tar@Au composite material was the same as in Example 2;

[0088] The preparation method of the HP-aZIF@Tar@Au sensor was the same as that in Example 3, except that the loading amount of the HP-aZIF@Tar@Au composite material on the AAO substrate was 0.045 mg / cm 2 .

[0089] The detection and uniformity performance of the sensors prepared in Example 3 and Comparative Examples 3-4 were tested. The HP-aZIF@Tar@Au sensors prepared in Example 3 and Comparative Example 3 were tested for 10 -5 The Raman test results of M tyrosine are shown in Figure Figure 6 , the HP-aZIF@Tar@Au sensor prepared in Example 3 and Comparative Example 4 is 10 -

[0090] 5 The relative standard deviation (RSD) chart of 20 test points of M tyrosine is shown in the following table: Figure 7 , by Figure 6 , Figure 7 It can be seen that reducing the amount of tartaric acid added and changing the loading amount of HP-aZIF@Tar@Au composite material on the AAO substrate will cause the test uniformity of the sensor to decrease.

[0091] The above merely provides the preferred embodiments of the present application, but the protection scope of the present application is not limited thereto, and any changes or substitutions easily conceived by those skilled in the art within the technical scope disclosed by the present application shall be covered within the protection scope of the present application. Therefore, the protection scope of the present application shall be subject to the protection scope of the claims.

Claims

1. Application of HP-aZIF@Tar@Au sensor in detection of human sweat tyrosine, characterized in that, The tyrosine in the human sweat attached to the surface of the HP-aZIF@Tar@Au sensor was measured by a Raman spectrometer, and the detection limit was 10 -8 M; The HP-aZIF@Tar@Au sensor comprises an AAO substrate and HP-aZIF@Tar@Au composite material loaded on the surface of the AAO substrate, and raw materials for preparing the HP-aZIF@Tar@Au composite material comprise HP-ZIF-67, HAuCl4 and L-tartaric acid; The loading amount of the HP-aZIF@Tar@Au composite material on the AAO substrate is 0.075 mg / cm 2 ; The preparation method of the HP-aZIF@Tar@Au sensor comprises the following steps: HP-ZIF-67 is dispersed with a solvent, then HAuCl4 solution is added and mixed and stirred, the product is centrifuged, then dispersed with a solvent again, NaBH4 solution is added and mixed and stirred in an ice water bath to obtain a mixed solution, the mixed solution is centrifuged and dried to obtain HP-ZIF-67@Au; The HP-ZIF-67@Au is mixed and ground with L-tartaric acid, then refluxed in toluene, cooled, centrifuged and dried to obtain HP-aZIF@Tar@Au composite material; The HP-aZIF@Tar@Au composite material is dispersed with DMF, then added dropwise to a hydrophilic treated AAO substrate to obtain the HP-aZIF@Tar@Au sensor; The molar ratio of the HP-ZIF-67@Au to L-tartaric acid is 1:1.2; The AAO substrate is hydrophilic treated with 2-amino terephthalic acid.

2. Use of the HP-aZIF@Tar@Au sensor according to claim 1 for detecting human sweat tyrosine, characterized in that, The molar ratio of the HP-ZIF-67 and HAuCl4 was 2:1, and the molar ratio of Au 3+ The molar ratio of the HP-ZIF-67 and HAuCl4 was 2:1, and the molar ratio of Au 3+ The molar ratio of the HP-ZIF-67 and HAuCl4 was 2:1, and the molar ratio of Au 3+ The molar ratio of the HP 3. Use of the HP-aZIF@Tar@Au sensor according to claim 1 for detecting human sweat tyrosine, characterized in that, The concentration of the HAuCl4 solution is 0.025M, and the concentration of the NaBH4 solution is 0.1M.

4. Use of the HP-aZIF@Tar@Au sensor according to claim 1 for detecting human sweat tyrosine, characterized in that, The concentration of the HP-aZIF@Tar@Au composite material after being dispersed with DMF is 2mg / mL.

5. Use of the HP-aZIF@Tar@Au sensor according to claim 1 for detecting human sweat tyrosine, characterized in that, HP-ZIF-67 is dispersed with a solvent, then HAuCl4 solution is added, mixed and stirred at room temperature for 3h, and NaBH4 solution is added and mixed and stirred in an ice water bath for 5h.

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