Zr-MOF / Fe3O4 composite material, preparation method and application

Abstract

The invention belongs to the field of in-vitro diagnosis, and particularly relates to a Zr-MOF / Fe3O4 composite material, a preparation method and application, the preparation method of the Zr-MOF / Fe3O4 composite material is provided, FeNPs is prepared through a solvothermal method, 2D MOF nanosheets are prepared, ZMF (Zr-MOF / FeNPs) nanosheets are prepared through an in-situ growth method, and Zr-MOF flowers are prepared through a dielectric barrier discharge method (DBD). And finally, preparing the ZMF flower, namely the Zr-MOF / Fe3O4 composite material, by adopting an in-situ growth method. The Zr-MOF / Fe3O4 composite material has a stable structure, and the structure also has light energy and heat energy binding functions, so that the detection intensity and comprehensiveness of signals can be ensured, and the application of metabolite detection can be widened.

Description

Technical Field

[0001] The present invention belongs to the field of in vitro diagnosis, and specifically relates to a Zr-MOF / Fe 3 O 4 Composite materials, preparation methods and applications. Background Art

[0002] Heart failure (HF) and heart failure complicated by myocardial infarction (HFMI) are common cardiac diseases with high morbidity and mortality. In the treatment context of acute or chronic HF and HFMI patients, clinicians need to implement different treatment strategies quickly and accurately to improve survival. However, the similarity of symptoms often complicates clinical judgment. Common non-invasive diagnostic indicators, such as cardiac troponin (cTn), B-type brain natriuretic peptide (BNP), and electrocardiogram (ECG), although highly sensitive, often lack specificity, resulting in a high false positive rate. This not only leads to unnecessary misdiagnosis, but also increases the risk of delaying critical treatment. Although cardiac ultrasound and angiography can improve diagnostic accuracy, their widespread application is limited by factors such as experience, invasiveness, and cost. Blood metabolites reflect physiological status and show good prospects as potential biomarkers for disease diagnosis. Metabolomics focuses on disease phenotypes and provides the potential for diagnosing complex diseases based on metabolic signature analysis of body fluids. However, conventional metabolic detection methods (such as nuclear magnetic resonance, high-performance liquid chromatography and tandem mass spectrometry) face limitations in sensitivity, complexity, cost and throughput, which requires addressing the problem of high-throughput, low-cost and accurate analysis of body fluid metabolic signatures.

[0003] Since the Nobel Prize for Nano-Assisted Laser Desorption / Ionization of Mass Spectrometry (MALDI-MS), laser desorption ionization mass spectrometry (LDI-MS) has attracted widespread attention due to its high throughput and simple operation. In recent years, methods that use nanomatrices rather than organic matrices for small molecule metabolite detection have flourished to reduce matrix background interference. The rational engineering design of the nanomatrix and its interaction with the target small molecule determine the detection performance. At present, the rational design of nanomatrices based on the LDI-MS mechanism is mainly focused on improving ionization efficiency and enhancing the desorption process. Strategies include introducing nanoscale adjustments such as tips, heterojunctions, and precious metals to enhance ionization intensity, constructing metal centers, or introducing high photothermal components to enhance thermal desorption capacity. Although these methods have made some progress, further improvements are still needed to fully utilize nanomatrices for comprehensive analysis of large amounts of complex biological fluids. This includes solving problems such as weak ionization caused by insufficient light absorption, decreased desorption capacity caused by rapid heat dissipation, and simplifying the complex preparation of nanomatrices; therefore, a nanomatrix and its preparation method that can overcome these shortcomings are urgently needed for accurate metabolite analysis and detection of various heart diseases. Summary of the invention

[0004] In view of the shortcomings of the prior art, the present invention provides a Zr-MOF / Fe 3 O 4 The method for preparing the composite material comprises the following steps:

[0005] S1: FeCl 3 6H 2 O, sodium citrate, sodium acetate and solvent are mixed and stirred to obtain a mixed liquid, subjected to an anaerobic high-pressure reaction in a reactor, and separated and washed by magnetic force to obtain FeNPs material;

[0006] S2: 1,3,5-benzenetricarboxylic acid and zirconium chloride are used as raw materials of organic ligands, added into a mixed solvent for mixing, and the mixed clarified liquid is loaded into a polytetrafluoroethylene reactor for anaerobic high-pressure reaction, and after the reaction, centrifuged and washed to obtain 2D MOF nanosheets;

[0007] S3: adding the 2D MOF nanosheets obtained in S2 to a mixed solution of ethylene glycol containing ferric chloride, 1,6-hexanediamine, and sodium acetate, stirring at room temperature, reacting at high temperature in the absence of oxygen, and then performing magnetic separation and washing to obtain 2D ZMF nanosheets;

[0008] S4: After mixing 1,3,5-benzenetricarboxylic acid and zirconium chloride as raw materials of organic ligands with a mixed solvent, the clarified liquid is loaded into a dielectric barrier discharge reactor, and after centrifugation and washing, a 3D MOF flower material is obtained;

[0009] S5: The Zr-MOF flower obtained in S4 is added to a mixed solution of ethylene glycol containing ferric chloride, 1,6-hexanediamine, and sodium acetate, stirred at room temperature, reacted in an oxygen-free high-pressure environment, and subjected to magnetic separation and washing to obtain ZMF flower, namely Zr-MOF / Fe 3 O 4 Composite materials.

[0010] Furthermore, the solvent in S1 is ethylene glycol, the stirring time is 1 hour, the high-pressure anaerobic reaction time is 12 hours, the temperature is 200° C., and the washing liquid is ethanol and deionized water.

[0011] Furthermore, the high-pressure anaerobic reaction time in S2 is 12 hours, the temperature is 120° C., and the washing liquid is DMF, ethanol and deionized water.

[0012] Furthermore, the stirring time in S3 is 1 hour, the high pressure oxygen-free reaction time is 6 hours, and the temperature is 200°C.

[0013] Furthermore, the dielectric barrier discharge reaction process in S4 is 20 min, the washing liquid is DMF, ethanol and deionized water; and the mixed solvent is N,N-dimethylformamide, deionized water and formic acid.

[0014] Furthermore, the stirring time described in S5 is 1 hour, the high-pressure anaerobic reaction time is 6 hours, the temperature is 200° C., and the washing liquid is ethanol and deionized water.

[0015] A Zr-MOF / Fe 3 O 4 Composite material, the Zr-MOF / Fe 3 O 4 The composite material has a three-dimensional flower-like Zr-MOF nano-skeleton structure.

[0016] A Zr-MOF / Fe 3 O 4 Application of the composite material, the Zr-MOF / Fe 3 O 4 The composite materials are used in the preparation of nanochips for metabolite detection of diseases.

[0017] Beneficial Effects

[0018] The Zr-MOF / Fe 3 O 4 The preparation method of the composite material can achieve Fe 3 O 4 In situ growth of MOF to obtain a Zr-MOF / Fe 3 O 4 The composite material has a stable structure, which also provides light energy and thermal energy binding function, which can reduce the overall heat loss and ensure the signal detection intensity and comprehensiveness; and can be widely used in the metabolite detection of heart diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] In order to more clearly illustrate the technical solutions of the exemplary embodiments of the present invention, the following is a brief introduction to the drawings required for use in the embodiments. It should be understood that the following drawings only illustrate certain embodiments of the present invention and should not be regarded as limiting the scope. For those of ordinary skill in the art, other relevant drawings can be obtained based on these drawings without creative work. In the drawings:

[0020] Figure 1 This is a diagram of the preparation process of Example 1 of the present invention;

[0021] Figure 2The FeNPs, ZMF sheets, and ZMF flowers (Zr-MOF / Fe 3 O 4 Composite material) morphology SEM image;

[0022] Figure 3 This is the standard metabolite intensity diagram of Example 1 of the present invention;

[0023] Figure 4 It is the matrix background image of Example 1 of the present invention;

[0024] Figure 5 This is a graph of the intensity of mixed standard metabolites under high protein and high salt conditions in Example 1 of the present invention;

[0025] Figure 6 This is a test diagram of different sample serums in Example 1 of the present invention. DETAILED DESCRIPTION

[0026] The following will be combined with Example 1 of the present invention and the attached Figures 1 to 6 , the technical solution of the present invention is clearly and completely described. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0027] Combination Figure 1 The specific preparation method of the present invention can be known from the preparation process diagram.

[0028] Example 1

[0029] S1: Prepared by solvothermal method, the reaction raw materials include 2.43g of ferric chloride (FeCl 3 6H 2 O), 0.45g of sodium citrate and 3.6g of sodium acetate, and 60ml of ethylene glycol as a solvent, the raw materials are mixed and stirred at room temperature for 1 hour, and then the mixed liquid is charged into a polytetrafluoroethylene reactor, and reacted at 200°C under high pressure and oxygen-free conditions for 12 hours; after the reaction, the FeNPs material is obtained by magnetic separation and washing with ethanol and deionized water;

[0030] S2: 12.3 mg of 1,3,5-benzenetricarboxylic acid (H 3 BTB) and 10.1 mg of zirconium chloride (ZrCl 4) as the raw material of the organic ligand; the mixed solvent is 5 ml of N,N-dimethylformamide (DMF), 0.06 ml of deionized water and 0.9 ml of formic acid; the mixed clarified liquid is loaded into a polytetrafluoroethylene reactor and subjected to high pressure reaction at 120°C in an oxygen-free state for 12 hours; after the reaction, the material is centrifuged and washed with DMF, ethanol and deionized water to obtain 2D MOF nanosheets;

[0031] S3: ZMF nanosheets were prepared by an in-situ growth method. 30 mg of 2D MOF nanosheets were added to a 18 ml ethylene glycol mixed solution containing 90 mg ferric chloride, 324 mg 1,6-hexanediamine and 360 mg sodium acetate. The mixture was stirred at room temperature for 1 hour and then reacted at 200 °C in a high pressure and oxygen-free environment for 6 hours. The 2D ZMF nanosheets were obtained by magnetic separation and washing with ethanol and deionized water.

[0032] S4: The Zr-MOF flower was prepared by the discharge arc method, with 12.3 mg of 1,3,5-benzenetricarboxylic acid and 10.1 mg of zirconium chloride as raw materials for organic ligands, and a mixed solvent of 5 ml of N,N-dimethylformamide, 0.06 ml of deionized water and 0.9 ml of formic acid; after mixing, the clarified liquid was loaded into a DBD reactor, and a sufficiently high AC voltage (37-50 V, 1.27-3 A) was applied between the two electrodes for 20 minutes to prepare the flower-like MOF material; after centrifugation and washing with DMF, ethanol and deionized water, the 3D MOF flower material was obtained.

[0033] S5: ZMF flowers were prepared by an in situ growth method. Initially, 30 mg of Zr-MOF flowers were added to a mixed solution of ethylene glycol (18 ml) containing 90 mg of ferric chloride, 324 mg of 1,6-hexanediamine, and 360 mg of sodium acetate; the mixture was stirred at room temperature for 1 hour, and then reacted at 200 °C for 6 hours in an oxygen-free high-pressure environment; the 3D ZMF flowers were obtained, and the obtained 3D ZMF flowers were separated by magnetic force and washed with ethanol and deionized water to obtain Zr-MOF / Fe 3 O 4 Composite materials.

[0034] Combination Figure 2 It can be seen that the morphology SEM images of FeNPs, ZMF sheets and ZMF flowers prepared by the present invention show that the material has been successfully produced. Figure 3It can be known that glucose, tryptophan, phenylalanine, arginine and lysine are used as standard samples. The intensity contrast between nanomatrices systematically explored the LDI enhancement. Five small molecule metabolites of glucose, tryptophan, phenylalanine, arginine and lysine were selected as standard samples to evaluate the performance of nanomaterials, and the ZMF flower had the highest intensity. Uniformly distributed two-dimensional sheets and three-dimensional flower-like Zr-MOF nanoskeletons can be clearly observed in the scanning electron microscope images, in which the nanosheets show independent distribution and the flower structure grows from multiple dispersed sheets. The thicknesses of the two nanoskeletons are 10nm and 5nm, respectively. FeNPs are regular spheres of 200 to 300nm, and ZMF nanosheets and ZMF flowers maintain the original two-dimensional and three-dimensional spatial structures. FeNPs are evenly distributed on the Zr-MOF nanoskeleton. The ZMF flower has a diameter of 5 microns and presents a regular flower shape.

[0035] Combination Figure 4 It can be seen that ZMF flower materials have no background interference peaks compared to traditional organic matrices. Figure 5 The intensity graph of mixed standard metabolites under high protein and high salt conditions can be obtained. ZMF flower not only obtains the highest signal intensity but also detects the most number of signals. Figure 6 The test charts of different sample serums show that ZMF flower can effectively detect metabolite signals in serum.

[0036] The steps for further making ZMF flowers into chips for metabolite detection include: sample preparation and LDI-MS analysis. First, standard metabolite solutions containing Glu, Arg, Try, Phe or Lys are prepared with deionized water. In order to improve protein and salt tolerance, BSA and NaCl are mixed with typical small metabolites Glu, Arg, Try, Phe and Lys, and the concentration of each metabolite is 1 ng / nL. All blood samples are punctured into vacuolar tubes by vein and coagulated within 1 hour at room temperature. The blood samples are centrifuged for 10 minutes, and the serum is collected and immediately stored at -80°C for further analysis. Different materials are prepared into 1 ng / nL water suspensions, in which Glu, Arg, Try, Phe or Lys are glucose, arginine, tryptophan, phenylalanine, and lysine, respectively.

[0037] LDI-MS analysis was performed using a MALDI-TOF / TOF mass spectrometer (Shimadzu, AXIMA-Performance) in positive ion mode equipped with a nitrogen laser (337 nm). The parameters were set, including 200 laser scans per sample, a laser intensity of 65%, and a molecular weight range of 100 to 1000 Da collected. The instrument was calibrated with standard samples before each use. Standard metabolite solutions or serum samples were pipetted onto the target plate. After drying at room temperature, the matrix was pipetted to cover the previously dried spots. After the samples were dried at room temperature, they were analyzed on the MALDI-TOF / TOF MS. For each sample, including small molecules and biological samples, 5 independent LDI-MS experiments were performed to ensure the reproducibility and reliability of the detection results.

[0038] The preparation method used in the present invention can achieve the in-situ growth of ferrosoferric oxide in MOF to obtain a stable structure. This structure also provides light energy and thermal energy binding functions, which can reduce overall heat loss and ensure signal detection intensity and comprehensiveness. The prepared ZMF flower can broaden the application in metabolite detection of heart diseases.

Claims

1. A method for preparing a Zr-MOF / Fe3O4 composite material, characterized in that: The following steps are involved: S1: FeCl3·6H2O, sodium citrate, sodium acetate and solvent are mixed and stirred to obtain a mixed liquid, and an anaerobic high-pressure reaction is carried out in a reactor, and FeNPs material is obtained by magnetic separation and washing; S2: 1,3,5-benzenetricarboxylic acid and zirconium chloride are used as raw materials of organic ligands, added into a mixed solvent for mixing, and the mixed clarified liquid is loaded into a polytetrafluoroethylene reactor for anaerobic high-pressure reaction, and after the reaction, centrifuged and washed to obtain 2D MOF nanosheets; S3: adding the 2D MOF nanosheets obtained in S2 to a mixed solution of ethylene glycol containing ferric chloride, 1,6-hexanediamine, and sodium acetate, stirring at room temperature, reacting at high temperature in the absence of oxygen, and then performing magnetic separation and washing to obtain 2D ZMF nanosheets; S4: After mixing 1,3,5-benzenetricarboxylic acid and zirconium chloride as raw materials of organic ligands with a mixed solvent, the clarified liquid is loaded into a dielectric barrier discharge reactor, and after centrifugation and washing, a 3D MOF flower material is obtained; S5: Add the Zr-MOF flower obtained in S4 to a mixed solution of ethylene glycol containing ferric chloride, 1,6-hexanediamine and sodium acetate, stir at room temperature, react under an oxygen-free high-pressure environment, and obtain ZMF flower, i.e., Zr-MOF / Fe3O4 composite material, after magnetic separation and washing.

2. The method for preparing a Zr-MOF / Fe3O4 composite material according to claim 1, characterized in that: The solvent in S1 is ethylene glycol, the stirring time is 1 hour, the high-pressure oxygen-free reaction time is 12 hours, the temperature is 200° C., and the washing liquid is ethanol and deionized water.

3. The method for preparing a Zr-MOF / Fe3O4 composite material according to claim 1, characterized in that: The high pressure anaerobic reaction time in S2 is 12 hours, the temperature is 120° C., and the washing liquid is DMF, ethanol and deionized water.

4. The method for preparing a Zr-MOF / Fe3O4 composite material according to claim 1, characterized in that: The stirring time in S3 is 1 hour, the high pressure oxygen-free reaction time is 6 hours, and the temperature is 200°C.

5. The method for preparing a Zr-MOF / Fe3O4 composite material according to claim 1, characterized in that: The dielectric barrier discharge reaction process in S4 is 20 min, the washing liquid is DMF, ethanol and deionized water; the mixed solvent is N,N-dimethylformamide, deionized water and formic acid.

6. The method for preparing a Zr-MOF / Fe3O4 composite material according to claim 1, characterized in that: The stirring time described in S5 is 1 hour, the high-pressure anaerobic reaction time is 6 hours, the temperature is 200° C., and the washing liquid is ethanol and deionized water.

7. A Zr-MOF / Fe3O4 composite material obtained by the preparation method according to any one of claims 1 to 6, characterized in that: The Zr-MOF / Fe3O4 composite material has a three-dimensional flower-like structure.

8. The use of a Zr-MOF / Fe3O4 composite material according to claim 7, characterized in that: The Zr-MOF / Fe3O4 composite material is used in preparing a nanochip for detecting metabolites of diseases.

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