Amorphous Fe-MOF / carbon-based composite nano matrix and preparation method thereof

Abstract

The invention discloses an amorphous Fe-MOF / carbon-based composite nano matrix and a preparation method thereof, and relates to the field of metabolism detection. The invention provides an amorphous Fe-MOF / carbon-based composite nano matrix for mass spectrum double-ion mode detection and a preparation method thereof. The amorphous Fe-MOF / carbon-based composite double-ion matrix integrates multiple physicochemical properties, and realizes non-targeted efficient detection of a biological sample in a double-ion mode. The MALDI-MS platform assisted by the amorphous Fe-MOF / carbon-based composite nanomaterial has the advantages of simple pretreatment, high throughput, rapid and comprehensive metabolism detection and the like, and meets the requirements of accurate medical treatment on rapid and accurate diagnosis.

Description

Technical Field

[0001] This invention relates to the field of metabolic detection, and more particularly to an amorphous Fe-MOF / carbon-based composite nanomatrix and its preparation method. Background Technology

[0002] Metabolites, as the end products of biological metabolic pathways, can directly reflect the real-time impact of genetic factors or environmental changes on the physiological state of an organism. They directly reflect the level of biochemical activity in the body and have a closer correlation with phenotype. In recent years, metabolomics analysis has received widespread attention in the field of disease diagnosis. By detecting metabolites in human body fluids, biomarkers closely related to diseases can be identified, thereby enabling early clinical diagnosis of diseases.

[0003] Current mainstream metabolic detection technologies include nuclear magnetic resonance spectroscopy (NMR), chromatography-mass spectrometry (GC-MS / LC-MS), and matrix-assisted laser desorption / ionization mass spectrometry (MALDI-MS). Among them, MALDI-MS has shown great potential in large-scale clinical screening due to its significant advantages such as simple sample pretreatment, high throughput, and low sample consumption.

[0004] In MALDI-MS detection, metabolites, due to their structural differences, typically exhibit a tendency to ionize in either positive or negative ion modes. Dual-ion mode detection holds promise for comprehensive coverage of metabolites in biological fluids (such as serum), thereby improving the accuracy of disease diagnosis. Notably, the matrix plays a crucial role in MALDI-MS detection, directly influencing ionization efficiency and detection sensitivity by promoting the desorption and ionization processes of the analyte.

[0005] However, existing matrices have significant limitations: 1) Traditional commercial organic matrices generate strong background interference in the low molecular weight region (m / z < 500 Da), severely limiting the detection of small molecule metabolites; 2) Most developed inorganic nanomatrices only exhibit high desorption and ionization efficiencies in a single positive or negative ion mode. Although a few dual-ion matrices have been reported, these matrices are mainly suitable for the targeted metabolite detection of standards or biological samples, and generally suffer from insufficient detection sensitivity and poor selectivity in complex biological samples.

[0006] Therefore, those skilled in the art are dedicated to developing an amorphous Fe-MOF / carbon-based composite nanomatrix and its preparation method. This integrates multiple physicochemical properties to achieve non-targeted, high-efficiency detection of biological samples using a dual-ion mode; the preprocessing is simple and high-throughput, meeting the needs of precision medicine for rapid and accurate diagnosis. Summary of the Invention

[0007] In view of the above-mentioned deficiencies of the prior art, the technical problem to be solved by the present invention is the insufficient sensitivity and poor selectivity of dual-ion matrices in complex biological samples.

[0008] To achieve the above objectives, the present invention provides an amorphous Fe-MOF / carbon-based composite nanomatrix, comprising a heterojunction structure, multifunctional group synergy, and a rough surface, for non-targeted detection of biological samples in dual-ion mode; The heterojunction structure facilitates the separation of electrons and holes; The synergistic effect of multiple functional groups, including carboxyl / hydroxyl functional groups and pyridine nitrogen, enables the matrix to have bidirectional proton transport capabilities, achieving dual-mode ionization. Rough surfaces: selectively and efficiently enrich small molecule metabolites.

[0009] This invention also provides a method for preparing amorphous Fe-MOF / carbon-based composite nanomaterials, comprising the following steps: Step 1: Synthesize Fe-MOF / carbon-based composite nanomaterials using an improved hydrothermal method; Step 2: Centrifuge, wash and disperse the Fe-MOF / carbon-based composite nanomaterials obtained in Step 1 in 10 mL of methanol; Step 3: The product obtained in Step 2 is amorphized by ligand exchange strategy, centrifuged, washed and dried to obtain amorphous Fe-MOF / carbon-based composite nanomaterials.

[0010] Further, in step 1, a mixed solution of ferric chloride and F127 is prepared, and acetic acid, aminoterephthalic acid and carbon-based nanomaterials are added sequentially to obtain a reaction mixture. The reaction mixture is then transferred to a high-pressure reactor for reaction.

[0011] Further, the concentration of ferric chloride is 10-20 mg / mL; the mass ratio of F127 to ferric chloride is 0.89:1; the volume of acetic acid is 0.6-0.8 mL; the concentration of aminoterephthalic acid is 3-6 mg / mL; and the concentration of carbon-based nanomaterials is 1-3 mg / mL.

[0012] Furthermore, in step 1, the reaction temperature is 90-110℃ and the reaction time is 12-24 hours.

[0013] Further, in step 2, the centrifugation speed is 8000 rpm and the time is 3 min; the mixture is washed continuously with deionized water and methanol and centrifuged 3 times, and then dispersed in 10 mL of methanol.

[0014] Further, in step 3, an aqueous solution of dimethylimidazole is added to the dispersion and reacted at 25±5℃ for 10 min.

[0015] Further, the concentration of the aqueous solution of the dimethylimidazole is 0.7-1.0 mg / mL.

[0016] Further, in step 3, the centrifugation speed is 8000 rpm and the time is 3 min; wash with deionized water and centrifuge 3 times, and dry in a vacuum drying oven at 40℃ for 12-36 hours.

[0017] This invention also provides an application of amorphous Fe-MOF / carbon-based composite nanomaterials, using the amorphous Fe-MOF / carbon-based composite nanomatrix to extract metabolic information from serum samples in a positive and negative dual-ion mode, including the following steps: Step 1: Dilute the serum sample 10 times with deionized water; Step 2: Prepare a matrix solution of amorphous Fe-MOF / carbon-based composite nanomaterials at a concentration of 1 mg / mL using deionized water; Step 3: Sample preparation on the mass spectrometry target plate. First, drop 1 μL of serum sample onto the mass spectrometry target plate. After the solution dries, the sample-loaded 384Polish target plate is obtained. Step 4: Load the matrix onto the 384Polish target plate after loading the sample obtained in Step 3. Drop 1 μL of matrix solution onto the dried blood sample and allow it to dry naturally at room temperature to obtain the matrix-loaded 384Polish target plate. Step 5: Using matrix-assisted laser desorption / ionization mass spectrometry, the 384Polish target plate loaded with the matrix obtained in step 4 is detected in positive and negative dual-ion mode to obtain a positive and negative dual-ion serum metabolic fingerprint.

[0018] Existing dual-ion matrices cannot achieve efficient, non-targeted detection of biological samples in both positive and negative ion modes: although dual-ion matrices have been developed, their insufficient sensitivity and selectivity cannot meet the dual-mode coverage requirements of non-targeted metabolomics in biological samples. This invention pioneers an amorphous Fe-MOF / carbon-based composite dual-ion matrix for non-targeted detection of biological samples in dual-ion modes, such as... Figure 1 As shown. The heterojunction structure of this invention promotes the separation of electrons and holes, improving the ionization and desorption efficiency in both positive and negative ion modes; multi-functional group synergy: carboxyl / hydroxyl functional groups and pyridine nitrogen endow the matrix with bidirectional proton transport capabilities, achieving efficient ionization in both modes; rough surface: selectively and efficiently enriches small molecule metabolites, such as... Figure 2 As shown.

[0019] Existing non-targeted matrices for biological samples exhibit excellent performance only in a single ion mode, limiting their application to large cohorts requiring high-throughput analysis. This invention, utilizing an amorphous Fe-MOF / carbon-based composite nanomaterial-assisted MALDI-MS platform, enables non-targeted analysis of 1 μL of biological sample in a dual-ion mode within 1 minute. Furthermore, this invention's amorphous Fe-MOF / carbon-based composite dual-ion matrix integrates multiple physicochemical properties, achieving non-targeted metabolomics detection of single samples using both positive and negative ions.

[0020] Compared with the prior art, the present invention has the following obvious substantive features and significant advantages: 1. This invention is the first to integrate multiple physicochemical properties into an amorphous Fe-MOF / carbon-based composite dual-ion matrix, enabling non-targeted and efficient detection of biological samples in a dual-ion mode. It can simultaneously detect 432 characteristic peaks of metabolites in a single sample (190 positive ions and 242 negative ions), and improve the coverage of serum metabolites by 2.1 times.

[0021] 2. The MALDI-MS platform assisted by the amorphous Fe-MOF / carbon-based composite nanomaterials of this invention exhibits advantages such as simple preprocessing, high throughput, and rapid and comprehensive metabolic detection, meeting the needs of precision medicine for rapid and accurate diagnosis.

[0022] The following will further explain the concept, specific structure, and technical effects of the present invention in conjunction with the accompanying drawings, so as to fully understand the purpose, features, and effects of the present invention. Attached Figure Description

[0023] Figure 1 This is a scanning electron microscope image of amorphous Fe-MOF / carbon-based composite nanomaterials according to a preferred embodiment of the present invention; Figure 2 This is standard data collected in a preferred embodiment of the present invention under a positive and negative dual-ion mode; Figure 3 This is a preferred embodiment of the dual-ion serum metabolic fingerprint spectrum of the present invention. Detailed Implementation

[0024] The following description, with reference to the accompanying drawings, illustrates several preferred embodiments of the present invention to make its technical content clearer and easier to understand. The present invention can be embodied in many different forms, and the scope of protection of the present invention is not limited to the embodiments mentioned herein.

[0025] In the accompanying drawings, components with the same structure are indicated by the same numerical designation, and components with similar structures or functions are indicated by similar numerical designations. The dimensions and thicknesses of each component shown in the drawings are arbitrary, and the present invention does not limit the dimensions and thicknesses of each component. To make the illustrations clearer, the thickness of some components has been appropriately exaggerated in the drawings.

[0026] This invention provides a method for preparing amorphous Fe-MOF / carbon-based composite nanomaterials for dual-ion mode detection, comprising the following steps: Step 1: Synthesize Fe-MOF / carbon-based composite nanomaterials using an improved hydrothermal method; Step 2: Centrifuge, wash and disperse the product obtained in Step 1 in 10 mL of methanol to obtain Fe-MOF / carbon-based composite nanomaterials.

[0027] Step 3: The product obtained in Step 2 is amorphized by ligand exchange strategy. The product is centrifuged, washed and dried to obtain amorphous Fe-MOF / carbon-based composite nanomaterials.

[0028] A mixed solution of ferric chloride and F127 was prepared, and acetic acid, aminoterephthalic acid and carbon-based nanomaterials were added sequentially to obtain a reaction mixture. The reaction mixture was then transferred to a high-pressure reactor for high-temperature reaction.

[0029] The concentration range of ferric chloride is 10-20 mg / mL; the mass ratio of F127 to ferric chloride is 0.89:1; the volume of acetic acid is 0.6-0.8 mL; the concentration range of aminoterephthalic acid is 3-6 mg / mL; and the concentration range of carbon-based nanomaterials is 1-3 mg / mL.

[0030] The reaction temperature is 90-110℃, and the reaction time is 12-24 hours.

[0031] In step 2, the centrifugation speed was 8000 rpm and the time was 3 min; the precipitate was washed continuously with deionized water and methanol and centrifuged 3 times to obtain Fe-MOF / carbon-based composite nanomaterials; the Fe-MOF / carbon-based composite nanomaterials were dispersed in 10 mL of methanol for later use.

[0032] An aqueous solution of dimethylimidazole was added to the dispersion of Fe-MOF / carbon-based composite nanomaterials, and the mixture was reacted at 25±5℃ for 10 min.

[0033] The concentration range of dimethylimidazole solution is 0.7-1.0 mg / mL.

[0034] In step 3, the centrifugation speed is 8000 rpm and the time is 3 min; the precipitate is washed with deionized water and centrifuged 3 times to obtain amorphous Fe-MOF / carbon-based composite nanomaterials; the amorphous Fe-MOF / carbon-based composite nanomaterials are dried in a vacuum drying oven at 40℃ for 12-36 hours.

[0035] This invention also discloses an application of an LDI-MS platform assisted by amorphous Fe-MOF / carbon-based composite nanomaterials for the dual-ion mode detection of serum metabolites.

[0036] Metabolic information of serum samples was extracted using amorphous Fe-MOF / carbon-based composite nanomaterials as a matrix in a dual-ion mode, including the following steps: Step 1: Dilute the serum sample 10 times with deionized water; Step 2: Prepare a matrix solution of amorphous Fe-MOF / carbon-based composite nanomaterials at a concentration of 1 mg / mL using deionized water.

[0037] Step 3: Sample preparation on the mass spectrometry target plate. First, drop 1 μL of serum sample onto the mass spectrometry target plate. After the solution dries, the sample-loaded 384Polish target plate is obtained. Step 4: Load the matrix onto the 384Polish target plate after loading the sample obtained in Step 3. Drop 1 μL of the matrix solution prepared above onto the dried blood sample and allow it to dry naturally at room temperature to obtain the matrix-loaded 384Polish target plate. Step 5: Using matrix-assisted laser desorption / ionization mass spectrometry (MADS), the matrix-loaded 384Polish target plate obtained in Step 4 was analyzed in dual-ion mode to obtain a dual-ion serum metabolic fingerprint, such as... Figure 3 As shown.

[0038] The preferred embodiments of the present invention have been described in detail above. It should be understood that those skilled in the art can make numerous modifications and variations based on the concept of the present invention without creative effort. Therefore, all technical solutions that can be obtained by those skilled in the art based on the concept of the present invention through logical analysis, reasoning, or limited experimentation on the basis of existing technology should be within the scope of protection defined by the claims.

Claims

1. An amorphous Fe-MOF / carbon-based composite nanomatrix, characterized in that, Including heterojunction structures, multifunctional group synergy, and rough surfaces, for non-targeted detection of biological samples in dual-ion mode; The heterojunction structure facilitates the separation of electrons and holes; The synergistic effect of multiple functional groups, including carboxyl / hydroxyl functional groups and pyridine nitrogen, enables the matrix to have bidirectional proton transport capabilities, achieving dual-mode ionization. Rough surfaces: selectively and efficiently enrich small molecule metabolites.

2. A method for preparing amorphous Fe-MOF / carbon-based composite nanomaterials, characterized in that, Includes the following steps: Step 1: Synthesize Fe-MOF / carbon-based composite nanomaterials using an improved hydrothermal method; Step 2: Centrifuge, wash and disperse the Fe-MOF / carbon-based composite nanomaterials obtained in Step 1 in 10 mL of methanol; Step 3: The product obtained in Step 2 is amorphized by ligand exchange strategy, centrifuged, washed and dried to obtain amorphous Fe-MOF / carbon-based composite nanomaterials.

3. The method for preparing amorphous Fe-MOF / carbon-based composite nanomaterials as described in claim 2, characterized in that, In step 1, a mixed solution of ferric chloride and F127 is prepared, and acetic acid, aminoterephthalic acid and carbon-based nanomaterials are added sequentially to obtain a reaction mixture. The reaction mixture is then transferred to a high-pressure reactor for reaction.

4. The method for preparing amorphous Fe-MOF / carbon-based composite nanomaterials as described in claim 3, characterized in that, The concentration of ferric chloride is 10-20 mg / mL; the mass ratio of F127 to ferric chloride is 0.89:1; the volume of acetic acid is 0.6-0.8 mL; the concentration of aminoterephthalic acid is 3-6 mg / mL; and the concentration of carbon-based nanomaterials is 1-3 mg / mL.

5. The method for preparing amorphous Fe-MOF / carbon-based composite nanomaterials as described in claim 3, characterized in that, In step 1, the reaction temperature is 90-110℃ and the reaction time is 12-24 hours.

6. The method for preparing amorphous Fe-MOF / carbon-based composite nanomaterials as described in claim 2, characterized in that, In step 2, the centrifugation speed is 8000 rpm and the time is 3 min; the mixture is washed continuously with deionized water and methanol and centrifuged 3 times, and then dispersed in 10 mL of methanol.

7. The method for preparing amorphous Fe-MOF / carbon-based composite nanomaterials as described in claim 2, characterized in that, In step 3, an aqueous solution of dimethylimidazole is added to the dispersion and reacted at 25±5℃ for 10 min.

8. The method for preparing amorphous Fe-MOF / carbon-based composite nanomaterials as described in claim 7, characterized in that, The concentration of the aqueous solution of the dimethylimidazole is 0.7-1.0 mg / mL.

9. The method for preparing amorphous Fe-MOF / carbon-based composite nanomaterials as described in claim 2, characterized in that, In step 3, the centrifugation speed is 8000 rpm and the time is 3 min; wash with deionized water and centrifuge 3 times, and dry in a vacuum drying oven at 40℃ for 12-36 hours.

10. An application of an amorphous Fe-MOF / carbon-based composite nanomaterial, characterized in that, Metabolic information of serum samples was extracted using an amorphous Fe-MOF / carbon-based composite nanomatrix in a dual-ion mode, including the following steps: Step 1: Dilute the serum sample 10 times with deionized water; Step 2: Prepare a matrix solution of amorphous Fe-MOF / carbon-based composite nanomaterials at a concentration of 1 mg / mL using deionized water; Step 3: Sample preparation on the mass spectrometry target plate. First, drop 1 μL of serum sample onto the mass spectrometry target plate. After the solution dries, the sample-loaded 384Polish target plate is obtained. Step 4: Load the matrix onto the 384Polish target plate after loading the sample obtained in Step 3. Drop 1 μL of matrix solution onto the dried blood sample and allow it to dry naturally at room temperature to obtain the matrix-loaded 384Polish target plate. Step 5: Using matrix-assisted laser desorption / ionization mass spectrometry, the 384Polish target plate loaded with the matrix obtained in step 4 is detected in positive and negative dual-ion mode to obtain a positive and negative dual-ion serum metabolic fingerprint.

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