MXene / ferroferric oxide / AuNPs, preparation method and application

Abstract

The application provides a MXene / ferroferric oxide / AuNPs, a preparation method and application, and comprises the following steps: step 1, dissolving LiF and Ti3AlC2·MAX in hydrochloric acid, and fully reacting to obtain MXene, and freeze-drying for standby; step 2, dispersing MXene in ethylene glycol, adding FeCl3·6H2O, 1,6-hexanediamine and NaAc with a molar ratio of 1:8:13, and fully reacting to obtain MXene / ferroferric oxide; step 3, preparing AuNPs; and step 4, dispersing MXene / ferroferric oxide in the AuNPs solution, and fully reacting to obtain MXene / ferroferric oxide / AuNPs. The preparation method can combine ferroferric oxide and MXene in the form of in-situ growth, can reduce heat loss, improve stability, and improve ionization desorption efficiency, and the prepared nanochip can be applied in metabolite detection.

Description

Technical Field

[0001] This invention relates to the field of in vitro diagnostics, specifically to an MXene / Fe3O4 / AuNPs, its preparation method, and its applications. Background Technology

[0002] Currently, several challenges remain for researchers in the biomedical field, among which the accurate diagnosis of multiple diseases occurring in the same organ, particularly the liver, is a significant challenge. For liver disease detection, metabolite testing methods are crucial for the accurate diagnosis and differentiation of liver diseases. The composition and dynamic changes of metabolites can serve as accurate, real-time indicators of the liver's physiological and pathological dynamics.

[0003] Existing technologies, such as nuclear magnetic resonance (NMR), have inherent limitations in detection sensitivity, while tandem mass spectrometry (MS / MS) is cumbersome and costly. Regarding the matrix, traditional organic matrices exhibit significant signal overlap in the low molecular weight range, interfering with qualitative analysis by MALDI-MS; simultaneously, uneven crystallization can also affect the accurate quantitative analysis of metabolic molecules.

[0004] For nanomatrices, the hydrophobicity and complex preparation processes of some nanomatrices hinder their further application. For instance, nanomatrix-assisted LDI-MS in metabolic detection suffers from poor ionization efficiency, high thermal conductivity which prevents effective desorption, and optical absorption bias. Therefore, there is an urgent need for a nanochip method that can overcome these drawbacks to accurately analyze and detect various liver diseases using metabolites. Summary of the Invention

[0005] In view of the prior art, the present invention provides MXene / Fe3O4 / AuNPs, its preparation method and application.

[0006] The technical solution adopted in this invention is as follows:

[0007] Step 1: LiF and Ti3AlC2·MAX are dissolved in HCl and reacted completely to obtain MXene, which is then lyophilized for later use;

[0008] Step 2: Disperse MXene in ethylene glycol using ultrasound, add FeCl3·6H2O, 1,6-hexanediamine and sodium acetate to the MXene suspension, and react fully to obtain MXene / Fe3O4.

[0009] Step 3: Preparation of AuNPs;

[0010] Step 4: MXene / Fe3O4 is dispersed in AuNPs solution and reacted completely to obtain MXene / Fe3O4 / AuNPs.

[0011] Furthermore, the reaction process in step 1 includes stirring, centrifugation, and washing; the stirring time is 24h to 36h, the temperature is 35℃, the washing is carried out until the pH is 5 to 7, and the centrifugation rate is 3500rpm.

[0012] Furthermore, the reaction process in step 2 includes stirring, preservation, and cleaning; the stirring time is 1 hour; the temperature during preservation is 200°C, and the preservation time is 6–8 hours.

[0013] Furthermore, in step 3, AuCl3·HCl·4H2O and sodium citrate were mixed in a mass ratio of 1:1, stored in boiling water, and then cooled to obtain AuNPs.

[0014] Furthermore, the reaction process in step 4 includes sonication, shaking, separation, and cleaning; the sonication time is 15–45 min, and the shaking time is 8–16 h.

[0015] A nanoparticle made of MXene / Fe3O4 / AuNPs, wherein the nanochip includes MXene, AuNPs and Fe3O4 nanoparticles; the MXene is stacked in a layered structure, the AuNPs are dispersed on the surface or interstitial regions of the MXene, and the Fe3O4 nanoparticles are grown in situ on the MXene.

[0016] Application of MXene / Fe3O4 / AuNPs nanoparticles in the fabrication of nanochips.

[0017] An application of a nanochip, wherein the nanochip is used for metabolite detection.

[0018] Beneficial effects

[0019] (1) The IMG structure obtained by this invention, namely MXene / Fe3O4 / AuNPs, has the function of multi-channel proton transport path, which can reduce the overall heat loss and ensure the strength and comprehensiveness of subsequent signal detection. Because Fe3O4 and MXene are combined by growing MXene in situ, the IMG has good stability.

[0020] (2) The MXene / Fe3O4 / AuNPs obtained in this invention can reduce the heat dissipation of nanochips, thereby achieving strong thermally driven desorption and strong ionization.

[0021] (3) The MXene / Fe3O4 / AuNPs obtained in this invention, from the perspective of microstructure, have abundant metabolite capture contact sites and a sheet-like structure that facilitates metabolite desorption, which to some extent improves the ionization desorption efficiency. Attached Figure Description

[0022] Figure 1 This is a diagram illustrating the preparation process of MXene / Fe3O4 / AuNPs in Example 1 of the present invention.

[0023] Figure 2 The image shows the SEM images of MXene, IM, and IMG in Embodiment 1 of the present invention.

[0024] Figure 3 This is a standard metabolite intensity diagram of Example 1 of the present invention.

[0025] Figure 4 This is an intensity diagram of mixed standard metabolites under high protein and high salt conditions in Example 1 of the present invention.

[0026] Figure 5 This is a serum test image of different samples using IMG-assisted LDI-MS in Example 1 of the present invention. Detailed Implementation

[0027] The following will describe in conjunction with embodiments 1-3 of the present invention and appendices. Figure 1-5 The technical solutions of the present invention have been clearly and completely described. It is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of them. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0028] The preparation method of the present invention mainly includes the following steps:

[0029] Step 1: Dissolve LiF and Ti3AlC2·MAX in HCl, stir magnetically at 35℃ for 24h, centrifuge at 3500rpm, wash with deionized water until pH is 5-7, centrifuge at the same speed and collect MXene, freeze dry for later use.

[0030] Step 2: Disperse MXene in ethylene glycol by ultrasonication for 30 min, add FeCl3·6H2O, 1,6-hexanediamine and sodium acetate to the MXene suspension and stir for 1 h, transfer the resulting mixture to a reaction vessel and store at 200℃ for 7 h, and obtain ferric oxide / MXene by magnetic separation and washing;

[0031] Step 3: Mix AuCl3·HCl·4H2O and sodium citrate, dissolve in 50mL boiling water, store at 100℃ for 10min, and cool to room temperature to obtain AuNPs;

[0032] Step 4: Disperse ferric oxide / MXene in AuNPs solution, sonicate for 15-45 min, gently shake for 12 h, obtain ferric oxide / MXene / AuNPs by magnetic separation, wash several times with ethanol and deionized water, and disperse in deionized water for subsequent use.

[0033] The steps for further fabricating ferric oxide / MXene / AuNPs particles 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 using deionized water. To improve protein and salt tolerance, BSA and NaCl are mixed with typical small metabolites Glu, Arg, Try, Phe, and Lys at a concentration of 1 ng / nL for each metabolite. All blood samples are inserted into vacuolated tubes via venipuncture and allowed to clot within 1 hour at room temperature. Blood samples are centrifuged for 10 min, and serum is collected and immediately stored at -80℃ for further analysis. Different materials are prepared into 1 ng / nL aqueous suspensions, where Glu, Arg, Try, Phe, or Lys represents glucose, arginine, tryptophan, phenylalanine, and lysine, respectively.

[0034] LDI-MS analysis was performed using MALDI-TOF / TOF mass spectrometry (Shimadzu, AXIMA-Performance) in positive ion mode, equipped with a nitrogen laser (337 nm). Parameters were set including 200 laser scans per sample, a laser intensity of 65%, and a molecular weight range of 100 to 1000 Da. 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 transferred to cover the previously dried spots. The matrix included IMG, IM, etc. Samples were dried at room temperature before analysis on MALDI-TOF / TOF ms. For each sample, including small molecules and biological samples, five independent LDI-MS experiments were performed to ensure the reproducibility and reliability of the results. IMG consisted of iron(III) oxide / MXene / AuNPs, and IM consisted of iron(III) oxide / MXene.

[0035] Individual MXene particles are approximately 2 mm thick, Au nanoparticles are approximately 40 mm thick, and magnetite is approximately 20 mm thick. Magnetite and MXene are combined through in-situ MXene growth, providing excellent stability. Au nanoparticles are bonded to the magnetite / MXene through electrostatic interactions. In stability studies, after one year of storage at room temperature, the surface potential and morphology remained unchanged. ICP-OES analysis of the supernatant showed no leakage of Au or Fe elements, demonstrating the material's long-term storage stability. The novel structure formed by magnetite / MXene / AuNPs utilizes a multi-channel proton transport pathway. The addition of Fe reduces overall heat loss, ensuring signal detection strength and comprehensiveness. Finally, the synergistic surface plasmon effect of Au nanoparticles and MXene enables rapid energy conversion and heat generation. The Au to Fe ratio was further optimized using computer analysis, ultimately set at 2:1.

[0036] Example 1

[0037] Step 1: Dissolve LiF and Ti3AlC2·MAX in HCl, stir magnetically at 35℃ for 24h, centrifuge at 3500rpm, wash with deionized water until pH=5, centrifuge at the same speed and collect MXene, freeze dry for later use;

[0038] Step 2: Disperse MXene in ethylene glycol by ultrasonication for 30 min, add FeCl3·6H2O, 1,6-hexanediamine and sodium acetate to the MXene suspension and stir for 1 h, transfer the resulting mixture to a reaction vessel and store at 200℃ for 6 h, and obtain ferric oxide / MXene by magnetic separation and washing;

[0039] Step 3: Mix AuCl3·HCl·4H2O and sodium citrate, dissolve in 50mL boiling water, store at 100℃ for 10min, and cool to room temperature to obtain AuNPs;

[0040] Step 4: Disperse ferric oxide / MXene in AuNPs solution, sonicate for 15 min, gently shake for 8 h, obtain ferric oxide / MXene / AuNPs by magnetic separation, wash several times with ethanol and deionized water, and disperse in deionized water for subsequent use.

[0041] Example 2

[0042] Step 1: Dissolve LiF and Ti3AlC2·MAX in HCl, stir magnetically at 35℃ for 30h, centrifuge at 3500rpm, wash with deionized water until pH=6, centrifuge at the same speed and collect MXene, freeze dry for later use;

[0043] Step 2: Disperse MXene in ethylene glycol by ultrasonication for 30 min, add FeCl3·6H2O, 1,6-hexanediamine and sodium acetate to the MXene suspension and stir for 1 h, transfer the resulting mixture to a reaction vessel and store at 200℃ for 7 h, and obtain ferric oxide / MXene by magnetic separation and washing;

[0044] Step 3: Mix AuCl3·HCl·4H2O and sodium citrate, dissolve in 50mL boiling water, store at 100℃ for 10min, and cool to room temperature to obtain AuNPs;

[0045] Step 4: Disperse ferric oxide / MXene in AuNPs solution, sonicate for 30 min, gently shake for 12 h, obtain ferric oxide / MXene / AuNPs by magnetic separation, wash several times with ethanol and deionized water, and disperse in deionized water for subsequent use.

[0046] Example 3

[0047] Step 1: Dissolve LiF and Ti3AlC2·MAX in HCl, stir magnetically at 35℃ for 36h, centrifuge at 3500rpm, wash with deionized water until pH=7, centrifuge at the same speed and collect MXene, freeze dry for later use;

[0048] Step 2: Disperse MXene in ethylene glycol by ultrasonication for 30 min, add FeCl3·6H2O, 1,6-hexanediamine and sodium acetate to the MXene suspension and stir for 1 h, transfer the resulting mixture to a reaction vessel and store at 200℃ for 8 h, and obtain ferric oxide / MXene by magnetic separation and washing;

[0049] Step 3: Mix AuCl3·HCl·4H2O and sodium citrate, dissolve in 50mL boiling water, store at 100℃ for 10min, and cool to room temperature to obtain AuNPs;

[0050] Step 4: Disperse ferric oxide / MXene in AuNPs solution, sonicate for 45 min, gently shake for 16 h, obtain ferric oxide / MXene / AuNPs by magnetic separation, wash several times with ethanol and deionized water, and disperse in deionized water for subsequent use.

[0051] Combination Figure 1 and Figure 2 As shown, Figure 2From left to right are SEM images of MXene, IM, and IMG. MXene is stacked in a layered structure, with AuNPs dispersed on the surface or interstitial regions of MXene. Ferric oxide nanoparticles are grown in situ on MXene. There are many electron-hole sources in MXene, resulting in a large number of electron-hole complexes originating from the retrograde high points. AuNPs establish a potential barrier on MXene, constructing a proton transport pathway, which to some extent reduces the electron-hole complexing efficiency of MXene. Furthermore, the large number of homogeneous charges accumulated on the surface of AuNPs can promote the desorption of ionized samples to some extent. Finally, FeNPs are integrated into MXene / AuNPs, where MXene provides electron-hole sources, AuNPs provide rapid energy conversion, and FeNPs provide efficient thermal resistance.

[0052] Combination Figure 3 In the text, a, b, c, d, and e represent the intensity comparison between standard samples and IMG, respectively, for glucose, tryptophan, phenylalanine, arginine, and lysine. Figure 4 As shown, LDI enhancement was systematically explored using traditional organic matrices, including DHB, CHCA, DHAP, P25 semiconductor, MXene, AuNPs, FeNPs, IG, IM, MG, and IMG nanochips. Five small molecule metabolites—glucose, tryptophan, phenylalanine, arginine, and lysine—were selected as standard samples to evaluate the performance of the nanomaterials. IMG was found to have the highest strength. Figure 5 As shown, IMG can assist LDI-MS in testing different serum samples.

[0053] The preparation method used in this invention enables the in-situ growth of iron oxide in MXene to obtain a stable structure. This structure also provides a multi-channel proton transport pathway, which can reduce overall heat loss and ensure signal detection strength and comprehensiveness. The fabricated nanochip can broaden the application of metabolite detection for multiple diseases in the same organ, especially liver diseases.

Claims

1. A method for preparing MXene / Fe3O4 / AuNPs, characterized in that, The specific steps are as follows: Step 1: Dissolve LiF and Ti3AlC2·MAX in hydrochloric acid, react completely to obtain MXene, and freeze-dry for later use; Step 2: Disperse MXene in ethylene glycol, add FeCl3·6H2O and 1,6-hexanediamine and NaAc in a molar ratio of 1:8:13, and react fully to obtain MXene / Fe3O4. Step 3: Prepare AuNPs solution; Step 4: MXene / Fe3O4 is dispersed in AuNPs solution and subjected to ultrasonic treatment for 15-45 min and shaking reaction for 8-16 h to obtain MXene / Fe3O4 / AuNPs nanoparticles for laser desorption / ionization mass spectrometry detection of small molecule metabolites. The nanoparticles include MXene, AuNPs, and iron oxide nanoparticles; the MXene is stacked in a layered structure, the AuNPs are dispersed on the surface or interstitial regions of the MXene, and the iron oxide nanoparticles are grown in situ on the MXene.

2. The preparation method of MXene / Fe3O4 / AuNPs as described in claim 1, characterized in that, The reaction in step 1 includes stirring, centrifugation, and washing; the stirring time is 24 h to 36 h, and the temperature is 35℃; the washing is performed until the pH is 5 to 7, and the centrifugation rate is 3500 rpm.

3. The preparation method of MXene / Fe3O4 / AuNPs as described in claim 1, characterized in that, The reaction in step 2 includes stirring, preservation, and cleaning; the stirring time is 1 h; the temperature during preservation is 200℃, and the preservation time is 6~8 h.

4. The preparation method of MXene / Fe3O4 / AuNPs as described in claim 1, characterized in that, In step 3, AuCl3·HCl·4H2O and sodium citrate are mixed in a mass ratio of 1:1, stored in boiling water, and then cooled to obtain AuNPs.

5. An application of the MXene / Fe3O4 / AuNPs nanoparticles as described in claim 1, characterized in that, The nanoparticles are used to fabricate nanochips.

6. The application of a nanochip as described in claim 5, characterized in that, The nanochip is used for metabolite detection.

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