Preparation method of zearalenone-14-glucoside self-assembly gel

Abstract

The present invention discloses a preparation method of a self-assembled zearalenone-14-glucoside gel, comprising mixing zearalenone-14-glucoside and methanol in a mass ratio of (10-20): 1 to obtain a zearalenone-14-glucoside methanol solution; adding distilled water to the zearalenone-14-glucoside methanol solution in a volume ratio of 2: 8 to obtain a zearalenone-14-glucoside methanol-water mixed solution, stirring thoroughly, heating at 50-90° C. for 1-2 hours, and then standing and cooling at room temperature to obtain a uniform translucent gel. The present invention uses a glycoside-type hidden fungal toxin with an estrogen-like effect as a gelling substance and a methanol-water mixed solvent to construct a natural self-assembled supramolecular gel with gel properties such as biocompatibility, sustained release, and molecular targeting.

Description

Technical Field

[0001] The present invention relates to the technical field of gel preparation, in particular to a method for preparing a zearalenone-14-glucoside self-assembly gel. Background Art

[0002] Zearalenone (ZEN) is an estrogenic mycotoxin. As a common mycotoxin in food and feed, it is widespread and highly toxic. ZEN is most likely to contaminate corn, but crops such as barley, oats, wheat, sorghum, millet, and rice are also susceptible. Zearalenone-14-glucoside (Z14G), one of the most prominent cryptic mycotoxins of ZEN, has attracted widespread attention from researchers because it can be completely converted to ZEN through hydrolysis by intestinal microorganisms. The estrogenic activity of ZEN and Z14G in environmental exposure is additive, with studies showing that Z14G is present in 60% of samples.

[0003] Natural self-assembling supramolecular gels are used in a variety of fields, including biomedicine, chemical synthesis, and materials engineering, due to their excellent biocompatibility, bioactivity, sustained release, molecular targeting, variability, multi-responsiveness, and appropriate viscoelasticity. However, current research on Z14G is limited to the single-molecule level, while the chemical structure and toxicity of its spontaneous supramolecular structure in natural liquids have not been studied. Therefore, a method for preparing self-assembling gels of zearalenone-14-glucoside is needed. Summary of the Invention

[0004] The purpose of the present invention is to solve the problems of the prior art and to provide a method for preparing a self-assembled gel of zearalenone-14-glucoside.

[0005] In order to achieve the above object, the present invention adopts the following technical solutions:

[0006] The present invention includes

[0007] A. Mixing zearalenone-14-glucoside and methanol in a mass ratio of (10-20):1 to obtain a zearalenone-14-glucoside methanol solution;

[0008] B. adding distilled water to the zearalenone-14-glucoside methanol solution at a volume ratio of 2:8 to obtain a zearalenone-14-glucoside methanol-water mixed solution;

[0009] C. The zearalenone-14-glucoside methanol-water mixed solution is fully stirred and heated at 50-90° C. for 1-2 hours, and then allowed to stand and cool at room temperature to obtain a uniform translucent gel.

[0010] Furthermore, the concentration of the translucent gel is 2.0-4.0 g / L.

[0011] Furthermore, in step C, the heating temperature may be 70°C.

[0012] In a second aspect, the zearalenone-14-glucoside self-assembled gel obtained according to the preparation method is used as a delivery carrier, and the application as a delivery carrier includes but is not limited to any one of the following: (1) as a drug delivery carrier;

[0013] (2) As a tool for studying the delivery performance of guest molecules; in the above aspect (2), the guest molecule is one including but not limited to a drug, a model drug or a pathogen; wherein the model drug is a drug used to achieve a disease modeling effect, and the pathogen is one including but not limited to bacteria, fungi, viruses, and bacteriophages.

[0014] Compared with the prior art, the present invention has the following beneficial effects:

[0015] The present invention uses glycoside-type concealed fungal toxins with estrogen-like effects as a gelling agent and a methanol-water mixed solvent to construct a natural self-assembled supramolecular toxin gel with gel properties such as biocompatibility, sustained release and molecular targeting, filling the gap in the field of natural self-assembled gels in toxicology and laying the foundation for the subsequent preparation and research of natural self-assembled toxin gels. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 Schematic diagram of the preparation method of the zearalenone-14-glucoside self-assembled gel of the present invention;

[0017] Figure 2 Schematic diagram of an optical microscope image of Z14G-gel in Example 1 of the present invention;

[0018] Figure 3 Schematic diagram of cryo-TEM of Z14G-gel in Example 1 of the present invention;

[0019] Figure 4 TEM diagram of the dried Z14G gel powder of Example 1 of the present invention;

[0020] Figure 5 Schematic diagram of AFM of dried Z14G gel powder according to Example 1 of the present invention;

[0021] Figure 6 Schematic diagram of SEM of dried Z14G gel powder of Example 1 of the present invention;

[0022] Figure 7Schematic diagram of the strain-dependent oscillatory shear rheology of Z14G-gel at a fixed frequency of 10 rad s-1 in Example 1 of the present invention;

[0023] Figure 8 Schematic diagram of dynamic time scanning of Z14G-gel in Example 1 of the present invention at 0.1% strain and 10 rad s-1 frequency; DETAILED DESCRIPTION

[0024] The present invention will be further described below with reference to specific examples. The exemplary embodiments and descriptions of the present invention are intended to explain the present invention but are not intended to limit the present invention.

[0025] The user experience effect evaluation method and system based on ergonomic analysis of the present invention include the following steps:

[0026] like Figure 1 As shown, in this embodiment, the following steps are included:

[0027] A weighed 4.0 mg of Z14G powder and dissolved in 0.2 mL of methanol;

[0028] B. Add 0.8 mL of distilled water to the Z14G methanol solution;

[0029] The mixture was stirred thoroughly at C and heated at 70°C to obtain a uniform solution. The Z14G methanol-water mixed solution was allowed to cool at room temperature until a uniform translucent gel with a gel concentration of 4.0 mg / mL was obtained. The Z14G-gel was lyophilized to obtain Z14G gel powder.

[0030] In this embodiment, the gel concentration is 4.0 mg / mL.

[0031] Example 2

[0032] A. Mixing zearalenone-14-glucoside and methanol in a mass ratio of 10:1 to obtain a zearalenone-14-glucoside methanol solution;

[0033] B. adding distilled water to the zearalenone-14-glucoside methanol solution at a volume ratio of 2:8 to obtain a zearalenone-14-glucoside methanol-water mixed solution;

[0034] C. The zearalenone-14-glucoside methanol-water mixed solution is fully stirred and heated at 50° C. for 1-2 hours, and then allowed to stand and cool at room temperature to obtain a uniform translucent gel.

[0035] In this embodiment, the gel concentration is 2.0 mg / mL.

[0036] Example 3

[0037] A. Mixing zearalenone-14-glucoside and methanol in a mass ratio of 20:1 to obtain a zearalenone-14-glucoside methanol solution;

[0038] B. adding distilled water to the zearalenone-14-glucoside methanol solution at a volume ratio of 2:8 to obtain a zearalenone-14-glucoside methanol-water mixed solution;

[0039] C. The zearalenone-14-glucoside methanol-water mixed solution is fully stirred and heated at 80° C. for 1-2 hours, and then allowed to stand and cool at room temperature to obtain a uniform translucent gel.

[0040] In this embodiment, the gel concentration is 3.8 mg / mL.

[0041] The Z14G-gel was morphologically characterized by optical microscopy, SEM, TEM, cryo-TEM, and AFM experiments; OM images of Z14G gel were taken by a DM 2500 optical microscope (Leica, Germany), and the dried Z14G gel was sputter-coated with a thin layer of gold before observation. SEM images were acquired by a field-emission scanning electron microscope (JSM-6700F, JEOL, Japan) at 10 mA, 20 kV. Dried Z14G gel samples were prepared by drop-casting the solution on a carbon-coated copper grid and then dried in air. TEM images were acquired using a FEI Talos F200X instrument. Dried Z14G gel powder was placed on a freshly cleaved mica plate, and AFM images were acquired on a Bruker Dension Icon (Bruker AXS). Cryo-TEM images were acquired on a field-emission cryo-transmission electron microscope (FEI Talos F200C).

[0042] Taking the morphological characterization and self-assembly properties of Z14G in Example 1 as the main observation objects, a thermoresponsive natural supramolecular self-assembly gel based on Z14G was obtained by keeping it stationary in an inverted vial, which can remain stable at room temperature.

[0043] The Z14G-gel was confirmed by rheological testing;

[0044] Rheological studies were performed on a rotational rheometer (Anton Paar MCR92, Austria). The strain-dependent modulus was measured from 0.1 to 100% strain. Dynamic time sweeps were measured at 0.1% strain and set to 30 min. All measurements were performed at a Z14G gel concentration of 4 mg / mL and 25 °C.

[0045] The cross-linked fibrous network can be directly observed in Z14G-gel by optical microscopy and cryo-TEM. Figure 2 and attached Figure 3 As shown in Figure 2, TEM and AFM of Z14G-gel showed a large number of interlaced fibers formed by Z14G molecules with a diameter of about 20 nm. Figure 4 and attached Figure 5 In addition, SEM images also show that Z14G can self-assemble into nanofibers as shown in the attached Figure 6 As shown, based on the above results, it can be inferred that Z14G molecules can first self-assemble into one-dimensional nanofibers, further extend and cross-link together to produce a three-dimensional fiber network, and finally fix the solvent molecules to form a gel.

[0046] The rheological behavior of Z14G-gel was determined by oscillatory shear rheology. A gel must meet the rheological definition, that is, the storage modulus (G') value is higher than the loss modulus (G") value. The strain-dependent oscillatory shear rheological results of Z14G-gel showed that G' was 10 times larger than G", indicating that the sample remained in the gel state until the strain exceeded about 30%, at which point G' decreased significantly and then even fell below G", indicating a clear phase transition of Z14G from gel to solution as shown in the attached figure. Figure 7 In addition, the results under dynamic time scanning (G'>G" in the text process) also confirmed the excellent solid-like properties of Z14G gel as shown in the attached figure. Figure 8 shown.

[0047] In the present invention, Z14G can form Z14G-gel under certain conditions, with a minimum gel concentration of 2.0 mg / mL and an optimal gel concentration of 4.0 mg / mL. Images collected by optical microscopy, SEM, TEM, cryo-TEM and AFM show that Z14G-gel can form a stable three-dimensional fiber network. Finally, rheological measurements of Z14G-gel also confirmed the gel properties of Z14G-gel.

[0048] The zearalenone-14-glucoside self-assembled gel is used as a delivery carrier

[0049] Hydrophilic small molecules (thioflavin T, rhodamine B and doxorubicin) and / or hydrophobic small molecules (Nile red and paclitaxel) are dissolved in the above solution according to the designed content (Mg) and composition. After the solvent evaporates, a self-assembled gel loaded with guest molecules is obtained.

[0050] The guest molecule content in the self-assembled gel was determined by weighing and UV-visible spectroscopy. The mass of the self-assembled gel was determined by drying 200 μL of the oligomer solution and averaging the mass of three samples (MFF-GA). For the guest molecule-loaded sample, 200 μL of water was added to the dried sample and shaken. After one day, the sample was centrifuged at 8000 rpm for 5 minutes. The supernatant was collected and analyzed by UV-visible spectroscopy.

[0051] When the initial concentration of the guest molecule was 1 mg / mL, the adsorption efficiency of hydrophilic Thioflavin T was approximately 94.5%, and the loading efficiency was approximately 17.4%. In contrast, the adsorption efficiency of hydrophobic Nile Red was 100%, and the loading efficiency was 18.3%. When both hydrophilic Thioflavin T and hydrophobic Nile Red were added simultaneously, the adsorption efficiency reached approximately 100%, and the loading efficiency was approximately 30.9%, significantly superior to conventional nanoparticle drug carriers (which struggle to simultaneously encapsulate both hydrophilic and hydrophobic guest molecules, and whose loading efficiency rarely exceeds 20%). When the initial concentration was increased to 2 mg / mL, the Nile Red loading efficiency increased to 30.9%, demonstrating that the encapsulation capacity of the guest molecule can be modified by varying the initial concentration of the self-assembling gel.

[0052] The above description is only a preferred specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any technician familiar with the technical field, within the technical scope disclosed by the present invention, who makes equivalent replacements or changes based on the technical solution and inventive concept of the present invention, should be covered by the scope of protection of the present invention.

Claims

1. A method for preparing a self-assembled gel of zearalenone-14-glucoside, characterized in that: The steps include: A. Mixing zearalenone-14-glucoside and methanol in a mass ratio of (10-20):1 to obtain a zearalenone-14-glucoside methanol solution; B. adding distilled water to the zearalenone-14-glucoside methanol solution at a volume ratio of 2:8 to obtain a zearalenone-14-glucoside methanol-water mixed solution; C. The zearalenone-14-glucoside methanol-water mixed solution is fully stirred and heated at 50-90° C. for 1-2 hours, and then allowed to stand and cool at room temperature to obtain a uniform translucent gel.

2. The method for preparing the zearalenone-14-glucoside self-assembled gel according to claim 1, wherein: The concentration of the translucent gel is 2.0-4.0 g / L.

3. The method for preparing the zearalenone-14-glucoside self-assembled gel according to claim 1, wherein: In step C, the heating temperature was 70°C.

4. Use of the zearalenone-14-glucoside self-assembled gel obtained by the preparation method according to any one of claims 1 to 3 as a delivery carrier, characterized in that: The application as a delivery carrier includes but is not limited to any of the following: (1) as a drug delivery carrier; (2) As a tool for studying the delivery performance of guest molecules; in the above aspect (2), the guest molecule is one including but not limited to a model drug or a pathogen; wherein the model drug is a drug used to achieve a disease modeling effect, and the pathogen is one including but not limited to bacteria, fungi, and viruses.

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