Method for embedding vitamin acetate in prolamin
By dissolving alcohol-soluble proteins and vitamin acetate in glacial acetic acid, stable nanoparticles are formed using hydrogen bonds and hydrophobic interactions, solving the problems of poor encapsulation effect and rapid release rate of alcohol-soluble proteins, and achieving high encapsulation rate and stable delivery of vitamin acetate.
Patent Information
- Application Number
- CN202511401760.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-09-28
- Publication Date
- 2025-12-19
AI Technical Summary
Existing methods for encapsulating vitamin acetate with alcohol-soluble proteins have problems such as poor encapsulation effect and rapid release rate, which affect its bioavailability.
Glacial acetic acid was used as a solvent to dissolve alcohol-soluble protein and vitamin acetate. Vitamin acetate was loaded into alcohol-soluble protein particles through hydrogen bonding and hydrophobic interactions. Stabilizers were used to keep the nanoparticles stable. The solvent was removed by dialysis to form stable nanoparticles.
It improved the encapsulation rate of vitamin acetate, slowed the release rate, enhanced the stability during storage, and improved oral bioavailability.
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Figure CN121154843A_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The present application belongs to the technical field of nano-encapsulation, and relates to embedding of vitamins by protein nanoparticles, in particular to a method for embedding vitamin acetate ester by prolamin. BACKGROUND
[0002] The information disclosed in this BACKGROUND section is only for the purpose of increasing the understanding of the general background of the application and does not necessarily constitute an admission by the patent applicant(s) that this information constitutes prior art.
[0003] Vitamin A acetate and vitamin E acetate are ester derivatives of vitamin A and vitamin E (collectively referred to as vitamin acetate ester in the present application), which have little difference in properties from vitamin A and vitamin E, but have good chemical stability. Vitamin acetate ester has poor water solubility, and is therefore usually dissolved in vegetable oil or soybean oil and the like for human vitamin A and E supplementation, but the problem of low oral bioavailability of vitamin acetate ester has not been solved. In order to improve the oral bioavailability of vitamin acetate ester, it can be encapsulated in protein particles with biological safety and cost-effectiveness to solve the limitations of poor water solubility and / or solubility, low permeability, gastrointestinal tract chemical instability and poor epithelial permeability, thereby greatly improving its bioavailability.
[0004] Prolamin not only has strong hydrophobicity and hydrophilic affinity for embedding vitamin acetate ester, but also can spontaneously form a film layer and has an antioxidant effect. At the same time, prolamin can also be enzymatically degraded in the intestinal tract, and the released vitamins form mixed micelles with bile salts to promote the absorption of fat-soluble vitamins through the intestinal mucosa. Therefore, the use of prolamin is conducive to improving the oral bioavailability of vitamin acetate ester. However, the present application has found that different preparation methods often form particles with different structures, which in turn affect the embedding process. The distribution of active substances in prolamin also affects the embedding efficiency and release rate of active substances. The current method for embedding vitamin acetate ester with prolamin has problems such as poor embedding effect and fast release rate. SUMMARY
[0005] In order to solve the problems of the prior art, the purpose of the present application is to provide a method for embedding vitamin acetate ester with prolamin. The method provided by the present application can improve the embedding rate of vitamin acetate ester with prolamin, slow down the release rate, and enhance the stability during storage.
[0006] In order to achieve the above-mentioned purpose, the technical scheme of the present application is as follows: In a first aspect, a method for embedding vitamin acetate ester with prolamin comprises the following steps: dissolving the prolamin and the vitamin acetate ester in glacial acetic acid to obtain a glacial acetic acid phase; The stabilizer is dissolved in water to form a stabilizer phase; The stabilizer phase is added to the glacial acetic acid phase to obtain a mixed solution; the volume of the stabilizer phase is less than that of the glacial acetic acid phase; The mixed solution is added to water to form the prolamin particle loaded with vitamin acetate wrapped by the stabilizer; the volume of the mixed solution is less than that of the water.
[0007] The prolamin used in the present application has a plurality of amino acids, and the mass content of more than 45% of the amino acids is hydrophobic amino acid. The prolamin is usually dissolved in an alcohol solvent, such as an ethanol aqueous solution, a methanol aqueous solution, a glycol aqueous solution, etc. The present application researches and finds that the molecular cross-linking and molecular aggregation of the prolamin in the alcohol solvent are stronger than the cross-linking and aggregation in the glacial acetic acid, and therefore the glacial acetic acid is a more excellent solvent for unfolding the prolamin. The present application uses the glacial acetic acid as a solvent to dissolve the prolamin and the vitamin acetate at the same time, and through the synergistic effect of the dissolution and protonation of the glacial acetic acid, the prolamin is fully unfolded, the prolamin after full unfolding can expose its amino and carboxyl groups, the exposed amino and carboxyl groups can form hydrogen bonds with the ester groups of the vitamin acetate, and the prolamin and the vitamin acetate are loaded into the particle through the hydrogen bonds and the hydrophobic interaction force, the prolamin with high unfolding degree and the vitamin acetate interact with each other, and after the self-assembly induced by water, the prolamin can load the vitamin acetate into the relative inner layer of the particle.
[0008] In addition, the present application first dissolves the stabilizer in water to form a stabilizer phase, and then adds the stabilizer phase to the glacial acetic acid phase, so that the stabilizer, the prolamin and the vitamin acetate can be fully mixed and reacted, and the entire nanoparticle can be kept stable in the preparation process by using the stabilizer.
[0009] In the second aspect, a nanoparticle is obtained by the above method.
[0010] The present application has the following beneficial effects: The present application uses the prolamin-vitamin acetate to dissolve in the glacial acetic acid and form intermolecular interaction force, and induces the prolamin molecules to polymerize under the condition of reducing the proportion of acetic acid. The advantage is that the glacial acetic acid as a solvent promotes the prolamin to unfold, and is more conducive to the embedding of the vitamin acetate. The nanoparticle provided by the present application can be used for the protection and delivery of the vitamin acetate. The experiments show that the nanoparticle obtained by the method of the present application has higher embedding rate of the vitamin acetate, better slow-release effect and stronger stability in the storage process, thereby being conducive to improving the oral availability of the vitamin acetate. BRIEF DESCRIPTION OF DRAWINGS
[0011] The drawings constituting a part of the specification of the present application serve to provide a further understanding of the present application, and the schematic embodiments of the present application and the description thereof serve to explain the present application, and do not constitute an improper limitation on the present application.
[0012] Figure 1 Figure 3 is a graph showing the particle size analysis of prolamine in different concentrations of acetic acid or ethanol; (A) is acetic acid, and (B) is ethanol.
[0013] Figure 2 Figure 4 is a graph showing the microstructure of prolamine in different proportions of acetic acid; (a) and (b) are 90% (v / v) acetic acid, (c) and (d) are 60% (v / v) acetic acid, and (e) and (f) are 3% (v / v) acetic acid.
[0014] Figure 3 Figure 5 is a graph showing the embedding efficiency and storage stability of the prolamine particles in different embodiments; (A) is the embedding rate, and (B) is the storage retention rate.
[0015] Figure 4 Figure 6 is a graph showing the in vitro release of the active substance in the prolamine particles in different embodiments.
[0016] Figure 5 Figure 7 is a graph showing the prolamine particles stabilized by the vitamin A acetate carboxymethyl β-cyclodextrin obtained in Example 1; the scale of (A) is 500 nm, and the scale of (B) is 200 nm. DETAILED DESCRIPTION
[0017] It should be noted that the following detailed description is merely exemplary in nature and is intended to provide further description of the application. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.
[0018] It is also important to note that the terms "including", "comprising", and / or "having" as used herein are specifically intended to be open-ended and also to mean including, comprising, and / or having other components, in addition to the listed components. It is also important to note that the use of relational terms, if any, such as first, second, third, and / or the like, are used solely to distinguish one from another entity or action without necessarily implying an actual such relationship or order between such entities or actions.
[0019] In view of the problems of poor embedding effect and fast release rate of the existing method of embedding vitamin acetate ester with prolamine, the present application provides a method of embedding vitamin acetate ester with prolamine.
[0020] In a typical embodiment of the present application, a method of embedding vitamin acetate ester with prolamine is provided, which comprises the following steps: dissolving the prolamine and the vitamin acetate ester in glacial acetic acid to obtain a glacial acetic acid phase; dissolving the stabilizer in water to form a stabilizer phase; adding the mixed solution into water to mix, to form the prolamine particle loaded with vitamin acetate and wrapped by the stabilizer; the volume of the mixed solution is less than the volume of the water. adding the mixed solution into water to mix, to form the prolamine particle loaded with vitamin acetate and wrapped by the stabilizer; the volume of the mixed solution is less than the volume of the water.
[0021] The present application utilizes the solvent effect of glacial acetic acid to dissolve prolamine, which can protonate the prolamine molecules, and the prolamine after protonation has positive charge. Through the dissolution effect of glacial acetic acid, the spherical prolamine can be well unfolded by the positive charge repulsion in the molecule. The hydrophobic amino acid is turned outwards to promote the interaction force with vitamin acetate, thereby increasing the distribution of vitamin acetate in the prolamine particle, improving the embedding effect and reducing the release rate.
[0022] In some embodiments, the vitamin acetate is vitamin A acetate and / or vitamin E acetate.
[0023] In some embodiments, the mass ratio of prolamine to vitamin acetate is 1-4:0.1-0.5. Under this condition, the embedding of vitamin acetate is more favorable, and the loss of vitamin acetate is reduced.
[0024] In some embodiments, the mass fraction of prolamine in the glacial acetic acid phase is 1-4%. This condition is favorable for the full unfolding of prolamine.
[0025] The stabilizer according to the present application includes, but is not limited to, water-soluble polysaccharides, proteins, ester substances and surfactants. In some embodiments, the stabilizer is carboxymethyl beta cyclodextrin, sodium alginate or soy lecithin. Studies have shown that the effect of this stabilizer is better.
[0026] In some embodiments, the mass concentration of the stabilizer in the stabilizer phase is 0.2-3%. Under this condition, the formation of the prolamine particle loaded with vitamin acetate and wrapped by the stabilizer is more favorable.
[0027] In some embodiments, the volume ratio of the glacial acetic acid phase to the stabilizer phase is 20:1-5.
[0028] In some embodiments, during the process of adding the mixed solution into water to mix, the volume ratio of the mixed solution to water is 0.8-1.0:20. Under this condition, the formation of the prolamine particle loaded with vitamin acetate and wrapped by the stabilizer is more favorable.
[0029] In some embodiments, after adding the mixed solution into water to mix, dialysis is used to remove glacial acetic acid. The removal of glacial acetic acid makes the prolamine shrink as much as possible, further improving the embedding effect. Specifically, the dialysis time is 24-48 hours.
[0030] Another embodiment of the present application provides a nanoparticle obtained by the above method.
[0031] In order to enable those skilled in the art to more clearly understand the technical solutions of the present application, the technical solutions of the present application will be described in detail below in combination with specific embodiments.
[0032] Pre-experiment Gliadin was dissolved in acetic acid aqueous solution with volume ratio of 40% (v / v), 50% (v / v), 60% (v / v), 70% (v / v), 80% (v / v), and 90% (v / v) respectively; and in ethanol aqueous solution with volume ratio of 40% (v / v), 50% (v / v), 60% (v / v), 70% (v / v), 80% (v / v), and 90% (v / v) respectively. Dynamic light scattering (DLS) and transmission electron microscopy were used to analyze the particle size and characterize the microstructure of the dissolved gliadin (concentration of 45 mg / mL) respectively.
[0033] The particle size analysis results are shown in Figure 1 Figure 1 A shows that as the proportion of acetic acid increases from 40% (v / v) to 90% (v / v), the size of gliadin decreases from about 950 nm to 43 nm. Figure 1 B shows that as the proportion of ethanol increases from 50% (v / v) to 90% (v / v), the size changes first decrease and then increase, and the minimum size is about 130 nm. This indicates that high concentration of acetic acid helps to dissolve gliadin, and 80% (v / v) ethanol is more suitable for gliadin dissolution. Through comparison of different solvents, it is shown that acetic acid is a more suitable solvent for gliadin.
[0034] The microstructure results are shown in Figure 2 It is shown that in 90% (v / v) and 60% (v / v) acetic acid, the microstructure of gliadin is that 90% (v / v) acetic acid reduces intermolecular crosslinking and the size is smaller, while 60% (v / v) presents a loose large particle structure. In low concentration of 3% (v / v) acetic acid, gliadin particles are more compact.
[0035] In summary, acetic acid is a more excellent solvent for unfolding gliadin. The following examples are designed and implemented based on the results of the pre-experiment.
[0036] Example 1 A method for embedding vitamin acetate in gliadin, comprising the following steps: 1. Dissolve prolamine and retinyl acetate in glacial acetic acid, stir to dissolve and let stand for 4 hours to obtain a retinyl acetate-prolamine glacial acetic acid phase; wherein the concentration of prolamine is 45 mg / mL and the concentration of retinyl acetate is 3.5 mg / mL.
[0037] 2. Dissolve carboxymethyl β-cyclodextrin in water, stir overnight to fully hydrate to obtain a carboxymethyl β-cyclodextrin water phase; wherein the concentration of carboxymethyl β-cyclodextrin is 10 mg / mL.
[0038] 3. Mix 0.8 mL of the retinyl acetate-prolamine glacial acetic acid phase obtained in step 1 and 0.15 mL of the carboxymethyl β-cyclodextrin water phase obtained in step 2, let stand for 0.5 hours to obtain a mixture.
[0039] 4. Disperse the mixture obtained in step 3 into 20 mL of water at a stirring rate of 800 rpm to obtain carboxymethyl β-cyclodextrin-coated retinyl acetate-loaded prolamine particles.
[0040] 5. Dialyze the colloidal dispersion obtained in step 4 for 48 hours to remove acetic acid.
[0041] Example 2 A method of embedding a vitamin acetate ester with prolamine, comprising the following steps: 1. Dissolve prolamine and retinyl acetate in an ethanol solution with a volume fraction of 85%, stir to dissolve and let stand for 4 hours to obtain a retinyl acetate-prolamine ethanol phase; wherein the concentration of prolamine is 45 mg / mL and the concentration of retinyl acetate is 3.5 mg / mL.
[0042] 2. Dissolve carboxymethyl β-cyclodextrin in water, stir overnight to fully hydrate to obtain a carboxymethyl β-cyclodextrin water phase; wherein the concentration of carboxymethyl β-cyclodextrin is 10 mg / mL.
[0043] 3. Mix 0.8 mL of the retinyl acetate-prolamine ethanol phase obtained in step 1 and 0.15 mL of the carboxymethyl β-cyclodextrin water phase obtained in step 2, let stand for 0.5 hours to obtain a mixture.
[0044] 4. Disperse the mixture obtained in step 3 into 20 mL of water at a stirring rate of 800 rpm to obtain carboxymethyl β-cyclodextrin-coated retinyl acetate-loaded prolamine particles.
[0045] 5. Dialyze the colloidal dispersion obtained in step 4 for 48 hours to remove ethanol.
[0046] Example 3 A method of embedding a vitamin acetate ester with prolamine, comprising the following steps: 1. Disperse prolamine and vitamin A acetate in glacial acetic acid, stir to dissolve, and let stand for 4 hours to obtain a vitamin A acetate-prolamine glacial acetic acid phase; wherein the concentration of prolamine is 45 mg / mL and the concentration of vitamin A acetate is 3.5 mg / mL.
[0047] 2. Dissolve carboxymethyl β-cyclodextrin in water, stir overnight to fully hydrate, and obtain a carboxymethyl β-cyclodextrin water phase; wherein the concentration of carboxymethyl β-cyclodextrin is 20 mg / mL.
[0048] 3. Mix 0.8 mL of the vitamin A acetate-prolamine glacial acetic acid phase obtained in step 1 and 0.12 mL of the carboxymethyl β-cyclodextrin water phase obtained in step 2, and let stand for 0.5 hours to obtain a mixture.
[0049] 4. Disperse the mixture obtained in step 3 into 20 mL of water at a stirring rate of 800 rpm to obtain carboxymethyl β-cyclodextrin-coated vitamin A acetate-loaded prolamine particles.
[0050] 5. Dialyze the colloidal dispersion obtained in step 4 for 72 hours to remove acetic acid.
[0051] Example 4 A method for embedding vitamin acetate ester with prolamine, comprising the following steps: 1. Disperse prolamine and vitamin A acetate in an ethanol solution with a volume fraction of 85%, stir to dissolve, and let stand for 4 hours to obtain a vitamin A acetate-prolamine ethanol phase; wherein the concentration of prolamine is 45 mg / mL and the concentration of vitamin A acetate is 3.5 mg / mL.
[0052] 2. Dissolve carboxymethyl β-cyclodextrin in water, stir overnight to fully hydrate, and obtain a carboxymethyl β-cyclodextrin water phase; wherein the concentration of carboxymethyl β-cyclodextrin is 20 mg / mL.
[0053] 3. Mix 0.8 mL of the vitamin A acetate-prolamine ethanol phase obtained in step 1 and 0.12 mL of the carboxymethyl β-cyclodextrin water phase obtained in step 2, and let stand for 0.5 hours to obtain a mixture.
[0054] 4. Disperse the mixture obtained in step 3 into 20 mL of water at a stirring rate of 800 rpm to obtain carboxymethyl β-cyclodextrin-coated vitamin A acetate-loaded prolamine particles.
[0055] 5. Dialyze the colloidal dispersion obtained in step 4 for 72 hours to remove ethanol.
[0056] Example 5 A method for embedding vitamin acetate ester with prolamine, comprising the following steps: 1. Dissolve prolamine and vitamin E acetate in glacial acetic acid, stir to dissolve and let stand for 4 hours to obtain a vitamin A acetate-prolamine glacial acetic acid phase; wherein the concentration of prolamine is 10 mg / mL and the concentration of vitamin A acetate is 1 mg / mL.
[0057] 2. Dissolve sodium alginate in water, stir overnight to fully hydrate, to obtain a carboxymethyl beta cyclodextrin water phase; wherein the concentration of carboxymethyl beta cyclodextrin is 15 mg / mL.
[0058] 3. Mix 0.8 mL of the vitamin E acetate-prolamine glacial acetic acid phase obtained in step 1 and 0.2 mL of the sodium alginate water phase obtained in step 2, let stand for 0.5 hours to obtain a mixture.
[0059] 4. Disperse the mixture obtained in step 3 into 20 mL of water at a stirring rate of 900 rpm to obtain sodium alginate-encapsulated vitamin E acetate-loaded prolamine particles.
[0060] 5. Dialyze the colloidal dispersion obtained in step 4 for 24 hours to remove acetic acid.
[0061] Example 6 A method of embedding vitamin acetate ester with prolamine, comprising the following steps: 1. Dissolve prolamine and vitamin E acetate in an ethanol solution with a volume fraction of 85%, stir to dissolve and let stand for 4 hours to obtain a vitamin A acetate-prolamine ethanol phase; wherein the concentration of prolamine is 10 mg / mL and the concentration of vitamin A acetate is 1 mg / mL.
[0062] 2. Dissolve sodium alginate in water, stir overnight to fully hydrate, to obtain a sodium alginate water phase; wherein the concentration of sodium alginate is 15 mg / mL.
[0063] 3. Mix 0.8 mL of the vitamin E acetate-prolamine ethanol phase obtained in step 1 and 0.2 mL of the sodium alginate water phase obtained in step 2, let stand for 0.5 hours to obtain a mixture.
[0064] 4. Disperse the mixture obtained in step 3 into 20 mL of water at a stirring rate of 900 rpm to obtain sodium alginate-encapsulated vitamin E acetate-loaded prolamine particles.
[0065] 5. Dialyze the colloidal dispersion obtained in step 4 for 24 hours to remove ethanol.
[0066] Example 7 A method of embedding vitamin acetate ester with prolamine, comprising the following steps: 1. Dissolve the prolamine and vitamin A acetate in glacial acetic acid, stir to dissolve and let stand for 4 hours to obtain a vitamin A acetate-prolamine glacial acetic acid phase; wherein the concentration of the prolamine is 20 mg / mL and the concentration of the vitamin A acetate is 2 mg / mL.
[0067] 2. Dissolve the soy lecithin in water to obtain a soy lecithin water phase with a concentration of 25 mg / mL.
[0068] 3. Mix 0.8 mL of the vitamin A acetate-prolamine glacial acetic acid phase obtained in step 1 and 0.1 mL of the soy lecithin water phase obtained in step 2, and let stand for 0.5 hours to obtain a mixture.
[0069] 4. Disperse the mixture obtained in step 3 into 20 mL of water at a stirring rate of 700 rpm to obtain soy lecithin-coated vitamin A acetate-loaded prolamine particles.
[0070] 5. Dialyze the colloidal dispersion obtained in step 4 for 24 hours to remove the acetic acid.
[0071] Example 8 A method for embedding vitamin acetate ester in prolamine, comprising the following steps: 1. Dissolve the prolamine and vitamin A acetate in an ethanol solution with a volume fraction of 85%, stir to dissolve and let stand for 4 hours to obtain a vitamin A acetate-prolamine ethanol phase; wherein the concentration of the prolamine is 20 mg / mL and the concentration of the vitamin A acetate is 2 mg / mL.
[0072] 2. Dissolve the soy lecithin in water to obtain a soy lecithin water phase with a concentration of 25 mg / mL.
[0073] 3. Mix 0.8 mL of the vitamin A acetate-prolamine ethanol phase obtained in step 2 and 0.1 mL of the soy lecithin water phase obtained in step 2, and let stand for 0.5 hours to obtain a mixture.
[0074] 4. Disperse the mixture into 20 mL of water at a stirring rate of 700 rpm to obtain soy lecithin-coated vitamin A acetate-loaded prolamine particles.
[0075] 5. Dialyze the colloidal dispersion obtained in step 4 for 24 hours to remove the ethanol.
[0076] The embedding rate is determined by the ratio of the active substance loaded in the colloid to the total active substance input. The loaded active substance is dissolved in dimethyl sulfoxide supernatant, and then determined by HPLC. The calculation formula is as follows:
[0077] The storage stability test is to place the active substance-loaded colloidal dispersion at 4℃, and to measure the concentration of the active substance in the supernatant after 1 month. The storage retention rate of the active substance is obtained by the ratio of the concentration of the active substance after 1 month of storage to the initial (freshly prepared) concentration. The calculation formula is shown as follows:
[0078] In the above examples, examples 1, 3, 5, and 7 are nanoparticles of vitamin A acetate prepared using acetic acid as the solvent, and examples 2, 4, 6, and 8 are nanoparticles of vitamin A acetate prepared using ethanol as the solvent.
[0079] Figure 3 It is shown that the nanoparticles of vitamin A acetate prepared using acetic acid as the solvent can improve the embedding rate and storage stability.
[0080] Figure 4 It is shown that the nanoparticles of vitamin A acetate prepared using acetic acid as the solvent slow down the release rate of vitamin A acetate.
[0081] Figure 5 It is shown that the particles prepared in example 1 are core-shell structure, i.e., carboxymethyl β-cyclodextrin-wrapped alcohol-soluble protein particles loaded with vitamin A acetate.
[0082] The above only describes the preferred embodiments of the present application and is not intended to limit the present application. For those skilled in the art, the present application can have various modifications and changes. Any modification, equivalent replacement, improvement, etc. within the spirit and principles of the present application shall be included in the protection scope of the present application.
Claims
1. A method of embedding a vitamin acetate with a prolamin protein, characterized by, The method comprises the following steps: dissolving the prolamine and vitamin acetate in glacial acetic acid to obtain a glacial acetic acid phase; dissolving the stabilizer in water to obtain a stabilizer phase; adding the stabilizer phase into the glacial acetic acid phase and mixing to obtain a mixed solution; the volume of the stabilizer phase is less than that of the glacial acetic acid phase; adding the mixed solution into water and mixing to form the prolamine particles loaded with the vitamin acetate and wrapped by the stabilizer; the volume of the mixed solution is less than that of the water.
2. The method of claim 1 wherein, The vitamin acetate is vitamin A acetate and / or vitamin E acetate.
3. The method of claim 1 wherein, The mass ratio of the prolamine to the vitamin acetate is 1-4:0.1-0.
5. Or, the mass fraction of the prolamine in the glacial acetic acid phase is 1-4%.
4. The method of claim 1 wherein, The stabilizer is carboxymethyl β-cyclodextrin, sodium alginate or soybean lecithin.
5. The method of claim 1 wherein, The mass concentration of the stabilizer in the stabilizer phase is 0.2-3%.
6. The method of claim 1 wherein, The volume ratio of the glacial acetic acid phase to the stabilizer phase is 20:1-5.
7. The method of claim 1 wherein, During the process of adding the mixed solution into water and mixing, the volume ratio of the mixed solution to the water is 0.8-1.0:
20.
8. The method of claim 1 wherein, After the process of adding the mixed solution into water and mixing, the glacial acetic acid is removed by dialysis.
9. The method of claim 8 wherein, The dialysis time is 24-48 hours.
10. A nanoparticle characterized by, The method is obtained by any one of claims 1-9.