Preparation method of protein-polysaccharide ternary composite nanoparticles loaded with epigallocatechin gallate
By constructing ternary composite nanoparticles of zein-gum arabic-EGCG using an ultrasound-assisted process, the instability of EGCG during processing and storage was solved, achieving efficient encapsulation and long-term stability, making it suitable for functional foods and nutritional supplements.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- NORTHEAST AGRICULTURAL UNIVERSITY
- Filing Date
- 2026-01-28
- Publication Date
- 2026-04-10
AI Technical Summary
EGCG is easily degraded during processing and storage. Existing protein-polyphenol binary composite systems have limited effect on improving its long-term stability, and the nanoparticles have poor dispersion stability. There is a lack of effective methods for constructing ternary composite systems.
Ternary composite nanoparticles were constructed using zein, gum arabic, and EGCG via an ultrasound-assisted process. Ultrasound treatment was used to enhance the protein-polysaccharide-polyphenol interactions, and combined with the traditional antisolvent precipitation method, stable nanoparticles were formed.
It significantly improves the encapsulation efficiency of EGCG, minimizes the loss of active ingredients, and the nanoparticles maintain stability under simulated heat treatment and long-term storage conditions. They also have good dispersibility and are suitable for functional foods and nutritional supplements.
Smart Images
Figure CN121817472A_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The present application belongs to the technical field of functional food and health product active ingredient delivery, and particularly relates to a preparation method of a composite nanoparticle based on natural biological macromolecules, and especially relates to a zein-arabic gum ternary composite nanoparticle for loading and protecting epigallocatechin gallate (EGCG) and a preparation method thereof. BACKGROUND
[0002] EGCG is a natural polyphenol active substance with physiological functions such as antioxidant activity, and has application value in food and functional products. However, the chemical structure of EGCG has obvious defects in practical application: it is extremely sensitive to light, heat, pH change and oxidation environment, and is prone to degradation, isomerization or polymerization during processing and storage, resulting in rapid decline in biological activity and low utilization efficiency. This stability problem seriously restricts the full play of its efficacy and commercial application. Therefore, it is crucial to develop a delivery system that can efficiently load and stably protect EGCG for a long time.
[0003] Delivery systems based on natural biological macromolecules such as proteins and polysaccharides have attracted much attention due to their good biocompatibility, biodegradability and protection ability for active ingredients. Among them, zein, as a hydrophobic plant protein, can form nanoparticles by self-assembly under specific solvent conditions, providing an embedding carrier for hydrophobic or amphiphilic active substances. However, single zein nanoparticle has poor dispersion stability in aqueous environment and is prone to aggregation and precipitation, limiting its application. Arabic gum is a common anionic polysaccharide with rich hydrophilic groups and carboxyl groups in the molecular chain, which can combine with proteins through electrostatic interaction, hydrogen bonding, etc., forming a hydrophilic protective layer on the surface of protein particles, thereby significantly improving the colloidal stability and dispersibility of the composite system in water.
[0004] Currently, the loading research of EGCG mainly focuses on protein-polyphenol binary composite systems. Although such systems can provide some protection, the long-term stability of EGCG is limited, and the physical stability problem of the protein carrier itself has not been fundamentally solved. By introducing polysaccharide components to construct a "protein-polysaccharide-polyphenol" ternary composite system, the encapsulation ability of proteins and the stability function of polysaccharides can be combined to achieve synergistic effect. However, the existing technology lacks a systematic and mature solution for how to effectively construct such a ternary composite system, especially how to accurately regulate the interaction between components to optimize the particle structure, improve the loading efficiency and stability.
[0005] Ultrasound processing technology shows unique advantages in the preparation of nanomaterials due to its cavitation effect, strong shear force and shock wave. In the application of protein system, ultrasound can induce reversible or irreversible conformational changes of protein molecules, expose more hydrophobic regions and binding sites, and thus enhance the interaction force with polysaccharides or polyphenols. This provides the possibility for strengthening the construction of ternary composite system, improving the loading efficiency of polyphenols and improving the physicochemical properties of the final nanoparticles. However, how to effectively combine and optimize the ultrasound processing with the traditional anti-solvent precipitation and other nanoparticle preparation processes to form an integrated method that can simultaneously achieve high-efficiency loading of EGCG, stable structure of composite particles and performance improvement is still a technical problem to be solved in the field. SUMMARY
[0006] The present application aims to overcome the deficiencies of the prior art, and provides a corn zein-arabic gum ternary composite nanoparticle loaded with EGCG and a preparation method thereof, in view of the problems of poor stability of EGCG, easy aggregation of single protein carrier and unclear construction process and synergistic mechanism of ternary composite system.
[0007] The object of the present application is achieved by the following technical solutions.
[0008] A preparation method of a corn zein-arabic gum ternary composite nanoparticle loaded with EGCG, comprising the following steps:
[0009] Step one, preparing an arabic gum aqueous solution: adding arabic gum into deionized water and stirring until completely dissolved to obtain a clear arabic gum aqueous solution;
[0010] Step two, preparing a corn zein-ethanol solution: dissolving corn zein in an ethanol aqueous solution with a certain concentration, first performing high-speed shearing dispersion, and then continuously stirring to ensure that the protein is fully dissolved, to obtain a uniform corn zein-ethanol solution;
[0011] Step three, preparing a corn zein-arabic gum binary composite nanoparticle dispersion: under continuous stirring, slowly adding the arabic gum aqueous solution prepared in step one into the corn zein-ethanol solution prepared in step two to fully mix the two, and then removing ethanol from the mixed solution to induce the complex assembly of corn zein and arabic gum to form a binary composite nanoparticle dispersion;
[0012] Step four, loading of EGCG: adding EGCG into the binary composite nanoparticle dispersion obtained in step three, promoting the effective loading of EGCG by means of ultrasonic treatment, to obtain a ternary composite nanoparticle dispersion loaded with EGCG;
[0013] Step five, drying: the EGCG-loaded ternary composite nanoparticles dispersion obtained in step four is subjected to drying treatment to obtain the EGCG-loaded zein-arabic gum ternary composite nanoparticles.
[0014] Further limitation, the concentration of the arabic gum in step one is 0.5%-2.0% (w / v);
[0015] Further limitation, the concentration of the zein in step two is 1.0%-3.0% (w / v);
[0016] Further limitation, the high-speed shearing condition in step two is 4000-6000 r / min for 3-5 min;
[0017] Further limitation, the ultrasonic power in step four is 300-600 W;
[0018] Further limitation, the ultrasonic time in step four is 5-15 min;
[0019] Further limitation, the mass ratio of EGCG to arabic gum in step four is 1:20-1:5.
[0020] The present application discloses the following technical effects:
[0021] First, the present application creatively combines zein, arabic gum and EGCG through an ultrasonic-assisted process to construct a ternary composite nano delivery system. This method effectively improves the dispersion stability of zein nanoparticles in aqueous phase using arabic gum, avoiding aggregation. At the same time, the interaction between protein, polysaccharide and polyphenol is strengthened through ultrasonic treatment, significantly improving the encapsulation efficiency of EGCG, which can be as high as about 86.8%, and minimizing the loss of active ingredients during preparation.
[0022] Second, the ternary composite nanoparticles prepared by the present application exhibit excellent physical and chemical stability. Under simulated heat treatment and long-term storage conditions, the particle size is uniform, there is no obvious aggregation or precipitation, an effective protective barrier is formed for the internal EGCG, the degradation rate is greatly delayed, and the core problem of instability of EGCG in processing and storage is solved.
[0023] Third, the preparation method provided by the present application has simple process steps, controllable parameters and good reproducibility. The main raw materials (zein and arabic gum) used are natural, safe and food-grade materials, which meet the development trend of green food industry, and lay a reliable technical foundation for the practical application of the composite nanoparticles in the fields of functional food, nutritional health products, etc. BRIEF DESCRIPTION OF DRAWINGS
[0024] The accompanying drawings, which constitute a part of this specification, are included to provide a further understanding of the application, and are incorporated in and constitute a part of this specification. The embodiments of the application, and their
[0025] Figure 1 Process flow diagram for preparation of EGCG loaded zein-Arabia gum ternary complex nanoparticles of the present application.
[0026] Figure 2 Encapsulation efficiency graph of ternary complex nanoparticles prepared for Examples 1-5 and Comparative Examples.
[0027] Figure 3 Scanning electron microscope (SEM) images of ternary complex nanoparticles prepared for Examples 1-5 and Comparative Examples.
[0028] Figure 4 Particle size, polydispersity index (PDI) and zeta potential change graph of ternary complex nanoparticles prepared for Example 1 at different temperature treatment and different storage time. DETAILED DESCRIPTION
[0029] Various illustrative embodiments of the present application are described in detail below. This detailed description is not intended to restrict the application unless so indicated, but to provide a description of certain aspects, features, and / or embodiments of the application.
[0030] It should be understood that the terms used herein are merely descriptive, but that the application should not be limited thereto unless so indicated. In addition, with respect to numerical ranges, it is to be understood that every numerical value between the upper and lower limits of this range is specifically and explicitly disclosed. Each and every stated range of values and intermediate values thereof, whether or not recited, are expressly included in the present application. The upper and lower limits of these smaller ranges can independently be included or excluded in the range.
[0031] Unless defined otherwise, 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. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present application, the preferred methods and materials are described. All publications mentioned herein are incorporated by reference to disclose and describe the methods and / or materials in connection with which the publications are cited. In case of conflict, the content of the present specification will control.
[0032] Many modifications and variations of the present application described in the specification are possible without departing from the scope or spirit of the application. Other implementations of the application will be apparent to those skilled in the art from consideration of the specification and practice of the application disclosed herein. The specification and examples are illustrative only.
[0033] As used herein, the terms "comprises", "comprising", "includes", "including", "has", "having", "contains", "containing", or variations thereof, are intended to be open-ended terms that mean inclusion, but not limited to, the listed material or list of materials.
[0034] It should be noted that the present application does not describe in detail the conventional means of the art, and is not the focus of the present application.
[0035] The raw materials and reagents used in the embodiments of the present application are commercially available, and the purchase route does not affect the realization of the technical effect.
[0036] Example 1
[0037] A method for preparing EGCG-loaded zein-arabic gum ternary composite nanoparticles
[0038] Step one, preparing an arabic gum aqueous solution: 1.0 g of arabic gum was weighed and added to 50.0 mL of water, and magnetically stirred until completely dissolved to prepare an arabic gum aqueous solution, with a concentration of 2.0% (w / v);
[0039] Step two, 1.0 g of zein was weighed and added to 50.0 mL of 75% ethanol aqueous solution, and first subjected to high-speed shearing dispersion (5000 r / min, 3 min), and then magnetically stirred (600 r / min, 30 min) to fully dissolve the protein, to prepare a zein-ethanol solution with a concentration of 2.0% (w / v).
[0040] Step three, preparing a zein-arabic gum binary composite nanoparticle dispersion: 20.0 mL of the arabic gum aqueous solution was added dropwise to 20.0 mL of the zein-ethanol solution, and stirring was maintained during the dropwise addition to make the mixture fully uniform. The mixture was placed in a rotary evaporator to remove the ethanol in the mixture, to obtain a zein-arabic gum binary composite nanoparticle dispersion;
[0041] Step four: 0.01 g of EGCG was weighed and added to the above zein-arabic gum binary composite nanoparticle dispersion, with a mass ratio of EGCG to arabic gum of 1:20. Subsequently, the mixed system was subjected to ultrasonic treatment (600 W, 5 min), and after the ultrasonic treatment, an ultrasonic-assisted preparation of EGCG-loaded ternary composite nanoparticle dispersion was obtained;
[0042] Step five: The EGCG-loaded zein-Arabia gum ternary composite nanoparticle dispersion prepared in step four was dried under vacuum freezing condition for 24 h to obtain the EGCG-loaded ternary composite nanoparticle.
[0043] Example 2
[0044] A method for preparing an EGCG-loaded zein-Arabia gum ternary composite nanoparticle
[0045] According to the method of steps one to five in Example 1, the EGCG-loaded ternary composite nanoparticle was prepared.
[0046] Different from Example 1, the concentration of Arabia gum was 1.5% (w / v), the concentration of zein was 3.0% (w / v), the mass ratio of EGCG to Arabia gum was 1:15, and the ultrasonic treatment condition was 400 W for 15 min.
[0047] Example 3
[0048] A method for preparing an EGCG-loaded zein-Arabia gum ternary composite nanoparticle
[0049] According to the method of steps one to five in Example 1, the EGCG-loaded ternary composite nanoparticle was prepared.
[0050] Different from Example 1, the concentration of Arabia gum was 1.5% (w / v), the concentration of zein was 1.0% (w / v), the mass ratio of EGCG to Arabia gum was 1:10, and the ultrasonic treatment condition was 400 W for 10 min.
[0051] Example 4
[0052] A method for preparing an EGCG-loaded zein-Arabia gum ternary composite nanoparticle
[0053] According to the method of steps one to five in Example 1, the EGCG-loaded ternary composite nanoparticle was prepared.
[0054] Different from Example 1, the concentration of Arabia gum was 1.0% (w / v), the concentration of zein was 2.5% (w / v), the mass ratio of EGCG to Arabia gum was 1:5, and the ultrasonic treatment condition was 600 W for 10 min.
[0055] Example 5
[0056] A method for preparing an EGCG-loaded zein-Arabia gum ternary composite nanoparticle
[0057] Ternary composite nanoparticles loaded with EGCG were prepared according to steps one through five in Example 1.
[0058] Unlike Example 1, the concentration of gum arabic was 0.5% (w / v), the concentration of zein was 2.0% (w / v), the mass ratio of EGCG to gum arabic was 1:7, and the ultrasonic treatment conditions were 300W for 15 min.
[0059] Comparative Example
[0060] A method for preparing EGCG-loaded zein-gum arabic ternary composite nanoparticles
[0061] Ternary composite nanoparticles loaded with EGCG were prepared according to steps one through five in Example 1.
[0062] The difference from Example 1 is in step four: EGCG is added to the above-mentioned zein-gum arabic binary composite nanoparticle dispersion, wherein the mass ratio of EGCG to gum arabic is 1:20, and then the mixture is stirred (2h) to obtain a ternary composite nanoparticle dispersion loaded with EGCG.
[0063] Test case
[0064] The technical effects of the present invention are demonstrated through the following experiments, which are described in detail with reference to the accompanying drawings, embodiments, and comparative examples.
[0065] Figure 2 The encapsulation efficiency of EGCG by the ternary composite nanoparticles prepared in Examples 1-5 and the comparative examples is shown. Figure 2 It was found that the encapsulation efficiency of EGCG in the ternary composite nanoparticles prepared by ultrasonic treatment was higher than 70.0%, with the highest reaching 86.8%. In contrast, the encapsulation efficiency of the ternary composite nanoparticles in the comparative example without ultrasonic treatment was only 53.4%. These results indicate that the ultrasonic-assisted zein-gum arabic ternary composite system can effectively improve the encapsulation efficiency of EGCG in nanoparticles.
[0066] Figure 3 SEM images of the ternary composite nanoparticles prepared in Examples 1-5 and Comparative Example 1. Figure 3 As shown, the ternary composite nanoparticles prepared by ultrasonic treatment exhibit a relatively regular spherical structure and a more uniform particle distribution; while the nanoparticles in the comparative example that were not ultrasonically treated showed aggregation and uneven particle size distribution. This indicates that ultrasonic treatment is beneficial to improving the structural uniformity and morphological stability of the composite nanoparticles.
[0067] Figure 4(A, B) are the particle size, PDI, and Zeta potential of the ultrasonic-assisted preparation of EGCG-loaded zein-Arabia gum ternary composite nanoparticles prepared in Example 1 after incubation at different temperatures for 30 min. As can be seen from the figure, the composite nanoparticles can still maintain good dispersibility and structural stability within a wide temperature range.
[0068] Figure 4 (C, D) are the particle size, PDI, and Zeta potential of the ultrasonic-assisted preparation of EGCG-loaded zein-Arabia gum ternary composite nanoparticles prepared in Example 1 after storage at 4°C in the dark for 0, 7, 14, 21, and 28 d. As can be seen from the figure, the composite nanoparticles have a relatively uniform size distribution after 28 d of storage, still maintain dispersibility, and have good storage stability.
Claims
1. A method for preparing a zein-arabic gum ternary composite nanoparticle loaded with epigallocatechin gallate (EGCG), comprising the following steps: Step one: arabic gum is added to water and stirred until completely dissolved to obtain an arabic gum aqueous solution; Step two: zein is added to an aqueous ethanol solution, and is first subjected to high-speed shearing dispersion and then stirring to fully dissolve the zein, thereby obtaining a zein-ethanol solution; Step three: the arabic gum aqueous solution obtained in step one is slowly added to the zein-ethanol solution obtained in step two under stirring, and then the ethanol in the mixture is removed to obtain a zein-arabic gum binary composite nanoparticle dispersion; Step four: EGCG is added to the zein-arabic gum binary composite nanoparticle dispersion prepared in step three, and then subjected to ultrasonic treatment to load the EGCG, thereby obtaining a ternary composite nanoparticle dispersion loaded with EGCG; Step five: the ternary composite nanoparticle dispersion loaded with EGCG is dried to obtain the final ternary composite nanoparticle loaded with EGCG.
2. The production method according to claim 1, characterized by, In step one, the concentration of the arabic gum is 0.5%-2.0% (w / v).
3. The preparation method according to claim 1, characterized in that, In step two, the high-speed shearing is performed at a speed of 4000-6000 r / min for 3-5 min.
4. The method of claim 1, wherein, In step two, the concentration of the zein is 1.0%-3.0% (w / v).
5. The preparation method according to claim 1, characterized in that, In step four, the mass ratio of EGCG to arabic gum is 1:20-1:
5.
6. The method of claim 1, wherein, In step four, the ultrasonic power is 300-600 W, and the ultrasonic time is 5-15 min. 7.A zein-arabic gum ternary composite nanoparticle loaded with EGCG prepared by the method according to any one of claims 1-6.