Construction method and application of super-amphiphilic molecular nanoparticles under high solid condition

Abstract

The invention discloses a preparation method and application of super-amphiphilic molecular nanoparticles under a high solid condition, and belongs to the technical field of biology. The preparation method comprises the following steps: preparing a high-concentration curcumin-cyclodextrin inclusion compound by combining a heating stirring method with ultrasound; adding the inclusion compound into a high-concentration alkalized sodium caseinate solution, and continuously adding food-grade sodium hydroxide to maintain the pH value of the system; performing ultrasonic treatment on the mixture, acidifying, performing high-pressure homogenization, centrifuging, adding maltodextrin, and performing spray drying to obtain the curcumin nanoparticles. And redissolving the obtained nanoparticles, and adding sodium erythorbate, sodium hexametaphosphate, sucrose and polyglycerol fatty acid ester to obtain the beverage. According to the powder prepared by the method, the content of super-amphiphilic molecule solids is remarkably increased, so that the cost of large-scale spray drying is reduced; the beverage obtained by redissolving has relatively good physical and chemical stability and shelf life.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology, specifically relating to a method for preparing and applying superamphilic molecular nanoparticles with high solids content. Background Technology

[0002] Traditionally, amphiphilic molecules are defined as molecules containing hydrophilic and hydrophobic regions linked by covalent bonds. Hyperamphiphilic molecules, however, are amphiphilic molecules formed through non-covalent interactions. They can connect two modules with unique functions via non-covalent interactions, enabling the construction of topological structures and functions not found in traditional amphiphilic molecules. Hyperamphiphilic molecules have opened up a new route for colloidal preparation in traditional colloid chemistry. They have been widely applied in biomedical engineering, optical material preparation, electrical material preparation, and functional food manufacturing, exhibiting extremely superior properties. However, common hyperamphiphilic molecules used in functional food manufacturing often form polymers based on the self-assembly mechanism of amphiphilic molecules under low solids conditions. Self-assembly of amphiphilic molecules under high solids conditions is often negatively affected, leading to reduced performance and efficacy of the hyperamphiphilic molecules. Furthermore, low solids conditions hinder the industrialization of hyperamphiphilic molecules; for example, they significantly increase the spray-drying production costs for functional food companies. This makes increasing the solids content a key factor for the industrialization of hyperamphiphilic molecules.

[0003] Patent document CN 120283967 A, entitled "An Amphiphilic Dietary Fiber Supramolecular Aggregate and Its Preparation and Application," discloses an amphiphilic dietary fiber supramolecular aggregate and its preparation method. This method utilizes a combination of glucomannan, arginine, a nonionic surfactant, an alkaline solution, and ethyl acetate, followed by cross-linking treatment to obtain the supramolecular aggregate. However, its application is primarily focused on the adsorption of gastrointestinal toxins, rather than in functional foods; furthermore, its low solids content leads to high industrial production costs, further limiting its application in food.

[0004] Patent document CN 108440765 A, entitled "Nano-supramolecular co-assembly of amphiphilic cyclodextrin (CD) and amphiphilic calixarene (CA), preparation method and application", discloses a supra-amphiphilic molecular assembly of cyclodextrin and calixarene with a particle size of about 140 nm and a zeta potential of -60.6 mV. It has high stability and can effectively recognize peptides. However, it has a low solids content and introduces toxic organic reagents that are harmful to the human body. It does not have a high efficiency in targeting bioactive factors, which limits its application in functional foods.

[0005] The article "Insights into the supramolecular self-assembly of sodium caseinate and calixarene" prepared a supramolecular amphiphilic molecular body using sodium caseinate and calixarene as building blocks. It achieved a low average particle size, high cellular antioxidant activity, certain flavonoid encapsulation ability, and pH responsiveness. However, the extremely low solid content of sodium caseinate and calixarene significantly increased the cost of industrial production of this supramolecular amphiphilic molecular body. Furthermore, its encapsulation ability for hydrophobic bioactive substances was limited, which restricted its application in the food industry.

[0006] The article “A supramolecular assembly of sulfobutyl ether-β-cyclodextrin and viologen calix[4]resorcinol as the master key to sustainable and eco-friendly catalyst for paraoxon hydrolysis” uses cyclodextrin derivatives and calixarenes to prepare a superamphilic molecule with a special structure. This method is green and environmentally friendly and the preparation method is simple, but its polysaccharide solid content is low, which greatly limits the application of superamphilic molecules in commercial and food industries. In addition, cyclodextrin derivatives and calixarenes are not allowed to be used in the GB2760-2024 food additive use standard, which also leads to its limited application in the food field.

[0007] Currently, there are no reports on the application of superamphiphilic molecules with high solids content. Creating superamphiphilic molecule assemblies with high solids content that can be industrialized is a key problem that technical personnel urgently need to overcome. High solids content can give superamphiphilic molecules the potential for industrial application, such as in functional foods and beverages, and has extremely high bioaccessibility, physicochemical stability and production cost. Summary of the Invention

[0008] To address the technical limitations hindering the industrial application of highly stable superamphiphilic molecules, this invention provides a method for preparing and applying superamphiphilic molecular nanoparticles under high solids conditions. The technical principle of this invention is to neutralize the protons ionized from the side chains of sodium caseinate through prolonged and continuous alkalization treatment, thereby enhancing the interaction between sodium caseinate and β-cyclodextrin. Simultaneously, β-cyclodextrin reduces the aggregation effect of sodium caseinate interactions caused by high solids. The stability of the superamphiphilic molecules under high solids conditions has been shown to be significantly higher than that under low solids conditions. Furthermore, the beverage prepared from the reconstituted solid powder obtained by spray drying exhibits superior performance.

[0009] The preparation of the superamphilic molecular nanoparticles of the present invention is achieved through the following technical solution: (1) Add a high concentration of curcumin-ethanol solution to a high concentration of β-cyclodextrin aqueous solution, and stir continuously under heating conditions until curcumin is completely encapsulated. Then, use an ultrasonic cell disruptor to sonicate to obtain solution A. (2) The pH of a high-concentration sodium caseinate aqueous solution, which is continuously stirred at high speed at room temperature until the solution is uniform, is adjusted to a strongly alkaline pH to obtain solution B. The pH is continuously adjusted with food-grade sodium hydroxide for a long time to neutralize the protons ionized from the side chain of sodium caseinate and maintained for a period of time. Curcumin powder and solution A are added to solution B. The pH is again continuously adjusted with food-grade sodium hydroxide for a long time to neutralize the protons ionized from the side chain of sodium caseinate. The system is always kept at a stable strongly alkaline pH condition. After sonication with an ultrasonic cell disruptor, the solution is stirred at high speed until the liquid color is uniform and there is no undesirable precipitate to obtain solution C. (3) Adjust the pH of solution C to neutral, stir at high speed until the liquid color is uniform, obtain the supernatant by high pressure homogenization and centrifugation, add maltodextrin with high solid content to the system with continuous high-speed stirring to obtain solution D with high solid content. (4) The D liquid was spray-dried to obtain E, a superamphilic molecular nanoparticle solid powder with high solid content; Preferably, the concentration of the β-cyclodextrin aqueous solution in step (1) is 5%-7%.

[0010] Preferably, the heating and stirring conditions in step (1) are a temperature of 60 °C-70 °C and a stirring speed of 450 rpm-1000 rpm.

[0011] Preferably, step (2) requires the use of food-grade sodium hydroxide to continuously adjust the pH over a long period of time, and the resulting alkaline solution has a pH of 11.8-12.0, more preferably 12.

[0012] Preferably, the stirring speed of the solution in step (2) is 1500 rpm-2000 rpm.

[0013] Preferably, the concentration of the sodium caseinate aqueous solution in step (2) is 9%-13%.

[0014] Preferably, the alkaline treatment time of the pure sodium caseinate aqueous solution in step (2) is 0 min-30 min, and more preferably 30 min.

[0015] Preferably, the alkaline treatment time of the sodium caseinate aqueous solution after adding the inclusion complex in step (2) is 60 min-90 min, and more preferably 60 min.

[0016] Preferably, the stirring speed of the solution in step (3) is 1500 rpm-2000 rpm.

[0017] Preferably, the high-pressure homogenization pressure in step (3) is 0 bar to 500 bar.

[0018] Preferably, the amount of maltodextrin solids added in step (3) is 5%-10% of the total mass of the solution.

[0019] This invention also provides an application of the aforementioned high-solids-content superamphilic molecular nanoparticles in the field of beverage preparation.

[0020] Furthermore, the present invention provides a method for preparing a spray-dried superamphilic molecular-based neutral beverage with high solids content. Based on the above step (4), step (5) is added: after redissolving powder E in warm water, sodium isoascorbate, sodium hexametaphosphate, sucrose, and polyglycerol fatty acid ester are added and stirred to obtain a spray-dried superamphilic molecular-based neutral beverage with high stability and high solids content.

[0021] Preferably, the mass ratio of sodium isoascorbate to modular superamophilic molecular nanoparticle powder with high solids content in step (5) is (0~3):3.

[0022] Preferably, the mass ratio of sodium hexametaphosphate to modular superamphiphilic molecular nanoparticle powder with high solids content in step (5) is (0~3):3.

[0023] Preferably, the mass ratio of the polyglycerol fatty acid ester to the modular superamphiphilic molecular nanoparticle powder with high solids content in step (5) is (0~5):3.

[0024] Preferably, the mass ratio of sucrose to modular superamophilic molecular nanoparticle powder with high solids content in step (5) is (70~90):3.

[0025] This invention encapsulates a high concentration of β-cyclodextrin inclusion complex in a high concentration of sodium caseinate to form a high-solids superamphilic molecular system, thereby reducing the self-aggregation effect of sodium caseinate and improving the stability of the system.

[0026] This invention is based on a pH-driven method to enhance the water solubility of sodium caseinate under alkaline conditions. Prolonged and continuous alkalization neutralizes the protons continuously ionized from the side chains of sodium caseinate. After continuous alkalization, a β-cyclodextrin-curcumin inclusion complex with high solids content is introduced as a building block. Under ultrasonic disruption, the flexible open chain of sodium caseinate forms numerous hydrogen bonds with the rigid macrocyclic β-cyclodextrin, promoting the formation of hyperamphiphilic molecular aggregates through hydrophobic interactions. Due to the effect of prolonged and continuous alkalization, the interaction between sodium caseinate molecules is lower due to high solids content, further enhancing the interaction between sodium caseinate and β-cyclodextrin. Under high solids conditions, there is a significant inhibitory effect of β-cyclodextrin on protein aggregation, making the hyperamphiphilic molecular aggregates with high solids content more stable than those with low solids content, specifically manifested in significantly improved physicochemical stability, such as photothermal stability. This method not only improves the stability of the superamphiphilic molecule, but also greatly reduces the industrialization cost of the superamphiphilic molecule, broadens the application range of the superamphiphilic molecule, and solves the production problem of the superamphiphilic molecule being difficult to industrialize.

[0027] Compared with existing superamphiphilic molecular encapsulation technologies, the beneficial effects and advantages of this invention are as follows: (1) The superamphilic molecular spray drying precursor solution prepared by the present invention has a high solid content, which effectively reduces the spray drying cost of superamphilic molecules and enhances the application potential for industrialization. (2) The high solids content superamphilic molecules prepared by the present invention have higher physicochemical stability than conventional low solids content superamphilic molecules. Under alkaline treatment conditions, they can maintain high curcumin stability to improve curcumin encapsulation rate and reduce curcumin loss rate, and can be efficiently applied to the production of functional foods. (3) The superamphilic molecular-based beverage with high solids content prepared by the present invention has a long shelf life. After heating at 95°C for 120 min, only about 10% of curcumin decomposes. After being directly irradiated at close range with D65 simulated natural light for 120 h, only about 10% of curcumin decomposes. After 2 h of physical destructive testing, the TSI index is less than 0.5, which shows that it has extremely high physicochemical stability, which means that it has high commercial value and commercial competitiveness as a functional food. (4) The digestibility of the high solids content superamphilic molecular base beverage prepared by the present invention is about 10 times higher than that of free curcumin, and its bioavailability is about 10-13 times higher than that of free curcumin, and it has extremely high potential for functional food applications.

[0028] (5) The raw materials used in this invention all comply with the national food safety standard GB2760-2024. The manufacturing process is simple and low-cost, making it suitable for large-scale industrial production. Attached Figure Description

[0029] Figure 1 shows the relationship between heating and stirring time and curcumin loss rate in the preparation of curcumin-β-cyclodextrin inclusion complex, pH of sodium caseinate treatment with alkali with curcumin encapsulation rate and loss rate, protein treatment time with curcumin encapsulation rate, and curcumin treatment time with curcumin encapsulation rate and loss rate.

[0030] Figure 2 shows the front and top views of the superamphilic molecular nanoparticle powders prepared in Examples 1-2 and Comparative Examples 1-3 and their water resolution.

[0031] Figure 3 shows the (A) photostability, (B) thermal stability, and (C) physical stability of the superamphilic molecular nanoparticle powders prepared in Examples 1-2 and Comparative Examples 1-3 after reconstitution.

[0032] Figure 4 shows the differential scanning calorimetry curves for Examples 1-2.

[0033] Figure 5 shows the (A) Fourier transform infrared spectrum, (B) ultraviolet spectrum, and (C) Raman spectrum of Examples 1-2.

[0034] Figure 6 shows the atomic force microscopy test results of Examples 1-2.

[0035] Figure 7 shows the transmission electron microscope test results of Examples 1-2.

[0036] Figure 8 shows the (A) light stability, (B) thermal stability, and (C) physical stability of Comparative Example 1 under different concentrations of sodium hexametaphosphate.

[0037] Figure 9 shows the (A) photostability, (B) thermal stability, and (C) physical stability of Comparative Example 1 under different concentrations of sodium isoascorbate.

[0038] Figure 10 shows the (A) light stability, (B) thermal stability, and (C) physical stability of Comparative Example 1 under different concentrations of polyglycerol fatty acid esters.

[0039] Figure 11 shows the electronic tongue test of beverages after the superamphiphilic molecular nanoparticle powder prepared in Examples 1-2 was reconstituted.

[0040] Figure 12 shows the in vitro digestibility test of the beverage after the superamphiphilic molecular nanoparticle powder prepared in Examples 1-2 was reconstituted. Detailed Implementation

[0041] The following examples further illustrate the specific implementation of the present invention, but the implementation and protection scope of the present invention are not limited thereto.

[0042] The experimental methods used in this embodiment are all conventional methods; the materials and reagents used (sodium caseinate, β-cyclodextrin, maltodextrin, curcumin, etc.) are commercially available unless otherwise specified.

[0043] Example 1: Prepare a 6.6% aqueous solution of β-cyclodextrin. Stir continuously at 900 rpm for 60 minutes at 60 °C. Add 2.14 g of curcumin dissolved in 225 ml of ethanol dropwise to 100 ml of the β-cyclodextrin aqueous solution while stirring. Stir the mixture at 500 rpm for 16 hours at 60 °C with the lid open to evaporate the ethanol. Sonicate the mixture to obtain a cyclodextrin inclusion complex suspension. Stir 100 ml of 11% sodium caseinate aqueous solution continuously at 2000 rpm for 10 hours at room temperature. After hydration for 6 hours, adjust the pH to 12 using food-grade hydroxide solution. Stir continuously at 2000 rpm for 30 minutes. Then, add 4.45 g of curcumin powder and the β-cyclodextrin inclusion complex suspension to the sodium caseinate. Stir continuously at 2000 rpm for 60 minutes. Sonicate the mixture for 10 minutes using an ultrasonic cell disruptor. The mixture was acidified to pH 7 with citric acid and stirred at 2000 rpm for 6 h until no obvious floating matter was found. The mixture was centrifuged (10000 rpm, 10 min) to obtain the supernatant. The centrifuged composite system solution was gradually mixed with 10 g of maltodextrin under stirring (2000 rpm) to obtain the composite system for spray drying. The inlet air temperature of the spray drying was 150 °C, the outlet air temperature was 200 °C, and the feed rate was 7 rpm to obtain uniform ultraamphiphilic curcumin nanoparticle powder.

[0044] Comparative Example 1: The difference between Comparative Example 1 and Example 1 is that Comparative Example 1 is prepared with low solids content of superamphilic molecular nanoparticles. Specifically, the concentration of β-cyclodextrin solution is 0.66%, the concentration of sodium caseinate solution is 1.09%, maltodextrin accounts for 0.13% of the total mass of the solution, and the total curcumin accounts for 0.24% of the total mass of the solution. The remaining operation process and parameter settings are the same as in Example 1.

[0045] Example 2: Following the process scheme of Example 1, and with the addition of high-pressure homogenization treatment, experiments were conducted to prepare high-solids, amphiphilic molecular-based nanoparticles: Prepare a 6.6% aqueous solution of β-cyclodextrin. Stir continuously at 900 rpm for 60 minutes at 60 °C. Add 2.14 g of curcumin dissolved in 225 ml of ethanol dropwise to 100 ml of the β-cyclodextrin aqueous solution while stirring. Stir the mixture at 500 rpm for 16 hours at 60 °C with the lid open to evaporate the ethanol. Sonicate the mixture to obtain a cyclodextrin inclusion complex suspension. Stir 100 ml of 11% sodium caseinate aqueous solution continuously at 2000 rpm for 10 hours at room temperature. After hydration for 6 hours, adjust the pH to 12 using food-grade hydroxide solution. Stir continuously at 2000 rpm for 30 minutes. Then, add 4.45 g of curcumin powder and the β-cyclodextrin inclusion complex suspension to the sodium caseinate. Stir continuously at 2000 rpm for 60 minutes. Sonicate the mixture for 10 minutes using an ultrasonic cell disruptor. The mixture was acidified to pH 7 with citric acid and stirred at 2000 rpm for 6 h until no obvious floating matter was found. High-pressure homogenization was performed at 500 bar to obtain a homogeneous and stable solution. The solution was centrifuged (10000 rpm, 10 min) to obtain the supernatant. The centrifuged composite system solution was gradually mixed with 10 g of maltodextrin under stirring (2000 rpm) to obtain a composite system for spray drying. The inlet air temperature of the spray drying was 150°C, the outlet air temperature was 200°C, and the feed rate was 7 rpm to obtain uniform ultraamphiphilic curcumin nanoparticle powder.

[0046] Comparative Example 2: The difference between Comparative Example 2 and Examples 1 and 2 is that no superamphilic molecular nanoparticles were constructed. Only sodium caseinate was used to encapsulate curcumin. Specifically, a sodium caseinate aqueous solution (2%) was prepared, stirred at 2000 rpm for 1 h, hydrated for 6 h, and the pH was adjusted to 12 using food-grade NaOH solution. 20 mg of curcumin powder was added, stirred at 2000 rpm for 90 min, and the mixture was acidified to pH 7 with citric acid. The mixture was then centrifuged (10000 rpm, 10 min) and spray-dried with the following parameters: inlet air temperature 150 °C, outlet air temperature 200 °C, and feed rate 7 rpm.

[0047] Comparative Example 3: The difference between Comparative Example 3 and Examples 1 and 2 is that no superamphilic molecular nanoparticles were constructed. Only β-cyclodextrin was used to encapsulate curcumin. Specifically, a β-cyclodextrin aqueous solution (1.2%) was prepared and stirred at 500 rpm for 1 h. A curcumin-ethanol solution containing 20 mg of curcumin was added and stirred at 900 rpm for 16 h. The mixture was then centrifuged (10,000 rpm, 10 min) and spray-dried. The parameters were set as follows: inlet air temperature was 150 °C, outlet air temperature was 200 °C, and feed rate was 7 rpm.

[0048] Experimental Example 1: Optimization of Construction Conditions for High Solids Content Superamphilic Molecular Nanoparticles Using Examples 1-2 prepared under different construction conditions as test samples, the curcumin encapsulation efficiency and curcumin loss rate were tested, as shown in Figure 1. Under different heating and stirring times (0-24 h) for the curcumin-β-cyclodextrin inclusion complex, the curcumin loss rate increased significantly at heating times of 20 h and 24 h compared to heating times of less than 20 h. Therefore, the preferred heating time for curcumin-β-cyclodextrin is 16 h. Under different alkali treatment pH conditions (9-12), low pH resulted in low curcumin encapsulation efficiency. As pH increased, the curcumin encapsulation efficiency gradually increased, while the curcumin loss rate also increased slowly with increasing pH. Considering all factors, pH 12 was the preferred pH condition. Under different alkali treatment times (0-60 min) for pure sodium caseinate, the curcumin encapsulation efficiency was less than 80% at 0-15 min, but greater than 85% at 30-60 min, and at 30 min... After 30 min, there was no significant increase in curcumin encapsulation rate as the stirring time increased. The optimal time for alkali treatment of pure sodium caseinate was 30 min. Under different alkali treatment time conditions (0-90 min), the curcumin loss rate increased significantly with the extension of time. The curcumin encapsulation rate first increased and then decreased rapidly. Considering all factors, the optimal time for alkali treatment of curcumin was 30-60 min.

[0049] Experimental Example 2: Morphology and Reconstitution of Spray-Dried Powder from Superamphilic Molecular Nanoparticles According to the methods of Examples 1-2 and Comparative Examples 1-3, ultraamphiphilic molecular nanoparticles were prepared by spray drying. The appearance was observed as shown in Figure 2. Compared with Comparative Examples 1-3, the color of Sample 1-2 was more orange and the color was uniform. After reconstitution, the sample solutions of Examples 1-2 and Comparative Examples 1-3 did not have any undesirable precipitates, which proved that the samples had good reconstitution properties.

[0050] Experimental Example 3: Spray-dried powder parameters of superamphilic molecular nanoparticles According to the methods of Examples 1-2 and Comparative Examples 1-3, ultraamphiphilic molecular nanoparticles were prepared by spray drying, and the test parameters are as follows. Table 1. Spray-dried powder parameters of nanoparticles in Examples 1-2 and Comparative Examples 1-3

[0051] The results indicate that the hydrated particle size of the high-solids-content superamphiphilic nanoparticles spray-dried is slightly higher than that of the low-solids-content superamphiphilic nanoparticles spray-dried. High-pressure homogenization leads to a reduction in the particle size of the superamphiphilic nanoparticles, as well as a decrease in the absolute value of the Zeta potential, moisture absorption, Karl Fischer index, and Hausner ratio. This suggests that high-pressure homogenization can improve the performance of the superamphiphilic nanoparticle powder to some extent. Compared to Comparative Example 1, the curcumin loading rate of Examples 1-2 is significantly increased. Compared to Comparative Example 2, the hydrated particle size of Examples 1-2 is much smaller than that of Comparative Example 2. This is because the supramolecular interaction between β-cyclodextrin and sodium caseinate results in β-cyclodextrin inhibiting the aggregation of sodium caseinate. Compared to Comparative Example 3, the flow and cohesive properties of the superamphiphilic nanoparticles spray-dried in Examples 1-2 are superior to those in Comparative Example 3. In summary, the high-solids-content superamphiphilic nanoparticles spray-dried powder exhibits better overall performance, especially Example 2, which demonstrates superior powder characteristics.

[0052] Experimental Example 4: Physicochemical Stability of Spray-Dried Powder of Superamphiphilic Molecular Nanoparticles As shown in Figure 3, during the 0-120 h photodestructive test, Examples 1-2 exhibited excellent curcumin photostability. After 120 h of photodestructive treatment, Examples 1 and 2 showed curcumin retention rates of 67.9% and 77.8%, respectively, demonstrating superior photostability compared to Comparative Examples 1-2. During the 0-120 min high-temperature destructive test, Examples 1-2 exhibited excellent curcumin thermal stability. After 120 min of thermal destructive treatment, Examples 1 and 2 showed curcumin retention rates of 81.3% and 93.6%, respectively, demonstrating superior photostability compared to Comparative Examples 1-3. After the 7200 s physical destructive test, compared to Comparative Examples 1-3, Example 1 had a higher TSI index, while Example 2 had a lower TSI index. This indicates that high-pressure homogenization leads to increased physical stability of the superamphilic molecular system, and the samples of Example 2 and Comparative Example 2 exhibited higher physical stability.

[0053] Experimental Example 5: Differential Scanning Calorimetry Analysis As shown in Figure 4, free curcumin exhibits a sharp peak near 179°C, while Examples 1-2 only show broad peaks of protein denaturation near 100-125°C. This indicates that the superamphilic molecular nanoparticles of Examples 1-2 successfully encapsulated curcumin, binding the vibration of curcumin molecules and transforming them into an amorphous state, resulting in the disappearance of the sharp peak.

[0054] Experimental Example 6: Spectral Analysis As shown in Figure 5, the Fourier transform infrared (FTIR) spectrum reveals numerous sharp peaks in free curcumin, indicating molecular vibrations within the curcumin crystals. After encapsulation with the amphiphilic molecules from Examples 1-2, the peaks of free curcumin disappear, indicating that curcumin is encapsulated within the amphiphilic molecules, restricting its molecular vibrations. This suggests that the interaction between curcumin and the amphiphilic molecular wall material is not a simple physical mixture. Following high-pressure homogenization, some characteristic protein peaks from Example 2 shift, indicating changes in the protein's secondary structure during homogenization, leading to structural changes in the amphiphilic molecules. The UV-Vis spectroscopy shows that after encapsulation with the amphiphilic molecules, the characteristic absorption peak of curcumin at 427 nm blue-shifts to 409 nm, further demonstrating that curcumin is bound to the amphiphilic molecules through hydrogen bonding rather than simple physical mixing. The Raman spectroscopy indicates that the characteristic peak of curcumin at 1626 cm⁻¹... -1 Shifting to higher wavenumbers to 1634 cm -1 This is due to the reduction in the conjugation of C=C and C=O double bonds, which proves that the phenolic hydroxyl groups of curcumin interact with the hydroxyl groups of β-cyclodextrin through hydrogen bonds, meaning that curcumin is successfully included in β-cyclodextrin. At 1319 cm⁻¹ -1 The characteristic peak of curcumin at [location], namely the stretching vibration of the CCH group in the middle of the curcumin molecule, shifted to 1308 cm⁻¹ in Examples 1-2. -1 and 1307 cm -1 The hydrophobic cavity of β-cyclodextrin does not affect the state of the central CCH group, which suggests the molecular interaction between the hydrophobic residues of sodium caseinate and the CCH group in the middle of the curcumin molecule. Spectroscopic analysis comprehensively proves the successful formation of the superamphilic molecular structure under high solids conditions and the successful encapsulation of curcumin by superamphilic molecular nanoparticles.

[0055] Experiment 7 Microscopic observation Using samples from Examples 1-2 as experimental samples, as shown in Figure 6 using atomic force microscopy, the samples from Examples 1-2 exhibited a uniformly dispersed spherical structure with some aggregation, demonstrating that the supra-amphiphilic molecules formed spherical nanoparticles. Using samples from Examples 1-2 as experimental samples, as shown in Figure 7, the system of Examples 1-2 showed a non-uniformly sized spherical structure with small spheres embedded in larger spheres, a typical spherical-encased supra-amphiphilic molecular structure, where the smaller β-cyclodextrin portion entered the sodium caseinate particles. This demonstrates that even with high solids content, the sodium caseinate and β-cyclodextrin inclusion complex can still generate good supramolecular interactions to form supra-amphiphilic curcumin nanoparticles.

[0056] Experimental Example 8: Optimization of the effect of sodium hexametaphosphate concentration on the stability of reconstituted solutions of superamphiphilic molecular nanoparticle powders As shown in Figure 8, curcumin gradually decomposes with increasing light exposure time. The solution of the superamphiphilic nanoparticles without sodium hexametaphosphate shows the fastest decomposition. After adding sodium hexametaphosphate, the photostability of the superamphiphilic molecules increases significantly, but there is no significant increase in photostability with increasing concentration. Sodium hexametaphosphate can reduce the decomposition of curcumin during high-temperature treatment to a lesser extent. Regarding physical stability, the TSI index is high without sodium hexametaphosphate. When a small amount of sodium hexametaphosphate is added (0.1%-0.3%), the TSI index decreases significantly, while when a large amount of sodium hexametaphosphate is added (0.3%-0.5%), the TSI index rises again. Based on the three stability tests, a concentration of 0.1% sodium hexametaphosphate is preferred as the stability enhancer.

[0057] Experimental Example 9: Optimization of the effect of sodium isoascorbate concentration on the stability of reconstituted solutions of superamphiphilic molecular nanoparticle powders As shown in Figure 9, with the increase of light exposure time, the photostability of the superamphiphilic molecules increased significantly by about 20% upon the addition of sodium isoascorbate, indicating that sodium isoascorbate improves the photostability of the reconstituted solution of superamphiphilic molecular nanoparticles, but there was no significant increase in photostability with increasing concentration. Similarly, sodium isoascorbate can significantly reduce the decomposition of curcumin during high-temperature treatment, and the thermal stability of the superamphiphilic molecules increased significantly by about 10%. In addition, after a 7200 s physical destructive test, the TSI index decreased significantly upon the addition of sodium isoascorbate, proving that the antioxidant sodium isoascorbate significantly improves physical stability. Based on the three stability tests, a concentration of 0.1% sodium isoascorbate is preferred as the stability enhancer.

[0058] Experimental Example 10: Optimization of the effect of polyglyceric acid fatty acid ester concentration on the stability of reconstituted solutions of superamphiphilic molecular nanoparticle powders. As shown in Figure 10, with increasing light exposure time, the addition of polyglycerol fatty acid ester significantly increases the photostability of the superamphiphilic molecules by approximately 15%-20%, demonstrating that polyglycerol fatty acid ester enhances the photostability of superamphiphilic molecules. Maximum photostability is achieved with the addition of 0.75% polyglycerol fatty acid ester. Regarding thermal stability, polyglycerol fatty acid ester can reduce curcumin decomposition during high-temperature treatment to a certain extent. The addition of 0.75% polyglycerol fatty acid ester significantly increases the thermal stability of the superamphiphilic molecules by approximately 10%. Polyglycerol fatty acid ester can significantly reduce the TSI index, demonstrating its significant improvement in physical stability; however, this improvement is not correlated with concentration. Based on the three stability tests, a concentration of 0.5%-1% polyglycerol fatty acid ester is preferred as the stability enhancer.

[0059] Experiment Example 11: Beverage Application 1200 mg of powder was dissolved in 200 ml of warm water, sodium hexametaphosphate (400 mg), sodium isoascorbate (400 mg), sucrose (32 g), and polyglycerol fatty acid ester solution (1%) were added. The mixture was stirred to obtain the beverage. As shown in Figure 11, the flavor of Examples 1-2 was tested by electronic tongue and it was found that the taste of Examples 1-2 was similar. The sodium caseinate-cyclodextrin superamphiphilic molecular system can effectively reduce the bitterness and astringency caused by the cyclodextrin system, thereby improving the commercial value of the beverage.

[0060] Using free curcumin as the experimental sample in Examples 1-2, the bioavailability of curcumin in the reconstituted beverage made from ultraamphiphilic molecular nanoparticles was tested by in vitro digestion (the test method can refer to the in vitro digestion method in Jin. Z. et al. [J]. Food Chemistry, 2025, 488: 144859). The bioavailability of the digested curcumin in Examples 1-2 was about 10-13 times higher than that of the digested free curcumin. Specifically, the ultraamphiphilic molecular nanoparticles significantly improved the digestive stability of curcumin during in vitro digestion, by about 10 times.

[0061] Experiment Example 12: Beverage Shelf Life Test 1200 mg of the powder from Examples 1-2 was dissolved in 200 ml of warm water. Sodium hexametaphosphate (400 mg), sodium isoascorbate (400 mg), sucrose (32 g), and polyglycerol fatty acid ester solution (1%) were added. The mixture was stirred to obtain a beverage. The beverage was placed at 4°C, 25°C, and 45°C for 60 days, and samples were taken every 15 days to test the average particle size, zeta potential, color parameters, curcumin retention rate, pH, and centrifugal stability of the beverage for kinetic and thermodynamic parameter calculations. The shelf life of the beverage was calculated using the Arrhenius equation.

[0062] Table 2 Storage kinetics test parameters of Example 1

[0063] Table 3 Storage kinetics test parameters of Example 2

[0064] Choose a kinetic equation with a good fit and substitute it into the Arrhenius formula for thermodynamic calculations.

[0065] Table 4. Arrhenius equation parameters for Examples 1-2

[0066] By screening the Arrhenius equations corresponding to each indicator, it was found that the Arrhenius equations for centrifugal stability in Examples 1-2 had the highest fitting degree and were common indicators for both samples. Therefore, centrifugal stability was selected as the key factor for shelf-life prediction. Combining the Arrhenius equations, activation energy, and kinetic equations, the shelf-life equation and predicted shelf-life are as follows: Table 5 Shelf life equations for Examples 1-2 and shelf life at various temperatures

[0067] In the shelf life equation, y represents the shelf life instability point of the centrifugal stability parameter. According to literature and experimental tests, the shelf life instability point of centrifugal stability is about 1.42-1.47. Here, the average value of this range, 1.445, is taken as the shelf life instability point. It can be calculated that the shelf life of Examples 1-2 at 4 ℃ can reach 178 days and 410 days, respectively, that is, the beverage after reconstitution of the superamphiphilic molecular nanoparticles under the high solids condition has a good shelf life.

[0068] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, and all such modifications or substitutions should be covered within the scope of the claims of the present invention.

Claims

1. A method for preparing superamphilic molecular nanoparticles under high solids conditions, characterized in that, The high-solids β-cyclodextrin-curcumin inclusion complex was ultrasonically mixed with high-concentration sodium caseinate after alkali treatment and stirred. After acidification and adjustment back to neutrality, the system was induced to form superamphilic molecular nanoparticles. Stable liquid water-soluble curcumin nanoparticles were formed by high-pressure homogenization. Free curcumin was removed by centrifugation, and maltodextrin was added. After stirring and spray drying, powder was obtained.

2. The method for preparing superamphilic molecular nanoparticles with high solids content according to claim 1, characterized in that, Specifically, it includes the following steps. (1) Add a high concentration of curcumin-ethanol solution to a high concentration of β-cyclodextrin aqueous solution, and stir continuously under heating conditions until curcumin is completely encapsulated. Then, sonicate the solution using an ultrasonic cell disruptor to obtain solution A. (2) The pH of a high-concentration sodium caseinate aqueous solution, which is continuously stirred at high speed at room temperature until the solution is uniform, is adjusted to a strong alkaline state to obtain solution B. The pH of solution B is continuously adjusted by food-grade sodium hydroxide for a long time to neutralize the protons ionized from the side chain of sodium caseinate and maintained for a period of time. Curcumin powder and solution A are added to solution B. The pH is again continuously adjusted by food-grade sodium hydroxide for a long time to neutralize the protons ionized from the side chain of sodium caseinate. The system is always kept at a stable strong alkaline pH condition. After sonication by an ultrasonic cell disruptor, the solution is stirred at high speed until the liquid color is uniform and there is no undesirable precipitate to obtain solution C. (3) Adjust the pH of solution C to neutral, stir at high speed until the liquid color is uniform, obtain the supernatant by high pressure homogenization and centrifugation, add maltodextrin with high solid content to the system of continuous high-speed stirring to obtain solution D with high solid content. (4) D liquid is spray dried to obtain superamphilic molecular nanoparticle solid powder E with high solid content.

3. The method for preparing superamphilic molecular nanoparticles with high solids content according to claim 2, characterized in that, The concentration of the β-cyclodextrin aqueous solution in step (1) is 5%-7%; the heating and stirring conditions in step (1) are a temperature of 60 °C-70 °C and a stirring speed of 450 rpm-1000 rpm.

4. The method for preparing superamphilic molecular nanoparticles under high solids conditions according to claim 2, characterized in that, Step (2) requires the use of food-grade sodium hydroxide to continuously adjust the pH over a long period of time, resulting in an alkaline solution with a pH of 11.8-12.

0.

5. The method for preparing superamphilic molecular nanoparticles with high solids content according to claim 2, characterized in that, The solution stirring speed in step (2) is 1500 rpm-2000 rpm.

6. The method for preparing superamphilic molecular nanoparticles under high solids conditions according to claim 2, characterized in that, The concentration of sodium caseinate aqueous solution in step (2) is 9%-13%; the alkaline treatment time of pure sodium caseinate aqueous solution is 0 min-30 min; the alkaline treatment time of sodium caseinate aqueous solution after adding inclusion complex in step (2) is 60 min-90 min.

7. The method for preparing superamphilic molecular nanoparticles with high solids content according to claim 2, characterized in that, The solution stirring speed in step (3) is 1500 rpm-2000 rpm; the high pressure homogenization pressure in step (3) is 0 bar-500 bar; the amount of maltodextrin solids added in step (3) is 5%-10% of the total mass of the solution.

8. The superamphilic molecular nanoparticles with high solids content prepared according to any one of the preceding claims.

9. The application of the high solids-content superamphilic molecular nanoparticles as described in claim 8 in the field of beverage preparation.

10. A method for preparing a spray-dried, ultra-amphiphilic molecular-based neutral beverage with high solids content, characterized in that, Based on step (4) of claim 2, powder E is redissolved in warm water and then sodium isoascorbate, sodium hexametaphosphate, sucrose, and polyglycerol fatty acid ester are added and stirred to obtain a spray-dried superamphilic molecular-based neutral beverage with high stability and high solids content. The mass ratio of sodium isoascorbate to modular superamphiphilic molecular nanoparticle powder with high solids content is (0~3):3; the mass ratio of sodium hexametaphosphate to modular superamphiphilic molecular nanoparticle powder with high solids content is (0~3):3; the mass ratio of polyglycerol fatty acid ester to modular superamphiphilic molecular nanoparticle powder with high solids content is (0~5):3; and the mass ratio of sucrose to modular superamphiphilic molecular nanoparticle powder with high solids content is (70~90):3.

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