A nutritional composition for improving concentration, its preparation method and application

Abstract

This invention belongs to the field of nutritional composition technology, specifically disclosing a nutritional composition for improving concentration, its preparation method, and its application. The composition comprises the following components: 8-15 parts of a fermentation complex, 6-12 parts of a directed enzymatic hydrolysis complex, 1-3 parts of phosphatidylserine, 1-2 parts of N-acetylneuraminic acid, and 2-5 parts of walnut peptide. The preparation method includes: encapsulating a suspension of surface-functionalized nanoparticles to obtain microencapsulated nanoparticle powder, mixing it with walnut peptide and food-grade excipients, adding water, homogenizing, vacuum degassing, and sterilizing to obtain the finished nutritional composition for improving concentration. This application provides a nutritional composition for improving concentration with high active ingredient content, good stability, and high bioavailability by extracting and enriching, fermenting and transforming, gently enzymatically hydrolyzing to retain glycosylation sites, sequentially adding and co-assembling core-shell nanoparticles, and encapsulating with sodium caseinate microcapsules to achieve gastric acid protection and intestinal sustained release.

Description

Technical Field

[0001] This invention belongs to the field of nutritional composition technology, specifically relating to a nutritional composition for improving concentration, its preparation method, and its application. Background Technology

[0002] Nutritional compositions designed to improve focus are dietary supplements that exist in the form of functional foods or health products. They are primarily targeted at adolescents and individuals with high cognitive loads, aiming to improve concentration, extend focus duration, and enhance learning efficiency. These products typically combine neurotrophic factors such as N-acetylneuraminic acid, phosphatidylserine, and milk fat globule membrane protein with medicinal and edible ingredients like jujube seed, poria cocos, and lily bulb. After oral administration and absorption, they act on the central nervous system.

[0003] In practical applications, existing nutritional compositions for improving concentration have several shortcomings. First, most existing products simply mix the components physically, allowing the food-grade ingredients and neurotrophic factors to be absorbed independently in the gastrointestinal tract. This lack of a synergistic delivery carrier prevents the formation of a combination of absorption time and target site, resulting in unsatisfactory overall bioavailability.

[0004] Secondly, many existing products using jujube seed, poria cocos, and lily bulb, which are both medicinal and edible ingredients, employ water extraction. While this process is simple, the low solubility of fat-soluble active ingredients such as jujube seed saponins, poria cocos triterpenoids, and lily bulb steroidal saponins in water results in significant residues remaining in the residue, leading to raw material waste and insufficient product efficacy. Some products use alcohol extraction instead of water extraction, improving the extraction rate of fat-soluble active ingredients. However, alcohol extracts still contain large amounts of high-molecular-weight polysaccharides and proteins with high molecular weights and unsatisfactory lipid-water partition coefficients, limiting their efficiency in crossing intestinal epithelial cell biomembranes.

[0005] Furthermore, during conventional processing, milk fat globule membrane protein powder undergoes high-temperature spray drying or heat treatment, which disrupts its natural phospholipid bilayer structure, denatures and inactivates the membrane protein, and breaks down the glycosylation sites on the glycoprotein surface. This not only results in the loss of its natural ability to encapsulate active ingredients but also in its ability to recognize and bind to intestinal epithelial cells. Other technologies use enzymatic hydrolysis of milk fat globule membrane protein to prepare bioactive peptides, but the hydrolysis conditions are not designed to protect the glycosylation sites, leading to the destruction of glycoprotein domains and the loss of its function as a targeted delivery carrier.

[0006] Furthermore, phosphatidylserine, an important phospholipid component of nerve cell membranes, is easily degraded by gastric acid and digestive enzymes when it enters the gastrointestinal tract after oral administration, and is also oxidized when exposed to oxygen, resulting in a low proportion of the effective component actually entering the bloodstream. Existing methods directly add N-acetylneuraminic acid as an ingredient to the composition, but it is unevenly dispersed in a liquid environment and does not form a chemical bond with the carrier. After entering the body, it is rapidly metabolized like ordinary monosaccharides, making it difficult to exert a sustained supply effect on ganglioside synthesis.

[0007] To address the aforementioned shortcomings, existing technologies have proposed several improvement schemes. Some studies have used sodium caseinate or gelatin to microencapsulate phosphatidylserine, which to some extent slows down gastric acid degradation. However, the lack of molecular structural similarity between the microcapsule wall material and phosphatidylserine leads to unstable encapsulation efficiency, and the microcapsule particle size is mostly over several hundred micrometers, making it difficult for them to enter the bloodstream through endocytosis by intestinal epithelial cells. Other technologies involve enzymatic hydrolysis of milk fat globule membrane proteins, but these often employ alkaline proteases or trypsin under high pH conditions. These conditions severely damage the N-glycosylation domains of glycoproteins, resulting in insufficient density of retained glycosylation sites and an inability to provide enough binding sites for N-acetylneuraminic acid.

[0008] Therefore, there is a need to design a nutritional composition that can improve concentration, as well as its preparation method and application. Summary of the Invention

[0009] To overcome the shortcomings of the prior art, a nutritional composition for improving concentration, its preparation method, and its application are provided.

[0010] To achieve the above objectives, the present invention provides the following technical solution: A nutritional composition for improving concentration, comprising, by weight, the following components: 8-15 parts of fermentation complex, 6-12 parts of targeted enzymatic hydrolysis complex, 1-3 parts of phosphatidylserine, 1-2 parts of N-acetylneuraminic acid, and 2-5 parts of walnut peptide.

[0011] The preparation method of the fermentation complex includes the following steps: drying jujube seed, poria cocos, and lily bulb separately to a moisture content of ≤8%, coarsely pulverizing them, passing them through a 40-60 mesh sieve, mixing the obtained coarse jujube seed powder, coarse poria cocos powder, and coarse lily bulb powder, adding an ethanol aqueous solution for reflux extraction, filtering and collecting the filtrate, repeating the extraction once with the filter residue, combining the filtrates, and vacuum concentrating them at 60-70℃ to a relative density of 1.05-1.15, recovering the ethanol, and obtaining the alcohol extract. The alcohol extract was dispersed in deionized water to prepare a suspension with a mass concentration of 5%-10%. 2%-5% glucose was added as a carbon source to the suspension. The mixture was stirred evenly and inoculated with Lactobacillus plantarum for anaerobic fermentation. After fermentation, the mixture was inactivated at 85-95℃ for 5-15 minutes. Subsequently, it was dried at 60-70℃ until the moisture content was ≤7%, and then pulverized through an 80-100 mesh sieve to obtain the fermentation complex.

[0012] The mass ratio of the crude jujube seed powder, crude poria cocos powder, and crude lily powder is 3-5:2-3:1; the volume fraction of the ethanol aqueous solution used in the reflux extraction is 60%-75%, the material-to-liquid ratio is 1:8-1:12, the extraction temperature is 50-60℃, and the extraction time is 2-3 hours; the inoculum amount of Lactobacillus plantarum in the anaerobic fermentation is 3%-7% of the total mass of the fermentation substrate, and the anaerobic fermentation is carried out at a temperature of 35-42℃ for 24-48 hours.

[0013] Water extraction results in significant loss of lipid-soluble active ingredients. Even with alcohol extraction, saponins and triterpenes in the resulting extract are mostly in glycoside-bound form, with large molecular weights that make them difficult to embed into lipid carriers.

[0014] This application employs reflux extraction of crude jujube seed powder, crude poria cocos powder, and crude lily powder using an ethanol-water solution. Utilizing the solubility properties of ethanol for saponins and triterpenes, the lipid-soluble active ingredients are selectively transferred from the plant matrix to the liquid phase. After ethanol recovery from the ethanol extract, the extract is dispersed in deionized water with glucose added as a carbon source, and then inoculated with *Lactobacillus plantarum* for anaerobic fermentation. Under sufficient energy supply, *Lactobacillus plantarum* metabolizes stably, secreting glycosidases and esterases. In an anaerobic environment, glycosidases hydrolyze large glycoside-bound saponins into smaller aglycones, and esterases convert bound triterpenic acids into free forms, reducing molecular weight and optimizing the lipid-water partition coefficient. Simultaneously, *Lactobacillus plantarum* produces short-chain fatty acids such as lactic acid and acetic acid. These metabolites can regulate the intestinal flora structure via the gut-brain axis, indirectly affecting central nervous system function. After fermentation, the extract is inactivated and dried to obtain a fermentation complex. The active ingredients in this fermentation complex exist in a low molecular weight form, making it easier to cross biomembranes, and it contains bioactive small molecules produced by microbial metabolism, resulting in higher overall bioavailability than the unfermented ethanol extract.

[0015] The preparation method of the directional enzymatic hydrolysis complex includes the following steps: dispersing milk fat globule membrane concentrate in deionized water to prepare a suspension with a mass concentration of 5%-10%, adjusting the pH to 6.5-7.5, adding neutral protease for enzymatic hydrolysis, then maintaining at 90-95℃ for 5-10 min to inactivate the enzyme, cooling to room temperature, and removing unhydrolyzed solid residue by microfiltration through a ceramic membrane with a pore size of 0.45-1.0 μm under a pressure of 0.2-0.5 MPa, collecting the filtrate, concentrating it under vacuum to a solid content of 15%-25%, and spray drying at an inlet air temperature of 150-160℃ and an outlet air temperature of 75-85℃ to obtain the directional enzymatic hydrolysis complex powder.

[0016] The total phospholipid content of the milk fat globule membrane concentrate is ≥6%, and the glycoprotein content is ≥15%; the amount of neutral protease added is 0.3%-1.5% of the protein mass in the milk fat globule membrane concentrate, and enzymatic hydrolysis is carried out at a temperature of 45-55℃ for 1-3 hours.

[0017] Milk fat globule membrane concentrate is the natural membrane structure encapsulating milk fat globules. Rich in phospholipids and glycoproteins, it possesses a bilayer structure similar to cell membranes, making it an ideal carrier for active ingredients. However, current enzymatic hydrolysis techniques for milk fat globule membrane proteins often employ alkaline proteases or trypsin at high pH conditions. This strongly alkaline environment causes deglycosylation of the N-glycosylation domains of glycoproteins or peptide chain breakage, resulting in the loss of numerous glycosylation sites and thus the loss of their ability to recognize and bind to intestinal epithelial cells. Furthermore, excessive enzymatic hydrolysis disintegrates the phospholipid-protein complex, failing to provide a stable loading space for hydrophobic active ingredients.

[0018] This application involves dispersing milk fat globule membrane concentrate, adjusting the pH to near neutral, and then performing mild enzymatic hydrolysis using a neutral protease. This pH range avoids the chemically unstable region of glycoprotein N-glycosylation sites. The neutral protease preferentially cleaves peptide bonds in non-glycosylated regions, while glycosylated domains are preserved due to steric hindrance and charge shielding effects of glycan chains. After hydrolysis, glycoprotein fragments retain intact glycosylated domains, and phospholipid fragments form hydrophobic regions similar to cell membranes. Unhydrolyzed solid residues are removed by ceramic membrane microfiltration, followed by vacuum concentration and spray drying to obtain a directional enzymatic hydrolysis complex powder. This powder maintains high levels of glycoprotein and phospholipid content, with sufficient density of glycosylation sites for subsequent N-acetylneuraminic acid binding. The phospholipids retain their natural bilayer structure characteristics, forming a hydrophobic core that provides embedding space for lipid-soluble components and phosphatidylserine in the fermentation complex.

[0019] The walnut peptides have an average molecular weight of 500-3000 Da, a peptide content of ≥75%, and a protein content of ≥80%.

[0020] A method for preparing a nutritional composition to improve concentration includes the following steps: co-assembling and surface functionalizing the fermentation complex, the directed enzymatic hydrolysis complex, phosphatidylserine, and N-acetylneuraminic acid to form a surface-functionalized nanoparticle suspension; then treating the surface-functionalized nanoparticle suspension with sodium caseinate microencapsulation to obtain microencapsulated nanoparticle powder; mixing the microencapsulated nanoparticle powder with walnut peptide and food-grade excipients; adding deionized water to prepare a mixture with a solid content of 15%-25%; homogenizing twice at a temperature of 50-60℃ and a pressure of 20-30MPa; and then vacuum degassing and ultra-high temperature instantaneous sterilization at 135℃ for 5 seconds to obtain the finished nutritional composition to improve concentration.

[0021] The process of forming a surface-functionalized nanoparticle suspension by co-assembling and surface functionalizing the fermentation complex, the directed enzymatic hydrolysis complex, phosphatidylserine, and N-acetylneuraminic acid specifically includes the following steps: dissolving phosphatidylserine in anhydrous ethanol to prepare a solution with a mass concentration of 10-20 mg / mL; dissolving N-acetylneuraminic acid in deionized water to prepare a solution with a mass concentration of 15-25 mg / mL; dispersing the directed enzymatic hydrolysis complex powder in deionized water to prepare a suspension with a mass concentration of 25-45 mg / mL; and dispersing the fermentation complex in a 30%-volume ... Prepare a suspension with a mass concentration of 15-25 mg / mL using a 50% ethanol aqueous solution. Under the conditions of stirring speed of 400-600 r / min and temperature of 35-45℃, first add the fermentation complex ethanol suspension dropwise to the directional enzymatic hydrolysis complex suspension and stir for 30-60 min. Then, add the phosphatidylserine ethanol solution dropwise to the mixture and stir for 30-60 min. Finally, add the N-acetylneuraminic acid aqueous solution dropwise to the mixture and stir for 30-60 min. After the addition is complete, continue stirring for 1-2 h to form a suspension of surface-functionalized nanoparticles.

[0022] Current technologies involve a simple physical mixing of fermented medicinal and edible components with neurotrophic factors such as phosphatidylserine and N-acetylneuraminic acid. Each component is independently dispersed in the liquid phase and absorbed independently after entering the gastrointestinal tract, failing to achieve synergy in absorption time and target site. This lack of structural synergistic design results in uncontrollable release of active ingredients in the intestines, low efficiency in crossing the blood-brain barrier, and difficulty in synchronous release to produce multi-target effects once in the brain.

[0023] In this application, the fermentation complex is dispersed in an aqueous ethanol solution, the directional enzymatic hydrolysis complex powder is dispersed in deionized water, phosphatidylserine is dissolved in anhydrous ethanol, and N-acetylneuraminic acid is dissolved in deionized water, and the mixture is added dropwise in sequence under gentle stirring conditions.

[0024] First, an ethanol suspension of the fermentation complex was added dropwise to the suspension of the directed enzymatic hydrolysis complex. Small aglycones and free triterpenic acids from the fermentation complex entered the hydrophobic region of the phospholipids in the directed enzymatic hydrolysis complex through hydrophobic interactions, forming a core load. Then, an ethanol solution of phosphatidylserine was added. The phosphate groups and fatty acid chains of phosphatidylserine are similar in structure to the phospholipids of the milk fat globule membrane, both possessing amphiphilic molecular structures. Driven by hydrophobic interactions and van der Waals forces, phosphatidylserine spontaneously intercalates into the phospholipid layer, forming a molecular recognition effect with the milk fat globule membrane phospholipids, enhancing the stability of phosphatidylserine and participating in membrane structure reconstruction. Finally, an aqueous solution of N-acetylneuraminic acid was added. The carboxyl and acetylamino groups in N-acetylneuraminic acid bind to the N-glycosylation sites retained on the surface of the glycoprotein of the directed enzymatic hydrolysis complex through hydrogen bonds, positioning it on the outer layer of the particles. This layer-by-layer construction method from the inside out allows the components to form a spatially ordered distribution at the nanoscale, resulting in a suspension of surface-functionalized nanoparticles.

[0025] In the aforementioned co-assembly process, saponins and triterpenes in the fermentation complex embed into the hydrophobic core of the phospholipids in the directed enzymatic hydrolysis complex through hydrophobic interactions. Phosphatidylserine embeds into the phospholipid layer through hydrophobic interactions, and N-acetylneuraminic acid binds to the glycosylation sites of the glycoprotein in the directed enzymatic hydrolysis complex via hydrogen bonds, thereby forming core-shell structured surface-functionalized nanoparticles. In this suspension, the active ingredient of the fermentation complex is located in the core, phosphatidylserine is embedded in the intermediate phospholipid layer, and N-acetylneuraminic acid is distributed on the surface of the outer shell glycoprotein, with each component forming an organic whole in structure.

[0026] The surface-functionalized nanoparticle suspension is encapsulated with sodium caseinate to obtain microencapsulated nanoparticle powder. The encapsulation process specifically includes the following steps: using the surface-functionalized nanoparticle suspension as the core material, dissolving sodium caseinate in deionized water to prepare an outer wall material solution with a mass concentration of 20-40 mg / mL, and maintaining a mass ratio of the surface-functionalized nanoparticle suspension (core material) to the outer wall material solution of 1:2-1:4. The surface-functionalized nanoparticles are then encapsulated under stirring conditions of 200-400 r / min and a temperature of 35-45℃. The rice particle suspension was added dropwise to a sodium caseinate solution and stirred for 30-60 minutes. Then, the pH was adjusted to 4.0-5.0 with 1 mol / L citric acid. The sodium caseinate underwent a coagulation reaction, forming a dense gel layer that encapsulated the nanoparticles. Stirring was continued for another 30-60 minutes. The mixture was collected by centrifugation, washed 2-3 times with deionized water, and redispersed in deionized water to prepare a suspension with a mass concentration of 10%-20%. Then, it was spray-dried at an inlet air temperature of 150-170℃ and an outlet air temperature of 70-85℃ to obtain microencapsulated nanoparticle powder.

[0027] When surface-functionalized nanoparticle suspensions are directly exposed to the acidic environment of the stomach, the phospholipid layer is easily attacked by pepsin, N-acetylneuraminic acid becomes less stable under strong acid conditions, and the core active ingredient may be released prematurely in the stomach. In existing microencapsulation technologies, the encapsulation effect is uncontrollable. Spray drying results in large fluctuations in the feed solids content, leading to unstable product quality.

[0028] This application uses a surface-functionalized nanoparticle suspension as the core material and sodium caseinate dissolved in deionized water to prepare an outer wall material solution.

[0029] The core material was dropwise added to the wall material solution under gentle stirring, allowing sodium caseinate to be uniformly adsorbed onto the surface of the nanoparticles. Subsequently, citric acid was used to adjust the pH to near the isoelectric point of sodium caseinate, neutralizing the protein molecules' charge, drastically reducing their solubility, and inducing a coagulation reaction that deposited a dense gel layer on the nanoparticle surface. By strictly controlling the mass ratio of the core material suspension to the wall material solution and slowly adjusting the pH, a sufficiently dense and uniform gel layer could be obtained. This gel layer, with its dense structure in the acidic gastric environment, can block pepsin and oxygen from contacting the core active ingredient, thus providing gastric acid protection. Upon entering the intestine, the ambient pH rises to neutral, causing the sodium caseinate gel layer to swell and rupture, releasing the internal nanoparticles. The nanoparticles promote endocytosis and absorption through the affinity of their surface N-acetylneuraminic acid with intestinal epithelial cells. After centrifugation and washing, the nanoparticles were redispersed in deionized water at a controlled concentration and then spray-dried to ensure that the particle size and wall material thickness of the microencapsulated nanoparticle powder were within a controllable range. This microencapsulated nanoparticle powder has gastric acid protection and intestinal sustained-release properties. When treated in simulated gastric juice, the core material release rate is low, but after being transferred to simulated intestinal juice, the cumulative release rate is high, achieving the targeted release of active ingredients in the intestine.

[0030] Application of a nutritional composition for improving concentration, said nutritional composition for manufacturing functional foods or health products that improve concentration.

[0031] In existing technologies, auxiliary ingredients such as walnut peptides are often simply mixed with the main active ingredient without homogenization and stabilization treatment, leading to phase separation or precipitation in the liquid phase of the product. If the sterilization process is not properly selected, heat-sensitive active ingredients are prone to degradation during prolonged heating. The lack of a final composition molding step results in a break in the technological chain from intermediate products to end products.

[0032] This application involves mixing microencapsulated nanoparticle powder with walnut peptides and food-grade excipients, adding deionized water to form a mixture, and then homogenizing under high pressure to ensure uniform dispersion of the components, forming a stable emulsion system. The average molecular weight of the walnut peptides falls within the oligopeptide range, allowing for direct absorption into the bloodstream through the intestines, promoting the expression of brain-derived neurotrophic factor, and enhancing nerve fiber growth and synaptic plasticity. Subsequently, ultra-high temperature instantaneous sterilization is employed to kill microorganisms in a very short time, while preserving the structure and activity of heat-sensitive active ingredients due to the short heating time. Finally, the product is filled or spray-dried to obtain the functional food or health product. The entire technology chain, from raw material processing to the end product, forms a complete closed loop.

[0033] The nutritional composition obtained in this application, through the aforementioned multi-level delivery structure, allows saponins, triterpenes, phosphatidylserine, and N-acetylneuraminic acid from the fermentation complex to be simultaneously released and absorbed in the intestine, while walnut peptides rapidly enter the bloodstream to promote nerve growth factor expression. The small-molecule aglycones in the fermentation complex inhibit excessive microglial activation and reduce neuroinflammation; poria cocos triterpenoids regulate the balance of γ-aminobutyric acid and glutamate, improving nerve excitability; N-acetylneuraminic acid participates in the glycosylation modification of gangliosides and glycoproteins, maintaining synaptic structural stability; phosphatidylserine embeds into the phospholipid bilayer of the nerve cell membrane, participating in membrane synthesis and improving nerve signal transduction efficiency; and walnut peptides promote the expression of brain-derived neurotrophic factor, enhancing nerve fiber growth and synaptic plasticity. This multi-dimensional approach inhibits neuroinflammation, balances neurotransmitters, maintains synaptic structure, and protects against neuropathic damage, thus meeting the needs of adolescents and individuals with high cognitive load for improved concentration.

[0034] Compared with the prior art, the advantages and beneficial effects of the present invention are as follows: 1. This application employs reflux extraction of jujube seed, poria cocos, and lily bulb using an ethanol-water solution to selectively transfer lipid-soluble active ingredients to the liquid phase. Subsequently, glucose is added as a carbon source, and *Lactobacillus plantarum* is inoculated for anaerobic fermentation. Under sufficient energy supply, *Lactobacillus plantarum* exhibits stable metabolism, secreting glycosidases and esterases to hydrolyze large glycoside-bound saponins into smaller aglycones, while simultaneously converting bound triterpenic acids into free forms, thus reducing the molecular weight of the active ingredients and optimizing the lipid-water partition coefficient. The short-chain fatty acids produced during fermentation can indirectly affect central nervous system function. The resulting fermentation complex contains active ingredients in a low molecular weight form, making it easier to cross biological membranes and resulting in higher bioavailability than the unfermented ethanol extract.

[0035] 2. This application describes a mild enzymatic hydrolysis of milk fat globule membrane concentrate using a neutral protease under near-neutral pH conditions. This pH range avoids the chemically unstable region of glycoprotein N-glycosylation sites. The neutral protease preferentially cleaves peptide bonds in non-glycosylated regions, while glycosylated domains are preserved due to steric hindrance and charge shielding effects of the glycan chains. In the resulting directional hydrolysis complex powder, glycoprotein fragments maintain intact glycosylated domains, and phospholipid fragments form hydrophobic regions similar to cell membranes. This structure provides binding sites for subsequent N-acetylneuraminic acid and also provides intercalation space for lipid-soluble components and phosphatidylserine in the fermentation complex, thus constituting a natural carrier of active ingredients.

[0036] 3. In this application, a fermentation complex, a targeted enzymatic hydrolysis complex, phosphatidylserine, and N-acetylneuraminic acid are sequentially added and mixed. This allows small molecule aglycones and free triterpenic acids to enter the hydrophobic core of the phospholipid in the targeted enzymatic hydrolysis complex through hydrophobic interactions. Phosphatidylserine embeds itself into the phospholipid layer through intermolecular hydrophobic interactions, while N-acetylneuraminic acid binds to N-glycosylation sites retained on the glycoprotein surface via hydrogen bonds, forming a spatially ordered suspension of surface-functionalized nanoparticles. Subsequently, sodium caseinate is used for microencapsulation, with strict control over the mass ratio of the core suspension to the wall solution, forming a dense gel layer near the isoelectric point. This dense gel layer, structurally sound in the acidic gastric environment, can prevent pepsin and oxygen from contacting the core active ingredient, thus providing gastric acid protection. Upon entering the intestine, the pH rises to neutral, causing the gel layer to swell and rupture, releasing the internal nanoparticles and achieving targeted release and sustained supply of the active ingredient in the intestine.

[0037] 4. This application mixes microencapsulated nanoparticle powder with walnut peptides and food-grade excipients, homogenizes under high pressure to form a stable emulsion system, and then sterilizes at ultra-high temperature to retain the activity of heat-sensitive active ingredients. Walnut peptides can be directly absorbed by the intestines and enter the bloodstream, promoting the expression of brain-derived neurotrophic factor and enhancing nerve fiber growth and synaptic plasticity. The small-molecule aglycones in the fermentation complex can inhibit excessive activation of microglia; poria triterpenoid acid can regulate neurotransmitter balance; N-acetylneuraminic acid can participate in the glycosylation modification of gangliosides and glycoproteins to maintain synaptic structural stability; and phosphatidylserine can embed into the phospholipid bilayer of nerve cell membranes to improve nerve signal transduction efficiency. These multiple components are spatially ordered within the nanoparticles, and upon entering the intestines, they are released simultaneously and absorbed in conjunction, addressing the need to improve concentration from multiple dimensions, including inhibition of neuroinflammation, neurotransmitter balance, synaptic structure maintenance, and neuroprotection. Attached Figure Description

[0038] Figure 1 Results of fermentation complex index testing Figure 2 Results of targeted enzymatic hydrolysis complex index test Figure 3 This is a process flow diagram of a method for preparing a nutritional composition for improving concentration according to the present invention. Detailed Implementation

[0039] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0040] In the specific embodiments of this application, the sources of various main raw materials are briefly described as follows: Ethanol: Purchased from Sinopharm Chemical Reagent Co., Ltd., CAS No. 64-17-5 Glucose: Purchased from Shandong Xiwang Sugar Industry Co., Ltd., CAS No. 50-99-7 Lactobacillus plantarum: purchased from China Industrial Microbial Culture Collection Center, strain number CICC 20022 Neutral protease: purchased from Nanning Pangbo Biotechnology Co., Ltd., with an enzyme activity of 200,000 U / g. Milk fat globule membrane concentrate: purchased from Jiangsu Caiwei Biotechnology Co., Ltd., food grade. Phosphatidylserine: Purchased from Guangdong Mingtong Biotechnology Co., Ltd., CAS No. 51446-62-9, food grade. N-Acetylneuraminic acid: Purchased from Jiangsu Caiwei Biotechnology Co., Ltd., CAS No. 131-48-6, purity 98%. Sodium caseinate: purchased from Zhejiang Yicun Biotechnology Co., Ltd., CAS No. 9005-46-3 Citric acid: purchased from Shandong Yingxuan Industrial Co., Ltd., CAS No. 77-92-9 Walnut peptides: purchased from Wuhan Tiantianhao Biological Products Co., Ltd. The technical solution of this application is as follows: A nutritional composition for improving concentration, comprising, by weight, the following components: 8-15 parts of fermentation complex, 6-12 parts of targeted enzymatic hydrolysis complex, 1-3 parts of phosphatidylserine, 1-2 parts of N-acetylneuraminic acid, and 2-5 parts of walnut peptide.

[0041] The preparation method of the fermentation complex includes the following steps: drying jujube seed, poria cocos, and lily bulb separately to a moisture content of ≤8%, coarsely pulverizing them, passing them through a 40-60 mesh sieve, mixing the obtained coarse jujube seed powder, coarse poria cocos powder, and coarse lily bulb powder, adding an ethanol aqueous solution for reflux extraction, filtering and collecting the filtrate, repeating the extraction once with the filter residue, combining the filtrates, and vacuum concentrating them at 60-70℃ to a relative density of 1.05-1.15, recovering the ethanol, and obtaining the alcohol extract. The alcohol extract was dispersed in deionized water to prepare a suspension with a mass concentration of 5%-10%. 2%-5% glucose was added as a carbon source to the suspension. The mixture was stirred evenly and inoculated with Lactobacillus plantarum for anaerobic fermentation. After fermentation, the mixture was inactivated at 85-95℃ for 5-15 minutes. Subsequently, it was dried at 60-70℃ until the moisture content was ≤7%, and then pulverized through an 80-100 mesh sieve to obtain the fermentation complex.

[0042] The mass ratio of the crude jujube seed powder, crude poria cocos powder, and crude lily powder is 3-5:2-3:1; the volume fraction of the ethanol aqueous solution used in the reflux extraction is 60%-75%, the material-to-liquid ratio is 1:8-1:12, the extraction temperature is 50-60℃, and the extraction time is 2-3 hours; the inoculum amount of Lactobacillus plantarum in the anaerobic fermentation is 3%-7% of the total mass of the fermentation substrate, and the anaerobic fermentation is carried out at a temperature of 35-42℃ for 24-48 hours.

[0043] The preparation method of the directional enzymatic hydrolysis complex includes the following steps: dispersing milk fat globule membrane concentrate in deionized water to prepare a suspension with a mass concentration of 5%-10%, adjusting the pH to 6.5-7.5, adding neutral protease for enzymatic hydrolysis, then maintaining at 90-95℃ for 5-10 min to inactivate the enzyme, cooling to room temperature, and removing unhydrolyzed solid residue by microfiltration through a ceramic membrane with a pore size of 0.45-1.0 μm under a pressure of 0.2-0.5 MPa, collecting the filtrate, concentrating it under vacuum to a solid content of 15%-25%, and spray drying at an inlet air temperature of 150-160℃ and an outlet air temperature of 75-85℃ to obtain the directional enzymatic hydrolysis complex powder.

[0044] The total phospholipid content of the milk fat globule membrane concentrate is ≥6%, and the glycoprotein content is ≥15%; the amount of neutral protease added is 0.3%-1.5% of the protein mass in the milk fat globule membrane concentrate, and enzymatic hydrolysis is carried out at a temperature of 45-55℃ for 1-3 hours.

[0045] The walnut peptides have an average molecular weight of 500-3000 Da, a peptide content of ≥75%, and a protein content of ≥80%.

[0046] A method for preparing a nutritional composition to improve concentration, such as... Figure 3As shown, the method includes the following steps: the fermentation complex, the directional enzymatic hydrolysis complex, phosphatidylserine, and N-acetylneuraminic acid are co-assembled and surface-functionalized to form a surface-functionalized nanoparticle suspension. Then, the surface-functionalized nanoparticle suspension is treated with sodium caseinate microencapsulation to obtain microencapsulated nanoparticle powder. The microencapsulated nanoparticle powder is mixed with walnut peptide and food-grade excipients, and deionized water is added to prepare a mixture with a solid content of 15%-25%. The mixture is homogenized twice at a temperature of 50-60℃ and a pressure of 20-30MPa, and then vacuum degassed and sterilized at 135℃ for 5 seconds to obtain the finished product of a nutritional composition for improving concentration.

[0047] The process of forming a surface-functionalized nanoparticle suspension by co-assembling and surface functionalizing the fermentation complex, the directed enzymatic hydrolysis complex, phosphatidylserine, and N-acetylneuraminic acid specifically includes the following steps: dissolving phosphatidylserine in anhydrous ethanol to prepare a solution with a mass concentration of 10-20 mg / mL; dissolving N-acetylneuraminic acid in deionized water to prepare a solution with a mass concentration of 15-25 mg / mL; dispersing the directed enzymatic hydrolysis complex powder in deionized water to prepare a suspension with a mass concentration of 25-45 mg / mL; and dispersing the fermentation complex in a 30%-volume ... Prepare a suspension with a mass concentration of 15-25 mg / mL using a 50% ethanol aqueous solution. Under the conditions of stirring speed of 400-600 r / min and temperature of 35-45℃, first add the fermentation complex ethanol suspension dropwise to the directional enzymatic hydrolysis complex suspension and stir for 30-60 min. Then, add the phosphatidylserine ethanol solution dropwise to the mixture and stir for 30-60 min. Finally, add the N-acetylneuraminic acid aqueous solution dropwise to the mixture and stir for 30-60 min. After the addition is complete, continue stirring for 1-2 h to form a suspension of surface-functionalized nanoparticles.

[0048] The surface-functionalized nanoparticle suspension is encapsulated with sodium caseinate to obtain microencapsulated nanoparticle powder. The encapsulation process specifically includes the following steps: using the surface-functionalized nanoparticle suspension as the core material, dissolving sodium caseinate in deionized water to prepare an outer wall material solution with a mass concentration of 20-40 mg / mL, and maintaining a mass ratio of the surface-functionalized nanoparticle suspension (core material) to the outer wall material solution of 1:2-1:4. The surface-functionalized nanoparticles are then encapsulated under stirring conditions of 200-400 r / min and a temperature of 35-45℃. The rice particle suspension was added dropwise to a sodium caseinate solution and stirred for 30-60 minutes. Then, the pH was adjusted to 4.0-5.0 with 1 mol / L citric acid. The sodium caseinate underwent a coagulation reaction, forming a dense gel layer that encapsulated the nanoparticles. Stirring was continued for another 30-60 minutes. The mixture was collected by centrifugation, washed 2-3 times with deionized water, and redispersed in deionized water to prepare a suspension with a mass concentration of 10%-20%. Then, it was spray-dried at an inlet air temperature of 150-170℃ and an outlet air temperature of 70-85℃ to obtain microencapsulated nanoparticle powder.

[0049] Application of a nutritional composition for improving concentration, said nutritional composition for manufacturing functional foods or health products that improve concentration.

[0050] The present invention will be described in detail below through examples and comparative examples, but the scope of protection of the present invention is not limited to these examples. Unless otherwise specified, the chemical reagents and raw materials used in the following examples and comparative examples are all conventional commercially available products.

[0051] Example 1 Sour jujube seed, Poria cocos, and lily bulb were dried to a moisture content of 8%, coarsely pulverized, and passed through a 60-mesh sieve. 50g of coarsely powdered sour jujube seed, 25g of coarsely powdered Poria cocos, and 10g of coarsely powdered lily bulb were weighed, mixed, and then added to a 75% (v / v) ethanol aqueous solution at a material-to-liquid ratio of 1:10. The mixture was refluxed at 60℃ for 2.5h, filtered, and the filtrate was collected. The residue was extracted once more with the same volume of ethanol aqueous solution. The filtrates were combined and concentrated under vacuum at 70℃ to a relative density of 1.15. The ethanol was recovered to obtain an alcohol extract. The alcohol extract was dispersed in deionized water to prepare a 10% (w / w) suspension. 5% (w / w) of glucose was added as a carbon source, and the mixture was stirred thoroughly. *Lactobacillus plantarum* was inoculated at an inoculum size of 7% of the total substrate mass. Anaerobic fermentation was carried out at 35℃ for 48h. After fermentation, the mixture was inactivated at 95℃ for 5min, then dried at 70℃ to a moisture content of 7%, pulverized, and passed through a 90-mesh sieve to obtain the fermentation complex.

[0052] Milk fat globule membrane concentrate was dispersed in deionized water to prepare a 5% (w / w) suspension. The pH was adjusted to 7.5, and neutral protease was added at a concentration of 0.9% of the protein mass in the milk fat globule membrane concentrate. Enzymatic hydrolysis was performed at 45°C for 3 hours, followed by enzyme inactivation at 95°C for 5 minutes. After cooling to room temperature, the undigested solid residue was removed by microfiltration through a 0.45 μm ceramic membrane at a pressure of 0.35 MPa. The filtrate was collected, concentrated under vacuum to a solid content of 25%, and spray-dried at an inlet air temperature of 155°C and an outlet air temperature of 75°C to obtain a directional enzymatic hydrolysis complex powder. Due to the significant reduction in the size of phospholipid and glycoprotein fragments after directional hydrolysis with neutral protease, they can permeate through the ceramic membrane, while undigested macromolecular fibers and insoluble polysaccharides are retained, thus achieving effective separation.

[0053] Phosphatidylserine was dissolved in anhydrous ethanol to prepare a solution with a mass concentration of 20 mg / mL. N-acetylneuraminic acid was dissolved in deionized water to prepare a solution with a mass concentration of 20 mg / mL. The directional enzymatic hydrolysis complex powder was dispersed in deionized water to prepare a suspension with a mass concentration of 25 mg / mL. The fermentation complex was dispersed in a 50% (v / v) ethanol aqueous solution to prepare a suspension with a mass concentration of 20 mg / mL. Under the conditions of stirring speed of 400 r / min and temperature of 45℃, the ethanol suspension of the fermentation complex was first added dropwise to the suspension of the directional enzymatic hydrolysis complex and stirred for 45 min. Then, the ethanol solution of phosphatidylserine was added dropwise to the mixture and stirred for 45 min. Finally, the aqueous solution of N-acetylneuraminic acid was added dropwise to the mixture and stirred for 30 min. After the addition was completed, stirring was continued for 1 h to form a suspension of surface-functionalized nanoparticles.

[0054] A surface-functionalized nanoparticle suspension was used as the core material. Sodium caseinate was dissolved in deionized water to prepare an outer wall material solution with a mass concentration of 40 mg / mL. The mass ratio of the core material suspension to the outer wall material solution was 1:3. Under the conditions of stirring speed of 200 r / min and temperature of 45℃, the surface-functionalized nanoparticle suspension was added dropwise to the sodium caseinate solution and stirred for 45 min. Then, the pH was adjusted to 4.0 with 1 mol / L citric acid. Sodium caseinate underwent a coagulation reaction to form a dense gel layer that encapsulated the nanoparticles. Stirring was continued for 60 min. After centrifugation, the nanoparticles were washed three times with deionized water and redispersed in deionized water to prepare a suspension with a mass concentration of 15%. Then, spray drying was performed with an inlet air temperature of 150℃ and an outlet air temperature of 85℃ to obtain microencapsulated nanoparticle powder.

[0055] 11.5 parts of fermentation complex, 6 parts of directional enzymatic hydrolysis complex, 3 parts of phosphatidylserine, 1.5 parts of N-acetylneuraminic acid, 3.5 parts of walnut peptide, and food-grade excipients were weighed. The average molecular weight of the walnut peptide was 1750 Da. The microencapsulated nanoparticle powder was mixed with the walnut peptide and food-grade excipients, and deionized water was added to prepare a mixture with a solid content of 25%. The mixture was homogenized twice at a temperature of 55℃ and a pressure of 20 MPa. After vacuum degassing and ultra-high temperature instantaneous sterilization at 135℃ for 5 seconds, the liquid beverage was aseptically filled.

[0056] Example 2 In this embodiment, the similarities to those in Embodiment 1 will not be repeated, and the differences are as follows: Sour jujube seed, Poria cocos, and lily bulb were dried to a moisture content of 5%, coarsely pulverized, and passed through a 60-mesh sieve. 30g of coarsely powdered sour jujube seed, 20g of coarsely powdered Poria cocos, and 10g of coarsely powdered lily bulb were weighed, mixed, and then added to a 75% (v / v) ethanol aqueous solution at a material-to-liquid ratio of 1:10. The mixture was refluxed at 50℃ for 3 hours. The filtrate was collected, and the residue was extracted once more with the same volume of ethanol aqueous solution. The filtrates were combined and concentrated under vacuum at 60℃ to a relative density of 1.05. The ethanol was recovered to obtain an alcohol extract. The alcohol extract was dispersed in deionized water to prepare a 7.5% (w / w) suspension. 2% (w / w) of glucose was added as a carbon source, and the mixture was stirred thoroughly. *Lactobacillus plantarum* was inoculated at 7% (w / w) of the total substrate mass. Anaerobic fermentation was carried out at 38.5℃ for 24 hours. After fermentation, the mixture was inactivated at 95℃ for 10 minutes, then dried at 60℃ to a moisture content of 7%, pulverized, and passed through a 100-mesh sieve to obtain the fermentation complex.

[0057] Milk fat globule membrane concentrate was dispersed in deionized water to prepare a 7.5% (w / w) suspension. The pH was adjusted to 6.5, and neutral protease was added at a concentration of 1.5% of the protein mass in the milk fat globule membrane concentrate. The mixture was enzymatically hydrolyzed at 50°C for 1 hour, followed by enzyme inactivation at 90°C for 10 minutes. After cooling to room temperature, the unhydrolyzed solid residue was removed by microfiltration through a ceramic membrane with a pore size of 0.725 μm at a pressure of 0.5 MPa. The filtrate was collected, concentrated under vacuum to a solid content of 15%, and spray-dried at an inlet air temperature of 160°C and an outlet air temperature of 80°C to obtain a directional enzymatic hydrolysis complex powder.

[0058] Phosphatidylserine was dissolved in anhydrous ethanol to prepare a solution with a mass concentration of 10 mg / mL. N-acetylneuraminic acid was dissolved in deionized water to prepare a solution with a mass concentration of 25 mg / mL. The directional enzymatic hydrolysis complex powder was dispersed in deionized water to prepare a suspension with a mass concentration of 35 mg / mL. The fermentation complex was dispersed in a 30% (v / v) ethanol aqueous solution to prepare a suspension with a mass concentration of 25 mg / mL. Under the conditions of stirring speed of 500 r / min and temperature of 35℃, the ethanol suspension of the fermentation complex was first added dropwise to the suspension of the directional enzymatic hydrolysis complex and stirred for 60 min. Then, the ethanol solution of phosphatidylserine was added dropwise to the mixture and stirred for 30 min. Finally, the aqueous solution of N-acetylneuraminic acid was added dropwise to the mixture and stirred for 60 min. After the addition was completed, stirring was continued for 1.5 h to form a suspension of surface-functionalized nanoparticles.

[0059] A surface-functionalized nanoparticle suspension was used as the core material. Sodium caseinate was dissolved in deionized water to prepare an outer wall material solution with a mass concentration of 20 mg / mL. The mass ratio of the core material suspension to the outer wall material solution was 1:4. Under the conditions of stirring speed of 300 r / min and temperature of 35℃, the surface-functionalized nanoparticle suspension was added dropwise to the sodium caseinate solution and stirred for 60 min. Then, the pH was adjusted to 4.5 with 1 mol / L citric acid. Sodium caseinate underwent a coagulation reaction to form a dense gel layer that encapsulated the nanoparticles. Stirring was continued for 30 min. After centrifugation, the nanoparticles were washed twice with deionized water and redispersed in deionized water to prepare a suspension with a mass concentration of 20%. Then, spray drying was performed with an inlet air temperature of 160℃ and an outlet air temperature of 70℃ to obtain microencapsulated nanoparticle powder.

[0060] Eight parts of fermentation complex, 12 parts of directional enzymatic hydrolysis complex, 2 parts of phosphatidylserine, 1 part of N-acetylneuraminic acid, 5 parts of walnut peptide, and food-grade excipients were weighed out. The average molecular weight of the walnut peptide was 1750 Da. The microencapsulated nanoparticle powder was mixed with the walnut peptide and food-grade excipients, and deionized water was added to prepare a mixture with a solid content of 20%. The mixture was homogenized twice at 50℃ and 30MPa, then vacuum degassed and sterilized at 135℃ for 5 seconds. Finally, it was spray-dried to obtain a powdered solid beverage.

[0061] Example 3 In this embodiment, the similarities to those in Embodiment 1 will not be repeated, and the differences are as follows: Sour jujube seed, Poria cocos, and lily bulb were dried to a moisture content of 6.5%, coarsely pulverized, and passed through a 40-mesh sieve. 50g of coarsely powdered sour jujube seed, 25g of coarsely powdered Poria cocos, and 10g of coarsely powdered lily bulb were weighed, mixed, and then added to a 60% (v / v) ethanol aqueous solution at a material-to-liquid ratio of 1:12. The mixture was refluxed at 55℃ for 2 hours, the filtrate was collected, and the residue was extracted once more with the same volume of ethanol aqueous solution. The filtrates were combined and concentrated under vacuum at 65℃ to a relative density of 1.10. The ethanol was recovered to obtain an alcohol extract. The alcohol extract was dispersed in deionized water to prepare a 10% (w / w) suspension. 3.5% (w / w) of glucose was added as a carbon source, and the mixture was stirred thoroughly. *Lactobacillus plantarum* was inoculated at an inoculum size of 3% of the total substrate mass. Anaerobic fermentation was carried out at 42℃ for 36 hours. After fermentation, the mixture was inactivated at 85℃ for 15 minutes, then dried at 65℃ to a moisture content of 7%, pulverized, and passed through an 80-mesh sieve to obtain the fermentation complex.

[0062] Milk fat globule membrane concentrate was dispersed in deionized water to prepare a 10% (w / w) suspension. The pH was adjusted to 7.0, and neutral protease was added at a concentration of 0.3% of the protein mass in the milk fat globule membrane concentrate. Enzymatic hydrolysis was carried out at 55°C for 2 hours, followed by enzyme inactivation at 92°C for 7 minutes. After cooling to room temperature, the undigested solid residue was removed by microfiltration through a ceramic membrane with a pore size of 1.0 μm at a pressure of 0.2 MPa. The filtrate was collected, concentrated under vacuum to a solid content of 20%, and spray-dried at an inlet air temperature of 150°C and an outlet air temperature of 85°C to obtain a directional enzymatic hydrolysis complex powder.

[0063] Phosphatidylserine was dissolved in anhydrous ethanol to prepare a solution with a mass concentration of 15 mg / mL. N-acetylneuraminic acid was dissolved in deionized water to prepare a solution with a mass concentration of 15 mg / mL. The directional enzymatic hydrolysis complex powder was dispersed in deionized water to prepare a suspension with a mass concentration of 45 mg / mL. The fermentation complex was dispersed in a 40% (v / v) ethanol aqueous solution to prepare a suspension with a mass concentration of 15 mg / mL. Under the conditions of stirring speed of 600 r / min and temperature of 40℃, the ethanol suspension of the fermentation complex was first added dropwise to the suspension of the directional enzymatic hydrolysis complex and stirred for 30 min. Then, the ethanol solution of phosphatidylserine was added dropwise to the mixture and stirred for 45 min. Finally, the aqueous solution of N-acetylneuraminic acid was added dropwise to the mixture and stirred for 30 min. After the addition was completed, stirring was continued for 2 h to form a suspension of surface-functionalized nanoparticles.

[0064] A surface-functionalized nanoparticle suspension was used as the core material. Sodium caseinate was dissolved in deionized water to prepare an outer wall material solution with a mass concentration of 30 mg / mL. The mass ratio of the core material suspension to the outer wall material solution was 1:2. Under the conditions of stirring speed of 400 r / min and temperature of 40℃, the surface-functionalized nanoparticle suspension was added dropwise to the sodium caseinate solution and stirred for 30 min. Then, the pH was adjusted to 5.0 with 1 mol / L citric acid. Sodium caseinate underwent a coagulation reaction to form a dense gel layer that encapsulated the nanoparticles. Stirring was continued for 45 min. After centrifugation, the nanoparticles were washed three times with deionized water and redispersed in deionized water to prepare a suspension with a mass concentration of 10%. Then, spray drying was performed with an inlet air temperature of 170℃ and an outlet air temperature of 77.5℃ to obtain microencapsulated nanoparticle powder.

[0065] Weigh out 15 parts of fermentation complex, 9 parts of directional enzymatic hydrolysis complex, 1 part of phosphatidylserine, 2 parts of N-acetylneuraminic acid, 3.5 parts of walnut peptide, and food-grade excipients. The average molecular weight of the walnut peptide is 500 Da. Mix the microencapsulated nanoparticle powder with the walnut peptide and food-grade excipients, add deionized water to prepare a mixture with a solid content of 15%, homogenize twice at 60℃ and 25MPa, then vacuum degas, sterilize at 135℃ for 5 seconds, and aseptically fill to obtain the liquid beverage.

[0066] Example 4 In this embodiment, the similarities to those in Embodiment 1 will not be repeated, and the differences are as follows: Sour jujube seed, Poria cocos, and lily bulb were dried to a moisture content of 6%, coarsely pulverized, and passed through a 60-mesh sieve. 40g of coarsely powdered sour jujube seed, 25g of coarsely powdered Poria cocos, and 10g of coarsely powdered lily bulb were weighed, mixed, and then added to a 70% (v / v) ethanol aqueous solution at a material-to-liquid ratio of 1:10. The mixture was refluxed at 55℃ for 2.5h, the filtrate was collected, and the residue was extracted once more with the same volume of ethanol aqueous solution. The filtrates were combined and concentrated under vacuum at 65℃ to a relative density of 1.10. The ethanol was recovered to obtain an alcohol extract. The alcohol extract was dispersed in deionized water to prepare an 8% (w / w) suspension. 3.5% (w / w) of glucose was added as a carbon source, and the mixture was stirred thoroughly. *Lactobacillus plantarum* was inoculated at 5% (w / w) of the total substrate mass. Anaerobic fermentation was carried out at 38℃ for 36h. After fermentation, the mixture was inactivated at 90℃ for 10min, then dried at 65℃ to a moisture content of 7%, pulverized, and passed through a 90-mesh sieve to obtain the fermentation complex.

[0067] Milk fat globule membrane concentrate was dispersed in deionized water to prepare an 8% (w / w) suspension. The pH was adjusted to 7.0, and neutral protease was added at a concentration of 1.0% of the protein mass in the milk fat globule membrane concentrate. Enzymatic hydrolysis was carried out at 50°C for 2 hours, followed by enzyme inactivation at 92°C for 8 minutes. After cooling to room temperature, the undigested solid residue was removed by microfiltration through a ceramic membrane with a pore size of 0.8 μm at a pressure of 0.35 MPa. The filtrate was collected, concentrated under vacuum to a solid content of 20%, and spray-dried at an inlet air temperature of 155°C and an outlet air temperature of 80°C to obtain a directional enzymatic hydrolysis complex powder.

[0068] Phosphatidylserine was dissolved in anhydrous ethanol to prepare a solution with a mass concentration of 15 mg / mL. N-acetylneuraminic acid was dissolved in deionized water to prepare a solution with a mass concentration of 20 mg / mL. The directional enzymatic hydrolysis complex powder was dispersed in deionized water to prepare a suspension with a mass concentration of 35 mg / mL. The fermentation complex was dispersed in a 40% (v / v) ethanol aqueous solution to prepare a suspension with a mass concentration of 20 mg / mL. Under the conditions of stirring speed of 500 r / min and temperature of 40℃, the ethanol suspension of the fermentation complex was first added dropwise to the suspension of the directional enzymatic hydrolysis complex, and stirred for 45 min. Then, the ethanol solution of phosphatidylserine was added dropwise to the mixture, and stirred for 45 min. Finally, the aqueous solution of N-acetylneuraminic acid was added dropwise to the mixture, and stirred for 45 min. After the addition was completed, stirring was continued for 1.5 h to form a suspension of surface-functionalized nanoparticles.

[0069] A surface-functionalized nanoparticle suspension was used as the core material. Sodium caseinate was dissolved in deionized water to prepare an outer wall material solution with a mass concentration of 30 mg / mL. The mass ratio of the core material suspension to the outer wall material solution was 1:3. Under the conditions of stirring speed of 300 r / min and temperature of 40℃, the surface-functionalized nanoparticle suspension was added dropwise to the sodium caseinate solution and stirred for 45 min. Then, the pH was adjusted to 4.5 with 1 mol / L citric acid. Sodium caseinate underwent a coagulation reaction to form a dense gel layer that encapsulated the nanoparticles. Stirring was continued for 45 min. After centrifugation, the nanoparticles were washed three times with deionized water and redispersed in deionized water to prepare a suspension with a mass concentration of 15%. Then, spray drying was performed with an inlet air temperature of 160℃ and an outlet air temperature of 80℃ to obtain microencapsulated nanoparticle powder.

[0070] Weigh out 12 parts of fermentation complex, 10 parts of directional enzymatic hydrolysis complex, 2 parts of phosphatidylserine, 1.5 parts of N-acetylneuraminic acid, 4 parts of walnut peptide, and food-grade excipients. The average molecular weight of the walnut peptide is 1500 Da. Mix the microencapsulated nanoparticle powder with the walnut peptide and food-grade excipients, add deionized water to prepare a mixture with a solid content of 20%, homogenize twice at 55℃ and 25MPa, degas under vacuum, sterilize at 135℃ for 5 seconds, and spray dry to obtain a powdered solid beverage.

[0071] Comparative Example 1 Compared with Example 1, the difference is that glucose is not added as a carbon source in the fermentation step, while the other steps and parameters are the same as in Example 1, and a liquid beverage is finally obtained.

[0072] Comparative Example 2 Compared with Example 2, the difference is that alkaline protease is used instead of neutral protease in the preparation step of the directional enzymatic hydrolysis complex, and the enzymatic hydrolysis pH is 8.5. The remaining steps and parameters are the same as in Example 2, and the final product is a powdered solid beverage.

[0073] Comparative Example 3 Compared with Example 3, the difference is that in the co-assembly step, the fermentation complex ethanol suspension, phosphatidylserine ethanol solution, N-acetylneuraminic acid aqueous solution and the directional enzymatic hydrolysis complex suspension are simultaneously poured into the mixing container, instead of being added dropwise. The remaining steps and parameters are the same as in Example 3, and a liquid beverage is finally obtained.

[0074] Comparative Example 4 Compared with Example 4, the difference is that the sodium caseinate microcapsule encapsulation treatment is not performed. Instead, the surface-functionalized nanoparticle suspension is directly mixed with walnut peptides and food-grade excipients. The remaining steps and parameters are the same as in Example 4, and a powdered solid beverage is finally obtained.

[0075] Comparative Example 5 Compared with Example 1, the difference is that the four raw materials are poured in and mixed at the same time in the co-assembly step, and the average molecular weight of the walnut peptide used is 5000 Da. The remaining steps and parameters are the same as in Example 1, and a liquid beverage is finally obtained.

[0076] Performance Test Results and Analysis The finished products obtained from each embodiment and comparative example were processed uniformly. All finished products were prepared into test solutions with a solid content of 15%. If the solid content of liquid beverage samples was higher than 15%, they were diluted with deionized water to 15%; powdered solid beverage samples were reconstituted with deionized water to prepare test solutions with a solid content of 15%. All physicochemical tests were performed on the test solutions of the above uniform concentration, or on the original samples of intermediate products from the preparation process.

[0077] The total saponin content of the fermentation complex was determined using the vanillin-perchloric acid colorimetric method; the content of small molecule aglycones was determined using high-performance liquid chromatography (HPLC); and the content of short-chain fatty acids was determined using gas chromatography (GC). The density of glycosylation sites in the directed enzymatic hydrolysis complex was determined using the m-hydroxybiphenyl method; and the content of glycoproteins and phospholipids was determined using the Kjeldahl method. The particle size and zeta potential of nanoparticles in the final product test solution were determined using dynamic light scattering (LPS); and the surface density of N-acetylneuraminic acid was determined using the WGA lectin affinity method. The microcapsule encapsulation efficiency was determined using centrifugation; the wall thickness was determined using scanning electron microscopy (SEM); and the microcapsule release rate was determined using in vitro simulated digestion in simulated gastric and intestinal fluids. The stability of the final product was determined using accelerated centrifugation; the product appearance and taste were evaluated using sensory evaluation; and the retention rate of active ingredients after digestion was determined using in vitro simulated digestion combined with HPLC. Specific test results are shown in the table below. Figure 1 , Figure 2 As shown in Table 1-3.

[0078] Figure 1 The results of the fermentation complex index test are provided by Figure 1 It can be seen that the total saponin content, small molecule aglycone content, and short-chain fatty acid content in the fermentation complexes of Examples 1 to 4 were all maintained at a high level. In Comparative Example 1, due to the absence of glucose as a carbon source, *Lactobacillus plantarum* had insufficient metabolic energy, resulting in low secretion of glycosidases and esterases, leading to incomplete conversion of macromolecular saponins and triterpenes. Consequently, the total saponin content, small molecule aglycone content, and short-chain fatty acid content were significantly lower than in the examples. Although Comparative Examples 2 to 5 changed subsequent steps or individual raw material parameters, the fermentation steps were the same as the corresponding examples, and their fermentation complex indicators were close to those of the examples, but still lower than all examples. This is because process fluctuations in subsequent treatments had a certain impact on the intermediate product testing. Overall, the fermentation process using glucose as a carbon source can effectively promote the metabolism of *Lactobacillus plantarum* and improve the conversion efficiency of active ingredients.

[0079] Figure 2 The results of the targeted enzymatic hydrolysis complex index tests show that the glycosylation site density, glycoprotein content, and phospholipid content of the targeted enzymatic hydrolysis complexes in Examples 1 to 4 are all at relatively high levels. Comparative Example 2 used alkaline protease for enzymatic hydrolysis at pH 8.5. The strongly alkaline environment caused deglycosylation of the N-glycosylation domains of the glycoprotein, resulting in the breakage of a large number of glycosylation sites. Therefore, the glycosylation site density, glycoprotein content, and phospholipid content were significantly lower than those of the examples. Comparative Examples 1, 3, 4, and 5 did not change the enzymatic hydrolysis steps, and their targeted enzymatic hydrolysis complex indexes were close to those of the corresponding examples, but all were lower than those of all examples, indicating that the near-neutral pH mild enzymatic hydrolysis conditions used in this application have a positive effect on preserving glycosylation sites.

[0080] Table 1. Test results of surface-functionalized nanoparticles.

[0081] Table 1 shows the test results of the surface-functionalized nanoparticles. As can be seen from the data in Table 1, the surface-functionalized nanoparticles in Examples 1 to 4 have a uniform particle size distribution, a large absolute value of the Zeta potential, and a high surface density of N-acetylneuraminic acid. In Comparative Examples 3 and 5, because the components were added simultaneously rather than sequentially, the components could not achieve spatially ordered distribution through hydrophobic interactions and hydrogen bonds, resulting in disordered self-assembly and thus larger nanoparticle sizes, smaller absolute values ​​of the Zeta potential, and lower surface density of N-acetylneuraminic acid. In Comparative Example 2, the enzymatic hydrolysis conditions destroyed the glycosylation sites, resulting in a lack of binding sites for N-acetylneuraminic acid, significantly reducing the surface density and increasing the particle size due to weak interfacial bonding. In Comparative Example 1, due to the slightly lower quality of the fermentation complex, the nanoparticle indicators were slightly lower than those of the examples. Although the co-assembly steps were not changed in Comparative Example 4, the lack of microencapsulation resulted in the nanoparticles being subjected to mechanical shearing and thermal shock during subsequent homogenization and sterilization processes. This led to some aggregation and detachment of surface groups, resulting in an increase in nanoparticle size, a decrease in the absolute value of the Zeta potential, and a decrease in the surface density of N-acetylneuraminic acid in the final product.

[0082] Table 2 Test results of microencapsulated nanoparticles Example 1 120 84 3.5 18 78 Example 2 135 83 3.2 20 76 Example 3 110 86 4.0 15 82 Example 4 100 89 4.5 12 85 Comparative Example 1 140 82 3.0 21 75 Comparative Example 2 155 80 2.8 22 72 Comparative Example 3 170 78 2.5 25 68 Comparative Example 4 - - - - - Comparative Example 5 180 77 2.2 28 65 Table 2 shows the test results of the microencapsulated nanoparticles. It can be seen that the microencapsulated nanoparticles in Examples 1 to 4 have high encapsulation rates and moderate wall thickness, exhibiting low release rates in simulated gastric fluid but high cumulative release rates in simulated intestinal fluid. The microencapsulated nanoparticles in Comparative Examples 1, 2, 3, and 5 have lower encapsulation rates and thinner wall thicknesses due to differences in core material quality or assembly methods, resulting in poor gastric acid protection and lower intestinal fluid release rates. Comparative Example 4 did not undergo microencapsulation treatment, and no data related to microencapsulated nanoparticles were available. During subsequent direct processing, the active ingredients in its surface-functionalized nanoparticle suspension were significantly degraded in the gastric fluid environment, making it impossible to determine effective microencapsulation release indicators.

[0083] Table 3 Final Product Performance Test Results Example 1 2.5 88 75 Example 2 3.0 85 72 Example 3 2.0 90 78 Example 4 1.5 92 82 Comparative Example 1 4.8 76 62 Comparative Example 2 5.2 74 60 Comparative Example 3 6.5 70 56 Comparative Example 4 8.5 65 50 Comparative Example 5 7.0 68 55 As shown in Table 3, the final products of Examples 1 to 4 exhibited low centrifugal sedimentation rates, high sensory scores, and high in vitro digestion retention rates. Comparative Example 1, due to insufficient fermentation, had low active ingredient content and poor stability, resulting in a high sedimentation rate and low sensory scores and retention rates. Comparative Example 2 suffered from poor quality of the directional enzymatic hydrolysis complex, leading to poor nanoparticle assembly and decreased product stability. Comparative Examples 3 and 5, due to improper co-assembly methods, resulted in large nanoparticle sizes and poor dispersibility, leading to increased product sedimentation rates and decreased sensory quality and active ingredient retention rates. Comparative Example 4, lacking microencapsulation, experienced significant release and degradation of active ingredients in simulated gastric juice, resulting in the lowest in vitro digestion retention rate, and its sensory score was also reduced due to poor product stability. In summary, the multi-step process employed in this application—including fermentation with added sugar, gentle enzymatic hydrolysis to retain glycosylation sites, sequential dropwise co-assembly, and sodium caseinate microencapsulation—effectively improves the active ingredient content, stability, and bioavailability of the nutritional composition.

[0084] This application describes a concentration-enhancing nutritional composition and its preparation method that utilizes ethanol reflux extraction and enrichment, fermentation and transformation with Lactobacillus plantarum, mild enzymatic hydrolysis with neutral protease to retain glycosylation sites, sequential dropwise addition and co-assembly to form core-shell nanoparticles, encapsulation with sodium caseinate microcapsules to achieve gastric acid protection and intestinal sustained release, and synergistic enhancement with walnut peptides.

[0085] The above description represents the preferred embodiments of the present invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principles of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A nutritional composition for improving concentration, characterized in that, The composition comprises, by weight, the following components: 8-15 parts of fermentation complex, 6-12 parts of directional enzymatic hydrolysis complex, 1-3 parts of phosphatidylserine, 1-2 parts of N-acetylneuraminic acid, and 2-5 parts of walnut peptide.

2. The nutritional composition for improving concentration according to claim 1, characterized in that, The preparation method of the fermentation complex includes the following steps: drying jujube seed, poria cocos, and lily bulb separately to a moisture content of ≤8%, coarsely pulverizing them, passing them through a 40-60 mesh sieve, mixing the obtained coarse jujube seed powder, coarse poria cocos powder, and coarse lily bulb powder, adding an ethanol aqueous solution for reflux extraction, filtering and collecting the filtrate, repeating the extraction once with the filter residue, combining the filtrates, and vacuum concentrating them at 60-70℃ to a relative density of 1.05-1.15, recovering the ethanol, and obtaining the alcohol extract. The alcohol extract was dispersed in deionized water to prepare a suspension with a mass concentration of 5%-10%. 2%-5% glucose was added as a carbon source to the suspension. The mixture was stirred evenly and inoculated with Lactobacillus plantarum for anaerobic fermentation. After fermentation, the mixture was inactivated at 85-95℃ for 5-15 minutes. Subsequently, it was dried at 60-70℃ until the moisture content was ≤7%, and then pulverized through an 80-100 mesh sieve to obtain the fermentation complex.

3. The nutritional composition for improving concentration according to claim 2, characterized in that, The mass ratio of the crude jujube seed powder, crude poria cocos powder, and crude lily powder is 3-5:2-3:1; the volume fraction of the ethanol aqueous solution used in the reflux extraction is 60%-75%, the material-to-liquid ratio is 1:8-1:12, the extraction temperature is 50-60℃, and the extraction time is 2-3 hours; the inoculum amount of Lactobacillus plantarum in the anaerobic fermentation is 3%-7% of the total mass of the fermentation substrate, and the anaerobic fermentation is carried out at a temperature of 35-42℃ for 24-48 hours.

4. The nutritional composition for improving concentration according to claim 1, characterized in that, The preparation method of the directional enzymatic hydrolysis complex includes the following steps: dispersing milk fat globule membrane concentrate in deionized water to prepare a suspension with a mass concentration of 5%-10%, adjusting the pH to 6.5-7.5, adding neutral protease for enzymatic hydrolysis, then maintaining at 90-95℃ for 5-10 min to inactivate the enzyme, cooling to room temperature, and removing unhydrolyzed solid residue by microfiltration through a ceramic membrane with a pore size of 0.45-1.0 μm under a pressure of 0.2-0.5 MPa, collecting the filtrate, concentrating it under vacuum to a solid content of 15%-25%, and spray drying at an inlet air temperature of 150-160℃ and an outlet air temperature of 75-85℃ to obtain the directional enzymatic hydrolysis complex powder.

5. The nutritional composition for improving concentration according to claim 4, characterized in that, The total phospholipid content of the milk fat globule membrane concentrate is ≥6%, and the glycoprotein content is ≥15%; the amount of neutral protease added is 0.3%-1.5% of the protein mass in the milk fat globule membrane concentrate, and enzymatic hydrolysis is carried out at a temperature of 45-55℃ for 1-3 hours.

6. The nutritional composition for improving concentration according to claim 1, characterized in that, The walnut peptides have an average molecular weight of 500-3000 Da, a peptide content of ≥75%, and a protein content of ≥80%.

7. A method for preparing a nutritional composition for improving concentration as described in any one of claims 1-6, characterized in that, The method includes the following steps: the fermentation complex, the directional enzymatic hydrolysis complex, phosphatidylserine, and N-acetylneuraminic acid are co-assembled and surface-functionalized to form a surface-functionalized nanoparticle suspension. The surface-functionalized nanoparticle suspension is then treated with sodium caseinate microencapsulation to obtain microencapsulated nanoparticle powder. The microencapsulated nanoparticle powder is mixed with walnut peptide and food-grade excipients, and deionized water is added to prepare a mixture with a solid content of 15%-25%. The mixture is homogenized twice at a temperature of 50-60℃ and a pressure of 20-30MPa, and then vacuum degassed and sterilized at 135℃ for 5 seconds to obtain the finished product of a nutritional composition for improving concentration.

8. The method for preparing a nutritional composition for improving concentration according to claim 7, characterized in that, The process of forming a surface-functionalized nanoparticle suspension by co-assembling and surface functionalizing the fermentation complex, the directed enzymatic hydrolysis complex, phosphatidylserine, and N-acetylneuraminic acid specifically includes the following steps: dissolving phosphatidylserine in anhydrous ethanol to prepare a solution with a mass concentration of 10-20 mg / mL; dissolving N-acetylneuraminic acid in deionized water to prepare a solution with a mass concentration of 15-25 mg / mL; dispersing the directed enzymatic hydrolysis complex powder in deionized water to prepare a suspension with a mass concentration of 25-45 mg / mL; and dispersing the fermentation complex in a 30%-volume ... Prepare a suspension with a mass concentration of 15-25 mg / mL using a 50% ethanol aqueous solution. Under the conditions of stirring speed of 400-600 r / min and temperature of 35-45℃, first add the fermentation complex ethanol suspension dropwise to the directional enzymatic hydrolysis complex suspension and stir for 30-60 min. Then, add the phosphatidylserine ethanol solution dropwise to the mixture and stir for 30-60 min. Finally, add the N-acetylneuraminic acid aqueous solution dropwise to the mixture and stir for 30-60 min. After the addition is complete, continue stirring for 1-2 h to form a suspension of surface-functionalized nanoparticles.

9. A method for preparing a nutritional composition for improving concentration according to claim 7, characterized in that, The surface-functionalized nanoparticle suspension is encapsulated with sodium caseinate to obtain microencapsulated nanoparticle powder. The encapsulation process specifically includes the following steps: using the surface-functionalized nanoparticle suspension as the core material, dissolving sodium caseinate in deionized water to prepare an outer wall material solution with a mass concentration of 20-40 mg / mL, and maintaining a mass ratio of the surface-functionalized nanoparticle suspension (core material) to the outer wall material solution of 1:2-1:

4. The surface-functionalized nanoparticles are then encapsulated under stirring conditions of 200-400 r / min and a temperature of 35-45℃. The rice particle suspension was added dropwise to a sodium caseinate solution and stirred for 30-60 minutes. Then, the pH was adjusted to 4.0-5.0 with 1 mol / L citric acid. The sodium caseinate underwent a coagulation reaction, forming a dense gel layer that encapsulated the nanoparticles. Stirring was continued for another 30-60 minutes. The mixture was collected by centrifugation, washed 2-3 times with deionized water, and redispersed in deionized water to prepare a suspension with a mass concentration of 10%-20%. Then, it was spray-dried at an inlet air temperature of 150-170℃ and an outlet air temperature of 70-85℃ to obtain microencapsulated nanoparticle powder.

10. The application of a nutritional composition for improving concentration as described in any one of claims 1-6, characterized in that, The nutritional composition is used to manufacture functional foods or health products that improve concentration.

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