Antioxidant composition based on aronia and use in dry eye prevention products

Abstract

This invention provides an antioxidant composition based on maquist and its application in dry eye prevention products. By weight, the composition includes: 8-18 parts of maquist anthocyanin extract, 4-9 parts of hyaluronic acid-chitosan grafted phospholipids, 12-22 parts of phosphatidylcholine, 3-6 parts of liposome stabilizer, 2-4 parts of synergistic antioxidant, 3-8 parts of osmotic pressure regulator, 0.1-0.3 parts of preservative, 2-5 parts of sodium hyaluronate, 1-3 parts of taurine, 0.5-1.5 parts of vitamin B6, 0.3-0.8 parts of lutein, and 30-60 parts of solvent. This invention provides an antioxidant composition and dry eye prevention products with good stability, safety, and practicality.

Description

Technical Field

[0001] This invention relates to the field of dry eye prevention products technology, specifically to an antioxidant composition based on maquiberry and its application in dry eye prevention products. Background Technology

[0002] Dry eye is a common ocular surface disease, and its occurrence is closely related to increased oxidative stress on the ocular surface, abnormal tear quality, and ocular surface inflammation. Therefore, protecting ocular surface cells and improving tear performance through antioxidant means has become an important direction for dry eye prevention. As a natural fruit rich in anthocyanins, the extract of maquiberry has been proven to have excellent antioxidant activity. Existing technologies have begun to use maquiberry-related components in the preparation of antioxidant compositions. However, for the specific ocular application scenario of dry eye prevention, further optimization is needed to improve the stability of the active ingredients, ocular surface retention, and overall efficacy of these compositions.

[0003] While existing maquilli-based antioxidant compositions can exert some free radical scavenging effects, their application in dry eye prevention products has significant limitations. Firstly, the anthocyanins and other active ingredients in maquilli are prone to degradation during storage and use, leading to a loss of antioxidant activity. Furthermore, the compositions have weak adhesion to the ocular surface, making it difficult to sustain their effect on ocular tissues. Secondly, some compositions focus only on a single antioxidant function, lacking synergistic effects with anti-inflammatory and tear film performance improvements. Additionally, the osmotic pressure of some products is incompatible with human tears, easily irritating sensitive ocular surfaces. These problems significantly reduce the applicability and effectiveness of existing compositions in dry eye prevention, failing to meet the requirements for stability, safety, and comprehensive efficacy in ophthalmic products. Summary of the Invention

[0004] To address the problems existing in the prior art, the present invention provides an antioxidant composition based on maquist and its application in dry eye prevention products, thereby obtaining an antioxidant composition and dry eye prevention products with good stability, safety and practicality.

[0005] To achieve the above objectives, the present invention provides the following technical solution:

[0006] This application discloses an antioxidant composition based on maquiberry, comprising, by weight, the following raw materials: 8-18 parts of maquiberry anthocyanin extract, 4-9 parts of hyaluronic acid-chitosan grafted phospholipids, 12-22 parts of phosphatidylcholine, 3-6 parts of liposome stabilizer, 2-4 parts of synergistic antioxidant, 3-8 parts of osmotic pressure regulator, 0.1-0.3 parts of preservative, 2-5 parts of sodium hyaluronate, 1-3 parts of taurine, 0.5-1.5 parts of vitamin B6, 0.3-0.8 parts of lutein, and 30-60 parts of solvent.

[0007] By implementing the above technical solutions, the masquilli anthocyanin extract can exert antioxidant effects, which helps to scavenge reactive oxygen species to protect ocular surface cells; hyaluronic acid-chitosan-grafted phospholipids can enhance the encapsulation effect of the masquilli anthocyanin extract, reduce its degradation, and prolong the adhesion and retention time of the composition on the ocular surface; phosphatidylcholine can form a liposome carrier to encapsulate the active ingredients and prevent their oxidative degradation; liposome stabilizers can maintain the integrity of the liposome membrane structure; synergistic antioxidants can help enhance the overall antioxidant effect; and osmotic pressure regulators can make the osmotic pressure of the composition compatible with the ocular surface environment. The ingredients are well-matched and reduce irritation; preservatives inhibit microbial growth and ensure the stability of the composition during storage; sodium hyaluronate enhances the moisturizing effect on the ocular surface and further prolongs adhesion time; taurine protects ocular surface epithelial cells and alleviates cell damage; vitamin B6 participates in the regulation of ocular surface cell metabolism and helps maintain normal cell function; lutein protects the retinal tissue of the ocular surface and filters harmful light; the solvent ensures uniform dispersion of each component and guarantees the stability of the composition system. The synergistic effect of each component can achieve antioxidant protection of the ocular surface, alleviate discomfort, and maintain the normal physiological state of the ocular surface.

[0008] Preferably, the liposome stabilizer is cholesterol or stearoyl lysophosphatidylcholine, the synergistic antioxidant is vitamin E or ascorbate palmitate, the osmotic pressure regulator is sorbitol or mannitol, and the solvent is water for injection (high-purity sterile water specifically used for pharmaceutical production or medical device preparation).

[0009] By setting up the above technical solutions, the liposome stabilizer (cholesterol or stearoyl lysophosphatidylcholine) can maintain the integrity of the liposome membrane structure, prevent the liposome particle size from increasing during storage, and ensure the stable encapsulation effect of the liposome on the active ingredient; the synergistic antioxidant (vitamin E or ascorbate palmitate) can embed itself in the liposome membrane to scavenge free radicals, inhibit the oxidative degradation of the liposome membrane, prolong the half-life of the liposome, and help maintain the stability of the active ingredient in the composition; the osmotic pressure regulator (sorbitol or mannitol) can adjust the osmotic pressure of the composition to a range that matches the tear film, avoid irritation to the ocular surface during use, and improve the ophthalmic safety of the composition; the solvent (water for injection) meets the sterility requirements of ophthalmic preparations and can be used as a dispersion medium to ensure the uniform dispersion of the liposome stabilizer, synergistic antioxidant, osmotic pressure regulator and other components, ensuring the overall dispersion uniformity and safety of the composition.

[0010] The preferred extraction method for marijuana anthocyanin extract is as follows:

[0011] (1) Take fresh Chilean Patagonian maquis fruits (maturity ≥90%), wash and crush them to a particle size of 3-5 mm, add 3%-5% of the weight of the crushed maquis fruits with mixed microbial agent, and anaerobic ferment at 37-40℃ for 8-12 h.

[0012] The mixed microbial agent is composed of *Lactobacillus plantarum* bacterial solution and *Saccharomyces cerevisiae* bacterial solution at a volume ratio of 2:1, with the *Lactobacillus plantarum* bacterial solution having a viable count ≥1×10⁻⁶. 9 CFU / mL, viable count of Saccharomyces cerevisiae in the culture ≥1×10⁻⁶ 8 CFU / mL;

[0013] (2) Add water for injection to the fermentation material obtained in (1) at a mass-volume ratio of 1g:18-25mL, and then add 0.5%-1.0% of the weight of crushed fresh maquist fruit of compound enzyme. Under ultra-high pressure of 500-600MPa and 35-45℃ for 15-25min, and then under ultrasonic treatment of 250-300W for 15-20min.

[0014] (3) Filter the extract obtained in (2) through a sterile microfiltration membrane of 0.20-0.22 μm, load the filtrate onto AB-8 macroporous resin, elute with 50%-70% ethanol solution, and collect the eluent;

[0015] (4) The eluent is vacuum concentrated to a solid content of 30%-35% at 45-50℃ and 0.08-0.09MPa, and then spray-dried at an inlet air temperature of 125-135℃ and an outlet air temperature of 65-75℃ to obtain the anthocyanin extract of macquip.

[0016] By setting up the above technical solution, anaerobic fermentation of highly mature fresh Maquistberries is carried out using a liquid of *Lactobacillus plantarum* with a specific number of viable bacteria and a liquid of *Saccharomyces cerevisiae* (mass ratio 2:1). This allows for the initial disruption of the Maquistberry cell wall by enzymes secreted by the microorganisms, creating conditions for anthocyanin release. Subsequently, combined with water for injection and a complex enzyme, the cavitation effect of 500-600 MPa ultra-high pressure and the synergistic effect of 250-300 W ultrasonic treatment further efficiently break down the cell wall, promoting the full dissolution of Maquistberry anthocyanins while reducing oxidative degradation of anthocyanins during extraction. Further purification using a 0.20-0.22 μm sterile microfiltration membrane and AB-8 macroporous resin removes impurities and improves the purity of the extract. Finally, vacuum concentration at 45-50℃ (avoiding high-temperature damage) and spray drying at specific inlet and outlet air temperatures effectively preserve the activity of Maquistberry anthocyanins while solidifying the extract, ensuring the extraction rate, purity, and activity of the Maquistberry anthocyanin extract.

[0017] Preferably, in step (2), the complex enzyme is composed of cellulase, pectinase and β-glucosidase in a mass ratio of (2-3):(1-2):1; in step (3), the flow rate of the filtrate is 1.5-2.5 BV / h (the volume of filtrate passing through the resin column per hour is 1.5-2.5 times the volume of the resin bed), and the flow rate of the ethanol solution is 0.8-1.2 BV / h.

[0018] By setting up the above technical solution, the compound enzyme is compounded with cellulase, pectinase, and β-glucosidase in a mass ratio of (2-3):(1-2):1. The three enzymes can specifically decompose the cellulose and pectin components of the cell wall of *Mallotus arvensis* and the glycosidic bonds binding anthocyanins. Under synergistic action, they can more fully destroy the cell wall structure and promote the release of bound anthocyanins, effectively improving the dissolution efficiency of anthocyanins. The filtrate is loaded onto AB-8 macroporous resin at a flow rate of 1.5-2.5 BV / h to ensure sufficient contact between the filtrate and the resin, so that the anthocyanins are efficiently adsorbed by the resin, avoiding insufficient adsorption due to excessive flow rate. Elution with 50%-70% ethanol solution at a flow rate of 0.8-1.2 BV / h can ensure that the ethanol and the anthocyanins adsorbed by the resin can fully interact to achieve complete elution, and can also reduce the amount of impurities eluted with the anthocyanins. The combined effect of the three can significantly improve the yield and purity of *Mallotus arvensis* anthocyanin extract.

[0019] Preferably, the raw materials for hyaluronic acid-chitosan grafted phospholipids, by weight, include: 5-7 parts hyaluronic acid, 3-5 parts chitosan, 10-12 parts phosphatidylcholine, 1-2 parts 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, and 50-60 parts water for injection.

[0020] By setting up the above technical solution, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide can effectively activate the carboxyl group of hyaluronic acid, creating conditions for the reaction between hyaluronic acid and chitosan. The hyaluronic acid-chitosan grafted phospholipid formed by the combination of this graft and phosphatidylcholine can enhance the binding ability to the ocular surface by taking advantage of the adhesive properties of hyaluronic acid, and enhance the encapsulation ability of active ingredients through the interaction between chitosan and phosphatidylcholine. Using water for injection as a solvent can ensure the uniform dispersion of each raw material, provide a stable environment for the reaction between each component, and ensure the smooth preparation of hyaluronic acid-chitosan grafted phospholipid. Using water for injection as a solvent can ensure the uniform dispersion of each raw material and meet the sterility requirements to ensure the purity of the product. Ultimately, the prepared hyaluronic acid-chitosan grafted phospholipid has the technical effect of enhancing the encapsulation rate of liposomes and the adhesion to the ocular surface.

[0021] Preferably, the preparation method of hyaluronic acid-chitosan grafted phospholipids includes the following steps:

[0022] 1) Dissolve hyaluronic acid in water for injection, add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, and then stir at 30-32℃ for 28-32 min;

[0023] 2) Add chitosan to the solution obtained in 1) to adjust the pH to 5.5-6.0, and react at a constant temperature of 34-36℃ for 6-8 hours to form hyaluronic acid-chitosan graft.

[0024] 3) Add phosphatidylcholine to the hyaluronic acid-chitosan graft and sonicate for 18-22 minutes at a temperature of 38-42℃ and a power of 180-220W to obtain the reaction solution;

[0025] 4) The reaction solution was filtered through a sterile microfiltration membrane of 0.20-0.22 μm, and then freeze-dried under vacuum at -45~-35℃ and 0.01-0.02 MPa for 8-12 hours. The solution was then pulverized and passed through a 120-125 mesh sieve to obtain hyaluronic acid-chitosan grafted phospholipids.

[0026] By setting up the above technical solution, step 1) can effectively activate the carboxyl group of hyaluronic acid, laying the foundation for subsequent reactions; step 2) can promote the full combination of hyaluronic acid and chitosan to form a stable hyaluronic acid-chitosan graft; in step 3), the ultrasonic action can make phosphatidylcholine uniformly grafted onto the surface of the graft, ensuring the grafting effect; in step 4), impurities are removed by filtration through a 0.20-0.22μm sterile microfiltration membrane, and then the product is freeze-dried at -45~-35℃ and 0.01-0.02MPa to retain the activity of the product and avoid structural damage. Finally, it is pulverized through a 120-125 mesh sieve to ensure uniform particle size. The whole process finally produces a hyaluronic acid-chitosan grafted phospholipid with stable structure and qualified performance, which can achieve the technical effect of enhancing the encapsulation rate of liposomes and the adhesion of ocular surface.

[0027] Preferably, the molecular weight of hyaluronic acid is 1000-1200kDa, and the degree of deacetylation of chitosan is ≥95%.

[0028] By setting up the above technical solution, hyaluronic acid with a molecular weight of 1000-1200kDa can retain its good adhesion properties, laying the foundation for improving the adhesion of the ocular surface in the future; chitosan with a degree of deacetylation ≥95% contains sufficient amino groups, which can easily form stable grafts with activated hyaluronic acid, and can also form hydrogen bonds with phosphatidylcholine to promote the grafting of phosphatidylcholine to the surface of the graft.

[0029] Preferably, the preservative is one of phenoxyethanol or sodium perborate.

[0030] By setting up the above technical solution, phenoxyethanol or sodium perborate can be used as preservatives to effectively inhibit the growth and reproduction of microorganisms in the antioxidant composition, ensuring the microbial stability of the composition during storage and use, and avoiding deterioration of the composition or eye infection caused by microbial contamination. At the same time, both are suitable for the application scenarios of ophthalmic preparations and will not cause significant irritation to the ocular surface tissue, ensuring the safety of the composition for ocular use.

[0031] Preferably, the method for preparing the berry-based antioxidant composition includes the following steps:

[0032] S1. Take phosphatidylcholine, liposome stabilizer, synergistic antioxidant, maquillia anthocyanin extract and hyaluronic acid-chitosan grafted phospholipids, add chloroform-methanol mixture, and rotary evaporate to form a uniform film at 45-50℃ and 0.07-0.08MPa, with a rotary evaporation speed of 50-60r / min.

[0033] The chloroform-methanol mixture is prepared by mixing chloroform and methanol in a volume ratio of 2:1 to 3:1, and the mass-volume ratio of the five raw materials to be dissolved to the chloroform-methanol mixture is 1g:8-12mL.

[0034] S2. Add an aqueous solution of osmotic pressure regulator to the material obtained in S1, hydrate by stirring in a water bath at 38-40℃ for 38-42 minutes, and homogenize under high pressure at 40-60MPa 4-6 times to obtain anthocyanin-modified liposomes.

[0035] The aqueous solution of the osmotic pressure regulator is composed of an osmotic pressure regulator and water for injection in a mass-to-volume ratio of 1g:15-25mL.

[0036] S3. Mix anthocyanin-modified liposomes with sodium hyaluronate, taurine, vitamin B6, lutein, and preservatives, add the remaining water for injection, and then stir at 580-620 r / min for 18-22 min. Filter through a 0.20-0.22 μm sterile microfiltration membrane and dispense under sterile conditions to obtain the anti-oxidative composition based on maquist.

[0037] By setting up the above technical solution, phosphatidylcholine, liposome stabilizers and other components are formed into a uniform liposome membrane through rotary evaporation, which effectively encapsulates the anthocyanin extract of masquillberry. Combined with high-pressure homogenization, anthocyanin-modified liposomes with uniform particle size are obtained, improving the dispersion stability of the composition. Sodium hyaluronate, taurine, vitamin B6, lutein and the core active components work synergistically to enhance the antioxidant, moisturizing and eye care effects of the composition. Preservatives can inhibit the growth of microorganisms and ensure the storage stability of the composition. Subsequent aseptic microfiltration and aseptic dispensing processes ensure that the composition meets the aseptic requirements of ophthalmic preparations, and finally, a stable, ophthalmic-based antioxidant composition is obtained.

[0038] This application also discloses the use of a maquiberry-based antioxidant composition in dry eye prevention products (such as ophthalmic gels or eye drops).

[0039] By setting up the above technical solution, when the antioxidant composition based on marijuana is applied to dry eye prevention products such as ophthalmic gels or eye drops, the dosage form of ophthalmic preparations can make the composition act evenly on the ocular surface. The marijuana anthocyanin extract contained therein can effectively remove reactive oxygen species on the ocular surface, inhibit corneal epithelial cell apoptosis, and reduce tear hyperosmolarity. It works synergistically with other components to exert anti-inflammatory, moisturizing, and ocular surface adhesion time-prolonging effects, thereby relieving ocular surface oxidative stress and inflammatory response, improving tear secretion and tear film stability, reducing the occurrence of dry eye-related discomfort symptoms, and achieving effective prevention of dry eye.

[0040] The beneficial effects of this invention are as follows:

[0041] Maquillic acid anthocyanin extract exerts antioxidant effects, helping to scavenge reactive oxygen species and protect ocular surface cells. Hyaluronic acid-chitosan-grafted phospholipids enhance the encapsulation effect of the maquillic acid anthocyanin extract, reducing its degradation and prolonging the adhesion and retention time of the composition on the ocular surface. Phosphatidylcholine forms a liposome carrier to encapsulate the active ingredients, preventing their oxidative degradation. Liposome stabilizers maintain the integrity of the liposome membrane structure. Synergistic antioxidants help enhance the overall antioxidant effect. Osmotic pressure regulators match the osmotic pressure of the composition with the ocular surface environment, reducing irritation. Preservatives inhibit microbial growth and ensure the storage stability of the composition. Sodium hyaluronate enhances the moisturizing effect of the ocular surface and further prolongs the adhesion time. Taurine protects ocular surface epithelial cells and alleviates cell damage. Vitamin B6 participates in the metabolic regulation of ocular surface cells and helps maintain normal cell function. Lutein protects the retinal tissue of the ocular surface and filters harmful light. Solvents ensure uniform dispersion of the components and maintain the stability of the composition system. The synergistic effect of the components achieves antioxidant protection of the ocular surface, alleviates discomfort, and maintains the normal physiological state of the ocular surface.

[0042] 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide can effectively activate the carboxyl group of hyaluronic acid, creating conditions for the reaction between hyaluronic acid and chitosan. The hyaluronic acid-chitosan grafted phospholipid formed by combining this graft with phosphatidylcholine can enhance its binding ability to the ocular surface by leveraging the adhesive properties of hyaluronic acid, and enhance the encapsulation ability of active ingredients through the interaction between chitosan and phosphatidylcholine. Using water for injection as a solvent can ensure the uniform dispersion of each raw material, providing a stable environment for the reaction between each component, and ensuring the smooth preparation of hyaluronic acid-chitosan grafted phospholipid. Using water for injection as a solvent can ensure the uniform dispersion of each raw material and meet the sterility requirements to ensure the purity of the product, ultimately enabling the prepared hyaluronic acid-chitosan grafted phospholipid to have the technical effect of enhancing the encapsulation rate of liposomes and the adhesion to the ocular surface. Detailed Implementation

[0043] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, 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.

[0044] Specific information on the raw materials used in the embodiments of this invention is shown in Table 1:

[0045] Table 1. Raw material names and sources

[0046] name Indicators such as components or parameters factory Maquilli anthocyanin extract The content of effective substances is 30%-35%. The preparation method is described below. Hyaluronic acid-chitosan grafted phospholipids 120-125 mesh The preparation method is described below. Phosphatidylcholine 99% purity Shanghai Yuanye Biotechnology Co., Ltd., Product No.: B27836-25mg Liposome stabilizers Cholesterol or stearoyl lysolecithin Cholesterol was purchased from Shanghai Maclean Biochemical Technology Co., Ltd., CAS No.: 57-88-5; Stearoyl lysophosphatidylcholine was purchased from Sigma-Aldrich, model S4403. Synergistic antioxidants Vitamin E or ascorbate palmitate Vitamin E was purchased from Zhejiang Medicine Co., Ltd., product code VE-98%; ascorbate palmitate was purchased from Shanghai Disano Pharmaceutical Group Co., Ltd., product code AP-99%. Osmotic pressure regulator Sorbitol (food grade) or mannitol (pharmaceutical grade) Sorbitol was purchased from Shandong Futian Pharmaceutical Co., Ltd., and mannitol was purchased from Roquette Lianyungang Co., Ltd. preservative Phenoxyethanol or sodium perborate Sodium perborate was purchased from Shanghai Maclean Biochemical Technology Co., Ltd., CAS No.: 122-99-6; and from Jiangsu Congzhong Chemical Co., Ltd., CAS No.: 10486-00-7. Sodium hyaluronate Molecular weight 500-800kDa Bloomage Biotechnology Co., Ltd. Taurine food grade Hangzhou Qianying Biotechnology Co., Ltd. Vitamin B6 Medical grade Zhejiang Xinhecheng Co., Ltd. Lutein Specifications: 5% CWS-S, 5% / 10% TAB-S Zhejiang Medicine Co., Ltd. solvent Water for injection (compliant with the standards of the 2020 edition of the Chinese Pharmacopoeia, Part II) Naipu (Guangzhou) Water Treatment Technology Co., Ltd. Hyaluronic acid Molecular weight 1000-1200kDa Bloomage Biotechnology Co., Ltd. Chitosan Deacetylation degree ≥ 95% Shandong Aokang Biotechnology Co., Ltd. 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide 98% purity Shanghai Yuanye Biotechnology Co., Ltd. Cellulase Model: Celluclast 1.5L Novozymes pectinase Model: Pectinex Ultra SP-L Novozymes β-glucosidase Model G0395 Sigma-Aldrich

[0047] Example 1:

[0048] This embodiment discloses an antioxidant composition based on maquiberry, which, by weight, comprises: 8 parts of maquiberry anthocyanin extract, 4 parts of hyaluronic acid-chitosan grafted phospholipids, 12 parts of phosphatidylcholine, 3 parts of cholesterol, 2 parts of ascorbate palmitate, 3 parts of sorbitol, 0.1 parts of phenoxyethanol, 2 parts of sodium hyaluronate, 1 part of taurine, 0.5 parts of vitamin B6, 0.3 parts of lutein, and 30 parts of water for injection.

[0049] The extraction method for malkiberella anthocyanin extract is as follows:

[0050] (1) Take fresh Maquistberries (maturity ≥90%), wash them, crush them to a particle size of 3mm, add 3% of the weight of the crushed fresh Maquistberries with mixed microbial agent, and anaerobic ferment at 37℃ for 8h.

[0051] The mixed microbial agent is composed of *Lactobacillus plantarum* bacterial solution and *Saccharomyces cerevisiae* bacterial solution at a volume ratio of 2:1, with the *Lactobacillus plantarum* bacterial solution having a viable count ≥1×10⁻⁶. 9 CFU / mL, viable count of Saccharomyces cerevisiae in the culture ≥1×10⁻⁶ 8 CFU / mL;

[0052] (2) Add water for injection to the fermentation material obtained in (1) at a mass-volume ratio of 1g:18mL, and then add 0.5% of the weight of the crushed fresh maquist fruit of the compound enzyme. Under ultra-high pressure treatment at 500MPa and 35℃ for 15min, and then under ultrasonic treatment at 250W for 15min.

[0053] The complex enzyme is composed of cellulase, pectinase and β-glucosidase in a mass ratio of 2:1:1.

[0054] (3) The extract obtained in (2) was filtered through a sterile microfiltration membrane of 0.2 μm. The filtrate was loaded onto AB-8 macroporous resin at a flow rate of 1.5 BV / h and eluted with 50% ethanol solution at a flow rate of 0.8 BV / h. The eluent was collected.

[0055] (4) The eluent was vacuum concentrated to a solid content of 30% at 45°C and 0.08 MPa, and then spray-dried at an inlet air temperature of 125°C and an outlet air temperature of 65°C to obtain the anthocyanin extract of macquip.

[0056] By weight, the raw materials for hyaluronic acid-chitosan grafted phospholipids include: 5 parts hyaluronic acid, 3 parts chitosan, 10 parts phosphatidylcholine, 1 part 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, and 50 parts water for injection; the molecular weight of hyaluronic acid is 1000 kDa, and the degree of deacetylation of chitosan is ≥95%.

[0057] The preparation method of hyaluronic acid-chitosan grafted phospholipids includes the following steps:

[0058] 1) Dissolve hyaluronic acid in water for injection, add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, and then stir at 30°C for 28 min;

[0059] 2) Add chitosan to the solution obtained in 1) to adjust the pH to 5.5, and react at 34℃ for 6 hours to form hyaluronic acid-chitosan graft.

[0060] 3) Add phosphatidylcholine to the hyaluronic acid-chitosan graft and sonicate for 18 minutes at 38°C and 180W to obtain the reaction solution.

[0061] 4) The reaction solution was filtered through a 0.2 μm sterile microfiltration membrane, and then freeze-dried under vacuum at -45℃ and 0.01 MPa for 8 h. The solution was then pulverized and passed through a 120-mesh sieve to obtain hyaluronic acid-chitosan grafted phospholipids.

[0062] The preparation method of the antioxidant composition based on maquilli includes the following steps:

[0063] S1. Take phosphatidylcholine, cholesterol, ascorbate palmitate, maquillia anthocyanin extract and hyaluronic acid-chitosan grafted phospholipids, add chloroform-methanol mixture, and rotary evaporate to form a uniform thin film at 45℃ and 0.07MPa. The rotary evaporation speed is 50r / min.

[0064] The chloroform-methanol mixture is made by mixing chloroform and methanol at a volume ratio of 2:1, and the mass-volume ratio of the five raw materials to be dissolved to the chloroform-methanol mixture is 1g:8mL.

[0065] S2. Add sorbitol aqueous solution to the material obtained in S1, hydrate by stirring in a water bath at 38°C for 38 min, and homogenize under high pressure at 40 MPa 4 times to obtain anthocyanin-modified liposomes.

[0066] The sorbitol aqueous solution is prepared by a mass-volume ratio of sorbitol to water for injection of 1g:15mL.

[0067] S3. Mix anthocyanin-modified liposomes with sodium hyaluronate, taurine, vitamin B6, lutein, and phenoxyethanol, add the remaining water for injection, stir at 580 r / min for 18 min, filter through a 0.20 μm sterile microfiltration membrane, and dispense under sterile conditions to obtain the anti-oxidative composition based on maquist.

[0068] This embodiment describes the application of a maquillberry-based antioxidant composition in an ophthalmic gel.

[0069] Example 2:

[0070] This embodiment discloses an antioxidant composition based on maquiberry, which, by weight, comprises: 18 parts of maquiberry anthocyanin extract, 9 parts of hyaluronic acid-chitosan grafted phospholipids, 22 parts of phosphatidylcholine, 6 parts of cholesterol, 4 parts of vitamin E, 8 parts of mannitol, 0.3 parts of sodium perborate, 5 parts of sodium hyaluronate, 3 parts of taurine, 1.5 parts of vitamin B6, 0.8 parts of lutein, and 60 parts of water for injection.

[0071] The extraction method for malkiberella anthocyanin extract is as follows:

[0072] (1) Take fresh Maquistberries (maturity ≥90%), wash them, crush them to a particle size of 5mm, add 5% of the weight of the crushed fresh Maquistberries with mixed microbial agent, and anaerobic ferment at 40℃ for 12h.

[0073] The mixed microbial agent is composed of *Lactobacillus plantarum* bacterial solution and *Saccharomyces cerevisiae* bacterial solution at a volume ratio of 2:1, with the *Lactobacillus plantarum* bacterial solution having a viable count ≥1×10⁻⁶. 9 CFU / mL, viable count of Saccharomyces cerevisiae in the culture ≥1×10⁻⁶ 8 CFU / mL;

[0074] (2) Add water for injection to the fermentation material obtained in (1) at a mass-volume ratio of 1g:25mL, and then add 1.0% of the weight of the crushed fresh maquist fruit of the compound enzyme. Under ultra-high pressure treatment at a pressure of 600MPa and a temperature of 45℃ for 25min, and then under ultrasonic treatment at a power of 300W for 20min.

[0075] The complex enzyme is composed of cellulase, pectinase and β-glucosidase in a mass ratio of 2:1:1.

[0076] (3) The extract obtained in (2) was filtered through a sterile microfiltration membrane of 0.22 μm. The filtrate was loaded onto AB-8 macroporous resin at a flow rate of 2.5 BV / h and eluted with 70% ethanol solution at a flow rate of 1.2 BV / h. The eluent was collected.

[0077] (4) The eluent was vacuum concentrated to a solid content of 35% at 50°C and 0.09 MPa, and then spray-dried at an inlet air temperature of 135°C and an outlet air temperature of 75°C to obtain the anthocyanin extract of macquip.

[0078] By weight, the raw materials for hyaluronic acid-chitosan grafted phospholipids include: 7 parts hyaluronic acid, 5 parts chitosan, 12 parts phosphatidylcholine, 2 parts 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, and 60 parts water for injection; the molecular weight of hyaluronic acid is 1200 kDa, and the degree of deacetylation of chitosan is ≥95%.

[0079] The preparation method of hyaluronic acid-chitosan grafted phospholipids includes the following steps:

[0080] 1) Dissolve hyaluronic acid in water for injection, add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, and then stir at 32°C for 32 min;

[0081] 2) Add chitosan to the solution obtained in 1) to adjust the pH to 6.0, and react at 36℃ for 8 hours to form hyaluronic acid-chitosan graft.

[0082] 3) Add phosphatidylcholine to the hyaluronic acid-chitosan graft and sonicate for 22 min at 42℃ and 220W to obtain the reaction solution;

[0083] 4) The reaction solution was filtered through a sterile microfiltration membrane of 0.22 μm, and then freeze-dried under vacuum at -35℃ and 0.02 MPa for 12 h. The solution was then pulverized and passed through a 125-mesh sieve to obtain hyaluronic acid-chitosan grafted phospholipids.

[0084] The preparation method of the antioxidant composition based on maquilli includes the following steps:

[0085] S1. Take phosphatidylcholine, cholesterol, vitamin E, maquillia anthocyanin extract and hyaluronic acid-chitosan grafted phospholipids, add chloroform-methanol mixture, and rotary evaporate to form a uniform film at 50℃ and 0.08MPa. The rotary evaporation speed is 60r / min.

[0086] The chloroform-methanol mixture is made by mixing chloroform and methanol in a volume ratio of 3:1, and the mass-volume ratio of the five raw materials to be dissolved to the chloroform-methanol mixture is 1g:12mL.

[0087] S2. Add mannitol aqueous solution to the material obtained in S1, hydrate by stirring in a water bath at 40°C for 42 min, and homogenize under high pressure at 60 MPa 6 times to obtain anthocyanin-modified liposomes.

[0088] The mannitol aqueous solution is prepared by a mass-volume ratio of mannitol to water for injection of 1g:25mL.

[0089] S3. Mix anthocyanin-modified liposomes with sodium hyaluronate, taurine, vitamin B6, lutein, and sodium perborate, add the remaining water for injection, stir at 620 r / min for 22 min, filter through a 0.22 μm sterile microfiltration membrane, and dispense under sterile conditions to obtain the anti-oxidative composition based on maquist.

[0090] This embodiment describes the application of a maquillberry-based antioxidant composition in eye drops.

[0091] Example 3:

[0092] This embodiment discloses an antioxidant composition based on maquiberry, which, by weight, comprises: 13 parts maquiberry anthocyanin extract, 6 parts hyaluronic acid-chitosan grafted phospholipids, 17 parts phosphatidylcholine, 4 parts cholesterol, 3 parts vitamin E, 5 parts mannitol, 0.2 parts phenoxyethanol, 3 parts sodium hyaluronate, 2 parts taurine, 1 part vitamin B6, 0.5 parts lutein, and 45 parts water for injection.

[0093] The extraction method for malkiberella anthocyanin extract is as follows:

[0094] (1) Take fresh Maquistberries (maturity ≥90%), wash them, crush them to a particle size of 4mm, add 4% of the weight of the crushed fresh Maquistberries with mixed microbial agent, and anaerobic ferment at 38℃ for 10h.

[0095] The mixed microbial agent is composed of *Lactobacillus plantarum* bacterial solution and *Saccharomyces cerevisiae* bacterial solution at a volume ratio of 2:1, with the *Lactobacillus plantarum* bacterial solution having a viable count ≥1×10⁻⁶. 9 CFU / mL, viable count of Saccharomyces cerevisiae in the culture ≥1×10⁻⁶ 8 CFU / mL;

[0096] (2) Add water for injection to the fermentation material obtained in (1) at a mass-volume ratio of 1g:21mL, and then add 0.7% of the weight of the crushed fresh maquist fruit of the compound enzyme. Under ultra-high pressure treatment at a pressure of 550MPa and a temperature of 40℃ for 20min, and then under ultrasonic treatment at a power of 275W for 17min.

[0097] The complex enzyme is composed of cellulase, pectinase and β-glucosidase in a mass ratio of 3:2:1.

[0098] (3) The extract obtained in (2) was filtered through a sterile microfiltration membrane of 0.21 μm. The filtrate was loaded onto AB-8 macroporous resin at a flow rate of 2 BV / h and eluted with 60% ethanol solution at a flow rate of 1 BV / h. The eluent was collected.

[0099] (4) The eluent was vacuum concentrated to a solid content of 32% at 47°C and 0.085 MPa, and then spray-dried at an inlet air temperature of 130°C and an outlet air temperature of 70°C to obtain the anthocyanin extract of macquip.

[0100] By weight, the raw materials for hyaluronic acid-chitosan grafted phospholipids include: 6 parts hyaluronic acid, 4 parts chitosan, 11 parts phosphatidylcholine, 1.5 parts 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, and 55 parts water for injection; the molecular weight of hyaluronic acid is 1100 kDa, and the degree of deacetylation of chitosan is ≥95%.

[0101] The preparation method of hyaluronic acid-chitosan grafted phospholipids includes the following steps:

[0102] 1) Dissolve hyaluronic acid in water for injection, add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, and then stir at 31°C for 30 min;

[0103] 2) Add chitosan to the solution obtained in 1) to adjust the pH to 5.8, and react at 35℃ for 7 hours to form hyaluronic acid-chitosan graft.

[0104] 3) Add phosphatidylcholine to the hyaluronic acid-chitosan graft and sonicate for 20 minutes at 40℃ and 200W to obtain the reaction solution;

[0105] 4) The reaction solution was filtered through a sterile microfiltration membrane of 0.21 μm, and then freeze-dried under vacuum at -40℃ and 0.015 MPa for 10 h. After being pulverized and passed through a 122-mesh sieve, hyaluronic acid-chitosan grafted phospholipids were obtained.

[0106] The preparation method of the antioxidant composition based on maquilli includes the following steps:

[0107] S1. Take phosphatidylcholine, cholesterol, vitamin E, maquillia anthocyanin extract and hyaluronic acid-chitosan grafted phospholipids, add chloroform-methanol mixture, and rotary evaporate to form a uniform film at 47℃ and 0.075MPa. The rotary evaporation speed is 55r / min.

[0108] The chloroform-methanol mixture is made by mixing chloroform and methanol in a volume ratio of 3:1, and the mass-volume ratio of the five raw materials to be dissolved to the chloroform-methanol mixture is 1g:10mL.

[0109] S2. Add mannitol aqueous solution to the material obtained in S1, hydrate by stirring in a water bath at 39°C for 40 min, and homogenize under high pressure at 50 MPa 5 times to obtain anthocyanin-modified liposomes.

[0110] The mannitol aqueous solution is prepared by a mass-volume ratio of mannitol to water for injection of 1g:20mL.

[0111] S3. Mix anthocyanin-modified liposomes with sodium hyaluronate, taurine, vitamin B6, lutein, and phenoxyethanol, add the remaining water for injection, stir at 600 r / min for 20 min, filter through a 0.21 μm sterile microfiltration membrane, and dispense under sterile conditions to obtain the anti-oxidative composition based on maquist.

[0112] This embodiment describes the application of a maquillberry-based antioxidant composition in eye drops.

[0113] Comparative Example 1:

[0114] An antioxidant composition based on maquiberry and its application in dry eye prevention products, differing from Example 3 only in that: hyaluronic acid-chitosan grafted phospholipids (HCP) are not added.

[0115] Comparative Example 2:

[0116] An antioxidant composition based on marijuana and its application in dry eye prevention products, differing from Example 3 only in that the marijuana is extracted using a "single ultrasonic method" (without fermentation and ultra-high pressure treatment, power 300-320W, time 40-45min).

[0117] Comparative Example 3:

[0118] An antioxidant composition based on maquist and its application in dry eye prevention products, differing from Example 3 only in that the maquist was not subjected to "double fermentation" during extraction (only crushing was performed, without fermentation pretreatment).

[0119] Comparative Example 4:

[0120] An antioxidant composition based on masquillberry and its application in dry eye prevention products differs from Example 3 only in that the masquillberry anthocyanin extract is not prepared as "anthocyanin-modified liposomes" (masquillberry anthocyanin extract is added directly).

[0121] Comparative Example 5:

[0122] An antioxidant composition based on maquist and its application in dry eye prevention products, differing from Example 3 only in that unmodified phosphatidylcholine is used instead of hyaluronic acid-chitosan-grafted phospholipids (HCP).

[0123] Comparative Example 6:

[0124] An antioxidant composition based on maquiberry and its application in dry eye prevention products, which differs from Example 3 only in that vitamin E (a synergistic antioxidant) is not added.

[0125] Comparative Example 7:

[0126] An antioxidant composition based on maquilli and its application in dry eye prevention products differs from Example 3 only in that the liposomes were not subjected to "high-pressure homogenization" during preparation (only stirring hydration was performed, without high-pressure treatment).

[0127] Comparative Example 8:

[0128] An antioxidant composition based on maquist and its application in dry eye prevention products, differing from Example 3 only in that: no chitosan was added during the preparation of hyaluronic acid-chitosan grafted phospholipid (HCP) (only hyaluronic acid-phospholipid grafting was used).

[0129] Comparative Example 9:

[0130] An antioxidant composition based on marijuana and its application in dry eye prevention products, differing from Example 3 only in that the ultra-high pressure extraction pressure is changed to 300 MPa.

[0131] Comparative Example 10:

[0132] An antioxidant composition based on maquist and its application in dry eye prevention products, which differs from Example 3 only in that phenoxyethanol (preservative) is not added.

[0133] Comparative Example 11:

[0134] An antioxidant composition based on maquist and its application in dry eye prevention products, which differs from Example 3 only in that mannitol (osmotic pressure regulator) is not added.

[0135] The antioxidant compositions based on maquilleria obtained in Examples 1-3 and Comparative Examples 1-11 were subjected to performance tests for total anthocyanin retention rate, liposome encapsulation rate, ROS scavenging rate, tear secretion, ocular surface IL-6 content, ocular surface adhesion time, and tear film breakup time (BUT). The test methods and standards for each performance are as follows:

[0136] 1. Total anthocyanin retention rate: Referring to GB 5009.149-2016 "Determination of anthocyanins in food", 0.5g of sample was dissolved in 10mL of 0.1% hydrochloric acid-methanol solution, sonicated for 30min, and filtered through a 0.22μm filter membrane; high performance liquid chromatography (HPLC) was used for detection. The chromatographic column was a C18 column (250mm×4.6mm), the mobile phase was methanol-0.1% formic acid water (40:60), the flow rate was 1.0mL / min, and the detection wavelength was 520nm; the 30-day retention rate was calculated as follows: Total anthocyanin retention rate (%) = (30-day sample anthocyanin content / initial sample anthocyanin content) × 100%.

[0137] 2. Liposome encapsulation efficiency: Ultrafiltration centrifugation-HPLC was used. 1 mL of liposome suspension was added to an ultrafiltration tube (molecular weight cutoff of 30 kDa), centrifuged at 8000 r / min for 15 min, and the filtrate was used to detect the free anthocyanin content (method is the same as for total anthocyanins); Encapsulation efficiency (%) = (1 - anthocyanin content in filtrate / total anthocyanin content in liposome suspension) × 100%.

[0138] 3. ROS scavenging rate: Human ocular surface epithelial cells (HCE-T) were cultured and oxidative stress model was established by inducing with 100 μmol / L H2O2 for 2 h; samples were added and incubated for 4 h, and stained with 10 μmol / L DCFH-DA for 30 min. Fluorescence intensity was detected by flow cytometry; ROS scavenging rate (%) = (1 - average fluorescence intensity of experimental group / average fluorescence intensity of model group) × 100%.

[0139] 4. Tear secretion: In accordance with the "Guidelines for the Preparation of Commonly Used Animal Models in Ophthalmic Research", a rat dry eye model was established using 0.1% benzalkonium chloride eye drops. The drug was administered twice daily for 14 consecutive days. Schirmer test strips (without surface anesthesia) were placed on the lower palpebral conjunctival sac of the rats, and the wet length of the test strips was measured after 5 minutes, which was the tear secretion (mm / 5min).

[0140] 5. Ocular surface IL-6 content: According to the ELISA kit instructions, rat ocular surface tissue homogenate was taken, centrifuged at 12000 r / min for 20 min at 4℃, and the supernatant was collected; the sample was added, incubated, and color developed according to the kit steps, and the absorbance was measured at 450 nm wavelength using an ELISA reader to calculate the IL-6 concentration (pg / mL).

[0141] 6. Adhesion time of ocular surface: The samples were labeled with sodium fluorescein. After the rabbit eyes were instilled with the drug, samples were taken at 0.5h, 1h and 2h respectively. The fluorescence intensity of the ocular surface was measured by a fluorescence spectrophotometer (excitation wavelength 490nm, emission wavelength 520nm). The adhesion time was defined as the time (h) when the fluorescence intensity dropped to 50% of the initial value.

[0142] 7. Tear film breakup time (BUT): After instilling medication into the rabbit eye, add 0.1% sodium fluorescein eye drops and observe the tear film breakup time under a slit-lamp microscope. The time from blinking to the appearance of the first dry spot on the tear film is the BUT (s).

[0143] The results are shown in Table 2.

[0144] Table 2. Total anthocyanin retention rate, liposome encapsulation rate, and ROS scavenging rate of the antioxidant compositions obtained in Examples 1-3 and Comparative Examples 1-11

[0145] Group Total anthocyanin retention rate (%) Liposome encapsulation efficiency (%) ROS clearance rate (%) Example 1 89 90 88 Example 2 93 94 92 Example 3 96 97 98 Comparative Example 1 73 76 90 Comparative Example 2 69 96 72 Comparative Example 3 76 97 77 Comparative Example 4 66 - 64 Comparative Example 5 79 83 88 Comparative Example 6 86 96 93 Comparative Example 7 91 86 92 Comparative Example 8 81 81 89 Comparative Example 9 83 96 89 Comparative Example 10 96 97 98 Comparative Example 11 96 97 98

[0146] Table 3. Tear secretion, ocular surface IL-6 content, ocular surface adhesion time, and tear film breakup time of the antioxidant compositions obtained in Examples 1-3 and Comparative Examples 1-11.

[0147] Group Tear secretion (mm / 5min) IL-6 levels on the ocular surface (pg / mL) Ocular surface adhesion time (h) Tear film breakup time (s) Example 1 11.2 23 4.6 40 Example 2 12.5 19 5.0 44 Example 3 14.0 16 5.5 50 Comparative Example 1 9.0 21 2.2 26 Comparative Example 2 7.0 36 5.3 29 Comparative Example 3 7.8 31 5.4 33 Comparative Example 4 6.2 40 1.6 23 Comparative Example 5 9.2 23 2.9 31 Comparative Example 6 12.2 18 5.1 41 Comparative Example 7 11.8 17 3.6 36 Comparative Example 8 9.0 22 2.6 28 Comparative Example 9 9.8 24 5.3 39 Comparative Example 10 13.9 16 5.5 50 Comparative Example 11 10.5 17 5.5 32

[0148] Using Example 3 as the control group, the performance differences and causes of Comparative Examples 1-11 are analyzed as follows:

[0149] Comparative Example 1 (without HCP): Total anthocyanin retention decreased from 96% to 73% (a decrease of 24.0%), liposome encapsulation efficiency decreased from 97% to 76% (a decrease of 21.65%), and ocular surface adhesion time decreased from 5.5 h to 2.2 h (a decrease of 60.0%). This is because the lack of hydrogen bond stabilization between the chitosan fragment and phospholipids from HCP leads to liposome aggregation and accelerated anthocyanin degradation; simultaneously, the loss of the binding site between the hyaluronic acid fragment and ocular surface mucin from HCP significantly reduces adhesion ability.

[0150] Comparative Example 2 (using "single ultrasound method" for extracting from maquistberries): The total anthocyanin retention rate decreased from 96% to 69% (a decrease of 28.1%), and the ROS scavenging rate decreased from 98% to 72% (a decrease of 26.5%). This is because single ultrasound cannot adequately disrupt the cellulose and pectin layers of the maquistberry cell wall, resulting in incomplete anthocyanin extraction; furthermore, prolonged ultrasound time exacerbates the oxidative degradation of delphinidin-type anthocyanins, leading to a double decrease in both the content and activity of active ingredients.

[0151] Comparative Example 3 (without "dual fermentation" during extracting from maquistberries): The total anthocyanin retention rate decreased from 96% to 76% (a decrease of 20.8%), and the ROS scavenging rate decreased from 98% to 77% (a decrease of 21.4%). This is because the synergistic effect of pectinase secreted by *Lactobacillus plantarum* and cellulase secreted by *Saccharomyces cerevisiae* resulted in insufficient cell wall disruption and reduced anthocyanin release. Furthermore, the organic acids produced during fermentation were not generated, failing to regulate the pH of the extraction system and further exacerbating anthocyanin degradation.

[0152] Comparative Example 4 (Maquilli anthocyanin extract not formulated as "anthocyanin-modified liposomes"): The total anthocyanin retention rate decreased from 96% to 66% (a decrease of 31.3%), and the ocular surface adhesion time decreased from 5.5 h to 1.6 h (a decrease of 70.9%). This is because maquilli anthocyanins are directly exposed to the aqueous environment without the protective barrier of a liposome bilayer, making them susceptible to oxidative degradation by oxygen free radicals. Furthermore, the lack of compatibility between liposomes and the tear lipid layer prevents the formation of a stable retention layer on the ocular surface. Additionally, anthocyanins not coated with liposomes are easily degraded by esterases in the tear film, further reducing the active ingredient's action time and significantly shortening the adhesion time.

[0153] Comparative Example 5 (using unmodified phosphatidylcholine instead of HCP): Total anthocyanin retention decreased from 96% to 79% (a decrease of 17.7%), and liposome encapsulation efficiency decreased from 97% to 83% (a decrease of 14.4%). This is because unmodified phosphatidylcholine cannot form hydrogen bonds with anthocyanins, resulting in decreased encapsulation capacity; furthermore, the lack of chitosan fragments from HCP inhibits liposome aggregation, leading to increased liposome size and reduced stability.

[0154] Comparative Example 6 (without added Vitamin E): Total anthocyanin retention decreased from 96% to 86% (a decrease of 10.4%), and tear film breakup time decreased from 50s to 41s (a decrease of 18.0%). This is because the lack of Vitamin E's free radical scavenging function within the liposome membrane accelerated oxidative degradation of the liposome membrane; furthermore, Vitamin E's repair function on the tear film lipid layer was absent, leading to decreased tear film stability.

[0155] Comparative Example 7 (liposomes were not homogenized under high pressure during preparation): the liposome encapsulation rate decreased from 97% to 86% (a decrease of 11.3%), and the ocular surface adhesion time decreased from 5.5 h to 3.6 h (a decrease of 34.5%). This was because the lack of high-pressure homogenization resulted in uneven liposome particle size, with some liposomes exceeding 300 nm in diameter (the standard particle size requirement for ocular liposomes is <200 nm), making uniform distribution on the ocular surface impossible. Furthermore, the excessively large particle size reduced the contact area with the ocular surface during adhesion, shortening the retention time.

[0156] Comparative Example 8 (HCP preparation without chitosan): Liposome encapsulation efficiency decreased from 97% to 81% (a decrease of 16.5%), and ocular surface adhesion time decreased from 5.5 h to 2.6 h (a decrease of 52.7%). This is because the lack of amino groups in chitosan prevents it from forming hydrogen bonds with the hydroxyl groups of phosphatidylcholine, thus reducing the encapsulation capacity of the liposomes; furthermore, the mucosal adhesion properties of chitosan are lost, reducing the binding force between HCP and ocular surface mucins.

[0157] Comparative Example 9 (with the ultra-high pressure extraction pressure changed to 300 MPa): The total anthocyanin retention rate decreased from 96% to 83% (a decrease of 13.5%), and tear secretion decreased from 14.0 mm / 5 min to 9.8 mm / 5 min (a decrease of 30.0%). This is because the 300 MPa pressure cannot generate sufficient cavitation effect (reduced number of cavitation bubbles), resulting in inadequate cell wall disruption and a lower anthocyanin extraction rate; furthermore, insufficient retention of active ingredients ultimately leads to a decreased effect on improving lacrimal gland function.

[0158] When no phenoxyethanol (preservative) was added to Comparative Example 10, no microbial contamination occurred in the composition during the short-term storage of 30 days. Therefore, the functional indicators such as total anthocyanin retention rate and liposome encapsulation rate were not significantly different from those of Example 3. However, long-term storage (such as more than 6 months) is prone to the growth of bacteria, fungi and other microorganisms, which can lead to product deterioration and failure, and the safety of eye use cannot be guaranteed.

[0159] Comparative Example 11 (without osmotic pressure regulator): Tear secretion decreased from 14.0 mm / 5 min to 10.5 mm / 5 min (a decrease of 25.0%), and tear film breakup time decreased from 50 s to 32 s (a decrease of 36.0%). This is because the absence of mannitol caused the composition's osmotic pressure to deviate from the physiological range (280-320 mOsm / kg), irritating the ocular surface mucosa and affecting tear secretion and tear film stability. Without mannitol (an osmotic pressure regulator), the composition's osmotic pressure deviated from the physiological range of human tears, irritating the delicate ocular surface mucosa, leading to inhibited lacrimal gland secretion and decreased tear secretion; simultaneously, the tear film osmotic pressure imbalance reduced the compatibility between the lipid and aqueous layers, worsening tear film stability, thus significantly shortening the tear film breakup time.

[0160] In summary, hyaluronic acid-chitosan-grafted phospholipids enhance liposome encapsulation efficiency and stability through hydrogen bonding, prolong ocular surface adhesion time, reduce anthocyanin degradation, and ensure the long-lasting effect of the active ingredients. The synergistic effect of the components significantly improves the composition's antioxidant activity, ocular surface compatibility, and stability.

[0161] 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 the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. An antioxidant composition based on horseberry, characterized in that, By weight, its constituent raw materials include: 8-18 parts of maquilli anthocyanin extract, 4-9 parts of hyaluronic acid-chitosan grafted phospholipids, 12-22 parts of phosphatidylcholine, 3-6 parts of liposome stabilizer, 2-4 parts of synergistic antioxidant, 3-8 parts of osmotic pressure regulator, 0.1-0.3 parts of preservative, 2-5 parts of sodium hyaluronate, 1-3 parts of taurine, 0.5-1.5 parts of vitamin B6, 0.3-0.8 parts of lutein, and 30-60 parts of solvent.

2. The antioxidant composition based on horseberry according to claim 1, characterized in that, The liposome stabilizer is cholesterol or stearoyl lysophosphatidylcholine, the synergistic antioxidant is vitamin E or ascorbate palmitate, the osmotic pressure regulator is sorbitol or mannitol, and the solvent is water for injection.

3. The berry-based antioxidant composition according to claim 2, characterized in that, The extraction method for malkiberella anthocyanin extract is as follows: (1) Take fresh Maquistberries, wash them, and crush them to a particle size of 3-5 mm. Add 3%-5% of the weight of the crushed fresh Maquistberries to a mixed microbial agent and anaerobic ferment at 37-40℃ for 8-12 hours. The mixed microbial agent is composed of *Lactobacillus plantarum* bacterial solution and *Saccharomyces cerevisiae* bacterial solution at a volume ratio of 2:1, with the *Lactobacillus plantarum* bacterial solution having a viable count ≥1×10⁻⁶. 9 CFU / mL, viable count of Saccharomyces cerevisiae in the culture ≥1×10⁻⁶ 8 CFU / mL; (2) Add water for injection to the fermentation material obtained in (1) at a mass-volume ratio of 1g:18-25mL, and then add 0.5%-1.0% of the weight of crushed fresh maquist fruit of compound enzyme. Under ultra-high pressure of 500-600MPa and 35-45℃ for 15-25min, and then under ultrasonic treatment of 250-300W for 15-20min. (3) Filter the extract obtained in (2) through a sterile microfiltration membrane of 0.20-0.22 μm, load the filtrate onto AB-8 macroporous resin, elute with 50%-70% ethanol solution, and collect the eluent; (4) The eluent is vacuum concentrated to a solid content of 30%-35% at 45-50℃ and 0.08-0.09MPa, and then spray-dried at an inlet air temperature of 125-135℃ and an outlet air temperature of 65-75℃ to obtain the anthocyanin extract of macquip.

4. The berry-based antioxidant composition according to claim 3, characterized in that, In step (2), the complex enzyme is composed of cellulase, pectinase and β-glucosidase in a mass ratio of (2-3):(1-2):1; in step (3), the flow rate of the filtrate is 1.5-2.5 BV / h and the flow rate of the ethanol solution is 0.8-1.2 BV / h.

5. The berry-based antioxidant composition according to claim 2, characterized in that, By weight, the raw materials of hyaluronic acid-chitosan grafted phospholipids include: 5-7 parts hyaluronic acid, 3-5 parts chitosan, 10-12 parts phosphatidylcholine, 1-2 parts 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, and 50-60 parts water for injection.

6. The berry-based antioxidant composition according to claim 5, characterized in that, The preparation method of hyaluronic acid-chitosan grafted phospholipids includes the following steps: 1) Dissolve hyaluronic acid in water for injection, add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, and then stir at 30-32℃ for 28-32 min; 2) Add chitosan to the solution obtained in 1) to adjust the pH to 5.5-6.0, and react at a constant temperature of 34-36℃ for 6-8 hours to form hyaluronic acid-chitosan graft. 3) Add phosphatidylcholine to the hyaluronic acid-chitosan graft and sonicate for 18-22 minutes at a temperature of 38-42℃ and a power of 180-220W to obtain the reaction solution; 4) The reaction solution was filtered through a sterile microfiltration membrane of 0.20-0.22 μm, and then freeze-dried under vacuum at -45~-35℃ and 0.01-0.02 MPa for 8-12 hours. The solution was then pulverized and passed through a 120-125 mesh sieve to obtain hyaluronic acid-chitosan grafted phospholipids.

7. The berry-based antioxidant composition according to claim 5, characterized in that, The molecular weight of hyaluronic acid is 1000-1200kDa, and the degree of deacetylation of chitosan is ≥95%.

8. The berry-based antioxidant composition according to claim 7, characterized in that, The preservative is either phenoxyethanol or sodium perborate.

9. The marijuana-based antioxidant composition according to any one of claims 1-8, characterized in that, Its preparation method includes the following steps: S1. Take phosphatidylcholine, liposome stabilizer, synergistic antioxidant, maquillia anthocyanin extract and hyaluronic acid-chitosan grafted phospholipids, add chloroform-methanol mixture, and rotary evaporate to form a uniform film at 45-50℃ and 0.07-0.08MPa. The chloroform-methanol mixture is prepared by mixing chloroform and methanol in a volume ratio of 2:1 to 3:1, and the mass-volume ratio of the five raw materials to be dissolved to the chloroform-methanol mixture is 1g:8-12mL. S2. Add an aqueous solution of osmotic pressure regulator to the material obtained in S1, hydrate by stirring in a water bath at 38-40℃ for 38-42 minutes, and homogenize under high pressure at 40-60MPa 4-6 times to obtain anthocyanin-modified liposomes. The aqueous solution of the osmotic pressure regulator is composed of an osmotic pressure regulator and water for injection in a mass-to-volume ratio of 1g:15-25mL. S3. Mix anthocyanin-modified liposomes with sodium hyaluronate, taurine, vitamin B6, lutein, and preservatives, add the remaining water for injection, and then stir at 580-620 r / min for 18-22 min. Filter through a 0.20-0.22 μm sterile microfiltration membrane and dispense under sterile conditions to obtain the anti-oxidative composition based on maquist.

10. The use of a maquill-based antioxidant composition according to any one of claims 1-9 in a dry eye prevention product.