Astaxanthin and collagen complex composition and preparation method thereof

Abstract

This invention belongs to the field of functional food technology, specifically relating to an astaxanthin and collagen complex composition and its preparation method. The composition comprises the following components: 5 to 20 parts of astaxanthin phospholipid composite nanoparticles, 60 to 110 parts of glycosylated collagen peptides, 2 to 6 parts of hyaluronic acid-collagen peptide grafts, and 1 to 3 parts of curcumin-encapsulated components. The preparation method of the composition is as follows: S1, preparing an activated aqueous phase base; S2, sequential mixing and primary stabilization; S3, integrating the encapsulated components and fine homogenization; S4, final blending and degassing; S5, solid particle preparation. This invention, by constructing a synergistic system containing multiple modified functional components such as astaxanthin phospholipid composite nanoparticles and glycosylated collagen peptides, and employing a refined compounding process, effectively improves the physical stability of the product and synergistically enhances the bioavailability of the active ingredients and the overall antioxidant support efficacy.

Description

Technical Field

[0001] This invention belongs to the field of functional food technology, specifically relating to a compound composition of astaxanthin and collagen and its preparation method. Background Technology

[0002] Astaxanthin and collagen are two core ingredients that have garnered significant attention in the field of functional foods and dietary supplements. Astaxanthin, a potent natural antioxidant derived from Haematococcus pluvialis, exhibits remarkable free radical scavenging capabilities. Collagen, especially its hydrolysate collagen peptides, is a key structural protein for maintaining the health and elasticity of connective tissues such as skin, bones, and joints. Combining these two ingredients to simultaneously achieve antioxidant defense and tissue repair has become an important direction in the development of oral beauty and joint health products.

[0003] However, the simple physical mixing or conventional compounding of astaxanthin and collagen peptides has revealed several specific technical bottlenecks that urgently need to be addressed in actual industrialization.

[0004] First, their physicochemical properties are fundamentally contradictory. Astaxanthin is a highly hydrophobic, fat-soluble substance, while collagen peptides are hydrophilic molecules. This difference in properties makes it difficult for them to coexist stably in liquid or solid final products. Common emulsions or mixtures are prone to phase separation, astaxanthin precipitation and crystallization, oxidative degradation, and other problems, leading to color changes, a significant decrease in the content of active ingredients, and the appearance of oil rings or sediment during the product's shelf life, resulting in poor stability.

[0005] Secondly, the bioavailability and absorption efficiency of each component face challenges. Unmodified astaxanthin (such as ordinary oleoresin) has limited solubility and bioavailability in the human gastrointestinal tract. Conventional collagen peptides have a wide molecular weight distribution, with excessively large fragments being difficult to absorb effectively, and excessively small fragments potentially having unclear physiological functions and unpleasant flavors. When these unoptimized raw materials are simply combined, their absorption kinetics in vivo are asynchronous, making it difficult to reach the concentration window required for synergistic effects in target tissues, and failing to achieve the ideal sequential synergistic effect of "first anti-oxidation to clear damage factors, then providing raw materials to promote repair."

[0006] Furthermore, current technologies lack sophisticated designs that enable functional synergy among different components at the molecular or supramolecular level. Most products merely represent the physical superposition of ingredients; astaxanthin, collagen peptides, and other potential functional components exist independently in the formulation, without employing technological means to construct a complex system that promotes co-absorption, targeted delivery, or intracellular synergistic effects. For example, there is a lack of delivery systems capable of simultaneously loading and protecting fat-soluble astaxanthin and water-soluble collagen peptides, and promoting their trans-intestinal epithelial cell transport.

[0007] Furthermore, existing production processes have relatively rudimentary control over the pretreatment of raw materials and the final compounding process. Although existing technologies have attempted to improve the defects of a single component through microencapsulation, enzymatic hydrolysis, and other methods, these improvements are usually isolated and fail to systematically solve the entire chain problem from deep modification of raw materials and compatibility compounding of multiple components to stable production.

[0008] Therefore, it is necessary to design a compound composition of astaxanthin and collagen and its preparation method. Summary of the Invention

[0009] To overcome the shortcomings of the prior art, an astaxanthin-collagen compound composition and its preparation method are provided.

[0010] To achieve the above objectives, the present invention provides the following technical solution: A compound composition of astaxanthin and collagen, comprising, by weight, the following components: 5 to 20 parts of astaxanthin phospholipid composite nanoparticles, 60 to 110 parts of glycosylated collagen peptides, 2 to 6 parts of hyaluronic acid-collagen peptide grafts, and 1 to 3 parts of curcumin inclusion complex.

[0011] The astaxanthin phospholipid composite nanoparticles were prepared by the following steps: (a) Combination: Astaxanthin oleoresin and refined soybean lecithin were dissolved together in anhydrous ethanol at a mass ratio of 1:3 to 1:6. The mixture was stirred and reacted at 60°C to 70°C for 2 to 3 hours. Then, the ethanol was completely removed by vacuum distillation to obtain the astaxanthin lecithin complex. (b) Nanoparticle formation: The astaxanthin phospholipid complex was added to a phosphate buffer solution at 55°C to 65°C and pre-dispersed for 3 minutes at 10,000 rpm using a high-speed shear mill. Then, it was transferred to a high-pressure microfluidic homogenizer and cyclically processed 4 to 6 times at a pressure of 100 MPa to 140 MPa to obtain a nanoparticle dispersion with an average particle size of 80 nm to 200 nm. After freeze-drying, astaxanthin phospholipid composite nanoparticles were obtained.

[0012] Both the glycosylated collagen peptides and the hyaluronic acid-collagen peptide grafts are prepared from collagen peptide solution as raw material. The collagen peptide solution is prepared by the following steps: fish skin collagen is dispersed in water, the pH is adjusted to 7.5 to 8.5, a complex protease is added, and enzymatic hydrolysis is carried out at 50°C to 55°C for 3 to 5 hours. Then, the temperature is raised to 90°C to inactivate the enzyme, and the solution is filtered to obtain a collagen peptide solution with a molecular weight distribution of 1500 Daltons to 5000 Daltons.

[0013] When preparing the collagen peptide solution, the fish skin collagen is dispersed at a concentration of 5% to 15% in water; The amount of the compound protease added is 1.0% to 3.0% of the dry weight of fish skin collagen; The complex protease consists of an alkaline protease and a flavor protease, wherein the mass ratio of the alkaline protease to the flavor protease is 3:1 to 5:1.

[0014] The preparation method of the glycosylated collagen peptide includes the following steps: take the collagen peptide solution, mix it with fructooligosaccharides at a peptide dry weight to sugar dry weight ratio of 6:1 to 12:1, adjust the pH value to 7.0 to 8.0, place it in a water bath at 75°C to 85°C and react for 40 to 80 minutes. After the reaction is completed, cool it, and spray dry the resulting reaction solution to obtain the glycosylated collagen peptide.

[0015] In the development of oral beauty and joint health products, combining astaxanthin with collagen peptides is a promising direction. However, astaxanthin is a highly hydrophobic, lipid-soluble molecule, while collagen peptides are hydrophilic molecules. Simply mixing the two directly, whether in liquid formulations or final solid products, easily leads to phase separation, astaxanthin precipitation and crystallization, and oxidative degradation, resulting in product deterioration in appearance and rapid loss of active ingredients during storage. Common solutions often focus on isolating single components, such as microencapsulating astaxanthin alone, failing to fundamentally address the interfacial compatibility issue between astaxanthin and the abundant collagen peptides present in the continuous aqueous phase. Simple physical encapsulation may introduce new problems, such as the capsule wall material potentially affecting astaxanthin release or causing undesirable interactions with the collagen peptide solution.

[0016] The technical solution proposed in this invention first modifies the raw materials at the molecular level to construct an endogenously stable complex system. Specifically, this involves two synergistic aspects: First, the preparation of astaxanthin-phospholipid composite nanoparticles. This is achieved by combining astaxanthin oleoresin with refined soybean phospholipids, followed by nano-sizing using high-pressure microfluidic homogenization technology. This process not only encapsulates astaxanthin with phospholipid molecules, significantly improving its surface hydrophilicity, but also results in nanoscale particles with high specific surface area and interfacial energy. Second, the glycosylation modification of collagen peptides to prepare glycosylated collagen peptides. Through a controlled Maillard reaction, fructooligosaccharides are grafted onto collagen peptide molecules. The introduced hydrophilic sugar chains alter the conformation and surface properties of the peptide chains. On the one hand, this reduces self-aggregation between peptide chains due to hydrophobic interactions; on the other hand, the introduction of sugar chains increases the interaction between molecules and water, providing a better interfacial compatibility basis for the astaxanthin-phospholipid composite nanoparticles, which also possess amphiphilic surfaces.

[0017] The combination of these two modified components enables the originally incompatible oil and water phase active ingredients to form a stable and uniform dispersion in the compound system, thereby providing a guarantee for the long-term stability of the product at the physical level.

[0018] The preparation method of the hyaluronic acid-collagen peptide graft comprises: mixing the collagen peptide solution with a sodium hyaluronate solution, wherein the dry basis mass ratio of sodium hyaluronate to collagen peptide is 1:8 to 1:12; adding 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride as a condensing agent, the amount of which is 0.8% to 1.2% of the mass of sodium hyaluronate; stirring and reacting at pH 5.0 to 6.0 and room temperature for 3 to 5 hours; after the reaction is completed, placing the mixture in a dialysis bag with a molecular weight cutoff of 3500 Daltons and dialyzing for 24 to 48 hours, and then freeze-drying to obtain the hyaluronic acid-collagen peptide graft.

[0019] Having initially resolved the physical compatibility issues, the next step is to ensure that these modified active ingredients can be efficiently absorbed by the human body and that absorption kinetics are coordinated. Untreated astaxanthin has low solubility in the intestines, while the absorption efficiency and rate of ordinary collagen peptides are greatly affected by their molecular weight distribution. If absorption is asynchronous, it is difficult to form an effective synergistic concentration window at the target tissue. In existing technologies, methods to improve bioavailability are usually carried out in isolation, such as using absorption enhancers or deep hydrolyzing peptides. However, these methods may introduce irritation or loss of functional peptides and do not consider the synergistic effect of the absorption processes of different components.

[0020] This invention addresses this problem through a series of targeted modifications. First, the small size of the astaxanthin phospholipid composite nanoparticles themselves facilitates their dispersion in gastrointestinal fluids. Phospholipids are a major component of cell membranes, and their presence may promote the transmembrane transport of astaxanthin by mimicking the formation of mixed micelles or by their affinity for intestinal epithelial cells.

[0021] Secondly, the modification of glycosylated collagen peptides alters their molecular properties. Grafting with fructooligosaccharides may not only provide new sites of action related to intestinal carbohydrate transport, but may also potentially regulate their absorption pathways and rates by affecting the peptide's folding state and thus altering the rate of degradation by intestinal peptidases or the efficiency of recognition by specific peptide transporters.

[0022] Furthermore, hyaluronic acid-collagen peptide grafts are introduced. Hyaluronic acid is a hydrophilic mucopolysaccharide with good biocompatibility and mucosal adhesion. By grafting it onto collagen peptides via chemical bonds, the resulting grafts may utilize the properties of hyaluronic acid to prolong its retention time on the intestinal mucosa and provide gentle adhesion and auxiliary delivery for other coexisting active ingredients. These modified components, when combined, can produce synergy in the digestive and absorptive environment, making it more likely that hydrophobic astaxanthin and hydrophilic collagen peptide derivatives will be delivered simultaneously or sequentially with high efficiency, creating conditions for subsequent synergistic physiological functions in vivo.

[0023] The composition also includes a compound excipient comprising the following components in parts by weight: 25 to 70 parts erythritol, 1 to 5 parts citric acid, 0.5 to 2 parts methylated catechin, and 3 to 15 parts microcrystalline cellulose. The methylated catechin was prepared by the following method: epigallocatechin gallate was dissolved in dimethyl sulfoxide, and dimethyl sulfate was added, wherein the molar amount of dimethyl sulfate was 1.0 to 1.5 times the estimated molar amount of phenolic hydroxyl groups in epigallocatechin gallate; the reaction was carried out at 35°C to 45°C for 4 to 6 hours under nitrogen protection; after the reaction was completed, the reaction solution was poured into ice water to precipitate the precipitate, filtered, the precipitate was washed with pure water until neutral, and then freeze-dried to obtain the methylated catechin.

[0024] The preparation method of the encapsulated curcumin includes the following steps: dissolving β-cyclodextrin in hot water at 55°C to 65°C to prepare a saturated β-cyclodextrin solution; dissolving curcumin in ethanol to obtain a curcumin ethanol solution; adding the curcumin ethanol solution dropwise to the saturated β-cyclodextrin solution under stirring, wherein the molar ratio of curcumin to β-cyclodextrin is 1:1.2 to 1:1.8; stirring continuously at 50°C to 60°C for 8 to 10 hours, then refrigerating at 4°C for more than 12 hours, filtering, collecting the precipitate, washing with cold water, and vacuum drying to obtain encapsulated curcumin.

[0025] Once active ingredients can coexist stably and be effectively delivered, maximizing their intrinsic biological efficacy, particularly the synergistic effect of antioxidation and tissue repair, becomes a crucial consideration. Single antioxidants have limited scope of action, and collagen supplementation requires an environment with controlled oxidative stress for more effective synthesis and deposition. Conventional products often simply add together several antioxidant ingredients, lacking a design to build a functional network within the dosage form. Each component may metabolize independently in the body, making it difficult to form sustained, multi-layered defense and repair support.

[0026] This invention enhances the overall antioxidant and anti-inflammatory support network of the system by introducing encapsulated curcumin and methylated catechins, combined with a compounding process. Encapsulated curcumin utilizes the cavity structure of β-cyclodextrin to embed curcumin molecules. This treatment improves the solubility and chemical stability of curcumin in aqueous systems, allowing it to be uniformly integrated into the compound composition and preventing its rapid degradation and inactivation.

[0027] Methylated catechins are products of methylation modification of epigallocatechin gallate. This modification alters their solubility and metabolic stability, potentially changing their time window of action in vivo.

[0028] In the preparation process, astaxanthin phospholipid composite nanoparticles are first dispersed in an aqueous substrate containing glycosylated collagen peptides and hyaluronic acid-collagen peptide grafts. Then, an aqueous solution of methylated catechins is added. This sequence facilitates the partial adsorption or interaction of methylated catechin molecules at the nanoparticle interface. Next, curcumin-encapsulated powder is added and homogenized. This step-by-step, orderly introduction method aims to allow different antioxidant active molecules (astaxanthin, curcumin, and methylated catechins) not only to be physically mixed, but also to potentially form a loose functional relationship or complementarity at the supramolecular level.

[0029] Once ingested, these antioxidant units, due to their different molecular structures, polarities, and sites of action, can counteract different types of reactive oxygen species or those generated in different cellular compartments, thus constructing a more comprehensive and hierarchical antioxidant defense system. This enhanced antioxidant network helps create a more favorable microenvironment for the utilization of collagen peptides and their tissue-repairing functions.

[0030] A method for preparing a compound composition of astaxanthin and collagen, the method comprising the following steps: S1. Preparation of activated aqueous phase base material: Dissolve glycosylated collagen peptides and hyaluronic acid-collagen peptide grafts in pure water at 48℃ to 52℃, and stir until completely dissolved to obtain activated aqueous phase base material; S2. Sequential mixing and primary stabilization: Cool the activated aqueous phase base to 32°C to 37°C; add the redispersible liquid of astaxanthin phospholipid composite nanoparticles under medium-speed stirring at 250 rpm to 400 rpm, and stir for 15 minutes to make it uniformly dispersed; then add the methylated catechin aqueous solution under stirring and stir for 10 minutes. S3. Integration of Inclusion Compounds and Fine Homogenization: Add inclusion compound curcumin powder to the system in step S2 and keep stirring for 20 minutes; then transfer the mixture to a high-pressure homogenizer and homogenize twice under a pressure of 65 MPa to 85 MPa. S4. Final preparation and degassing: Add erythritol, citric acid and microcrystalline cellulose to the homogenized liquid in step S3, and stir until completely dissolved and dispersed; finally, place the liquid in a degassing tank and degas for 20 minutes under a vacuum of 0.06 MPa to 0.08 MPa to obtain the composition liquid. S5. Preparation of solid particles: The composition liquid obtained in step S4 is mixed with sodium octenyl succinate starch at a dry matter mass ratio of 1:0.3 to 1:0.6. The powder collected by spray drying is then granulated in a fluidized bed. The resulting solid particles are the finished product of the astaxanthin and collagen compound composition.

[0031] In step S2, the redispersible solution of the astaxanthin phospholipid composite nanoparticles is prepared by redispersing the astaxanthin phospholipid composite nanoparticles at a mass concentration of 8% to 12% in pure water containing 0.1% trehalose at 35°C to 40°C and then vortexing for 2 minutes. In step S2, the method for preparing the methylated catechin aqueous solution is as follows: dissolve the methylated catechin in pure water at 60°C to 70°C to prepare a solution with a mass concentration of 5% to 10%, and cool it to below 40°C. In step S5, the inlet air temperature of the spray dryer is 162°C to 168°C, and the outlet air temperature is 82°C to 88°C.

[0032] The present invention uses purified water containing trehalose for the redispersibility of astaxanthin nanoparticles to protect the nanostructure and promote its uniform distribution in the aqueous phase; the precise control of the spray drying temperature is to avoid overheating and damage to the heat-sensitive active ingredients while achieving efficient dehydration.

[0033] Compared with the prior art, the advantages and beneficial effects of the present invention are as follows: 1. This invention effectively solves the physical incompatibility problem between astaxanthin and collagen peptides by constructing astaxanthin-phospholipid composite nanoparticles and utilizing amphiphilic modification of glycosylated collagen peptides. After astaxanthin is composited with phospholipids and nano-sized, its surface hydrophilicity is improved, enabling it to be more stably dispersed in an aqueous environment dominated by collagen peptides. Simultaneously, the controllable glycosylation modification of the collagen peptides introduces hydrophilic glycan chains, which not only partially shields the hydrophobic interactions between peptide chains and reduces aggregation, but also enhances the interfacial affinity with the surface-active astaxanthin-phospholipid composite nanoparticles. This dual effect based on molecular-level modification allows the originally hydrophobic astaxanthin and hydrophilic collagen peptides to coexist stably for a long time in the final compound system, reducing the risks of phase separation, precipitation, and oxidative degradation of astaxanthin.

[0034] 2. The astaxanthin of this invention forms a phospholipid complex with an average particle size at the nanoscale. Its small size effect and phospholipid composition help promote its dispersion, dissolution, and absorption through intestinal epithelial cells in the gastrointestinal tract. Glycosylated collagen peptides, due to the grafting of fructooligosaccharides, undergo changes in molecular conformation and polarity, which may affect their recognition efficiency by intestinal peptide transporters and potentially optimize their absorption kinetics by utilizing sugar-related absorption pathways. Hyaluronic acid-collagen peptide grafts, by grafting hyaluronic acid with collagen peptides, not only retain the hydrophilic and moisturizing properties of hyaluronic acid, but their grafted structure may also facilitate interaction with the intestinal mucosa, providing a certain degree of auxiliary delivery for other coexisting active ingredients. These modified components contribute to synergy during absorption, allowing active substances with different properties to be delivered and absorbed within a similar timeframe, creating favorable conditions for subsequent synergistic physiological functions in vivo.

[0035] 3. This invention enhances the antioxidant and anti-inflammatory support network by introducing encapsulated curcumin and methylated catechins. Encapsulated curcumin improves the water dispersibility and stability of curcumin, allowing it to better integrate into the system. Methylated catechins, compared to their precursors, may have modified lipophilicity and metabolic stability. During preparation, the aqueous solution of methylated catechins is added after the astaxanthin nanoparticles are dispersed, and its molecules may partially act on the nanoparticle interface; while encapsulated curcumin is added during subsequent homogenization. This stepwise introduction method allows for potential physical interactions or functional complementarity between different antioxidant components. When this composition is ingested, antioxidant units such as astaxanthin, curcumin, and methylated catechins may construct a more hierarchical and efficient antioxidant defense by scavenging different types of free radicals or those located in different microenvironments.

[0036] 4. The present invention uses purified water containing trehalose for the redispersibility of astaxanthin nanoparticles to protect the nanostructure and promote its uniform distribution in the aqueous phase; the precise control of the spray drying temperature is to avoid overheating and damage to the heat-sensitive active ingredients while achieving efficient dehydration. Detailed Implementation

[0037] 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.

[0038] In the specific embodiments of this application, the sources of various main raw materials are briefly described as follows: Astaxanthin oleoresin, with an astaxanthin content of no less than 10%, is sourced from Yunnan Aierfa Biotechnology Co., Ltd.

[0039] Refined soybean lecithin, purchased from Hubei Haijia Biotechnology Co., Ltd.

[0040] 1-Ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride: purchased from Hubei Xingyan New Material Technology Co., Ltd., CAS: 25952-53-8, active ingredient content 99%.

[0041] Dimethyl sulfoxide: Shandong Fite Chemical Technology Co., Ltd., CAS: 67-68-5.

[0042] Dimethyl sulfate: Linyi Yuanbo Chemical Co., Ltd.

[0043] Fish skin collagen: Hainan Huayan Collagen Technology Co., Ltd. Alkaline protease, with an enzyme activity ≥200,000 U / g, was purchased from Jiangsu Ruiyang Biotechnology Co., Ltd.

[0044] Flavor protease, with an enzyme activity ≥100,000 U / g, was purchased from Jiangsu Ruiyang Biotechnology Co., Ltd.

[0045] Fructooligosaccharides, with a purity of not less than 95%, were purchased from Shandong Bailong Chuangyuan Biotechnology Co., Ltd.

[0046] Sodium hyaluronate, with a molecular weight range of 50,000 to 200,000 Daltons, was purchased from Bloomage Biotechnology Co., Ltd.

[0047] β-Cyclodextrin was purchased from Anhui Shanhe Pharmaceutical Excipients Co., Ltd.

[0048] Curcumin was purchased from Xi'an Tianyi Biotechnology Co., Ltd.

[0049] Epigallocatechin gallate, purchased from Chengdu Desite Biotechnology Co., Ltd.

[0050] Erythritol, purchased from Sichuan Kangbairui Biotechnology Co., Ltd.

[0051] Citric acid was purchased from Sichuan Tongchuanxing Chemical Co., Ltd.

[0052] Microcrystalline cellulose was purchased from Chengdu Jinran Chemical Co., Ltd.

[0053] Trehalose was purchased from Hubei Haijia Biotechnology Co., Ltd.

[0054] Sodium octenyl succinate starch was purchased from Hubei Haijia Biotechnology Co., Ltd.

[0055] Maltodextrin, purchased from Shandong Xiwang Sugar Industry Co., Ltd.

[0056] The technical solution of this application is as follows: A compound composition of astaxanthin and collagen, comprising, by weight, the following components: 5 to 20 parts of astaxanthin phospholipid composite nanoparticles, 60 to 110 parts of glycosylated collagen peptides, 2 to 6 parts of hyaluronic acid-collagen peptide grafts, and 1 to 3 parts of curcumin inclusion complex.

[0057] The composition also includes a compound excipient comprising the following components in parts by weight: 25 to 70 parts erythritol, 1 to 5 parts citric acid, 0.5 to 2 parts methylated catechin, and 3 to 15 parts microcrystalline cellulose. The methylated catechin was prepared by the following method: epigallocatechin gallate was dissolved in dimethyl sulfoxide, and dimethyl sulfate was added, wherein the molar amount of dimethyl sulfate was 1.0 to 1.5 times the estimated molar amount of phenolic hydroxyl groups in epigallocatechin gallate; the reaction was carried out at 35°C to 45°C for 4 to 6 hours under nitrogen protection; after the reaction was completed, the reaction solution was poured into ice water to precipitate the precipitate, filtered, the precipitate was washed with pure water until neutral, and then freeze-dried to obtain the methylated catechin.

[0058] The astaxanthin phospholipid composite nanoparticles were prepared by the following steps: (a) Combination: Astaxanthin oleoresin and refined soybean lecithin were dissolved together in anhydrous ethanol at a mass ratio of 1:3 to 1:6. The mixture was stirred and reacted at 60°C to 70°C for 2 to 3 hours. Then, the ethanol was completely removed by vacuum distillation to obtain the astaxanthin lecithin complex. (b) Nanoparticle formation: The astaxanthin phospholipid complex was added to a phosphate buffer solution at 55°C to 65°C and pre-dispersed for 3 minutes at 10,000 rpm using a high-speed shear mill. Then, it was transferred to a high-pressure microfluidic homogenizer and cyclically processed 4 to 6 times at a pressure of 100 MPa to 140 MPa to obtain a nanoparticle dispersion with an average particle size of 80 nm to 200 nm. After freeze-drying, astaxanthin phospholipid composite nanoparticles were obtained.

[0059] Both the glycosylated collagen peptides and the hyaluronic acid-collagen peptide grafts are prepared from collagen peptide solution as raw material. The collagen peptide solution is prepared by the following steps: fish skin collagen is dispersed in water, the pH is adjusted to 7.5 to 8.5, a complex protease is added, and enzymatic hydrolysis is carried out at 50°C to 55°C for 3 to 5 hours. Then, the temperature is raised to 90°C to inactivate the enzyme, and the solution is filtered to obtain a collagen peptide solution with a molecular weight distribution of 1500 Daltons to 5000 Daltons.

[0060] When preparing the collagen peptide solution, the fish skin collagen is dispersed at a concentration of 5% to 15% in water; The amount of the compound protease added is 1.0% to 3.0% of the dry weight of fish skin collagen; The complex protease consists of an alkaline protease and a flavor protease, wherein the mass ratio of the alkaline protease to the flavor protease is 3:1 to 5:1.

[0061] The preparation method of the glycosylated collagen peptide includes the following steps: take the collagen peptide solution, mix it with fructooligosaccharides at a peptide dry weight to sugar dry weight ratio of 6:1 to 12:1, adjust the pH value to 7.0 to 8.0, place it in a water bath at 75°C to 85°C and react for 40 to 80 minutes. After the reaction is completed, cool it, and spray dry the resulting reaction solution to obtain the glycosylated collagen peptide.

[0062] The preparation method of the hyaluronic acid-collagen peptide graft comprises: mixing the collagen peptide solution with a sodium hyaluronate solution, wherein the dry basis mass ratio of sodium hyaluronate to collagen peptide is 1:8 to 1:12; adding 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride as a condensing agent, the amount of which is 0.8% to 1.2% of the mass of sodium hyaluronate; stirring and reacting at pH 5.0 to 6.0 and room temperature for 3 to 5 hours; after the reaction is completed, placing the mixture in a dialysis bag with a molecular weight cutoff of 3500 Daltons and dialyzing for 24 to 48 hours, and then freeze-drying to obtain the hyaluronic acid-collagen peptide graft.

[0063] The preparation method of the encapsulated curcumin includes the following steps: dissolving β-cyclodextrin in hot water at 55°C to 65°C to prepare a saturated β-cyclodextrin solution; dissolving curcumin in ethanol to obtain a curcumin ethanol solution; adding the curcumin ethanol solution dropwise to the saturated β-cyclodextrin solution under stirring, wherein the molar ratio of curcumin to β-cyclodextrin is 1:1.2 to 1:1.8; stirring continuously at 50°C to 60°C for 8 to 10 hours, then refrigerating at 4°C for more than 12 hours, filtering, collecting the precipitate, washing with cold water, and vacuum drying to obtain encapsulated curcumin.

[0064] A method for preparing a compound composition of astaxanthin and collagen, the method comprising the following steps: S1. Preparation of activated aqueous phase base material: Dissolve glycosylated collagen peptides and hyaluronic acid-collagen peptide grafts in pure water at 48℃ to 52℃, and stir until completely dissolved to obtain activated aqueous phase base material; S2. Sequential mixing and primary stabilization: Cool the activated aqueous phase base to 32°C to 37°C; add the redispersible liquid of astaxanthin phospholipid composite nanoparticles under medium-speed stirring at 250 rpm to 400 rpm, and stir for 15 minutes to make it uniformly dispersed; then add the methylated catechin aqueous solution under stirring and stir for 10 minutes. S3. Integration of Inclusion Compounds and Fine Homogenization: Add inclusion compound curcumin powder to the system in step S2 and keep stirring for 20 minutes; then transfer the mixture to a high-pressure homogenizer and homogenize twice under a pressure of 65 MPa to 85 MPa. S4. Final preparation and degassing: Add erythritol, citric acid and microcrystalline cellulose to the homogenized liquid in step S3, and stir until completely dissolved and dispersed; finally, place the liquid in a degassing tank and degas for 20 minutes under a vacuum of 0.06 MPa to 0.08 MPa to obtain the composition liquid. S5. Preparation of solid particles: The composition liquid obtained in step S4 is mixed with sodium octenyl succinate starch at a dry matter mass ratio of 1:0.3 to 1:0.6. The powder collected by spray drying is then granulated in a fluidized bed. The resulting solid particles are the finished product of the astaxanthin and collagen compound composition.

[0065] In step S2, the redispersible solution of the astaxanthin phospholipid composite nanoparticles is prepared by redispersing the astaxanthin phospholipid composite nanoparticles at a mass concentration of 8% to 12% in pure water containing 0.1% trehalose at 35°C to 40°C and then vortexing for 2 minutes. In step S2, the method for preparing the methylated catechin aqueous solution is as follows: dissolve the methylated catechin in pure water at 60°C to 70°C to prepare a solution with a mass concentration of 5% to 10%, and cool it to below 40°C. In step S5, the inlet air temperature of the spray dryer is 162°C to 168°C, and the outlet air temperature is 82°C to 88°C.

[0066] This invention constructs a synergistic system comprising multiple modified functional components such as astaxanthin phospholipid composite nanoparticles and glycosylated collagen peptides, and employs a refined compounding process to effectively improve the physical stability of the product and synergistically enhance the bioavailability of the active ingredients and the overall antioxidant support efficacy.

[0067] 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. Example 1

[0068] This embodiment provides a compound composition of astaxanthin and collagen and its preparation method.

[0069] The composition, by weight, comprises the following components: 20 parts astaxanthin phospholipid composite nanoparticles, 60 parts glycosylated collagen peptides, 2 parts hyaluronic acid-collagen peptide grafts, and 3 parts curcumin-encapsulated compounds. The composition also includes a composite excipient comprising the following components by weight: 70 parts erythritol, 1 part citric acid, 2 parts methylated catechins, and 3 parts microcrystalline cellulose.

[0070] Preparation of the methylated catechin: Epigallocatechin gallate was dissolved in dimethyl sulfoxide, and dimethyl sulfate was added. The molar amount of dimethyl sulfate was 1.5 times the estimated molar amount of the phenolic hydroxyl groups in the epigallocatechin gallate. The reaction was carried out at 45°C for 4 hours under nitrogen protection. After the reaction was completed, the reaction solution was poured into ice water to precipitate the precipitate. The precipitate was filtered, washed with pure water until neutral, and then freeze-dried to obtain the methylated catechin.

[0071] The preparation of the astaxanthin phospholipid composite nanoparticles includes the following steps: A compounding step: Astaxanthin oleoresin and refined soybean phospholipids are dissolved together in anhydrous ethanol at a mass ratio of 1:6. The mixture is stirred and reacted at 70°C for 2 hours, followed by vacuum distillation to completely remove the ethanol, yielding the astaxanthin phospholipid complex. A nano-sizing step: The astaxanthin phospholipid complex is added to phosphate buffer at 65°C and pre-dispersed for 3 minutes using a high-speed shear mill at 10,000 rpm. It is then transferred to a high-pressure microfluidic homogenizer and circulated four times at 140 MPa to obtain a nanoparticle dispersion with an average particle size of approximately 200 nm. After freeze-drying, the astaxanthin phospholipid composite nanoparticles are obtained.

[0072] The collagen peptide solution used to prepare glycosylated collagen peptides and hyaluronic acid-collagen peptide grafts is prepared by the following steps: Fish skin collagen is dispersed in water at a concentration of 5%, the pH is adjusted to 8.5, and a complex protease is added at an amount of 3.0% of the dry weight of the fish skin collagen. This complex protease consists of an alkaline protease and a flavor protease in a mass ratio of 5:1. Enzymatic hydrolysis is performed at 55°C for 3 hours, followed by enzyme inactivation at 90°C. The solution is then filtered to obtain a collagen peptide solution with a molecular weight distribution ranging from 1500 Daltons to 5000 Daltons.

[0073] Preparation of the glycosylated collagen peptide: Take the above collagen peptide solution and mix it with fructooligosaccharides at a peptide dry weight to sugar dry weight ratio of 12:1. Adjust the pH value to 8.0 and place it in an 85℃ water bath for 40 minutes. After the reaction is completed, cool it and spray dry the resulting reaction solution to obtain the glycosylated collagen peptide.

[0074] Preparation of the hyaluronic acid-collagen peptide graft: The above-mentioned collagen peptide solution was mixed with a sodium hyaluronate solution, wherein the dry basis mass ratio of sodium hyaluronate to collagen peptide was 1:12. 1-Ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride was added as a condensing agent, with an amount equal to 1.2% of the mass of sodium hyaluronate. The reaction was stirred at pH 6.0 and room temperature for 3 hours. After the reaction was completed, the mixture was dialyzed for 48 hours in a dialysis bag with a molecular weight cutoff of 3500 Daltons, and then freeze-dried to obtain the hyaluronic acid-collagen peptide graft.

[0075] Preparation of the encapsulated curcumin: β-cyclodextrin was dissolved in hot water at 65°C to prepare a saturated β-cyclodextrin solution. Curcumin was dissolved in ethanol to obtain a curcumin ethanol solution. The curcumin ethanol solution was added dropwise to the saturated β-cyclodextrin solution with stirring, and the molar ratio of curcumin to β-cyclodextrin was 1:1.8. The mixture was stirred continuously at 60°C for 8 hours, then refrigerated at 4°C for 12 hours. After filtration, the precipitate was collected, washed with cold water, and vacuum dried to obtain the encapsulated curcumin.

[0076] The preparation method of the astaxanthin and collagen complex includes the following steps: S1. Preparation of activated aqueous phase base material: Dissolve the glycosylated collagen peptide and hyaluronic acid-collagen peptide graft prepared above in pure water at 52°C, and stir until completely dissolved to obtain activated aqueous phase base material.

[0077] S2. Sequential Mixing and Primary Stabilization: The activated aqueous phase base is cooled to 37°C. Under medium-speed stirring at 400 rpm, a redispersible solution of astaxanthin phospholipid composite nanoparticles is added, and the mixture is stirred for 15 minutes to ensure uniform dispersion. The redispersible solution is prepared by redispersing the astaxanthin phospholipid composite nanoparticles at a mass concentration of 12% in purified water containing 0.1% trehalose at 40°C, followed by vortexing for 2 minutes. Subsequently, a methylated catechin aqueous solution is added under stirring, and the mixture is stirred for 10 minutes. The methylated catechin aqueous solution is prepared by dissolving methylated catechin in purified water at 70°C to prepare a 10% mass concentration solution, and then cooling it to below 40°C.

[0078] S3. Integration and Homogenization of Inclusion Compounds: Add the inclusion compound curcumin powder to the system from step S2 and stir for 20 minutes. Then transfer the mixture to a high-pressure homogenizer and homogenize twice at a pressure of 85 MPa.

[0079] S4. Final Preparation and Degassing: Add the prescribed amounts of erythritol, citric acid, and microcrystalline cellulose to the homogenized liquid from step S3, and stir until completely dissolved and dispersed. Finally, place the liquid in a degassing tank and degas for 20 minutes under a vacuum of 0.08 MPa to obtain the composite liquid.

[0080] S5. The composition liquid obtained in step S4 is mixed with sodium octenyl succinate starch at a dry matter mass ratio of 1:0.6, and then dried in a spray drying tower. The inlet air temperature of the spray dryer is 168°C, and the outlet air temperature is 88°C. The collected powder is then subjected to fluidized bed granulation, and the resulting solid particles are the finished product of the astaxanthin and collagen compound composition of this embodiment. Example 2

[0081] In this embodiment, the similarities to those in Embodiment 1 will not be repeated, and the differences are as follows: This embodiment provides another astaxanthin and collagen compound composition and its preparation method.

[0082] The composition, by weight, comprises the following components: 5 parts astaxanthin phospholipid composite nanoparticles, 110 parts glycosylated collagen peptides, 6 parts hyaluronic acid-collagen peptide grafts, and 1 part curcumin-encapsulated compound. The composition also includes a composite excipient comprising the following components by weight: 25 parts erythritol, 5 parts citric acid, 0.5 parts methylated catechin, and 15 parts microcrystalline cellulose.

[0083] In the preparation of the methylated catechin, the molar amount of dimethyl sulfate is 1.0 times the estimated molar amount of the phenolic hydroxyl group in epigallocatechin gallate, and the reaction is carried out at 35°C for 6 hours.

[0084] In the preparation of the astaxanthin phospholipid composite nanoparticles, the mass ratio of astaxanthin oleoresin to refined soybean phospholipids was 1:3, and the reaction was carried out by stirring at 60°C for 3 hours. In the nano-sizing step, the composite was added to phosphate buffer at 55°C and cyclically treated 6 times under a pressure of 100 MPa to obtain a nanoparticle dispersion with an average particle size of approximately 80 nanometers.

[0085] In the preparation of collagen peptide solution, the concentration of fish skin collagen dispersion is 15%, the pH is adjusted to 7.5, the amount of compound protease added is 1.0% of the dry basis mass, the mass ratio of alkaline protease to flavor protease in the compound protease is 3:1, and enzymatic hydrolysis is carried out at 50℃ for 5 hours.

[0086] In the preparation of the glycosylated collagen peptide, the ratio of peptide dry weight to sugar dry weight is 6:1, the pH is adjusted to 7.0, and the reaction is carried out in a 75°C water bath for 80 minutes.

[0087] In the preparation of the hyaluronic acid-collagen peptide graft, the dry basis mass ratio of sodium hyaluronate to collagen peptide is 1:8. The amount of condensing agent is 0.8% of the mass of sodium hyaluronate. The reaction is carried out by stirring at pH 5.0 for 5 hours, followed by dialysis for 24 hours.

[0088] In the preparation of the curcumin-encapsulated curcumin, β-cyclodextrin is dissolved in hot water at 55°C, the molar ratio of curcumin to β-cyclodextrin is 1:1.2, and the mixture is stirred continuously at 50°C for 10 hours and then refrigerated for more than 12 hours.

[0089] In the preparation method, the dissolution temperature in step S1 is 48℃, and in step S2, the activated aqueous phase base is cooled to 32℃, with a stirring speed of 250 rpm. The mass concentration of the astaxanthin phospholipid composite nanoparticle redispersible solution is 8%, and the dispersion medium temperature is 35℃. The mass concentration of the methylated catechin aqueous solution is 5%, and the dissolution temperature is 60℃. The homogenization pressure in step S3 is 65 MPa. The degassing vacuum degree in step S4 is 0.06 MPa. In step S5, the dry matter mass ratio of the composite solution to sodium octenyl succinate starch is 1:0.3, the spray drying inlet air temperature is 162℃, and the outlet air temperature is 82℃. Example 3

[0090] In this embodiment, the similarities to those in Embodiment 1 will not be repeated, and the differences are as follows: This embodiment provides another astaxanthin and collagen compound composition and its preparation method.

[0091] The composition, by weight, comprises the following components: 12.5 parts astaxanthin phospholipid composite nanoparticles, 85 parts glycosylated collagen peptides, 4 parts hyaluronic acid-collagen peptide grafts, and 2 parts curcumin-encapsulated compounds. The composition also includes a composite excipient comprising the following components by weight: 47.5 parts erythritol, 3 parts citric acid, 1.25 parts methylated catechin, and 9 parts microcrystalline cellulose.

[0092] In the preparation of the methylated catechin, the molar amount of dimethyl sulfate is 1.25 times the estimated molar amount of the phenolic hydroxyl group in epigallocatechin gallate, and the reaction is carried out at 40°C for 5 hours.

[0093] In the preparation of the astaxanthin phospholipid composite nanoparticles, the mass ratio of astaxanthin oleoresin to refined soybean phospholipids was 1:4.5, and the reaction was carried out at 65°C with stirring for 2.5 hours. In the nano-sizing step, the composite was added to phosphate buffer at 60°C and cyclically treated 5 times under a pressure of 120 MPa to obtain a nanoparticle dispersion with an average particle size of approximately 140 nm.

[0094] In the preparation of collagen peptide solution, the concentration of fish skin collagen dispersion was 10%, the pH was adjusted to 8.0, the amount of compound protease added was 2.0% of the dry basis mass, the mass ratio of alkaline protease to flavor protease in the compound protease was 4:1, and enzymatic hydrolysis was carried out at 52.5℃ for 4 hours.

[0095] In the preparation of the glycosylated collagen peptide, the ratio of peptide dry weight to sugar dry weight is 9:1, the pH is adjusted to 7.5, and the mixture is placed in an 80°C water bath for 60 minutes.

[0096] In the preparation of the hyaluronic acid-collagen peptide graft, the dry basis mass ratio of sodium hyaluronate to collagen peptide is 1:10. The amount of condensing agent is 1.0% of the mass of sodium hyaluronate. The reaction is carried out by stirring at pH 5.5 for 4 hours, followed by dialysis for 36 hours.

[0097] In the preparation of the curcumin-encapsulated curcumin, β-cyclodextrin is dissolved in hot water at 60°C, the molar ratio of curcumin to β-cyclodextrin is 1:1.5, and the mixture is stirred continuously at 55°C for 9 hours and then refrigerated for more than 12 hours.

[0098] In the preparation method, the dissolution temperature in step S1 is 50℃, and in step S2, the activated aqueous phase base is cooled to 34.5℃, with a stirring speed of 325 rpm. The mass concentration of the astaxanthin phospholipid composite nanoparticle redispersible solution is 10%, and the dispersion medium temperature is 37.5℃. The mass concentration of the methylated catechin aqueous solution is 7.5%, and the dissolution temperature is 65℃. The homogenization pressure in step S3 is 75 MPa. The degassing vacuum degree in step S4 is 0.07 MPa. In step S5, the dry matter mass ratio of the composite solution to sodium octenyl succinate starch is 1:0.45, the spray drying inlet air temperature is 165℃, and the outlet air temperature is 85℃. Comparative Example 1

[0099] In this comparative example, the similarities with Example 3 will not be repeated, and the differences are as follows: No core raw materials were modified. Specifically: untreated astaxanthin oleoresin was used instead of astaxanthin phospholipid composite nanoparticles; commercially available fish collagen peptide powder was used instead of glycosylated collagen peptides and hyaluronic acid-collagen peptide grafts. These commercially available collagen peptides were obtained by direct spray drying of the same fish skin raw material after similar enzymatic hydrolysis, without glycosylation or graft modification; commercial curcumin powder was used instead of incorporated curcumin; methylated catechins were not used, and the composite excipients did not contain this component. The preparation method involves dry mixing all the above raw material powders (astaxanthin oleoresin needs to be pre-dispersed with a small amount of emulsifier) ​​and composite excipients in a mixer at one time, and then directly dispensing the solid mixture product. Comparative Example 2

[0100] In this comparative example, the similarities with Example 3 will not be repeated, and the differences are as follows: The astaxanthin phospholipid complex nanoparticles were missing. Specifically, the astaxanthin phospholipid complex nanoparticles in the formulation of Example 3 were replaced with an equal mass of ordinary astaxanthin powder (obtained by spray drying after encapsulation with astaxanthin oleoresin via maltodextrin), and the preparation method was adjusted accordingly. In step S2, the ordinary astaxanthin powder was directly added and mixed with other ingredients. Comparative Example 3

[0101] In this comparative example, the similarities with Example 3 will not be repeated, and the differences are as follows: The glycosylation step was missing. Specifically, ordinary collagen peptide powder obtained by direct spray drying from the same collagen peptide solution but without fructooligosaccharide glycosylation was used instead of the glycosylated collagen peptides in Example 3. The hyaluronic acid-collagen peptide graft was prepared using the original method. Comparative Example 4

[0102] In this comparative example, the similarities with Example 3 will not be repeated, and the differences are as follows: The preparation order of the key raw materials was changed. Specifically, in step S2 of the preparation method, the order of adding the methylated catechin aqueous solution and the astaxanthin phospholipid composite nanoparticle redispersible solution was interchanged. That is, the methylated catechin aqueous solution was added first, and after stirring for 10 minutes, the astaxanthin phospholipid composite nanoparticle redispersible solution was added. Comparative Example 5

[0103] In this comparative example, the similarities with Example 3 will not be repeated, and the differences are as follows: The preparation method for curcumin inclusion complex differs. Specifically, a simple physical grinding and mixing method is used to prepare the curcumin complex. Curcumin powder and β-cyclodextrin powder are directly added to a ball mill in the same mass ratio (calculated by molar ratio) as in Example 3, and dry-grind and mix for 2 hours. The mixture is then sieved to obtain a physical mixture, which replaces the curcumin inclusion complex in Example 3. In this physical mixture, curcumin is not effectively included. Comparative Example 6

[0104] In this comparative example, the similarities with Example 3 will not be repeated, and the differences are as follows: The types and proportions of complex proteases differ. Specifically, when preparing collagen peptide solution, a single alkaline protease is used, flavor proteases are not used, and the total amount added is maintained at 2.0% of the dry weight of fish skin collagen. The collagen peptide solution obtained in this way is then used to prepare glycosylated collagen peptides and hyaluronic acid-collagen peptide grafts. Performance Test Results and Analysis

[0105] To verify the effectiveness of the present invention, a series of performance tests were conducted on the products obtained in Examples 1-3 and Comparative Examples 1-6. Before the tests, all solid samples were equilibrated for 24 hours under the same conditions (temperature 25±2℃, relative humidity 60±5%). Liquid intermediate products (such as the composition liquid obtained in step S4) were sampled and tested immediately after preparation or subjected to accelerated stability tests.

[0106] Test 1: Product Physical Stability Assessment The physical stability of the liquid system (sample solution in step S4) was assessed using the accelerated sedimentation method. 10 mL of sample was placed in a graduated centrifuge tube and centrifuged at 4000 rpm for 30 minutes. The layering was observed and recorded, and the volume of the precipitate or oil phase was measured. Simultaneously, the solid powder was placed in a stoppered glass bottle and stored in a constant temperature and humidity chamber at 40°C and 75% relative humidity for 30 days. Changes in appearance, such as clumping or discoloration, were observed.

[0107] Test 2: Determination of Active Ingredient Retention Rate Accelerated shelf-life simulation test. Solid finished products were packaged in commercially available aluminum-plastic bags and placed in a constant temperature and humidity chamber at 40℃ and 75% relative humidity. Samples were taken at 0 days, 30 days, and 60 days. Referring to the Chinese Pharmacopoeia and relevant national standards, the contents of astaxanthin and curcumin in the samples were determined by high-performance liquid chromatography (HPLC). The changes in collagen peptide content were indirectly determined by the Kjeldahl method or an amino acid analyzer. The retention rate relative to the content at day 0 was calculated.

[0108] Test 3: Simulated in vitro digestion and intestinal cell absorption assessment A static in vitro simulated gastrointestinal digestion model was used. Samples equivalent to a certain amount of active ingredients were accurately weighed and sequentially placed in simulated gastric juice (containing pepsin, pH 2.0, shaken at 37°C for 2 hours) and simulated intestinal juice (containing pancreatic enzymes and bile salts, pH 7.0, shaken at 37°C for 4 hours). After digestion, the digestive fluid was centrifuged at 15,000 rpm for 20 minutes, and the supernatant was collected and filtered through a 0.22-micron filter membrane. The concentrations of lipid-soluble components such as astaxanthin and curcumin in the filtrate were determined using high-performance liquid chromatography (HPLC), and the concentrations of soluble peptides in the filtrate were determined using the Folin-Ciocalteu method or the BCA method. The bioavailability of each component (i.e., the proportion released from the solid phase into the digestive fluid) was calculated.

[0109] Test 4: In vitro antioxidant activity assay The antioxidant activity of the samples was comprehensively evaluated using three commonly used methods: DPPH radical scavenging capacity, ABTS cationic radical scavenging capacity, and oxygen radical absorption capacity. Equal masses of each sample were weighed and dissolved or dispersed in solvents to prepare a series of concentration gradients. The absorbance was measured at specific wavelengths according to the operating procedures of each assay method, and the overall antioxidant capacity of the samples (μmol TE / g) was calculated. The specific test results are shown in Table 1.

[0110] Table 1 Analysis of Test Results

[0111] As shown in Table 1, the products of Examples 1-3 exhibit excellent physical stability. They showed almost no stratification after centrifugation and maintained good appearance under high temperature and humidity conditions. This indicates that the physical incompatibility between astaxanthin and collagen peptides was effectively solved by constructing astaxanthin-phospholipid composite nanoparticles and glycosylation modification of collagen peptides. The surface hydrophilicity of the astaxanthin-phospholipid composite nanoparticles and the enhanced interfacial affinity between them and glycosylated collagen peptides ensured the homogeneity and stability of the entire compound system.

[0112] In contrast, Comparative Example 1, lacking any modification to the raw materials, exhibited severe incompatibility between the astaxanthin oleoresin and the hydrophilic environment, leading to intense oil-water separation and rapid deterioration of the product's appearance. Comparative Example 2, using ordinary astaxanthin powder, showed better stability than Comparative Example 1, but still significantly worse than the examples, indicating that simple physical encapsulation is less effective than the systematic compounding of phospholipid composite nanoparticles. Comparative Example 3, lacking a glycosylation step, also showed slightly lower stability than the examples, demonstrating that glycosylation modification plays a positive role in reducing collagen peptide aggregation and enhancing compatibility with lipid nanoparticles.

[0113] In the examples, the retention rates of astaxanthin and collagen peptides after accelerated storage were significantly higher than those in Comparative Examples 1 and 2. This further confirms the protective effect of the modification strategy of the present invention on the active ingredients. The structure of the astaxanthin phospholipid composite nanoparticles and the stable environment formed by the entire compound system effectively slowed down the oxidative degradation of astaxanthin. Comparative Example 5, which used a physically ground curcumin complex, also had a lower astaxanthin retention rate than the examples, indicating that the inclusion of curcumin may not only be stable on its own, but its form may also have a positive impact on the system's microenvironment, an effect that simple physical mixing cannot achieve. The retention rate of Comparative Example 6 was close to that of the examples, indicating that single enzymatic hydrolysis had a relatively small impact on the storage stability of the final product.

[0114] The bioavailability of astaxanthin in the example products was significantly higher than in all comparative examples, especially far higher than in Comparative Examples 1 and 2. This confirms that the small size effect of the astaxanthin phospholipid composite nanoparticles and the phospholipid component effectively promote its dissolution and micellization in a simulated intestinal environment. The bioavailability of peptides also remained at a high level in the examples. The bioavailability of peptides in Comparative Example 3 was slightly lower than that in the examples, suggesting that glycosylation modification may affect the solubility and stability of peptides during digestion by altering their molecular properties. Although the change in the order of addition in Comparative Example 4 had little impact on the stability of the final solid product, the data showed that its astaxanthin bioavailability was slightly lower than that in Example 3. This may be because the order of addition affected the pre-distribution of methylated catechins at the astaxanthin nanoparticle interface, thus subtly affecting its subsequent behavior in the digestive fluid. The data from Comparative Example 5 also showed that the effective inclusion of curcumin had a positive effect on its own and possibly on the solubilization of the overall lipophilic components.

[0115] The comprehensive antioxidant capacity data indicate that the product in the examples has the highest antioxidant value. This is not only due to the high retention rates of astaxanthin and curcumin, but may also be related to the introduction of methylated catechins and potential synergistic effects among the components. Comparative Example 1 has the lowest antioxidant capacity due to severe loss of active ingredients. The antioxidant capacities of Comparative Examples 2, 3, and 5 are successively lower than those of the examples, reflecting the negative impacts of the lack of nano-sized astaxanthin, the lack of potential synergistic support from glycosylated peptides, and the ineffective encapsulation and utilization of curcumin, respectively. The antioxidant capacity of Comparative Example 6 is relatively close to that of the examples, indicating that the difference between compound enzymatic hydrolysis and single enzymatic hydrolysis does not have a dominant effect on this indicator.

[0116] 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 compound composition of astaxanthin and collagen, characterized in that, The composition comprises, by weight, the following components: 5 to 20 parts of astaxanthin phospholipid complex nanoparticles, 60 to 110 parts of glycosylated collagen peptides, 2 to 6 parts of hyaluronic acid-collagen peptide grafts, and 1 to 3 parts of curcumin-encapsulated compounds.

2. The astaxanthin and collagen compound composition according to claim 1, characterized in that, The composition also includes a compound excipient comprising the following components in parts by weight: 25 to 70 parts erythritol, 1 to 5 parts citric acid, 0.5 to 2 parts methylated catechin, and 3 to 15 parts microcrystalline cellulose; The methylated catechin was prepared by the following method: epigallocatechin gallate was dissolved in dimethyl sulfoxide, and dimethyl sulfate was added, wherein the molar amount of dimethyl sulfate was 1.0 to 1.5 times the estimated molar amount of phenolic hydroxyl groups in epigallocatechin gallate; the reaction was carried out at 35°C to 45°C for 4 to 6 hours under nitrogen protection; after the reaction was completed, the reaction solution was poured into ice water to precipitate the precipitate, filtered, the precipitate was washed with pure water until neutral, and then freeze-dried to obtain the methylated catechin.

3. The astaxanthin and collagen compound composition according to claim 1, characterized in that, The astaxanthin phospholipid composite nanoparticles were prepared by the following method: (a) Combination: Astaxanthin oleoresin and refined soybean lecithin were dissolved together in anhydrous ethanol at a mass ratio of 1:3 to 1:

6. The mixture was stirred and reacted at 60°C to 70°C for 2 to 3 hours. Then, the ethanol was completely removed by vacuum distillation to obtain the astaxanthin lecithin complex. (b) Nanoparticle formation: The astaxanthin phospholipid complex was added to a phosphate buffer solution at 55°C to 65°C and pre-dispersed for 3 minutes at 10,000 rpm using a high-speed shear mill. Then, it was transferred to a high-pressure microfluidic homogenizer and cyclically processed 4 to 6 times at a pressure of 100 MPa to 140 MPa to obtain a nanoparticle dispersion with an average particle size of 80 nm to 200 nm. After freeze-drying, astaxanthin phospholipid composite nanoparticles were obtained.

4. The astaxanthin and collagen compound composition according to claim 1, characterized in that, Both the glycosylated collagen peptide and the hyaluronic acid-collagen peptide graft are prepared from collagen peptide liquid as raw material. The collagen peptide solution is prepared by the following steps: fish skin collagen is dispersed in water, the pH is adjusted to 7.5 to 8.5, a complex protease is added, and the solution is enzymatically hydrolyzed at 50°C to 55°C for 3 to 5 hours. Then the temperature is raised to 90°C to inactivate the enzyme, and the solution is filtered to obtain a collagen peptide solution with a molecular weight distribution of 1500 Daltons to 5000 Daltons.

5. The astaxanthin and collagen compound composition according to claim 4, characterized in that, When preparing the collagen peptide solution, the fish skin collagen is dispersed at a concentration of 5% to 15% in water; The amount of the compound protease added is 1.0% to 3.0% of the dry weight of fish skin collagen; The complex protease consists of an alkaline protease and a flavor protease, wherein the mass ratio of the alkaline protease to the flavor protease is 3:1 to 5:

1.

6. The astaxanthin and collagen compound composition according to claim 4, characterized in that, The preparation method of the glycosylated collagen peptide includes the following steps: take the collagen peptide solution, mix it with fructooligosaccharides at a peptide dry weight to sugar dry weight ratio of 6:1 to 12:1, adjust the pH value to 7.0 to 8.0, place it in a water bath at 75°C to 85°C and react for 40 to 80 minutes. After the reaction is completed, cool it, and spray dry the resulting reaction solution to obtain the glycosylated collagen peptide.

7. The astaxanthin and collagen compound composition according to claim 4, characterized in that, The preparation method of the hyaluronic acid-collagen peptide graft is as follows: the collagen peptide solution is mixed with a sodium hyaluronate solution, wherein the dry basis mass ratio of sodium hyaluronate to collagen peptide is 1:8 to 1:12; 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride is added as a condensing agent, the amount of which is 0.8% to 1.2% of the mass of sodium hyaluronate; the mixture is stirred and reacted at pH 5.0 to 6.0 and room temperature for 3 to 5 hours; after the reaction is completed, the mixture is placed in a dialysis bag with a molecular weight cutoff of 3500 Daltons and dialyzed for 24 to 48 hours, and then freeze-dried to obtain the hyaluronic acid-collagen peptide graft.

8. The astaxanthin and collagen compound composition according to claim 1, characterized in that, The preparation method of the encapsulated curcumin includes the following steps: dissolving β-cyclodextrin in hot water at 55°C to 65°C to prepare a saturated β-cyclodextrin solution; dissolving curcumin in ethanol to obtain a curcumin ethanol solution; adding the curcumin ethanol solution dropwise to the saturated β-cyclodextrin solution under stirring, wherein the molar ratio of curcumin to β-cyclodextrin is 1:1.2 to 1:1.8; stirring continuously at 50°C to 60°C for 8 to 10 hours, then refrigerating at 4°C for more than 12 hours, filtering, collecting the precipitate, washing with cold water, and vacuum drying to obtain encapsulated curcumin.

9. A method for preparing a compound composition of astaxanthin and collagen as described in any one of claims 1-8, characterized in that, The method includes the following steps: S1. Preparation of activated aqueous phase base material: Dissolve glycosylated collagen peptides and hyaluronic acid-collagen peptide grafts in pure water at 48℃ to 52℃, and stir until completely dissolved to obtain activated aqueous phase base material; S2. Sequential mixing and primary stabilization: Cool the activated aqueous phase base to 32°C to 37°C; add the redispersible liquid of astaxanthin phospholipid composite nanoparticles under medium-speed stirring at 250 rpm to 400 rpm, and stir for 15 minutes to make it uniformly dispersed; then add the methylated catechin aqueous solution under stirring and stir for 10 minutes. S3. Integration of Inclusion Compounds and Fine Homogenization: Add inclusion compound curcumin powder to the system in step S2 and keep stirring for 20 minutes; then transfer the mixture to a high-pressure homogenizer and homogenize twice under a pressure of 65 MPa to 85 MPa. S4. Final preparation and degassing: Add erythritol, citric acid and microcrystalline cellulose to the homogenized liquid in step S3, and stir until completely dissolved and dispersed; finally, place the liquid in a degassing tank and degas for 20 minutes under a vacuum of 0.06 MPa to 0.08 MPa to obtain the composition liquid. S5. Preparation of solid particles: The composition liquid obtained in step S4 is mixed with sodium octenyl succinate starch at a dry matter mass ratio of 1:0.3 to 1:0.

6. The powder collected by spray drying is then granulated in a fluidized bed. The resulting solid particles are the finished product of the astaxanthin and collagen compound composition.

10. The method for preparing an astaxanthin and collagen complex composition according to claim 9, characterized in that, In step S2, the redispersible solution of the astaxanthin phospholipid composite nanoparticles is prepared by redispersing the astaxanthin phospholipid composite nanoparticles at a mass concentration of 8% to 12% in pure water containing 0.1% trehalose at 35°C to 40°C and then vortexing for 2 minutes. In step S2, the method for preparing the methylated catechin aqueous solution is as follows: dissolve the methylated catechin in pure water at 60°C to 70°C to prepare a solution with a mass concentration of 5% to 10%, and cool it to below 40°C. In step S5, the inlet air temperature of the spray dryer is 162°C to 168°C, and the outlet air temperature is 82°C to 88°C.

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