Fish collagen and rose double-effect anti-aging composition and compound polysaccharide microencapsulation embedding preparation method thereof

Abstract

The invention belongs to the technical field of biological medicines, and discloses a fish collagen and rose double-effect anti-aging composition, and the core active ingredients of the fish collagen and rose double-effect anti-aging composition comprise fish collagen peptide, red roses with double petals, dried cranberries, Chinese wolfberry fruits, roxburgh rose puree and saffron. According to the preparation method, specific degradation of intestinal flora is realized by virtue of the targeting effect of the xylooligosaccharide on bifidobacteria through the compound polysaccharide microcapsules, so that collagen peptide and antioxidant components are accurately released in colon, gastric acid damage is effectively avoided, and meanwhile, the piperine nanoparticles inhibit the activity of UGT enzyme, so that the anti-oxidation effect of the collagen peptide on the colon is improved, and the anti-oxidation effect of the collagen peptide on the colon is improved. The bioavailability of crocin is increased to 45% or above from 2%, the absorption rate of rose polyphenol is increased by 3 times, the intestinal absorption efficiency of collagen peptide reaches 95%, a remarkable breakthrough is achieved compared with 60-80% of a traditional process, and an efficient delivery mode of targeted release-absorption enhancement is constructed.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and specifically relates to a fish collagen and rose dual-effect anti-aging composition and a preparation method of a composite polysaccharide microencapsulation embedding thereof. Background Art

[0002] Traditional anti-aging compositions face multiple technical bottlenecks in practical applications. First, the ingredients lack synergy. When fish collagen peptides are directly compounded with antioxidant ingredients such as rose polyphenols and crocin, their activity will be degraded due to intermolecular interactions. In addition, collagen peptides are easily destroyed by gastric acid during intestinal absorption, affecting their biological functions.

[0003] Secondly, the problem of low bioavailability is significant. Antioxidant plant ingredients such as proanthocyanidins and VC have the defect of poor water solubility and prominent first-pass metabolism. For example, the oral bioavailability of crocin is only 1-2%, and marine polysaccharides such as sea cucumber saponins and fucoidan are difficult to be effectively absorbed by the intestine due to their large molecular weight, which greatly limits the efficacy of the ingredients.

[0004] In addition, traditional formulas also have the problem of lack of targeting, and are unable to accurately deliver active ingredients to the skin and immune cells. For key problems such as decreased fibroblast activity and macrophage function decline caused by aging, the improvement efficiency is obviously insufficient. To address these pain points, the innovative breakthrough lies in the use of complex polysaccharide microencapsulation technology to construct a three-dimensional network carrier with brown algae polysaccharides, oligoxylose and sea cucumber polysaccharides, and combine it with a nano-scale piperine synergistic absorption system to form a "dual-effect targeted anti-aging delivery system", which achieves comprehensive improvement in the three dimensions of ingredient stability, absorption efficiency and cell targeting, and effectively solves the inherent defects of traditional technology. Summary of the Invention

[0005] The purpose of the present invention is to provide a fish collagen rose dual-effect anti-aging composition and a composite polysaccharide microencapsulation embedding preparation method thereof, so as to solve the problems raised in the above background technology.

[0006] In order to achieve the above-mentioned objectives, the present invention provides the following technical solutions: a fish collagen and rose dual-effect anti-aging composition, wherein the core active ingredients of the fish collagen and rose dual-effect anti-aging composition include fish collagen peptides, double-petal red roses, dried cranberries, wolfberries, sea buckthorn puree, and saffron, wherein the small molecule peptides in the fish collagen peptides are <3000Da; double-petal red roses: rich in anthocyanins and polyphenols, inhibit free radicals ROS; dried cranberries: contain proanthocyanidins PACs, inhibit the adhesion of pathogenic bacteria; wolfberries: enhance mitochondrial function and improve cellular energy metabolism; sea buckthorn puree: ultra-high vitamin C 2g / 100g, promotes collagen synthesis as a cofactor of prolyl hydroxylase; saffron: active ingredient crocin, inhibits the NF-κB inflammatory pathway, reduces IL-6 inflammatory factors, and protects nerve cells.

[0007] Preferably, the specific steps of the fish collagen rose dual-effect anti-aging composition and the composite polysaccharide microencapsulation embedding preparation method are as follows: S1: Targeted enzymatic hydrolysis and graded purification of fish collagen peptides: Deep-sea cod skin is defatted and then soaked in acetic acid solution to remove impurities and proteins. The fish collagen peptide powder is obtained by enzymatic hydrolysis with a composite protease, purification with an ultrafiltration membrane, and spray drying. S2: Low-temperature synergistic extraction of antioxidant plant components: Double red roses were mixed in proportion and ultrasonically extracted with an ethanol-water solution containing citric acid. Saffron stigmas were cold-extracted at low temperature and then concentrated under reduced pressure. S3: Enzymatic modification and activity retention of marine active substances: The body wall of sea cucumbers was homogenized and then subjected to enzymatic hydrolysis, enzyme inactivation, centrifugation, and ethanol precipitation to extract polysaccharides. Kelp powder was degraded and deiodinated using high-temperature energy conversion technology. S4: Construction of composite polysaccharide microcapsule carriers: Dissolve xylooligosaccharides in water and stir to form a sol, add chitosan to adjust the pH, mix with plant extracts and marine polysaccharide solution, add olive oil for emulsification, and then cross-link to form microcapsule precursors; S5: Targeted coupling of fish collagen peptide and microcapsule: The fish collagen peptide was dissolved in buffer, activated with EDC and NHS, and then reacted with the microcapsule suspension at a suitable temperature to form a complex through Schiff base reaction; S6: Encapsulation of piperine nanosynergistic system: dissolving black pepper extract in solution, adding poloxamer and homogenizing under high pressure to prepare nanoemulsion, mixing with the complex and spray freeze drying to obtain a composition; S7: Finished product homogenization and quality control: The microencapsulated composition was sieved, mixed with xylitol and Angelica dahurica extract, and then ultrafinely ground at low temperature to detect the embedding efficiency and particle size and conduct stability tests.

[0008] Preferably, the directed enzymatic hydrolysis and graded purification of fish collagen peptides in S1 refers to the following: raw material pretreatment: taking deep-sea cod skin, defatting it, and soaking it in 0.1M acetic acid solution at 4°C for 24 hours to remove impurities; enzymatic hydrolysis process: adding a composite protease, a neutral protease and collagenase mass ratio of 3:1, and enzymatic hydrolysis at 50°C and pH 7.0 for 8 hours to generate small molecule peptides with a molecular weight of <2000Da; graded purification: separating through an ultrafiltration membrane with a molecular weight cutoff of 1000Da, collecting the filtrate and spray drying it with an inlet air temperature of 180°C and an outlet air temperature of 80°C to obtain refined fish collagen peptide powder.

[0009] Preferably, the specific steps of low-temperature synergistic extraction of antioxidant plant components in S2 are as follows: Step 1: Mixed extraction system: double red rose petals, dried cranberries, wolfberries, and roxburghii purees were mixed in a ratio of 2:1:1:0.5, and 80% ethanol-water containing 0.5% citric acid was added to the mixed solution. Ultrasonic-assisted extraction was performed at 40°C, power 300W, and time 40min. Step 2: Saffron active enrichment: Take saffron stigma separately and perform low-temperature cold extraction with 50℃ deionized water, with a material-liquid ratio of 1:20, stirring speed of 200 rpm, and time of 3 hours. The extract is concentrated under reduced pressure at 45℃ and vacuum degree of -0.08MPa to 1 / 10 of the original volume.

[0010] Preferably, the specific steps of enzymatic modification and activity retention of the marine active substance in S3 are as follows: Step 1: Extraction of sea cucumber polysaccharides: After the sea cucumber body wall is homogenized, 1000 U / g of cellulase is added to the raw material and enzymatically hydrolyzed at 45°C for 4 hours. The enzyme is inactivated in a boiling water bath, and the mixture is centrifuged at 8000 rpm for 15 minutes. The supernatant is collected and ethanol precipitation is performed to obtain sea cucumber polysaccharides at a final concentration of 80%. Step 2: Degradation of laminarin: A mixture of kelp powder and deionized water was treated with a high-temperature energy conversion technology at 85°C and a pulse power of 200W, with a material-to-liquid ratio of 1:15, to degrade alginate into low-molecular-weight fucoidan with a molecular weight of <50kDa. The alginate was then deiodinated by anion exchange resin and set aside.

[0011] Preferably, the construction of the composite polysaccharide microcapsule carrier in S4 refers to the preparation of polysaccharide sol: 3% oligoxylose, 2% fucoidan, and 1% sea cucumber polysaccharide are dissolved in deionized water, magnetically stirred at 300 rpm at 60°C to form a uniform sol, 0.1% chitosan is added and the pH is adjusted to 5.5 to enhance the stability of the sol; mixed emulsification treatment: the plant extract and the marine polysaccharide liquid are mixed in a 1:1 ratio and then the polysaccharide sol is added, olive oil is added in a 1:2 volume ratio, and emulsified in a 12000 rpm high-speed homogenizer for 20 minutes to form a stable water-in-oil emulsion; cross-linking capsule reaction: 1% calcium chloride crosslinking agent is added to the emulsion, and the reaction is carried out at a low temperature of 4°C for 4 hours. The composite polysaccharide microcapsule precursor with a three-dimensional network structure is formed through the cross-linking effect of calcium ions and polysaccharide chains.

[0012] Preferably, the specific steps of the targeted coupling of fish collagen peptide and microcapsule in S5 are as follows: Step 1: Peptide activation: The fish collagen peptide in S1 was dissolved in 0.1 M phosphate buffer, pH 7.4, and 1% EDC, 1-ethyl-3-, 3-dimethylaminopropyl, carbodiimide and 0.5% NHS, N-hydroxysuccinimide were added and activated at room temperature for 30 min; Step 2: Coupling reaction: Add the activated collagen peptide dropwise to the S4 microcapsule suspension and react at 37°C, 150 rpm on a shaker for 6 hours. The collagen peptide is covalently coupled to the microcapsule surface through the Schiff base reaction to form a "collagen peptide-microcapsule" complex.

[0013] Preferably, the coating of the piperine nano-synergistic system in S6: Nanoemulsion preparation: dissolving a black pepper extract containing high-purity piperine in an ethanol-water mixed solution, adding 0.5% poloxamer 188 as an emulsifier, and subjecting the mixture to 5 high-pressure homogenization cycles at a pressure of 100 MPa to prepare a stable piperine nanoemulsion with a particle size of 50-100 nm; Composite encapsulation treatment: mixing the nanoemulsion with a "collagen peptide-microcapsule" complex in a volume ratio of 1:5, using liquid nitrogen pre-cooling spray freeze drying technology, and drying under a vacuum environment of -0.09 MPa to form a microencapsulated composition with piperine nanoparticles uniformly loaded on the surface, and controlling the water content to ≤3%.

[0014] Preferably, the finished product homogenization and quality control in S7 refers to a homogenization treatment: passing the dried microencapsulated composition through a 200-mesh sieve, adding 5% xylitol and 0.5% angelica extract, mixing evenly, and adopting low-temperature ultrafine grinding at -20°C for 10 minutes to a particle size D90 <10 μm; quality control indicators: detecting the microcapsule embedding rate, ≥90%, the particle size distribution of 1-5 μm, ≥85%, the active ingredient retention rate VC ≥95%, and crocin ≥90%, and conducting an accelerated stability test at 40°C / 75%RH for 1 month.

[0015] The beneficial effects of the present invention are as follows: 1. The present invention uses composite polysaccharide microcapsules with the help of oligoxylose to target bifidobacteria, achieving specific degradation of intestinal flora, allowing collagen peptides and antioxidant ingredients to be accurately released in the colon, effectively avoiding gastric acid damage. At the same time, piperine nanoparticles inhibit the activity of UGT enzymes, thereby significantly improving the absorption efficiency of fat-soluble components - the bioavailability of crocin jumps from 2% to more than 45%, the absorption rate of rose polyphenols increases by 3 times, and the intestinal absorption efficiency of collagen peptides reaches 95%, a significant breakthrough compared to the 60-80% of traditional processes, establishing a high-efficiency delivery model of "targeted release-enhanced absorption".

[0016] 2. The surface-coupled collagen peptides of the present invention can directly bind to the skin keratinocyte receptors, promote the synthesis of hyaluronic acid by 25%, reduce the transepidermal water loss (TEWL) by 30%, and significantly improve the skin barrier function. At the level of systemic metabolic regulation, the microencapsulated composition optimizes the intestinal flora, increases the abundance of bifidobacteria by 2 times, reduces the serum inflammatory factor TNF-α level by 40%, and increases the glutathione (GSH) content of hepatocytes by 35%, thereby enhancing the systemic antioxidant capacity from intestinal immunity to liver detoxification, achieving the dual benefits of skin repair and systemic metabolic regulation.

[0017] 3. The surface-coupled collagen peptides of the present invention can directly bind to the skin keratinocyte receptors, promote the synthesis of hyaluronic acid by 25%, reduce the transepidermal water loss (TEWL) by 30%, and significantly improve the skin barrier function. At the level of systemic metabolic regulation, the microencapsulated composition optimizes the intestinal flora, increases the abundance of bifidobacteria by 2 times, reduces the serum inflammatory factor TNF-α level by 40%, and increases the glutathione (GSH) content of hepatocytes by 35%, thereby enhancing the systemic antioxidant capacity from intestinal immunity to liver detoxification, achieving the dual benefits of skin repair and systemic metabolic regulation. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 This is a flow chart of the preparation method of the fish collagen and rose dual-effect anti-aging composition and its complex polysaccharide microencapsulation embedding method of the present invention. DETAILED DESCRIPTION

[0019] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0020] like Figure 1 As shown, an embodiment of the present invention provides a fish collagen rose dual-effect anti-aging composition, the core active ingredients of which include fish collagen peptides, double red roses, dried cranberries, wolfberries, sea buckthorn puree, and saffron, wherein the small molecule peptides in the fish collagen peptides are <3000Da; double red roses: rich in anthocyanins and polyphenols, inhibit free radicals ROS; dried cranberries: contain proanthocyanidins PACs, inhibit the adhesion of pathogenic bacteria; wolfberries: enhance mitochondrial function and improve cellular energy metabolism; sea buckthorn puree: ultra-high vitamin C 2g / 100g, promotes collagen synthesis as a cofactor of prolyl hydroxylase; saffron: active ingredient crocin, inhibits the NF-κB inflammatory pathway, reduces IL-6 inflammatory factors, and protects nerve cells.

[0021] The specific steps of the preparation method of the fish collagen rose dual-effect anti-aging composition and its composite polysaccharide microencapsulation embedding method are as follows: S1: Targeted enzymatic hydrolysis and graded purification of fish collagen peptides: Deep-sea cod skin is defatted and then soaked in acetic acid solution to remove impurities and proteins. The fish collagen peptide powder is obtained by enzymatic hydrolysis with a composite protease, purification with an ultrafiltration membrane, and spray drying. S2: Low-temperature synergistic extraction of antioxidant plant components: Double red roses were mixed in proportion and ultrasonically extracted with an ethanol-water solution containing citric acid. Saffron stigmas were cold-extracted at low temperature and then concentrated under reduced pressure. S3: Enzymatic modification and activity retention of marine active substances: The body wall of sea cucumbers was homogenized and then subjected to enzymatic hydrolysis, enzyme inactivation, centrifugation, and ethanol precipitation to extract polysaccharides. Kelp powder was degraded and deiodinated using high-temperature energy conversion technology. S4: Construction of composite polysaccharide microcapsule carriers: Dissolve xylooligosaccharides in water and stir to form a sol, add chitosan to adjust the pH, mix with plant extracts and marine polysaccharide solution, add olive oil for emulsification, and then cross-link to form microcapsule precursors; S5: Targeted coupling of fish collagen peptide and microcapsule: The fish collagen peptide was dissolved in buffer, activated with EDC and NHS, and then reacted with the microcapsule suspension at a suitable temperature to form a complex through Schiff base reaction; S6: Encapsulation of piperine nanosynergistic system: dissolving black pepper extract in solution, adding poloxamer and homogenizing under high pressure to prepare nanoemulsion, mixing with the complex and spray freeze drying to obtain a composition; S7: Finished product homogenization and quality control: The microencapsulated composition was sieved, mixed with xylitol and Angelica dahurica extract, and then ultrafinely ground at low temperature to detect the embedding efficiency and particle size and conduct stability tests.

[0022] Among them, the directed enzymatic hydrolysis and graded purification of fish collagen peptides in S1 refers to the following: raw material pretreatment: taking deep-sea cod skin, degreasing it, and soaking it in 0.1M acetic acid solution at 4°C for 24 hours to remove impurities; enzymatic hydrolysis process: adding composite protease, neutral protease and collagenase in a mass ratio of 3:1, and enzymatic hydrolysis for 8 hours at 50°C and pH 7.0 to generate small molecule peptides with a molecular weight of <2000Da; graded purification: separating through an ultrafiltration membrane with a molecular weight cutoff of 1000Da, collecting the filtrate and spray drying it with an inlet air temperature of 180°C and an outlet air temperature of 80°C to obtain refined fish collagen peptide powder.

[0023] Deep-sea cod skin was first pretreated and then degreased, then soaked in 0.1M acetic acid solution at 4°C for 24 hours to remove impurities. It was then enzymatically hydrolyzed with a composite protease (neutral protease to collagenase mass ratio of 3:1) at 50°C and pH 7.0 for 8 hours to generate small molecular peptides <2000Da. Finally, it was purified by a 1000Da ultrafiltration membrane and spray-dried (inlet air 180°C, outlet air 80°C) to obtain refined peptide powder.

[0024] The specific steps of low-temperature synergistic extraction of antioxidant plant components in S2 are as follows: Step 1: Mixed extraction system: double red rose petals, dried cranberries, wolfberries, and roxburghii purees were mixed in a ratio of 2:1:1:0.5, and 80% ethanol-water containing 0.5% citric acid was added to the mixed solution. Ultrasonic-assisted extraction was performed at 40°C, power 300W, and time 40min. Step 2: Saffron active enrichment: Take saffron stigma separately and perform low-temperature cold extraction with 50℃ deionized water, with a material-liquid ratio of 1:20, stirring speed of 200 rpm, and time of 3 hours. The extract is concentrated under reduced pressure at 45℃ and vacuum degree of -0.08MPa to 1 / 10 of the original volume.

[0025] Double red rose petals, dried cranberries, wolfberries, and sea buckthorn purees were mixed in a ratio of 2:1:1:0.5, and an 80% ethanol-water solution containing 0.5% citric acid was added. The antioxidant components were extracted by ultrasonic-assisted extraction at 40°C with a power of 300W for 40 minutes. The stigma of saffron was taken separately and cold-extracted at 50°C deionized water with a material-liquid ratio of 1:20 and a stirring speed of 200 rpm for 3 hours. The extract was then concentrated under reduced pressure at 45°C and a vacuum degree of -0.08MPa to 1 / 10 of the original volume to enrich active substances such as crocin.

[0026] The specific steps of enzymatic modification and activity retention of the marine active substances in S3 are as follows: Step 1: Extraction of sea cucumber polysaccharides: After the sea cucumber body wall is homogenized, 1000 U / g of cellulase is added to the raw material and enzymatically hydrolyzed at 45°C for 4 hours. The enzyme is inactivated in a boiling water bath, and the mixture is centrifuged at 8000 rpm for 15 minutes. The supernatant is collected and ethanol precipitation is performed to obtain sea cucumber polysaccharides at a final concentration of 80%. Step 2: Degradation of laminarin: A mixture of kelp powder and deionized water was treated with a high-temperature energy conversion technology at 85°C and a pulse power of 200W, with a material-to-liquid ratio of 1:15, to degrade alginate into low-molecular-weight fucoidan with a molecular weight of <50kDa. The alginate was then deiodinated by anion exchange resin and set aside.

[0027] After the sea cucumber body wall is homogenized, cellulase is added at 1000U / g of raw material, and enzymatic hydrolysis is carried out at 45°C for 4 hours. The enzyme is inactivated in a boiling water bath, and the supernatant is collected by centrifugation at 8000rpm for 15 minutes. The supernatant is then precipitated with ethanol at a final concentration of 80% to obtain sea cucumber polysaccharides. A high-temperature energy conversion technology of 85°C and a pulse power of 200W is used to treat a mixture of kelp powder and deionized water with a material-liquid ratio of 1:15 to degrade alginate into low-molecular-weight fucoidan with a molecular weight of <50kDa. Finally, the alginate is deiodinated by anion exchange resin for later use.

[0028] Among them, the construction of the composite polysaccharide microcapsule carrier in S4 refers to the preparation of polysaccharide sol: 3% oligoxylose, 2% fucoidan, and 1% sea cucumber polysaccharide are dissolved in deionized water, and magnetically stirred at 300 rpm at 60°C to form a uniform sol, 0.1% chitosan is added and the pH is adjusted to 5.5 to enhance the stability of the sol; mixed emulsification treatment: the plant extract and the marine polysaccharide liquid are mixed in a 1:1 ratio and then the polysaccharide sol is added, and olive oil is added in a 1:2 volume ratio, and emulsified in a 12000 rpm high-speed homogenizer for 20 minutes to form a stable water-in-oil emulsion; cross-linking capsule reaction: 1% calcium chloride crosslinker is added to the emulsion, and the reaction is carried out at a low temperature of 4°C for 4 hours. Through the cross-linking effect of calcium ions and polysaccharide chains, a composite polysaccharide microcapsule precursor with a three-dimensional network structure is formed.

[0029] 3% xylooligosaccharide, 2% fucoidan, and 1% sea cucumber polysaccharide were dissolved in water, stirred at 300 rpm at 60°C to form a sol, and 0.1% chitosan was added to adjust the pH to 5.5; the plant extract and marine polysaccharide solution were mixed in a 1:1 ratio, then added to the sol, and olive oil was added in a 1:2 ratio, and emulsified at 12,000 rpm for 20 minutes to form an emulsion; 1% calcium chloride was added and cross-linked at 4°C for 4 hours to form the initial body of the complex polysaccharide microcapsules.

[0030] The specific steps of the targeted coupling of fish collagen peptide and microcapsule in S5 are as follows: Step 1: Peptide activation: The fish collagen peptide in S1 was dissolved in 0.1 M phosphate buffer, pH 7.4, and 1% EDC, 1-ethyl-3-, 3-dimethylaminopropyl, carbodiimide and 0.5% NHS, N-hydroxysuccinimide were added and activated at room temperature for 30 min; Step 2: Coupling reaction: Add the activated collagen peptide dropwise to the S4 microcapsule suspension and react at 37°C, 150 rpm on a shaker for 6 hours. The collagen peptide is covalently coupled to the microcapsule surface through the Schiff base reaction to form a "collagen peptide-microcapsule" complex.

[0031] During peptide activation, the fish collagen peptide obtained from S1 was dissolved in 0.1M phosphate buffer (pH 7.4), and 1% EDC (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide) and 0.5% NHS (N-hydroxysuccinimide) were added. It was activated at room temperature for 30 minutes. In the coupling reaction, the activated collagen peptide was added dropwise to the microcapsule suspension of S4, and the reaction was carried out at 37°C and 150 rpm on a shaker for 6 hours. The collagen peptide was covalently coupled to the microcapsule surface through the Schiff base reaction to form a "collagen peptide-microcapsule" complex.

[0032] Among them, the coating of the piperine nano-synergistic system in S6: Nanoemulsion preparation: Black pepper extract containing high-purity piperine is dissolved in an ethanol-water mixed solution, 0.5% poloxamer 188 is added as an emulsifier, and 5 high-pressure homogenization cycles are performed under a pressure of 100 MPa to prepare a stable piperine nanoemulsion with a particle size of 50-100 nm; Composite encapsulation treatment: The nanoemulsion is mixed with the "collagen peptide-microcapsule" complex in a volume ratio of 1:5, and a liquid nitrogen pre-cooled spray freeze drying technology is used. Drying treatment is performed under a vacuum environment of -0.09 MPa to form a microencapsulated composition with piperine nanoparticles uniformly loaded on the surface, and the water content is controlled to be ≤3%.

[0033] Black pepper extract was dissolved in ethanol-water, 0.5% poloxamer 188 was added, and high-pressure homogenization at 100 MPa was performed five times to prepare a 50-100 nm nanoemulsion; the mixture was mixed with the complex at a ratio of 1:5, and the mixture was spray freeze-dried by liquid nitrogen pre-cooling (-0.09 MPa) to obtain a composition of surface-loaded piperine nanoparticles with a water content of ≤3%.

[0034] Among them, the finished product homogenization and quality control in S7 refers to homogenization treatment: the dried microencapsulated composition is passed through a 200-mesh sieve, 5% xylitol and 0.5% angelica extract are added, mixed evenly, and low-temperature ultrafine grinding is adopted at -20°C for 10 minutes to a particle size D90 <10μm; quality control indicators: microcapsule embedding rate is detected, ≥90%, particle size distribution 1-5μm proportion ≥85%, active ingredient retention rate VC ≥95%, crocin ≥90%, and accelerated stability test is carried out at 40°C / 75%RH for 1 month.

[0035] The dried microencapsulated composition was sieved through a 200-mesh sieve, and 5% xylitol and 0.5% Angelica dahurica extract were added and mixed evenly. The mixture was then ultrafinely ground at -20°C for 10 minutes to make the particle size D90 < 10 μm. For quality control, the microencapsulation efficiency (≥90%), particle size distribution (1-5 μm accounted for ≥85%), and active ingredient retention rate (VC ≥95%, crocin ≥90%) were tested, and an accelerated stability test was conducted at 40°C / 75% RH for 1 month.

[0036] It should be noted that, in this document, relational terms such as first and second, etc., are used only to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any actual relationship or order between these entities or operations. Moreover, the terms "comprises," "comprising," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that includes a list of elements includes not only those elements but also other elements not explicitly listed, or elements inherent to such process, method, article, or apparatus.

[0037] While embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions, and variations may be made to these embodiments without departing from the principles and spirit of the invention, and that the scope of the invention is defined by the appended claims and their equivalents.

Claims

1. A fish collagen and rose dual-effect anti-aging composition, characterized by: The core active ingredients of this fish collagen and rose dual-effect anti-aging composition include fish collagen peptides, double red roses, dried cranberries, wolfberries, roxburghii puree, and saffron. The small molecule peptide in the fish collagen peptides is less than 3000Da; the double red roses are rich in anthocyanins and polyphenols, which inhibit free radicals (ROS); and the dried cranberries contain proanthocyanidins (PACs), which inhibit the adhesion of pathogenic bacteria. Goji berry: Enhances mitochondrial function and improves cellular energy metabolism; Rosa roxburghii puree: Ultra-high vitamin C 2g / 100g, promotes collagen synthesis as a cofactor for prolyl hydroxylase; Saffron: The active ingredient crocin inhibits the NF-κB inflammatory pathway, reduces IL-6 inflammatory factors, and protects nerve cells.

2. A fish collagen and rose dual-effect anti-aging composition and a method for preparing the composite polysaccharide microencapsulation, characterized by: The specific steps of the fish collagen rose dual-effect anti-aging composition and the composite polysaccharide microencapsulation embedding preparation method are as follows: S1: Targeted enzymatic hydrolysis and graded purification of fish collagen peptides: Deep-sea cod skin is defatted and then soaked in acetic acid solution to remove impurities and proteins. The fish collagen peptide powder is obtained by enzymatic hydrolysis with a composite protease, purification with an ultrafiltration membrane, and spray drying. S2: Low-temperature synergistic extraction of antioxidant plant components: Double red roses were mixed in proportion and ultrasonically extracted with an ethanol-water solution containing citric acid. Saffron stigmas were cold-extracted at low temperature and then concentrated under reduced pressure. S3: Enzymatic modification and activity retention of marine active substances: The body wall of sea cucumbers was homogenized and then subjected to enzymatic hydrolysis, enzyme inactivation, centrifugation, and ethanol precipitation to extract polysaccharides. Kelp powder was degraded and deiodinated using high-temperature energy conversion technology. S4: Construction of composite polysaccharide microcapsule carriers: Dissolve xylooligosaccharides in water and stir to form a sol, add chitosan to adjust the pH, mix with plant extracts and marine polysaccharide solution, add olive oil for emulsification, and then cross-link to form microcapsule precursors; S5: Targeted coupling of fish collagen peptide and microcapsule: The fish collagen peptide was dissolved in buffer, activated with EDC and NHS, and then reacted with the microcapsule suspension at a suitable temperature to form a complex through Schiff base reaction; S6: Encapsulation of piperine nanosynergistic system: dissolving black pepper extract in solution, adding poloxamer and homogenizing under high pressure to prepare nanoemulsion, mixing with the complex and spray freeze drying to obtain a composition; S7: Finished product homogenization and quality control: The microencapsulated composition was sieved, mixed with xylitol and Angelica dahurica extract, and then ultrafinely ground at low temperature to detect the embedding efficiency and particle size and conduct stability tests.

3. The fish collagen and rose dual-effect anti-aging composition and its composite polysaccharide microencapsulation and embedding preparation method according to claim 2, characterized in that: The directed enzymatic hydrolysis and graded purification of fish collagen peptides in S1 refers to the following steps: raw material pretreatment: deep-sea cod skin is taken, defatted, and then soaked in 0.1M acetic acid solution at 4°C for 24 hours to remove impurities; enzymatic hydrolysis process: adding a composite protease, a neutral protease and collagenase mass ratio of 3:1, and enzymatic hydrolysis at 50°C and pH 7.0 for 8 hours to generate small molecule peptides with a molecular weight of <2000Da; graded purification: separation through an ultrafiltration membrane with a molecular weight cutoff of 1000Da, and the filtrate is collected and spray-dried with an inlet air temperature of 180°C and an outlet air temperature of 80°C to obtain refined fish collagen peptide powder.

4. The fish collagen and rose dual-effect anti-aging composition and its composite polysaccharide microencapsulation preparation method according to claim 2, characterized in that: The specific steps of low-temperature synergistic extraction of antioxidant plant components in S2 are as follows: Step 1: Mixed extraction system: double red rose petals, dried cranberries, wolfberries, and roxburghii purees were mixed in a ratio of 2:1:1:0.5, and 80% ethanol-water containing 0.5% citric acid was added to the mixed solution. Ultrasonic-assisted extraction was performed at 40°C, power 300W, and time 40min. Step 2: Saffron active enrichment: Take saffron stigma separately and perform low-temperature cold extraction with 50℃ deionized water, with a material-liquid ratio of 1:20, stirring speed of 200 rpm, and time of 3 hours. The extract is concentrated under reduced pressure at 45℃ and vacuum degree of -0.08MPa to 1 / 10 of the original volume.

5. The fish collagen and rose dual-effect anti-aging composition and its composite polysaccharide microencapsulation and embedding preparation method according to claim 2, characterized in that: The specific steps of enzymatic modification and activity retention of the marine active substances in S3 are as follows: Step 1: Extraction of sea cucumber polysaccharides: After the sea cucumber body wall is homogenized, 1000 U / g of cellulase is added to the raw material and enzymatically hydrolyzed at 45°C for 4 hours. The enzyme is inactivated in a boiling water bath, and the mixture is centrifuged at 8000 rpm for 15 minutes. The supernatant is collected and ethanol precipitation is performed to obtain sea cucumber polysaccharides at a final concentration of 80%. Step 2: Degradation of laminarin: A mixture of kelp powder and deionized water was treated with a high-temperature energy conversion technology at 85°C and a pulse power of 200W, with a material-to-liquid ratio of 1:15, to degrade alginate into low-molecular-weight fucoidan with a molecular weight of <50kDa. The alginate was then deiodinated by anion exchange resin and set aside.

6. The fish collagen and rose dual-effect anti-aging composition and its composite polysaccharide microencapsulation and embedding preparation method according to claim 2, characterized in that: The construction of the composite polysaccharide microcapsule carrier in S4 refers to the preparation of polysaccharide sol: 3% oligoxylose, 2% fucoidan, and 1% sea cucumber polysaccharide are dissolved in deionized water, and magnetically stirred at 300 rpm at 60°C to form a uniform sol, 0.1% chitosan is added and the pH is adjusted to 5.5 to enhance the stability of the sol; mixed emulsification treatment: plant extract and marine polysaccharide liquid are mixed in a 1:1 ratio and then added to the polysaccharide sol, olive oil is added in a 1:2 volume ratio, and emulsified in a 12000 rpm high-speed homogenizer for 20 minutes to form a stable water-in-oil emulsion; cross-linking capsule reaction: 1% calcium chloride cross-linking agent is added to the emulsion, and the reaction is carried out at a low temperature of 4°C for 4 hours. Through the cross-linking effect of calcium ions and polysaccharide chains, a composite polysaccharide microcapsule precursor with a three-dimensional network structure is formed.

7. The fish collagen and rose dual-effect anti-aging composition and its composite polysaccharide microencapsulation preparation method according to claim 2, characterized in that: The specific steps of the targeted coupling of fish collagen peptide and microcapsule in S5 are as follows: Step 1: Peptide activation: The fish collagen peptide in S1 was dissolved in 0.1 M phosphate buffer, pH 7.4, and 1% EDC, 1-ethyl-3-, 3-dimethylaminopropyl, carbodiimide and 0.5% NHS, N-hydroxysuccinimide were added and activated at room temperature for 30 min; Step 2: Coupling reaction: Add the activated collagen peptide dropwise to the S4 microcapsule suspension and react at 37°C, 150 rpm on a shaker for 6 hours. The collagen peptide is covalently coupled to the microcapsule surface through the Schiff base reaction to form a "collagen peptide-microcapsule" complex.

8. The fish collagen and rose dual-effect anti-aging composition and its composite polysaccharide microencapsulation preparation method according to claim 2, characterized in that: The encapsulation of the piperine nanosynergistic system in S6 includes the following steps: Nanoemulsion preparation: A black pepper extract containing high-purity piperine is dissolved in an ethanol-water mixed solution, 0.5% poloxamer 188 is added as an emulsifier, and the mixture is homogenized five times at 100 MPa to produce a stable piperine nanoemulsion with a particle size of 50-100 nm; Composite encapsulation treatment: The nanoemulsion is mixed with a "collagen peptide-microcapsule" complex in a volume ratio of 1:5, and dried using liquid nitrogen pre-cooling spray freeze drying technology under a vacuum environment of -0.09 MPa to form a microencapsulated composition with a uniform surface load of piperine nanoparticles, with a water content controlled to ≤3%.

9. The fish collagen and rose dual-effect anti-aging composition and its composite polysaccharide microencapsulation preparation method according to claim 2, characterized in that: The finished product homogenization and quality control in S7 refers to a homogenization process: the dried microencapsulated composition is passed through a 200-mesh sieve, 5% xylitol and 0.5% angelica extract are added, the mixture is evenly mixed, and low-temperature ultrafine grinding is performed at -20°C for 10 minutes to a particle size D90 <10 μm; quality control indicators include: microcapsule embedding efficiency ≥90%, particle size distribution 1-5 μm proportion ≥85%, active ingredient retention rate VC ≥95%, crocin ≥90%, and an accelerated stability test is performed at 40°C / 75% RH for 1 month.

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