An anti-aging active polypeptide composition, and a preparation method and application thereof

Abstract

This invention discloses an anti-aging active polypeptide composition, its preparation method, and its application. The anti-aging active polypeptide composition comprises the following components in parts by weight: 35-45 parts of collagen active tripeptide, 15-25 parts of artificially bred giant salamander peptide, 20-30 parts of ray peptide, and 15-25 parts of type III collagen peptide. Furthermore, the purity of each individual component of the collagen active tripeptide, artificially bred giant salamander peptide, ray peptide, and type III collagen peptide is ≥95%. This invention effectively solves the problems of low raw material purity, poor absorption, single efficacy, and insufficient stability in existing technologies by compounding the collagen active tripeptide, artificially bred giant salamander peptide, ray peptide, and type III collagen peptide in specific parts by weight and strictly controlling the purity of each individual component to be no less than 95%. This results in a high-purity, high-absorption, multi-target synergistic, and stable anti-aging polypeptide composition.

Description

Technical Field

[0001] This invention relates to the field of bioactive polypeptide technology, specifically to an anti-aging active polypeptide composition, its preparation method, and its application. Background Technology

[0002] With the improvement of people's living standards, anti-aging has become a core research topic in the fields of life sciences and health and beauty. Skin aging is a complex biological process, mainly manifested in collagen loss, elastic fiber breakage, free radical oxidative damage, and weakened skin barrier function, ultimately leading to problems such as fine lines, sagging, and dryness.

[0003] Currently, there are many types of anti-aging products on the market, among which peptides have attracted much attention due to their high bioactivity, well-defined targets, and good safety profile. However, existing anti-aging peptide products generally suffer from the following technical deficiencies: First, the purity of raw materials is insufficient, and impurities cause significant interference: Most commercially available peptide raw materials are produced using relatively crude processes, lacking systematic and refined purification. The raw materials often contain impurities such as large-molecule proteins, inorganic salts, heavy metals, and tissue residues, with the purity of individual components often below 85%. These impurities not only interfere with the bioactivity of the peptides themselves, reducing their efficacy, but also easily cause skin irritation, allergies, or gastrointestinal discomfort after oral administration, seriously affecting the product's safety and anti-aging effects.

[0004] Secondly, absorption efficiency is low and the onset of action is slow: Ordinary collagen peptides generally have a large molecular weight, mostly above 3000 Da. Large molecular weight peptides have difficulty effectively penetrating the skin's stratum corneum barrier, and when used in topical skincare products, they mostly remain on the skin's surface, unable to reach the dermis to exert their effects. In oral applications, large molecular weight peptides are easily broken down by digestive enzymes in the gastrointestinal tract, leading to a significant loss of active ingredients. The actual proportion that can be absorbed and utilized by the body is extremely low, resulting in negligible anti-aging effects and a very slow onset of action.

[0005] Third, their effects are singular and lack synergy: Most existing peptide products are single peptide ingredients or simple dual-component formulations, resulting in a single pathway of action. For example, some products only focus on surface hydration, while others only replenish superficial collagen. This singular mode of action cannot simultaneously meet the skin's multiple and comprehensive anti-aging needs, such as anti-oxidation, collagen regeneration, elasticity repair, barrier strengthening, and anti-photoaging. The lack of scientifically designed compounding systems means that the various components cannot produce synergistic effects, resulting in very limited overall anti-aging effects.

[0006] Fourth, insufficient component stability: Low-purity peptide raw materials contain various impurities, making their chemical properties more unstable and easily deactivated by environmental factors such as temperature, pH, light, and oxygen. This results in products made from such raw materials having a shorter shelf life, and during long-term use, the product's efficacy is poorly sustained, with a significant decline in anti-aging effects, failing to provide users with long-lasting and stable anti-aging protection.

[0007] Therefore, developing a high-purity, highly absorbable, multi-target synergistic, and stable anti-aging peptide product is a technical problem that urgently needs to be solved by those skilled in the art. Summary of the Invention

[0008] The purpose of this invention is to provide an anti-aging active polypeptide composition, its preparation method, and its application, aiming to improve the problems of low purity, poor absorption, single efficacy, and insufficient stability of existing anti-aging polypeptide products.

[0009] This invention is implemented as follows: According to a first aspect of the present invention, the present invention provides an anti-aging active polypeptide composition, the composition comprising the following components in parts by weight: 35-45 parts of collagen active tripeptide 15-25 parts of peptides from artificially bred giant salamanders 20-30 parts of ray peptides Type III collagen peptides, 15-25 parts; Furthermore, the purity of each individual component of the collagen active tripeptide, artificially bred giant salamander peptide, ray peptide, and type III collagen peptide is ≥95%.

[0010] In the technical solution of this invention, the four active peptides each undertake a specific anti-aging function, and produce a synergistic effect through compounding: Collagen active tripeptide: With an extremely small molecular weight (approximately 280 Da), it has a very strong transdermal absorption capacity and mainly acts on the epidermal layer of the skin to exert antioxidant, moisturizing and hydrating effects.

[0011] Artificially bred giant salamander peptides: rich in various active factors, which can activate skin cells, repair damaged skin barriers, and enhance the skin's own defense capabilities.

[0012] Ray peptides: Focused on repairing the elastic fibers of the dermis, effectively improving skin elasticity and reducing existing fine lines and wrinkles.

[0013] Type III collagen peptides are one of the main types of collagen that make up the reticular layer of the dermis. They can directly replenish the key collagen lost by the skin, reconstruct the dermal scaffold, and improve skin laxity from the root.

[0014] These four ingredients have distinct functions, acting on the epidermis, basement membrane, and dermis respectively, forming a complete chain of action for "layered anti-aging and targeted repair." Meanwhile, the guarantee of high purity (≥95%) eliminates the interference of impurities on efficacy and potential irritation, ensuring the product's safety and high efficacy.

[0015] Furthermore, the collagen active tripeptide has a molecular weight of 280 Da and a purity ≥98%; the artificially bred giant salamander peptide has a molecular weight ≤1000 Da and a purity ≥95%; the ray peptide has a molecular weight of 500-1000 Da and a purity ≥96%; and the type III collagen peptide has a molecular weight of 300-500 Da and a purity ≥97%. Using these specific molecular weights and purities of active peptides provides the technical basis for achieving high absorption, high activity, and high stability in anti-aging active polypeptide compositions.

[0016] According to a second aspect of the present invention, the present invention provides a method for preparing the above-mentioned anti-aging active polypeptide composition, comprising the following steps: S100, independently prepare collagen active tripeptides, artificially bred salamander peptides, ray peptides and type III collagen peptides of the required purity; S200. Weigh the four high-purity active peptide powders prepared in step S100 according to the specified ratio. S300. Mix the four weighed powders evenly under sterile, light-proof, and low-temperature conditions to obtain a mixed powder. S400. The mixed powder is subjected to secondary vacuum freeze-drying to obtain the finished anti-aging active polypeptide composition.

[0017] Furthermore, the specific steps of step S100 are as follows: S101. Raw material pretreatment: After cleaning, mincing and removing impurities from the corresponding raw materials, add purified water and stir to make a uniform slurry. S102, Low-temperature biological enzymatic hydrolysis: Add a complex protease to the homogenate and hydrolyze for 4-6 hours at pH 6.0-7.0 and temperature 40-45℃, then raise the temperature to 85-90℃ to inactivate the enzyme for 15 minutes. S103, Centrifugation to remove impurities: Centrifuge the enzyme hydrolysate after enzyme inactivation and collect the supernatant enzyme hydrolysate; S104, Multi-stage membrane separation and purification: The supernatant enzymatic hydrolysate is sequentially separated and purified by passing it through a microfiltration membrane, an ultrafiltration membrane, and a nanofiltration membrane to obtain a high-purity peptide solution; S105, Decolorization and Deodorization Refining: High-purity peptide liquid is adsorbed, decolorized and deodorized using activated carbon, and then filtered to obtain a colorless and odorless refined peptide liquid. S106. Low-temperature freeze drying: The refined peptide liquid is subjected to vacuum freeze drying to obtain a single active peptide powder with a moisture content of ≤5%.

[0018] Furthermore, in step S101, the corresponding raw materials for the collagen active tripeptide, artificially bred giant salamander peptide, ray peptide and type III collagen peptide are tilapia skin, artificially bred giant salamander muscle tissue, deep-sea ray cartilage and / or skin and tilapia scales, respectively; after adding purified water, the material-to-liquid ratio is 1:8 to 1:12, and the temperature is controlled at 4 to 8°C throughout the process to prevent the raw materials from deteriorating and becoming inactive.

[0019] Furthermore, in step S102, the complex protease includes a neutral protease and a flavor protease, and the mass ratio of the neutral protease to the flavor protease is 1:1 to 2:1; the amount of the complex protease added accounts for 0.8% to 1.2% of the total mass of the homogenate.

[0020] Furthermore, in step S103, the enzyme hydrolysate after enzyme inactivation is first cooled to room temperature, and then transferred to a low-temperature high-speed centrifuge at a speed of 8000 r / min and a temperature of 4℃ for 25 min to remove residues, fats and insoluble macromolecular impurities, and the supernatant enzyme hydrolysate is collected.

[0021] Furthermore, in step S104, the molecular weight cutoff of the ultrafiltration membrane is 1000 Da, and the multi-stage membrane separation and purification process is operated at 4-8℃; in step S105, the amount of activated carbon added is 0.5% of the peptide solution mass, and the stirring and adsorption time is not less than 30 min; in step S106, the parameters for vacuum freeze drying are: temperature -20℃, vacuum degree 10 Pa, and drying time 3-4 h.

[0022] Furthermore, in step S300, the mixing process is carried out in a Class 10,000 clean and sterile mixer, in complete darkness, with an ambient temperature ≤25℃. The mixer is first stirred at a speed of 20-30 r / min for no less than 15 min, and then stirred at a speed of 50-60 r / min for no less than 25 min.

[0023] According to a third aspect of the present invention, the present invention provides an anti-aging product comprising the above-mentioned anti-aging active polypeptide composition; the anti-aging product is a skin care product, an oral dietary supplement, or a medical repair product.

[0024] This invention also provides applications of the above-mentioned anti-aging active polypeptide composition: the anti-aging active polypeptide composition can be used to prepare products with anti-aging, antioxidant, collagen synthesis-promoting, skin moisturizing, or skin barrier-repairing functions. These products include, but are not limited to, skincare products, oral dietary supplements, or medical repair products.

[0025] Compared with the prior art, the beneficial effects of the present invention are: 1. High purity guaranteed, free from impurities: Through a proprietary preparation process, the purity of each of the four core peptides is ≥95%, effectively removing impurities such as macromolecular proteins, inorganic salts, heavy metals, and tissue residues. This not only significantly enhances peptide activity but also eliminates skin irritation and gastrointestinal discomfort caused by impurities, thus improving both safety and efficacy.

[0026] 2. Comprehensive anti-aging across the entire dermis, targeting every angle: Collagen active tripeptides protect the epidermis, providing antioxidant and hydrating benefits; artificially bred salamander peptides activate cells and repair the skin barrier; ray peptides repair dermal elastic fibers and reduce fine lines; type III collagen peptides replenish core collagen and reconstruct the dermal support structure. The four components have clearly defined roles and work synergistically to combat aging across all layers.

[0027] 3. Small molecules with high penetration and low absorption loss: All four active peptides are small molecule polypeptides with an average molecular weight of ≤650Da. In addition, they are high-purity and free from impurities. When used externally, they can quickly penetrate the stratum corneum of the skin and reach the dermis. When taken orally, they do not require gastrointestinal enzymatic decomposition and can be directly absorbed and utilized by the human body. The bioavailability is 2-3.5 times higher than that of ordinary low-purity polypeptides.

[0028] 4. High stability and long-lasting anti-aging: High-purity small molecule peptides form a stable polypeptide complex after compounding. It is resistant to acids and alkalis (active within the pH range of 4-8), and resistant to high and low temperatures (active remains intact at -20℃ to 60℃). It is not easily deactivated by light or oxygen. The shelf life can reach 24 months when sealed at room temperature away from light. The activity retention rate is ≥95% after accelerated stability testing. Attached Figure Description

[0029] Figure 1 This is a flowchart of the method for preparing anti-aging active polypeptide compositions provided by the present invention; Figure 2 This is a flowchart illustrating the specific method for step S100. Detailed Implementation

[0030] The following description, in conjunction with the accompanying drawings and specific embodiments, provides further details: Unless otherwise specified, the raw materials used in the following embodiments are all commercially available conventional products with the following specifications: Collagen active tripeptide (molecular weight 280 Da, purity ≥98%). Artificially bred giant salamander peptides (molecular weight ≤1000Da, purity ≥95%). Ray peptides (molecular weight 500-1000 Da, purity ≥96%) Type III collagen peptides (molecular weight 300-500 Da, purity ≥97%) Ordinary collagen peptides (molecular weight 1000-2000 Da, purity ≥95%). Bird's nest peptides (molecular weight 800-1200 Da, purity ≥95%) Elastin peptides (molecular weight 600-1000 Da, purity ≥96%).

[0031] Example 1 (Preferred formulation): The formulation of the anti-aging active peptide composition (parts by weight) is as follows: 40 parts collagen active tripeptide, 20 parts artificially bred giant salamander peptide, 25 parts ray peptide, and 20 parts type III collagen peptide. Furthermore, the purity of each individual component of the collagen active tripeptide, artificially bred giant salamander peptide, ray peptide, and type III collagen peptide is ≥95%.

[0032] like Figure 1 and Figure 2 As shown, the specific preparation method is as follows: S100, separately prepare collagen active tripeptides, artificially bred salamander peptides, ray peptides and type III collagen peptides of the required purity: S101. Raw material pretreatment: The corresponding raw materials (tilapia skin for collagen active tripeptide, muscle tissue of artificially bred giant salamander for artificially bred giant salamander peptide, cartilage and / or skin of deep-sea ray for ray peptide, and tilapia scale for type III collagen peptide) are washed, minced, and impurities are removed. Then, purified water is added, and the material-to-liquid ratio is controlled at 1:8 to 1:12. The temperature is controlled at 4 to 8℃ throughout the process. The mixture is stirred to form a uniform slurry to prevent the raw materials from deteriorating and becoming inactive.

[0033] S102. Low-temperature enzymatic hydrolysis: Add a complex protease, including neutral protease and flavor protease, to the above homogenate at a mass ratio of 1:1 to 2:1. The amount of complex protease added accounts for 0.8% to 1.2% of the total mass of the homogenate. Adjust the pH to 6.0 to 7.0 and control the temperature at 40 to 45°C for 4 to 6 hours of enzymatic hydrolysis. After the enzymatic hydrolysis is completed, rapidly raise the temperature to 85 to 90°C to inactivate the enzyme for 15 minutes and terminate the enzymatic hydrolysis reaction.

[0034] S103. Centrifugation to remove impurities: Cool the enzyme hydrolysate after enzyme inactivation to room temperature, transfer it to a low-temperature high-speed centrifuge, and centrifuge at 8000 r / min and 4℃ for 25 min to remove residues, fats and insoluble macromolecular impurities, and collect the supernatant enzyme hydrolysate.

[0035] S104. Multi-stage membrane separation and purification: The supernatant enzymatic hydrolysate is sequentially separated and purified by microfiltration, ultrafiltration, and nanofiltration. Microfiltration removes residual suspended solids and bacteria; ultrafiltration has a molecular weight cutoff of 1000 Da, retaining large protein molecules, polysaccharides, and other impurities while allowing the target small peptides to pass through; nanofiltration retains small inorganic salts, monosaccharides, and free amino acids, concentrating the target active peptides. The entire multi-stage membrane separation and purification process is operated at 4-8℃. After this step, a high-purity peptide solution is obtained, with the purity of a single peptide solution initially increased to over 95%.

[0036] S105. Decolorization and deodorization refining: Food-grade activated carbon is used to adsorb, decolorize, and deodorize the high-purity peptide solution. The amount of activated carbon added is 0.5% of the peptide solution mass. After stirring and adsorbing for 30 minutes, the activated carbon is removed by filtration to obtain a colorless and odorless high-purity peptide solution.

[0037] S106. Low-temperature freeze drying: The purified peptide solution is subjected to vacuum freeze drying. The parameters for vacuum freeze drying are: temperature -20℃, vacuum degree 10Pa, drying time 3~4h, to obtain a single active peptide powder with a moisture content ≤5%.

[0038] Tests showed that the purity of the four single active peptide powders prepared by the above methods was ≥95%, which met the requirements. After sealing, they were temporarily stored in a nitrogen environment for later use.

[0039] S200. Raw material weighing: According to the above formula ratio (40 parts of collagen active tripeptide, 20 parts of artificially bred salamander peptide, 25 parts of ray peptide, and 20 parts of type III collagen peptide), accurately weigh the four high-purity active peptide powders prepared in step S106, with a weighing error ≤0.1%.

[0040] S300. Aseptic Mixing: Transfer the weighed four powders into a Class 10,000 clean and sterile mixer. Keep the mixture away from light throughout the process. The ambient temperature should be ≤25℃. First, stir at 30r / min for 15min, then stir at 60r / min for at least 25min to achieve thorough physical mixing without layering, agglomeration, or segregation, resulting in a uniformly mixed composite peptide powder.

[0041] S400, Secondary Vacuum Freeze-drying: The above-mentioned composite peptide powder is subjected to secondary vacuum freeze-drying to remove trace amounts of moisture adsorbed during the mixing process. The parameters for vacuum freeze-drying are: temperature -20℃, vacuum degree 10Pa, drying time 3~4h, to further consolidate purity and stability. After freeze-drying, sterile nitrogen gas is quickly filled and sealed to obtain the finished anti-aging active peptide composition. Example 2

[0042] Formula composition (parts by weight): 35 parts collagen active tripeptide, 20 parts artificially bred giant salamander peptide, 25 parts ray peptide, and 20 parts type III collagen peptide.

[0043] Preparation method: Same as in Example 1, except that the amount of collagen active tripeptide added was adjusted to 35 parts. Example 3

[0044] Formula composition (parts by weight): 45 parts of collagen active tripeptide, 20 parts of artificially bred giant salamander peptide, 25 parts of ray peptide, and 20 parts of type III collagen peptide.

[0045] Preparation method: Same as in Example 1, except that the amount of collagen active tripeptide added was adjusted to 45 parts. Example 4

[0046] Formula composition (parts by weight): 40 parts of collagen active tripeptide, 15 parts of artificially bred giant salamander peptide, 25 parts of ray peptide, and 20 parts of type III collagen peptide.

[0047] Preparation method: Same as in Example 1, except that the amount of artificially bred giant salamander peptide added was adjusted to 15 parts. Example 5

[0048] Formula composition (parts by weight): 40 parts of collagen active tripeptide, 25 parts of artificially bred giant salamander peptide, 25 parts of ray peptide, and 20 parts of type III collagen peptide.

[0049] Preparation method: Same as in Example 1, except that the amount of artificially bred giant salamander peptide added was adjusted to 25 parts. Example 6

[0050] Formula composition (parts by weight): 40 parts of collagen active tripeptide, 20 parts of artificially bred giant salamander peptide, 20 parts of ray peptide, and 20 parts of type III collagen peptide.

[0051] Preparation method: Same as in Example 1, except that the amount of ray peptide added was adjusted to 20 parts. Example 7

[0052] Formula composition (parts by weight): 40 parts of collagen active tripeptide, 20 parts of artificially bred giant salamander peptide, 30 parts of ray peptide, and 20 parts of type III collagen peptide.

[0053] Preparation method: Same as in Example 1, except that the amount of ray peptide added was adjusted to 30 parts. Example 8

[0054] Formula composition (parts by weight): 40 parts of collagen active tripeptide, 20 parts of artificially bred giant salamander peptide, 25 parts of ray peptide, and 15 parts of type III collagen peptide.

[0055] Preparation method: Same as in Example 1, except that the amount of type III collagen peptides added was adjusted to 15 parts. Example 9

[0056] Formula composition (parts by weight): 40 parts of collagen active tripeptide, 20 parts of artificially bred giant salamander peptide, 25 parts of ray peptide, and 25 parts of type III collagen peptide.

[0057] Preparation method: Same as in Example 1, except that the amount of type III collagen peptides added was adjusted to 25 parts. Example 10

[0058] Formula composition (parts by weight): 38 parts of collagen active tripeptide, 18 parts of artificially bred giant salamander peptide, 22 parts of ray peptide, and 18 parts of type III collagen peptide.

[0059] Preparation method: Same as in Example 1, except that the amount of each component added is adjusted to the above intermediate value. Example 11

[0060] Formula composition (parts by weight): 42 parts of collagen active tripeptide, 22 parts of artificially bred giant salamander peptide, 28 parts of ray peptide, and 22 parts of type III collagen peptide.

[0061] Preparation method: Same as in Example 1, except that the amount of each component added is adjusted to the above intermediate value. Example 12

[0062] Formula composition (parts by weight): 40 parts of collagen active tripeptide, 15 parts of artificially bred giant salamander peptide, 20 parts of ray peptide, and 20 parts of type III collagen peptide.

[0063] Preparation method: Same as in Example 1, except that the amount of artificially bred giant salamander peptide was adjusted to 15 parts and the amount of ray peptide was adjusted to 20 parts. Example 13

[0064] Formula composition (parts by weight): 40 parts of collagen active tripeptide, 20 parts of artificially bred giant salamander peptide, 30 parts of ray peptide, and 25 parts of type III collagen peptide.

[0065] Preparation method: Same as in Example 1, except that the amount of ray peptide was adjusted to 30 parts and the amount of type III collagen peptide was adjusted to 25 parts.

[0066] Comparative Example To fully verify the superiority of the technical solution of this invention, the following comparative examples are provided. The preparation methods of each comparative example differ from those of Example 1 only in the composition and ratio of the active peptides; all other conditions remain the same.

[0067] Comparative Examples 1-4 (Single Core Peptide Groups) Comparative Example 1: Single collagen active tripeptide group.

[0068] Comparative Example 2: Single artificially bred giant salamander peptide group.

[0069] Comparative Example 3: Single ray peptide group.

[0070] Comparative Example 4: Single type III collagen peptide group.

[0071] Comparative Examples 5-6 Comparative Example 5: 50 parts of collagen active tripeptide, 10 parts of artificially bred giant salamander peptide, 10 parts of ray peptide, and 10 parts of type III collagen peptide.

[0072] Comparative Example 6: 30 parts of collagen active tripeptide, 30 parts of artificially bred giant salamander peptide, 15 parts of ray peptide, and 15 parts of type III collagen peptide.

[0073] Comparative ratio 7-9 (common single peptide groups on the market) Comparative Example 7: Single ordinary collagen peptide group.

[0074] Comparative Example 8: Single bird's nest peptide group.

[0075] Comparative Example 9: Single elastin peptide group.

[0076] Comparative ratio 10-12 (common dual-peptide combinations on the market) Comparative Example 10: Regular collagen peptides + bird's nest peptides, 1:1 ratio.

[0077] Comparative Example 11: Regular collagen peptides + elastin peptides, 1:1 ratio.

[0078] Comparative Example 12: Bird's Nest Peptide + Elastin Peptide, 1:1 ratio.

[0079] Performance testing and effect evaluation 1. Test Indicators and Methods DPPH free radical scavenging rate: The absorbance of the sample at 517 nm was detected by spectrophotometry, the scavenging rate was calculated, and the antioxidant capacity was evaluated.

[0080] Collagen synthesis promotion rate: The ELISA method was used to detect the promoting effect of the sample on collagen synthesis in human skin fibroblasts and the promotion rate was calculated.

[0081] Skin moisture retention rate: The sample was applied to the inside of the arm, and the skin moisture content was measured before and 24 hours after application using a skin moisture meter to calculate the retention rate.

[0082] Cell viability: The MTT assay was used to detect the effect of the samples on the activity of human skin fibroblasts (HDF) and to calculate cell viability.

[0083] 2. Test Results Table 1: Comparison results between the preferred formulation group (Example 1) and the single peptide group (Comparative Examples 1-4, 7-9)

[0084] As shown in Table 1, the preferred formulation group (Example 1) of the present invention has significantly better performance than all single peptide groups, which verifies that the combination of the four core peptides produces excellent synergistic effects.

[0085] Table 2: Comparison results of the optimized formulation group (Example 1) with the conventional compound group (Comparative Examples 5-6) and the commercially available single peptide group (Comparative Examples 7-9)

[0086] As shown in Table 2, the preferred formulation of the present invention (Example 1) has significantly better performance in all aspects than the conventional compound formulation and the common single peptide formulation on the market. This indicates that the scientific formulation of the present invention can better exert the synergistic effect between the components and has better effect than the simple compound products of the prior art.

[0087] Table 3: Comparison results of different formulation groups of the present invention (Examples 1-13) with single peptide groups and dual peptide combination groups (Comparative Examples 7-12)

[0088] As shown in Table 3, all indicators of the formulation groups of this invention (Examples 1-13) are comprehensively superior to those of the single peptide group and the popular dual peptide combination group on the market. Furthermore, whether it is the upper or lower limit of the formulation range for each component, the intermediate value, or the combination of multiple components taking the upper or lower limit simultaneously, the effect is significantly better than the comparative example. Moreover, the differences between the various example groups are within an acceptable range, verifying the scientific validity and stability of the formulation range of this invention and providing flexible adjustment space for industrial production.

[0089] 3. Core efficacy and purity verification Purity verification: According to the high performance liquid chromatography (HPLC) method used by the third-party testing agency, the purity of collagen active tripeptide in the finished product prepared by this process is 98%, artificially bred salamander peptide is 95%, ray peptide is 96%, and type III collagen peptide is 97.0%. The purity of the four single components is consistently up to standard.

[0090] Absorption efficiency: Transdermal absorption test shows that the transdermal absorption rate of the composition of the present invention reaches 82% in 30 minutes and 87% in 24 hours.

[0091] Human efficacy test: 35 volunteers aged 28-58 years were selected. After continuous external application of the product of this invention (Example 1) for 28 days, skin moisture increased by 34.2%, elasticity increased by 30.6%, fine lines faded by 41.3%, and collagen density increased by 20.8%.

[0092] Stability: After being stored at room temperature and protected from light for 24 months, the purity of the single component was still ≥95%, and the activity retention rate was ≥95.3%.

[0093] Safety: The skin irritation test result was level 0, with no irritation reaction, indicating high safety.

[0094] The above are merely preferred embodiments of the present invention and are not intended to limit the present invention. For those skilled in the art, the present invention can have various modifications and variations. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. An anti-aging active polypeptide composition, characterized in that, The composition comprises the following components in parts by weight: 35-45 parts of collagen active tripeptide 15-25 parts of peptides from artificially bred giant salamanders 20-30 parts of ray peptides Type III collagen peptides, 15-25 parts; Furthermore, the purity of each individual component of the collagen active tripeptide, artificially bred giant salamander peptide, ray peptide, and type III collagen peptide is ≥95%.

2. A method for preparing the anti-aging active polypeptide composition of claim 1, characterized in that, Includes the following steps: S100, independently prepare collagen active tripeptides, artificially bred salamander peptides, ray peptides and type III collagen peptides of the required purity; S200. Weigh the four high-purity active peptide powders prepared in step S100 according to the specified ratio. S300. Mix the four weighed powders evenly under sterile, light-proof, and low-temperature conditions to obtain a mixed powder. S400. The mixed powder is subjected to secondary vacuum freeze-drying to obtain the finished anti-aging active polypeptide composition.

3. The preparation method according to claim 2, characterized in that, The specific steps of step S100 are as follows: S101. Raw material pretreatment: After cleaning, mincing and removing impurities from the corresponding raw materials, add purified water and stir to make a uniform slurry. S102, Low-temperature biological enzymatic hydrolysis: Add a complex protease to the homogenate and hydrolyze for 4-6 hours at pH 6.0-7.0 and temperature 40-45℃, then raise the temperature to 85-90℃ to inactivate the enzyme for 15 minutes. S103, Centrifugation to remove impurities: Centrifuge the enzyme hydrolysate after enzyme inactivation and collect the supernatant enzyme hydrolysate; S104, Multi-stage membrane separation and purification: The supernatant enzymatic hydrolysate is sequentially separated and purified by passing it through a microfiltration membrane, an ultrafiltration membrane, and a nanofiltration membrane to obtain a high-purity peptide solution; S105, Decolorization and Deodorization Refining: High-purity peptide liquid is adsorbed, decolorized and deodorized using activated carbon, and then filtered to obtain a colorless and odorless refined peptide liquid. S106. Low-temperature freeze drying: The refined peptide liquid is subjected to vacuum freeze drying to obtain a single active peptide powder with a moisture content of ≤5%.

4. The preparation method according to claim 3, characterized in that, In step S101, the corresponding raw materials for the collagen active tripeptide, artificially bred giant salamander peptide, ray peptide and type III collagen peptide are tilapia skin, artificially bred giant salamander muscle tissue, deep-sea ray cartilage and / or skin and tilapia scales, respectively; after adding purified water, the material-to-liquid ratio is 1:8~1:12, and the temperature is controlled at 4~8℃ throughout the process to prevent the raw materials from deteriorating and becoming inactive.

5. The preparation method according to claim 3, characterized in that, In step S102, the complex protease includes a neutral protease and a flavor protease, and the mass ratio of the neutral protease to the flavor protease is 1:1 to 2:1; the amount of the complex protease added accounts for 0.8% to 1.2% of the total mass of the homogenate.

6. The preparation method according to claim 3, characterized in that, In step S103, the enzyme hydrolysate after enzyme inactivation is first cooled to room temperature, and then transferred to a low-temperature high-speed centrifuge at a speed of 8000 r / min and a temperature of 4℃ for 25 min to remove residues, fats and insoluble macromolecular impurities, and the supernatant enzyme hydrolysate is collected.

7. The preparation method according to claim 3, characterized in that, In step S104, the ultrafiltration membrane has a molecular weight cutoff of 1000 Da, and the multi-stage membrane separation and purification process is operated at 4-8℃; in step S105, the activated carbon addition is 0.5% of the peptide solution mass, and the stirring and adsorption time is not less than 30 min; in step S106, the parameters for vacuum freeze drying are: temperature -20℃, vacuum degree 10 Pa, and drying time 3-4 h.

8. The preparation method according to claim 3, characterized in that, In step S300, the mixing process is carried out in a Class 10,000 clean and sterile mixer, in complete darkness, with an ambient temperature ≤25℃. The mixer is first stirred at a speed of 20-30 r / min for no less than 15 min, and then stirred at a speed of 50-60 r / min for no less than 25 min.

9. The use of the anti-aging active polypeptide composition according to claim 1 in the preparation of products having anti-aging, antioxidant, collagen synthesis-promoting, skin moisturizing, or skin barrier-repairing functions.

10. An anti-aging product, characterized in that, The product comprises the anti-aging active polypeptide composition as described in claim 1; the anti-aging product is a skin care product, an oral dietary supplement, or a medical repair product.

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