A collagen peptide for delaying aging and a preparation method thereof

Abstract

The application discloses a collagen peptide for delaying aging and a preparation method thereof, wherein the collagen peptide for delaying aging is a low-moisture freeze-dried powder preparation which is composed of the following effective components in mass fraction: 55-75 parts of small-molecule fish skin collagen peptide, 8-15 parts of mitochondrial activating peptide, 6-12 parts of elastin repair peptide, 5-10 parts of ultra-micro broken aronia melanocarpa extract, 3-8 parts of sodium hyaluronate and 2-5 parts of vitamin E microcapsule; the moisture content of the collagen peptide freeze-dried powder finished product is less than or equal to 5%; the components of the application have synergistic effect, the anti-aging effect is comprehensive, a multi-active component compound system is adopted, the functions of the components are complementary and there is no antagonistic effect, and the skin aging problems can be improved in multiple dimensions such as cell metabolism, tissue repair, antioxidation and collagen supplement.

Description

Technical Field

[0001] This invention belongs to the field of bioactive peptide preparation technology, specifically a collagen peptide for delaying aging and its preparation method. Background Technology

[0002] Aging is a progressive degenerative disease that occurs in the body's cells, tissues, and organs with age. Externally, it manifests as loose skin, decreased elasticity, fine lines, and dull skin tone. Internally, it is characterized by reduced antioxidant capacity, weakened mitochondrial metabolic activity, and accelerated cell apoptosis. The core mechanism of skin aging lies in the significant loss of collagen in the dermis, the breakage and aging of elastic fibers, the decline of mitochondrial function, and the continuous oxidative stress damage caused by the excessive accumulation of free radicals. Collagen peptides, with their advantages of small molecular weight, easy absorption, high biocompatibility, and safety without irritation, are widely used in anti-aging health care and skincare.

[0003] Existing collagen peptide products are mostly made from single fish skin, pig skin, or bovine bone collagen peptides, with limited functional mechanisms. They can only replenish lost collagen and cannot address core aging issues such as declining mitochondrial function, damaged elastic fibers, and persistent oxidative stress damage. Furthermore, traditional preparation processes often involve high-temperature single-stage enzymatic hydrolysis, conventional stirring and mixing, and high-temperature concentration and drying, which easily leads to degradation and inactivation of active peptides, uneven mixing of components, poor system stability, and low bioavailability. In addition, existing anti-aging collagen peptides rarely combine multiple mechanisms of action such as mitochondrial activation, elastic fiber repair, and endogenous antioxidant activity, resulting in a one-sided and limited anti-aging effect, making it difficult to achieve comprehensive and long-lasting anti-aging benefits.

[0004] Based on this, a collagen peptide for delaying aging and its preparation method were designed. Summary of the Invention

[0005] In view of the above situation and to overcome the shortcomings of the prior art, the present invention provides an anti-aging collagen peptide and its preparation method, which effectively solves the problems mentioned in the background.

[0006] To achieve the above objectives, the present invention provides the following technical solution: a collagen peptide for delaying aging, wherein the collagen peptide is a low-moisture lyophilized powder formulation, composed of the following effective components in parts by weight: Small molecule fish skin collagen peptides 55-75 parts, mitochondrial activation peptides 8-15 parts, elastin repair peptides 6-12 parts, ultra-micro cell wall broken black chokeberry extract 5-10 parts, sodium hyaluronate 3-8 parts, vitamin E microcapsules 2-5 parts. The moisture content of the finished collagen peptide freeze-dried powder is ≤5%.

[0007] Preferably, the method for preparing the mitochondrial activating peptide includes the following steps: S11. Raw material pretreatment: Select fresh bovine myocardial tissue, remove fascia and fat, rinse with sterile water 3-5 times, freeze dry at low temperature, pulverize, and pass through an 80-100 mesh sieve to obtain bovine myocardial dry powder; add bovine myocardial dry powder to sterile purified water at a material-to-liquid ratio of 1:8-1:10 g / mL, stir evenly, and prepare myocardial suspension. S12. Low-temperature stepwise enzymatic hydrolysis: Adjust the pH of the myocardial suspension to 7.0-7.5, raise the temperature to 45-50℃, add neutral protease at a concentration of 1.2-1.8% of the substrate weight, and hydrolyze at a constant temperature with stirring for 2.5-3.5 hours at a stirring speed of 180-220 r / min; then adjust the pH to 8.0-8.5, raise the temperature to 50-55℃, add alkaline protease at a concentration of 0.8-1.2% of the substrate weight, and continue enzymatic hydrolysis for 1.5-2 hours; S13. Enzyme inactivation centrifugation: After the enzymatic hydrolysis is completed, the temperature is rapidly raised to 90-95℃ and the enzyme is inactivated at a constant temperature for 8-12 minutes. After cooling to room temperature, the enzyme is centrifuged at 8000-10000r / min for 15-20 minutes and the upper enzymatic hydrolysis supernatant is collected. S14. Gradient ultrafiltration purification: The supernatant is sequentially passed through 10kDa, 3kDa, and 1kDa ultrafiltration membranes for gradient ultrafiltration, and the filtrate in the 1-3kDa molecular weight range is collected. S15. Freeze-drying: Place the ultrafiltration filtrate in a vacuum freeze dryer, pre-freeze at -40~-35℃ for 4-6 hours, vacuum degree 10-15Pa, freeze-dry for 18-24 hours to obtain powdered mitochondrial activating peptides.

[0008] Preferably, the method for preparing the elastin repair peptide includes the following steps: S21. Raw material pretreatment: Select fresh porcine aortic elastin raw material, remove impurities and blood, wash with sterile water, drain at low temperature, pulverize and pass through a 100-120 mesh sieve to obtain coarse elastin powder; add coarse elastin powder to sterile water at a material-to-liquid ratio of 1:10-1:12 g / mL, soak for 12-16 hours to fully swell. S22. Compound enzymatic hydrolysis treatment: Adjust the pH of the swollen mixture to 6.0-6.5, stabilize the temperature at 48-52℃, add compound protease, wherein the compound protease is papain and flavor protease in a mass ratio of 2:1, and the total amount of enzyme added is 1.5-2.0% of the substrate mass. Stir at a constant temperature for 3-4 hours and stir at a speed of 200-240 r / min. S23. Enzyme inactivation and impurity removal: After the enzymatic hydrolysis is completed, the temperature is raised to 85-90℃ and the enzyme is inactivated at a constant temperature for 10-15 min. The temperature is then naturally cooled to 25-30℃. The mixture is centrifuged at 9000-11000 r / min for 20-25 min to remove precipitated impurities and the clear supernatant is collected. S24. Refining and purification: The supernatant is filtered using a 2kDa ultrafiltration membrane. After collecting the filtrate, it is decolorized and deodorized using macroporous adsorption resin. The elution flow rate is 2-3 BV / h and the elution time is 40-60 min. S25. Low-temperature freeze drying: The purified filtrate after decolorization and deodorization is subjected to vacuum freeze drying. The pre-freezing temperature is -45 to -40℃, the pre-freezing time is 5-7 hours, the vacuum degree is 8-12 Pa, and the freeze drying time is 20-26 hours to obtain highly active elastin repair peptide powder.

[0009] Preferably, the molecular weight of the small molecule fish skin collagen peptide is 800-1500 Da; The cell wall breaking particle size of the ultra-micro cell wall broken black chokeberry extract is 200-300 nm. The encapsulation rate of the vitamin E microcapsules is ≥92%.

[0010] A method for preparing collagen peptides that delay aging includes the following steps: S1. Preparation of base solution: Take 100-150 parts of purified water as the dissolving medium according to the process ratio, add it to a sterilized reaction vessel, heat to 35-40℃, stir at 150-180 r / min, add small molecule fish skin collagen peptides and sodium hyaluronate in sequence, stir at a constant temperature for 15-20 min until the materials are completely dissolved to obtain a uniform base mixture; the purified water is only used for process dissolution and is completely removed in subsequent processes and does not remain in the finished product. S2, Innovative Peptide Combination: Maintain a temperature of 35-40℃ and a stirring speed of 180-200r / min. Slowly add mitochondrial activating peptides and elastin repair peptides to the base mixture and stir at a uniform speed for 25-30min to fully disperse and blend the active peptides. S3. Addition of functional excipients: Cool to 28-32℃, reduce stirring speed to 120-150r / min, add ultra-fine cell wall broken black chokeberry extract and vitamin E microcapsules in sequence, stir at low speed for 18-22min to avoid high temperature destroying active ingredients; S4. Low-temperature homogenization treatment: Transfer the mixture to a high-pressure homogenizer and homogenize it 2-3 times under the conditions of 30-35MPa pressure and 25-30℃ temperature, with each homogenization time being 3-5min, so that the components are evenly mixed and the stability of the system is improved. S5. Sterilization and purification: The homogenized material is filtered and sterilized using a 0.22μm sterile microporous membrane to remove trace impurities and bacteria. S6. Finished product preparation: The sterilized filtrate is concentrated under vacuum at a low temperature of 40-45℃ and a vacuum degree of 15-20Pa until the solid content is 25-30%. Then, it is freeze-dried under vacuum to obtain powdered anti-aging collagen peptide finished product.

[0011] Preferably, in step S4, after homogenization is completed, the particle size of the material system is detected, and the average particle size is controlled at 500-800nm, and the system is uniform without stratification.

[0012] Preferably, in step S6, the freeze-drying parameters are: pre-freezing temperature -40 to -38°C, pre-freezing for 5 hours, vacuum degree 10-15 Pa, freeze-drying for 22-24 hours, and finally obtain a freeze-dried powder product with a moisture content ≤5%.

[0013] Compared with the prior art, the beneficial effects of the present invention are: 1. This invention features synergistic effects from its components, providing comprehensive anti-aging benefits. Utilizing a multi-active component compound system, each component has complementary functions without antagonistic effects, addressing skin aging from multiple dimensions including cell metabolism, tissue repair, anti-oxidation, and collagen replenishment. Specifically, mitochondrial activating peptides improve mitochondrial metabolism in skin fibroblasts, enhance cellular energy metabolism efficiency, reduce oxidative stress-induced cell damage and apoptosis, and improve skin aging defects caused by declining cell metabolism. Elastin repair peptides act on the elastic tissue of the dermis, improving the aging and breakage of elastic fibers, optimizing skin elasticity, and alleviating signs of aging such as skin laxity and fine lines. Ultra-micro cell wall breaking... Black chokeberry extract is rich in highly active anthocyanins and polyphenols. After cell wall disruption, its solubility and antioxidant activity are significantly enhanced, effectively scavenging free radicals, inhibiting oxidative stress, and reducing the oxidative degradation of collagen and elastin. Small molecule fish skin collagen peptides serve as a base component; their small molecular weight and easy absorption replenish exogenous collagen and stimulate endogenous collagen regeneration, increasing dermal collagen density. Sodium hyaluronate provides long-lasting hydration and repairs the skin barrier, offering a stable microenvironment for skin tissue repair. Vitamin E microcapsules, through encapsulation technology, prevent the active ingredients from oxidizing and becoming inactive, ensuring stable antioxidant, moisturizing, and repairing effects, and significantly improving product storage stability. 2. The process optimization of this invention improves the activity and stability of the product. It adopts a low-temperature stepwise enzymatic hydrolysis and gradient ultrafiltration purification process, which can accurately enrich highly active peptides and remove ineffective impurities and proteins, thereby improving the purity and efficacy of active peptides. The entire process adopts low-temperature stirring, low-temperature homogenization, and low-temperature concentration freeze-drying processes to avoid the damage of heat-sensitive components caused by high temperature and high shear force, and to retain the biological activity of each component to the greatest extent. This effectively solves the problems of easy inactivation of active ingredients, poor product stability, and low bioavailability in traditional processes. Detailed Implementation

[0014] The technical solution of the present invention will be clearly and completely described below with reference to specific embodiments of the present invention. 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.

[0015] This invention provides an anti-aging collagen peptide, which is a low-moisture lyophilized powder formulation composed of the following effective components in parts by weight: Small molecule fish skin collagen peptides 55-75 parts, mitochondrial activation peptides 8-15 parts, elastin repair peptides 6-12 parts, ultra-micro cell wall broken black chokeberry extract 5-10 parts, sodium hyaluronate 3-8 parts, vitamin E microcapsules 2-5 parts. The moisture content of the finished collagen peptide freeze-dried powder is ≤5%.

[0016] The method for preparing mitochondrial activating peptides includes the following steps: S11. Raw material pretreatment: Select fresh bovine myocardial tissue, remove fascia and fat, rinse with sterile water 3-5 times, freeze dry at low temperature, pulverize, and pass through an 80-100 mesh sieve to obtain bovine myocardial dry powder; add bovine myocardial dry powder to sterile purified water at a material-to-liquid ratio of 1:8-1:10 g / mL, stir evenly, and prepare myocardial suspension. S12. Low-temperature stepwise enzymatic hydrolysis: Adjust the pH of the myocardial suspension to 7.0-7.5, raise the temperature to 45-50℃, add neutral protease at a concentration of 1.2-1.8% of the substrate weight, and hydrolyze at a constant temperature with stirring for 2.5-3.5 hours at a stirring speed of 180-220 r / min; then adjust the pH to 8.0-8.5, raise the temperature to 50-55℃, add alkaline protease at a concentration of 0.8-1.2% of the substrate weight, and continue enzymatic hydrolysis for 1.5-2 hours; S13. Enzyme inactivation centrifugation: After the enzymatic hydrolysis is completed, the temperature is rapidly raised to 90-95℃ and the enzyme is inactivated at a constant temperature for 8-12 minutes. After cooling to room temperature, the enzyme is centrifuged at 8000-10000r / min for 15-20 minutes and the upper enzymatic hydrolysis supernatant is collected. S14. Gradient ultrafiltration purification: The supernatant is sequentially passed through 10kDa, 3kDa, and 1kDa ultrafiltration membranes for gradient ultrafiltration, and the filtrate in the 1-3kDa molecular weight range is collected. S15. Freeze-drying: Place the ultrafiltration filtrate in a vacuum freeze dryer, pre-freeze at -40~-35℃ for 4-6 hours, vacuum degree 10-15Pa, freeze-dry for 18-24 hours to obtain powdered mitochondrial activating peptides.

[0017] The preparation method of elastin repair peptides includes the following steps: S21. Raw material pretreatment: Select fresh porcine aortic elastin raw material, remove impurities and blood, wash with sterile water, drain at low temperature, pulverize and pass through a 100-120 mesh sieve to obtain coarse elastin powder; add coarse elastin powder to sterile water at a material-to-liquid ratio of 1:10-1:12 g / mL, soak for 12-16 hours to fully swell. S22. Compound enzymatic hydrolysis treatment: Adjust the pH of the swollen mixture to 6.0-6.5, stabilize the temperature at 48-52℃, add compound protease, wherein the compound protease is papain and flavor protease in a mass ratio of 2:1, and the total amount of enzyme added is 1.5-2.0% of the substrate mass. Stir at a constant temperature for 3-4 hours and stir at a speed of 200-240 r / min. S23. Enzyme inactivation and impurity removal: After the enzymatic hydrolysis is completed, the temperature is raised to 85-90℃ and the enzyme is inactivated at a constant temperature for 10-15 min. The temperature is then naturally cooled to 25-30℃. The mixture is centrifuged at 9000-11000 r / min for 20-25 min to remove precipitated impurities and the clear supernatant is collected. S24. Refining and purification: The supernatant is filtered using a 2kDa ultrafiltration membrane. After collecting the filtrate, it is decolorized and deodorized using macroporous adsorption resin. The elution flow rate is 2-3 BV / h and the elution time is 40-60 min. S25. Low-temperature freeze drying: The purified filtrate after decolorization and deodorization is subjected to vacuum freeze drying. The pre-freezing temperature is -45 to -40℃, the pre-freezing time is 5-7 hours, the vacuum degree is 8-12 Pa, and the freeze drying time is 20-26 hours to obtain highly active elastin repair peptide powder.

[0018] The molecular weight of small molecule fish skin collagen peptides is 800-1500 Da; The cell wall breakage particle size of the ultra-micro cell wall broken black chokeberry extract is 200-300nm; The encapsulation efficiency of vitamin E microcapsules is ≥92%.

[0019] A method for preparing collagen peptides that delay aging includes the following steps: S1. Preparation of base solution: Take 100-150 parts of purified water as the dissolving medium according to the process ratio, add it to a sterilized reaction vessel, heat to 35-40℃, stir at 150-180 r / min, add small molecule fish skin collagen peptides and sodium hyaluronate in sequence, stir at a constant temperature for 15-20 min until the materials are completely dissolved to obtain a uniform base mixture; the purified water is only used for process dissolution and is completely removed in subsequent processes and does not remain in the finished product. S2, Innovative Peptide Combination: Maintain a temperature of 35-40℃ and a stirring speed of 180-200r / min. Slowly add mitochondrial activating peptides and elastin repair peptides to the base mixture and stir at a uniform speed for 25-30min to fully disperse and blend the active peptides. S3. Addition of functional excipients: Cool to 28-32℃, reduce stirring speed to 120-150r / min, add ultra-fine cell wall broken black chokeberry extract and vitamin E microcapsules in sequence, stir at low speed for 18-22min to avoid high temperature destroying active ingredients; S4. Low-temperature homogenization treatment: Transfer the mixture to a high-pressure homogenizer and homogenize it 2-3 times under the conditions of 30-35MPa pressure and 25-30℃ temperature, with each homogenization time being 3-5min, so that the components are evenly mixed and the stability of the system is improved. S5. Sterilization and purification: The homogenized material is filtered and sterilized using a 0.22μm sterile microporous membrane to remove trace impurities and bacteria. S6. Finished product preparation: The sterilized filtrate is concentrated under vacuum at a low temperature of 40-45℃ and a vacuum degree of 15-20Pa until the solid content is 25-30%. Then, it is freeze-dried under vacuum to obtain powdered anti-aging collagen peptide finished product.

[0020] In step S4, after homogenization is completed, the particle size of the material system is detected. The average particle size is controlled at 500-800nm, and the system is uniform without stratification.

[0021] In step S6, the freeze-drying parameters are: pre-freezing temperature -40 to -38℃, pre-freezing for 5 hours, vacuum degree 10-15 Pa, freeze-drying for 22-24 hours, and finally obtain a freeze-dried powder product with a moisture content ≤5%.

[0022] Example 1: A collagen peptide for delaying aging, a lyophilized collagen peptide powder formulation, composed of the following effective components in parts by weight: Small molecule fish skin collagen peptides 65 parts, mitochondrial activation peptides 12 parts, elastin repair peptides 9 parts, ultra-micro cell wall broken black chokeberry extract 7 parts, sodium hyaluronate 5 parts, vitamin E microcapsules 3 parts. Among them, the molecular weight of small molecule fish skin collagen peptide is 800-1500 Da, the particle size of ultra-micro broken black chokeberry extract is 200-300 nm, and the encapsulation rate of vitamin E microcapsules is ≥92%.

[0023] The method for preparing mitochondrial activating peptides includes the following steps: S11. Raw material pretreatment: Select fresh bovine myocardial tissue, remove fascia and fat, rinse 4 times with sterile water, freeze dry at low temperature, pulverize, and pass through a 90-mesh sieve to obtain bovine myocardial dry powder; add bovine myocardial dry powder to sterile purified water at a material-to-liquid ratio of 1:9 g / mL, stir evenly, and prepare myocardial suspension; S12. Low-temperature stepwise enzymatic hydrolysis: Adjust the pH of the myocardial suspension to 7.2, raise the temperature to 48°C, add neutral protease at a concentration of 1.5% of the substrate weight, and hydrolyze at a constant temperature with stirring for 3 hours at a stirring speed of 200 r / min; then adjust the pH to 8.2, raise the temperature to 52°C, add alkaline protease at a concentration of 1.0% of the substrate weight, and continue enzymatic hydrolysis for 1.8 hours; S13. Enzyme inactivation centrifugation: After the enzymatic hydrolysis is completed, the temperature is rapidly raised to 92℃ and kept at a constant temperature for 10 min to inactivate the enzyme. After cooling to room temperature, the enzyme is centrifuged at 9000 r / min for 18 min using a high-speed centrifuge and the upper enzymatic hydrolysis supernatant is collected. S14. Gradient ultrafiltration purification: The supernatant is sequentially passed through 10kDa, 3kDa, and 1kDa ultrafiltration membranes for gradient ultrafiltration, and the filtrate in the 1-3kDa molecular weight range is collected. S15. Freeze-drying: Place the ultrafiltration filtrate in a vacuum freeze dryer, pre-freeze at -38℃ for 5 hours, vacuum at 12Pa, and freeze-dry for 22 hours to obtain powdered mitochondrial activating peptides.

[0024] The preparation method of elastin repair peptides includes the following steps: S21. Raw material pretreatment: Select fresh porcine aortic elastin raw material, remove impurities and blood, wash with sterile water, drain at low temperature, pulverize and pass through a 110-mesh sieve to obtain coarse elastin powder; add the coarse elastin powder to sterile water at a material-to-liquid ratio of 1:11 g / mL, soak for 14 hours to fully swell; S22. Compound enzymatic hydrolysis treatment: Adjust the pH of the swollen mixture to 6.2, stabilize the temperature at 50℃, add compound protease, wherein the compound protease is papain and flavor protease in a mass ratio of 2:1, and the total amount of enzyme added is 1.8% of the substrate mass. The mixture is kept at a constant temperature and stirred for 3.5 hours at a stirring speed of 220 r / min. S23. Enzyme inactivation and impurity removal: After the enzymatic hydrolysis is completed, the temperature is raised to 88℃ and the enzyme is inactivated at a constant temperature for 12 min. The temperature is then naturally cooled to 28℃ and centrifuged at 10000 r / min for 22 min to remove precipitated impurities. The clear supernatant is then collected. S24. Refining and purification: The supernatant was filtered through a 2kDa ultrafiltration membrane. After collecting the filtrate, it was decolorized and deodorized using D101 macroporous adsorption resin. The elution flow rate was 2.5 BV / h and the elution time was 50 min. S25. Low-temperature freeze drying: The purified filtrate after decolorization and deodorization is subjected to vacuum freeze drying. The pre-freezing temperature is -42℃, the pre-freezing time is 6 hours, the vacuum degree is 10Pa, and the freeze drying time is 24 hours to obtain highly active elastin repair peptide powder.

[0025] A method for preparing collagen peptides that delay aging includes the following steps: S1. Preparation of base solution: Take 120 parts of purified water and add it to a sterilized reaction vessel. Heat the temperature to 38°C and stir at 160r / min. Add small molecule fish skin collagen peptide and sodium hyaluronate in sequence. Stir at constant temperature for 18min until the materials are completely dissolved to obtain a uniform base solution. S2, Innovative Peptide Combination: Maintain a temperature of 38℃ and a stirring speed of 190r / min, slowly add the prepared mitochondrial activation peptide and elastin repair peptide to the base mixture, and stir at a uniform speed for 28min to fully disperse and fuse the active peptides. S3. Addition of functional excipients: Cool to 30℃, reduce stirring speed to 130r / min, add ultra-fine cell wall broken black chokeberry extract and vitamin E microcapsules in sequence, stir at low speed for 20min to avoid high temperature destroying active ingredients; S4. Low-temperature homogenization treatment: The mixture is transferred to a high-pressure homogenizer and homogenized twice at a pressure of 32MPa and a temperature of 28℃, each time for 4 minutes. After homogenization, the average particle size of the material is controlled at 600nm, and the system is uniform without stratification. S5. Sterilization and purification: The homogenized material is filtered and sterilized using a 0.22μm sterile microporous membrane to remove trace impurities and bacteria. S6. Finished product preparation: The sterilized filtrate is concentrated under vacuum at a low temperature of 42°C and a vacuum degree of 18Pa until the solid content is 28%. Then, it is freeze-dried under vacuum at a pre-freezing temperature of -39°C for 5 hours, a vacuum degree of 12Pa, and freeze-dried for 23 hours to obtain a powdered anti-aging collagen peptide product with a moisture content of ≤5%.

[0026] Example 2: A collagen peptide for delaying aging, a lyophilized collagen peptide powder formulation, composed of the following effective components in parts by weight: Small molecule fish skin collagen peptides 55 parts, mitochondrial activation peptides 15 parts, elastin repair peptides 12 parts, ultra-micro cell wall broken black chokeberry extract 10 parts, sodium hyaluronate 8 parts, vitamin E microcapsules 5 parts. Among them, the molecular weight of small molecule fish skin collagen peptide is 800-1500 Da, the particle size of ultra-micro broken black chokeberry extract is 200-300 nm, and the encapsulation rate of vitamin E microcapsules is ≥92%.

[0027] The method for preparing mitochondrial activating peptides includes the following steps: S11. Raw material pretreatment: Select fresh bovine myocardial tissue, remove fascia and fat, rinse 4 times with sterile water, freeze dry at low temperature, pulverize, and pass through a 90-mesh sieve to obtain bovine myocardial dry powder; add bovine myocardial dry powder to sterile purified water at a material-to-liquid ratio of 1:9 g / mL, stir evenly, and prepare myocardial suspension; S12. Low-temperature stepwise enzymatic hydrolysis: Adjust the pH of the myocardial suspension to 7.2, raise the temperature to 48°C, add neutral protease at a concentration of 1.5% of the substrate weight, and hydrolyze at a constant temperature with stirring for 3 hours at a stirring speed of 200 r / min; then adjust the pH to 8.2, raise the temperature to 52°C, add alkaline protease at a concentration of 1.0% of the substrate weight, and continue enzymatic hydrolysis for 1.8 hours; S13. Enzyme inactivation centrifugation: After the enzymatic hydrolysis is completed, the temperature is rapidly raised to 92℃ and kept at a constant temperature for 10 min to inactivate the enzyme. After cooling to room temperature, the enzyme is centrifuged at 9000 r / min for 18 min using a high-speed centrifuge and the upper enzymatic hydrolysis supernatant is collected. S14. Gradient ultrafiltration purification: The supernatant is sequentially passed through 10kDa, 3kDa, and 1kDa ultrafiltration membranes for gradient ultrafiltration, and the filtrate in the molecular weight range of 1-3kDa is collected. S15. Freeze-drying: Place the ultrafiltration filtrate in a vacuum freeze dryer, pre-freeze at -38℃ for 5 hours, vacuum at 12Pa, and freeze-dry for 22 hours to obtain powdered mitochondrial activating peptides.

[0028] The preparation method of elastin repair peptides includes the following steps: S21. Raw material pretreatment: Select fresh porcine aortic elastin raw material, remove impurities and blood, wash with sterile water, drain at low temperature, pulverize and pass through a 110-mesh sieve to obtain coarse elastin powder; add the coarse elastin powder to sterile water at a material-to-liquid ratio of 1:11 g / mL, soak for 14 hours to fully swell; S22. Compound enzymatic hydrolysis treatment: Adjust the pH of the swollen mixture to 6.2, stabilize the temperature at 50℃, add compound protease, wherein the compound protease is papain and flavor protease in a mass ratio of 2:1, and the total amount of enzyme added is 1.8% of the substrate mass. The mixture is kept at a constant temperature and stirred for 3.5 hours at a stirring speed of 220 r / min. S23. Enzyme inactivation and impurity removal: After the enzymatic hydrolysis is completed, the temperature is raised to 88℃ and the enzyme is inactivated at a constant temperature for 12 min. The temperature is then naturally cooled to 28℃ and centrifuged at 10000 r / min for 22 min to remove precipitated impurities. The clear supernatant is then collected. S24. Refining and purification: The supernatant was filtered through a 2kDa ultrafiltration membrane. After collecting the filtrate, it was decolorized and deodorized using D101 macroporous adsorption resin. The elution flow rate was 2.5 BV / h and the elution time was 50 min. S25. Low-temperature freeze drying: The purified filtrate after decolorization and deodorization is subjected to vacuum freeze drying. The pre-freezing temperature is -42℃, the pre-freezing time is 6 hours, the vacuum degree is 10Pa, and the freeze drying time is 24 hours to obtain highly active elastin repair peptide powder.

[0029] A method for preparing collagen peptides that delay aging includes the following steps: S1. Preparation of base solution: Take 150 parts of purified water and add it to a sterilized reaction vessel. Heat the temperature to 38°C and stir at 160r / min. Add small molecule fish skin collagen peptide and sodium hyaluronate in sequence. Stir at constant temperature for 18min until the materials are completely dissolved to obtain a uniform base solution. S2, Innovative Peptide Combination: Maintain a temperature of 38℃ and a stirring speed of 190r / min, slowly add the prepared mitochondrial activation peptide and elastin repair peptide to the base mixture, and stir at a uniform speed for 28min to fully disperse and fuse the active peptides. S3. Addition of functional excipients: Cool to 30℃, reduce stirring speed to 130r / min, add ultra-fine cell wall broken black chokeberry extract and vitamin E microcapsules in sequence, stir at low speed for 20min to avoid high temperature destroying active ingredients; S4. Low-temperature homogenization treatment: The mixture is transferred to a high-pressure homogenizer and homogenized twice at a pressure of 32MPa and a temperature of 28℃, each time for 4 minutes. After homogenization, the average particle size of the material is controlled at 600nm, and the system is uniform without stratification. S5. Sterilization and purification: The homogenized material is filtered and sterilized using a 0.22μm sterile microporous membrane to remove trace impurities and bacteria. S6. Finished product preparation: The sterilized filtrate is concentrated under vacuum at a low temperature of 42°C and a vacuum degree of 18Pa until the solid content is 28%. Then, it is freeze-dried under vacuum at a pre-freezing temperature of -39°C for 5 hours, a vacuum degree of 12Pa, and freeze-dried for 23 hours to obtain a powdered anti-aging collagen peptide product with a moisture content of ≤5%.

[0030] Example 3: A collagen peptide for delaying aging, a lyophilized collagen peptide powder formulation, composed of the following effective components in parts by weight: 75 parts of small molecule fish skin collagen peptides, 8 parts of mitochondrial activating peptides, 6 parts of elastin repair peptides, 5 parts of ultra-micro cell wall broken black chokeberry extract, 3 parts of sodium hyaluronate, and 2 parts of vitamin E microcapsules. Among them, the molecular weight of small molecule fish skin collagen peptide is 800-1500 Da, the particle size of ultra-micro broken black chokeberry extract is 200-300 nm, and the encapsulation rate of vitamin E microcapsules is ≥92%.

[0031] The method for preparing mitochondrial activating peptides includes the following steps: S11. Raw material pretreatment: Select fresh bovine myocardial tissue, remove fascia and fat, rinse 4 times with sterile water, freeze dry at low temperature, pulverize, and pass through a 90-mesh sieve to obtain bovine myocardial dry powder; add bovine myocardial dry powder to sterile purified water at a material-to-liquid ratio of 1:9 g / mL, stir evenly, and prepare myocardial suspension; S12. Low-temperature stepwise enzymatic hydrolysis: Adjust the pH of the myocardial suspension to 7.2, raise the temperature to 48°C, add neutral protease at a concentration of 1.5% of the substrate weight, and hydrolyze at a constant temperature with stirring for 3 hours at a stirring speed of 200 r / min; then adjust the pH to 8.2, raise the temperature to 52°C, add alkaline protease at a concentration of 1.0% of the substrate weight, and continue enzymatic hydrolysis for 1.8 hours; S13. Enzyme inactivation centrifugation: After the enzymatic hydrolysis is completed, the temperature is rapidly raised to 92℃ and kept at a constant temperature for 10 min to inactivate the enzyme. After cooling to room temperature, the enzyme is centrifuged at 9000 r / min for 18 min using a high-speed centrifuge and the upper enzymatic hydrolysis supernatant is collected. S14. Gradient ultrafiltration purification: The supernatant is sequentially passed through 10kDa, 3kDa, and 1kDa ultrafiltration membranes for gradient ultrafiltration, and the filtrate in the molecular weight range of 1-3kDa is collected. S15. Freeze-drying: Place the ultrafiltration filtrate in a vacuum freeze dryer, pre-freeze at -38℃ for 5 hours, vacuum at 12Pa, and freeze-dry for 22 hours to obtain powdered mitochondrial activating peptides.

[0032] The preparation method of elastin repair peptides includes the following steps: S21. Raw material pretreatment: Select fresh porcine aortic elastin raw material, remove impurities and blood, wash with sterile water, drain at low temperature, pulverize and pass through a 110-mesh sieve to obtain coarse elastin powder; add the coarse elastin powder to sterile water at a material-to-liquid ratio of 1:11 g / mL, soak for 14 hours to fully swell; S22. Compound enzymatic hydrolysis treatment: Adjust the pH of the swollen mixture to 6.2, stabilize the temperature at 50℃, add compound protease, wherein the compound protease is papain and flavor protease in a mass ratio of 2:1, and the total amount of enzyme added is 1.8% of the substrate mass. The mixture is kept at a constant temperature and stirred for 3.5 hours at a stirring speed of 220 r / min. S23. Enzyme inactivation and impurity removal: After the enzymatic hydrolysis is completed, the temperature is raised to 88℃ and the enzyme is inactivated at a constant temperature for 12 min. The temperature is then naturally cooled to 28℃ and centrifuged at 10000 r / min for 22 min to remove precipitated impurities. The clear supernatant is then collected. S24. Refining and purification: The supernatant was filtered through a 2kDa ultrafiltration membrane. After collecting the filtrate, it was decolorized and deodorized using D101 macroporous adsorption resin. The elution flow rate was 2.5 BV / h and the elution time was 50 min. S25. Low-temperature freeze drying: The purified filtrate after decolorization and deodorization is subjected to vacuum freeze drying. The pre-freezing temperature is -42℃, the pre-freezing time is 6 hours, the vacuum degree is 10Pa, and the freeze drying time is 24 hours to obtain highly active elastin repair peptide powder.

[0033] A method for preparing collagen peptides that delay aging includes the following steps: S1. Preparation of base solution: Take 100 parts of purified water and add it to a sterilized reaction vessel. Heat the water to 38°C and stir at 160 r / min. Add small molecule fish skin collagen peptide and sodium hyaluronate in sequence. Stir at a constant temperature for 18 min until the materials are completely dissolved to obtain a uniform base solution. S2, Innovative Peptide Combination: Maintain a temperature of 38℃ and a stirring speed of 190r / min, slowly add the prepared mitochondrial activation peptide and elastin repair peptide to the base mixture, and stir at a uniform speed for 28min to fully disperse and fuse the active peptides. S3. Addition of functional excipients: Cool to 30℃, reduce stirring speed to 130r / min, add ultra-fine cell wall broken black chokeberry extract and vitamin E microcapsules in sequence, stir at low speed for 20min to avoid high temperature destroying active ingredients; S4. Low-temperature homogenization treatment: The mixture is transferred to a high-pressure homogenizer and homogenized twice at a pressure of 32MPa and a temperature of 28℃, each time for 4 minutes. After homogenization, the average particle size of the material is controlled at 600nm, and the system is uniform without stratification. S5. Sterilization and purification: The homogenized material is filtered and sterilized using a 0.22μm sterile microporous membrane to remove trace impurities and bacteria. S6. Finished product preparation: The sterilized filtrate is concentrated under vacuum at a low temperature of 42°C and a vacuum degree of 18Pa until the solid content is 28%. Then, it is freeze-dried under vacuum at a pre-freezing temperature of -39°C for 5 hours, a vacuum degree of 12Pa, and freeze-dried for 23 hours to obtain a powdered anti-aging collagen peptide product with a moisture content of ≤5%.

[0034] Comparative Example 1 (Traditional Single Collagen Peptide Control Group) This invention uses commercially available small-molecule fish skin collagen peptides (molecular weight 800-1500 Da), without adding the mitochondrial activating peptides, elastin repair peptides, and ultra-micro cell-wall-breaking black chokeberry extract prepared in this invention. It consists only of single fish skin collagen peptides, conventional sodium hyaluronate, and common vitamin E as active ingredients, compounded with purified water as the process solvent. The preparation process employs traditional room-temperature high-speed stirring, high-temperature concentration, and conventional drying processes, without using the refined processes of this invention, such as low-temperature stepwise enzymatic hydrolysis, gradient ultrafiltration, low-temperature homogenization, and microencapsulation protection. All other basic preparation environment and equipment parameters remain completely consistent with Example 1, resulting in a conventional single anti-aging collagen peptide dry powder product, which serves as a blank control sample to highlight the technological advancements of this invention.

[0035] To intuitively and scientifically verify the antioxidant, anti-aging, and tissue repair properties of the product of this invention, parallel performance tests were conducted on three sets of example products and one set of comparative products using unified standards, environments, and testing equipment. All tests employed standardized in vitro experimental models to avoid the uncertainties of subjective human testing, ensuring repeatability and objectivity of the data. Specific testing items and standards are as follows: 1. Antioxidant performance: The DPPH free radical scavenging rate method was used to evaluate the product's in vitro antioxidant activity; 2. Anti-aging performance: A human skin fibroblast in vitro culture oxidative stress damage model was used to detect cell viability and cell aging inhibition rate; 3. Skin repair-related performance: The auxiliary effects of the samples on skin collagen repair and elasticity improvement were evaluated through in vitro simulated skin penetration and tissue repair-related index testing. Each group of samples was tested in parallel three times, and the average value was taken. Specific test results are shown in the table below:

[0036] The gradient test data in the table above clearly show that: Example 1, with its balanced ratio, offers comprehensive performance suitable for industrial mass production; Example 2, with its highly active functional component ratio, exhibits superior antioxidant and cell protection properties; and Example 3, with its high collagen base ratio, still demonstrates excellent anti-aging and repair effects, validating the overall effectiveness of the formulation's protective range. All in vitro test indicators for the three examples are significantly better than those of traditional single collagen peptide formulations, proving that the multi-component compound system of this invention has a significant synergistic effect, effectively improving the technical problems of traditional products having single anti-aging effects and insufficient performance. The formulation design is reasonable, and the process is stable and controllable.

[0037] It should be noted that, in this document, relational terms such as "first" and "second" are used only to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such process, method, article, or apparatus.

[0038] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A collagen peptide for delaying aging, characterized in that, The collagen peptides are a low-moisture lyophilized powder formulation, composed of the following effective components in parts by weight: Small molecule fish skin collagen peptides 55-75 parts, mitochondrial activation peptides 8-15 parts, elastin repair peptides 6-12 parts, ultra-micro cell wall broken black chokeberry extract 5-10 parts, sodium hyaluronate 3-8 parts, vitamin E microcapsules 2-5 parts. The moisture content of the finished collagen peptide freeze-dried powder is ≤5%.

2. The collagen peptide for delaying aging according to claim 1, characterized in that, The method for preparing the mitochondrial activating peptide includes the following steps: S11. Raw material pretreatment: Select fresh bovine myocardial tissue, remove fascia and fat, rinse with sterile water 3-5 times, freeze dry at low temperature, pulverize, and pass through an 80-100 mesh sieve to obtain bovine myocardial dry powder; add bovine myocardial dry powder to sterile purified water at a material-to-liquid ratio of 1:8-1:10 g / mL, stir evenly, and prepare myocardial suspension. S12. Low-temperature stepwise enzymatic hydrolysis: Adjust the pH of the myocardial suspension to 7.0-7.5, raise the temperature to 45-50℃, add neutral protease at a concentration of 1.2-1.8% of the substrate weight, and hydrolyze at a constant temperature with stirring for 2.5-3.5 hours at a stirring speed of 180-220 r / min; then adjust the pH to 8.0-8.5, raise the temperature to 50-55℃, add alkaline protease at a concentration of 0.8-1.2% of the substrate weight, and continue enzymatic hydrolysis for 1.5-2 hours; S13. Enzyme inactivation centrifugation: After the enzymatic hydrolysis is completed, the temperature is rapidly raised to 90-95℃ and the enzyme is inactivated at a constant temperature for 8-12 minutes. After cooling to room temperature, the enzyme is centrifuged at 8000-10000r / min for 15-20 minutes and the upper enzymatic hydrolysis supernatant is collected. S14. Gradient ultrafiltration purification: The supernatant is sequentially passed through 10kDa, 3kDa, and 1kDa ultrafiltration membranes for gradient ultrafiltration, and the filtrate in the molecular weight range of 1-3kDa is collected. S15. Freeze-drying: Place the ultrafiltration filtrate in a vacuum freeze dryer, pre-freeze at -40~-35℃ for 4-6 hours, vacuum degree 10-15Pa, freeze-dry for 18-24 hours to obtain powdered mitochondrial activating peptides.

3. The collagen peptide for delaying aging according to claim 1, characterized in that, The preparation method of the elastin repair peptide includes the following steps: S21. Raw material pretreatment: Select fresh porcine aortic elastin raw material, remove impurities and blood, wash with sterile water, drain at low temperature, pulverize and pass through a 100-120 mesh sieve to obtain coarse elastin powder; add coarse elastin powder to sterile water at a material-to-liquid ratio of 1:10-1:12 g / mL, soak for 12-16 hours to fully swell. S22. Compound enzymatic hydrolysis treatment: Adjust the pH of the swollen mixture to 6.0-6.5, stabilize the temperature at 48-52℃, add compound protease, wherein the compound protease is papain and flavor protease in a mass ratio of 2:1, and the total amount of enzyme added is 1.5-2.0% of the substrate mass. Stir at a constant temperature for 3-4 hours and stir at a speed of 200-240 r / min. S23. Enzyme inactivation and impurity removal: After the enzymatic hydrolysis is completed, the temperature is raised to 85-90℃ and the enzyme is inactivated at a constant temperature for 10-15 min. The temperature is then naturally cooled to 25-30℃. The mixture is centrifuged at 9000-11000 r / min for 20-25 min to remove precipitated impurities and the clear supernatant is collected. S24. Refining and purification: The supernatant is filtered using a 2kDa ultrafiltration membrane. After collecting the filtrate, it is decolorized and deodorized using macroporous adsorption resin. The elution flow rate is 2-3 BV / h and the elution time is 40-60 min. S25. Low-temperature freeze drying: The purified filtrate after decolorization and deodorization is subjected to vacuum freeze drying. The pre-freezing temperature is -45 to -40℃, the pre-freezing time is 5-7 hours, the vacuum degree is 8-12 Pa, and the freeze drying time is 20-26 hours to obtain highly active elastin repair peptide powder.

4. The collagen peptide for delaying aging according to claim 1, characterized in that, The molecular weight of the small molecule fish skin collagen peptide is 800-1500 Da; The cell wall breaking particle size of the ultra-micro cell wall broken black chokeberry extract is 200-300 nm. The encapsulation rate of the vitamin E microcapsules is ≥92%.

5. A method for preparing collagen peptides that delay aging, characterized in that, Includes the following steps: S1. Preparation of base solution: Take 100-150 parts of purified water as the dissolving medium according to the process ratio, add it to a sterilized reaction vessel, heat to 35-40℃, stir at 150-180 r / min, add small molecule fish skin collagen peptides and sodium hyaluronate in sequence, stir at a constant temperature for 15-20 min until the materials are completely dissolved to obtain a uniform base mixture; the purified water is only used for process dissolution and is completely removed in subsequent processes and does not remain in the finished product. S2, Innovative Peptide Combination: Maintain a temperature of 35-40℃ and a stirring speed of 180-200r / min. Slowly add mitochondrial activating peptides and elastin repair peptides to the base mixture and stir at a uniform speed for 25-30min to fully disperse and blend the active peptides. S3. Addition of functional excipients: Cool to 28-32℃, reduce stirring speed to 120-150r / min, add ultra-fine cell wall broken black chokeberry extract and vitamin E microcapsules in sequence, stir at low speed for 18-22min to avoid high temperature destroying active ingredients; S4. Low-temperature homogenization treatment: Transfer the mixture to a high-pressure homogenizer and homogenize it 2-3 times under the conditions of 30-35MPa pressure and 25-30℃ temperature, with each homogenization time being 3-5min, so that the components are evenly mixed and the stability of the system is improved. S5. Sterilization and purification: The homogenized material is filtered and sterilized using a 0.22μm sterile microporous membrane to remove trace impurities and bacteria. S6. Finished product preparation: The sterilized filtrate is concentrated under vacuum at a low temperature of 40-45℃ and a vacuum degree of 15-20Pa until the solid content is 25-30%. Then, it is freeze-dried under vacuum to obtain powdered anti-aging collagen peptide finished product.

6. The method for preparing an anti-aging collagen peptide according to claim 5, characterized in that, In step S4, after homogenization is completed, the particle size of the material system is detected. The average particle size is controlled at 500-800nm, and the system is uniform without stratification.

7. The method for preparing an anti-aging collagen peptide according to claim 5, characterized in that, In step S6, the freeze-drying parameters are: pre-freezing temperature -40 to -38℃, pre-freezing for 5 hours, vacuum degree 10-15 Pa, freeze-drying for 22-24 hours, and finally obtain a freeze-dried powder product with a moisture content ≤5%.

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