Anti-aging composition containing sea cucumber peptide and preparation method of anti-aging composition

Abstract

The invention provides an anti-aging composition containing sea cucumber peptide and a preparation method of the anti-aging composition, and belongs to the technical field of cosmetics. Mixing the sea cucumber peptide compound, the fermented smallanthus sonchifolius extract, the grape seed extract and the clam peptide at 23-25 DEG C for 10-15 minutes to obtain a mixture; the preparation method comprises the following steps: dispersing xanthan gum and sodium carboxymethyl cellulose in purified water at 85-90 DEG C, and stirring and mixing for 20-30 minutes to obtain a mixed system A; mixing the mixed system A, sodium hyaluronate, butanediol, hydroxyethyl urea, stearyl alcohol polyether-21 and glycosylglycerol at 50-55 DEG C for 10-15 minutes to obtain a mixed system B; and mixing the mixed system B and the mixture at 40-45 DEG C for 20-30 minutes to obtain the anti-aging composition. The sea cucumber peptide and the clam peptide jointly stimulate collagen synthesis through the synergistic effect, and good anti-aging and moisturizing effects are achieved through the synergistic effect of multiple components.

Description

Technical Field

[0001] The present invention relates to the technical field of cosmetics, and in particular to an anti-aging composition containing sea cucumber peptide and a preparation method thereof. Background Art

[0002] With the accelerating aging of the global population and the increasing pursuit of a higher quality of life, anti-aging has become a hot research topic in the fields of beauty, skincare, and health care. Consumer demand for anti-aging products is no longer limited to a single benefit; instead, consumers expect products to achieve multi-dimensional goals, such as delaying aging, improving skin condition, and enhancing physical function. Traditional anti-aging compositions often rely on chemically synthesized ingredients or common natural extracts. While these ingredients can meet market demand to a certain extent, long-term use may pose safety risks, and the durability and comprehensiveness of their efficacy are increasingly difficult to meet consumers' growing needs. Despite progress in the research of anti-aging compositions, numerous challenges remain. Many substances with potential anti-aging benefits are limited in source, and the extraction process is complex and costly, making large-scale production difficult. Even when successfully extracted, the active ingredients are easily damaged during the extraction process, resulting in reduced or even loss of their anti-aging activity. During formulation design, the interactions between different anti-aging ingredients are complex. Due to insufficient research on the synergistic or antagonistic effects between ingredients, many anti-aging compositions lack scientific formulations. This not only fails to fully utilize the strengths of each component, but can also cause adverse reactions such as allergies due to improper combination of ingredients. Furthermore, existing anti-aging compositions generally lack stability, being significantly affected by environmental factors such as temperature, light, and humidity. Active ingredients are susceptible to degradation or deterioration during storage and transportation, shortening the product's shelf life and significantly reducing its anti-aging effectiveness.

[0003] Chinese patent application CN109223651A discloses a skin care composition with whitening properties and its preparation method. The composition utilizes Scutellaria baicalensis to achieve whitening effects by inhibiting chronic inflammation, Centella asiatica to brighten the skin by increasing skin elasticity, and Rose and Emblica officinalis to synergize and inhibit tyrosinase activity, providing antioxidant and whitening effects. However, in actual use, only a small portion of these active ingredients penetrates the stratum corneum and penetrates deep into the skin, resulting in the active ingredients' full effectiveness not being fully realized.

[0004] Therefore, providing an anti-aging composition that can improve the skin care effect of active ingredients is an important problem to be solved in the art. Summary of the Invention

[0005] In view of the problems existing in the prior art, the present invention provides an anti-aging composition containing sea cucumber peptides and a preparation method thereof, which can improve the skin care effect of the active ingredients and show good anti-aging and moisturizing properties.

[0006] Specifically, the technical solution of the present invention includes the following contents: A method for preparing an anti-aging composition containing sea cucumber peptide, the preparation method comprising the following steps: The sea cucumber peptide complex, fermented yacon extract, grape seed extract, and clam peptide were mixed at 23-25° C. for 10-15 minutes to obtain a mixture; Xanthan gum and sodium carboxymethyl cellulose were dispersed in purified water at 85-90°C and stirred for 20-30 minutes to obtain a mixed system A; Mixing system A, sodium hyaluronate, butylene glycol, hydroxyethyl urea, steareth-21 and glyceryl glucoside at 50-55° C. for 10-15 minutes to obtain mixing system B; The mixed system B and the mixture are mixed at 40-45° C. for 20-30 minutes to prepare the anti-aging composition.

[0007] Furthermore, the preparation method of the sea cucumber peptide complex comprises the following steps: The chopped sea cucumber and the first phosphate buffer solution were mixed at a weight ratio of 1:8-10 and homogenized at 2-4°C for 10-15 minutes to obtain a homogenate solution; Neutral protease is added to the homogenate to perform enzymatic hydrolysis to obtain a primary enzymatic hydrolyzate; Flavor protease and aminopeptidase are added to the primary enzymatic hydrolysis solution for secondary enzymatic hydrolysis to obtain a secondary enzymatic hydrolysis solution; The secondary enzymatic hydrolysis solution and ammonium sulfate are mixed in a weight ratio of 10:2-3 and then allowed to stand, and the supernatant is collected to obtain sea cucumber peptide; Sea cucumber peptide and maleimide-PEG2-succinimide ester are reacted at a weight ratio of 3-5:0.01-0.02 at 23-25°C for 3-4 hours to obtain an intermediate; The intermediate and 1,2-dipalmitoyl-sn-glycero-3-phosphothioethanol are reacted at a weight ratio of 7-9:0.05-0.07 at 23-25° C. for 8-12 hours to obtain a modified sea cucumber peptide; Modified sea cucumber peptide, soybean lecithin and cholesterol are mixed in a weight ratio of 1.5-2:12-20:3-4.5 and then subjected to rotary evaporation to obtain a lipid film; First, phosphate buffer is added to the lipid film and subjected to swelling treatment at 45-50° C. for 2-4 hours to obtain a liposome suspension; chitosan oligosaccharide is dispersed in the second phosphate buffer at a material-liquid ratio of 8-10:50 to obtain a chitosan oligosaccharide solution; The liposome suspension and the chitosan oligosaccharide solution are reacted at a weight ratio of 10:10-15 at 23-25° C. for 3-4 hours to prepare the sea cucumber peptide complex.

[0008] Furthermore, the pH of the first phosphate buffer is 7.4.

[0009] Furthermore, the amount of the neutral protease used is 0.2% to 0.3% of the weight of the chopped sea cucumber.

[0010] Furthermore, the conditions for the primary enzymatic hydrolysis include an enzymatic hydrolysis pH of 6.8-7.2, an enzymatic hydrolysis temperature of 45-50° C., and an enzymatic hydrolysis time of 4-6 hours.

[0011] Furthermore, the amount of the flavor protease used is 0.4% to 0.5% of the weight of the chopped sea cucumber.

[0012] Furthermore, the dosage of the aminopeptidase is 0.15% to 0.2% of the weight of the chopped sea cucumber.

[0013] Furthermore, the conditions for the secondary enzymatic hydrolysis include an enzymatic hydrolysis pH of 7.2-7.6, an enzymatic hydrolysis temperature of 50-55° C., and an enzymatic hydrolysis time of 8-12 h.

[0014] Furthermore, the conditions for mixing the secondary enzymatic hydrolysis solution and ammonium sulfate include a mixing temperature of 2-4° C. and a mixing time of 1-2 h.

[0015] Furthermore, the standing conditions include a standing temperature of 2-4° C. and a standing time of 10-12 h.

[0016] Furthermore, the pH of the second phosphate buffer is 5.5.

[0017] Furthermore, the preparation method of the fermented yacon extract comprises the following steps: The yacon pulp minced meat is soaked in 80% ethanol by mass at a material-liquid ratio of 1:10-15 at 50-60°C for 1.5-2.5 hours, filtered and concentrated in sequence to obtain an extract, the extract is diluted with purified water to obtain a dilution, and then lactic acid bacteria is added for fermentation to obtain the fermented yacon extract.

[0018] Furthermore, the diluent is 15 to 20 times the weight of the extract.

[0019] Furthermore, the lactic acid bacteria include Lactobacillus fermentum or Lactobacillus plantarum.

[0020] Furthermore, the dosage of the lactic acid bacteria is 1% to 5% of the weight of the yacon pulp minced meat.

[0021] Furthermore, the fermentation treatment conditions include a treatment temperature of 37-40° C. and a treatment time of 72-96 h.

[0022] Furthermore, the weight ratio of the sea cucumber peptide complex, fermented yacon extract, grape seed extract, clam peptide, xanthan gum, sodium carboxymethyl cellulose, sodium hyaluronate, butylene glycol, hydroxyethyl urea, steareth-21, glyceryl glucoside and purified water is 3~5:1~2:1~3:0.5~1.5:0.6~1.2:0.3~1:0.2~0.5:2~4:0.5~1:0.8~1.5:0.3~1:65~85.

[0023] The invention discloses an anti-aging composition prepared by a method for preparing an anti-aging composition containing sea cucumber peptide.

[0024] Compared with the prior art, the present invention has the following beneficial effects: (1) The present invention adopts neutral protease, flavor protease and aminopeptidase to synergistically treat to obtain sea cucumber peptides with smaller molecular weight, then modifies the sea cucumber peptides so that they carry phospholipid groups to obtain modified sea cucumber peptides, reacts the modified sea cucumber peptides, soybean lecithin and cholesterol to obtain liposomes encapsulating sea cucumber peptides, and then encapsulates the liposomes with chitosan oligosaccharides to obtain sea cucumber peptide complexes; the modified sea cucumber peptides can protect the peptide segments from degradation by skin enzymes and improve the utilization rate of active ingredients; the chitosan oligosaccharides in the outer layer of the liposomes prolong the residence time through electrostatic adsorption, and the liposomes can carry the modified sea cucumber peptides into the lipid gaps of the stratum corneum and deliver them to the deep layer of the epidermis by fusion-release of small molecule peptides, thereby enhancing the skin care effect of the active ingredients while enhancing the anti-aging and moisturizing properties of the anti-aging composition.

[0025] (2) The sea cucumber peptide complex of the present invention can provide small molecule active peptides, which are easily absorbed by the skin, promote collagen synthesis, repair damaged cells, and enhance skin elasticity; the hydrophilic peptide segment in the sea cucumber peptide locks water, can repair the barrier and reduce water loss, and the biological activity of the yacon extract is enhanced after lactic acid bacteria fermentation, further improving the moisturizing effect; sodium hyaluronate, butylene glycol and hydroxyethyl urea have moisture absorption and water locking effects, glycerol glucoside can stimulate the synthesis of endogenous moisturizing factors, chitosan oligosaccharides and liposomes can repair and strengthen the skin barrier, and the biological activity of the fermented yacon extract is enhanced, further improving the moisturizing effect. Through the multi-dimensional synergy of the above various substances, a long-lasting moisturizing effect is achieved.

[0026] (3) The sea cucumber peptide and clam peptide in the present invention stimulate collagen synthesis through synergistic action. Grape seed extract effectively removes free radicals, has a strong antioxidant effect, strengthens the capillary wall, and improves microcirculation. Glycerol glucoside can activate the self-repair mechanism of skin cells and enhance cell vitality. Through the synergistic action of multiple components, a good anti-aging effect is achieved. DETAILED DESCRIPTION

[0027] The following will clearly and completely describe the technical solutions of the present invention through the embodiments of the present invention. Obviously, the embodiments described 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 those skilled in the art without making any creative efforts shall fall within the scope of protection of the present invention.

[0028] Unless otherwise specified, the raw materials and reagents used in the present invention are commercially available or can be prepared by known methods.

[0029] Preparation Example 1: The preparation method of the sea cucumber peptide complex comprises the following steps: Fresh sea cucumbers were washed with clean water, their internal organs were removed, and their body walls were taken out and cleaned with distilled water. The body walls were pulverized in a chopper for 3 minutes to obtain sea cucumber pieces. 10 parts by weight of the sea cucumber pieces were mixed with 80 parts by weight of a first phosphate buffer solution having a pH of 7.4, and the mixture was homogenized at 2°C for 10 minutes to obtain a homogenate. 0.02 parts by weight of neutral protease was added to the homogenate, the pH was adjusted to 6.8, the temperature was raised to 45° C., and the mixture was stirred for 4 hours to perform a primary enzymolysis. After the primary enzymolysis was completed, the mixture was heated to 90° C. and inactivated for 10 minutes to obtain a primary enzymolysis solution. The primary enzymolysis solution was heated to 50° C., 0.04 parts by weight of flavor protease and 0.015 parts by weight of aminopeptidase were added, the pH was adjusted to 7.2, the mixture was stirred for 8 hours to perform a secondary enzymolysis. After the secondary enzymolysis was completed, the mixture was heated to 90° C. and inactivated for 10 minutes to obtain a secondary enzymolysis solution. After 100 parts by weight of the secondary enzymatic hydrolysate was cooled to 2°C, 20 parts by weight of ammonium sulfate was added while stirring, and the mixture was stirred at 2°C for 1 hour. After the reaction was completed, the mixture was allowed to stand at 2°C for 10 hours, and the supernatant was collected by centrifugation at 8000 r / min for 20 minutes. The supernatant was filtered using a 1000Da molecular sieve ultrafiltration membrane to collect the polypeptide to obtain sea cucumber peptide; 3 parts by weight of sea cucumber peptide and 0.01 parts by weight of maleimide-PEG2-succinimide ester were dispersed in 50 parts by weight of a first phosphate buffer at pH 7.4, and stirred at 23° C. in a nitrogen atmosphere for 3 hours. After the reaction, ultrafiltration was performed to terminate the reaction and remove unreacted small molecules to obtain an intermediate; 7 parts by weight of the intermediate and 0.05 parts by weight of 1,2-dipalmitoyl-sn-glycero-3-phosphothioethanol were dispersed in 50 parts by weight of a first phosphate buffer at pH 7.4, and stirred at 23° C. in a nitrogen atmosphere for 8 hours. After the reaction, the dialyzate was dialyzed with a 3500Da dialysis bag for 24 hours to obtain a dialysate, which was sterilized with a 0.22μm filter membrane and freeze-dried to obtain a modified sea cucumber peptide; 1.5 parts by weight of modified sea cucumber peptide, 12 parts by weight of soybean lecithin, and 3 parts by weight of cholesterol were dispersed in 500 parts by weight of ethanol, transferred to an eggplant-shaped flask, heated to 40°C, and rotary evaporated to obtain a lipid film; 500 parts by weight of a first phosphate buffer solution with a pH of 7.4 preheated to 45°C was added, and the lipid film fell off when the eggplant-shaped flask was rotated, and the solution was allowed to swell at 45°C for 2 hours. After the swelling was completed, the solution was homogenized by ultrasonication to obtain a liposome suspension; 8 parts by weight of chitosan oligosaccharide are dispersed in 500 parts by weight of a second phosphate buffer solution with a pH of 5.5 to obtain a chitosan oligosaccharide solution; 10 parts by weight of a liposome suspension and 10 parts by weight of the chitosan oligosaccharide solution are mixed, stirred and reacted at 23° C. for 3 hours, and after the reaction is completed, dialyzed and freeze-dried to obtain a sea cucumber peptide complex.

[0030] Preparation Example 2: The preparation method of the sea cucumber peptide complex comprises the following steps: Fresh sea cucumbers were washed with clean water, their internal organs were removed, and their body walls were taken out and cleaned with distilled water. The body walls were pulverized in a chopper for 3.5 minutes to obtain sea cucumber pieces. 10 parts by weight of the sea cucumber pieces were mixed with 85 parts by weight of a first phosphate buffer solution with a pH of 7.4, and the mixture was homogenized at 2.5°C for 11 minutes to obtain a homogenate. 0.022 parts by weight of neutral protease was added to the homogenate, the pH was adjusted to 6.9, the temperature was raised to 46° C., and the mixture was stirred for 4.5 hours to perform a primary enzymatic hydrolysis. After the primary enzymatic hydrolysis was completed, the mixture was heated to 91° C. and inactivated for 11 minutes to obtain a primary enzymatic hydrolyzate. The primary enzymatic hydrolyzate was heated to 51° C., 0.042 parts by weight of flavor protease and 0.016 parts by weight of aminopeptidase were added, the pH was adjusted to 7.3, the mixture was stirred for 9 hours to perform a secondary enzymatic hydrolysis, and after the secondary enzymatic hydrolysis was completed, the mixture was heated to 91° C. and inactivated for 11 minutes to obtain a secondary enzymatic hydrolyzate. After 100 parts by weight of the secondary enzymatic hydrolysate was cooled to 2.5°C, 22 parts by weight of ammonium sulfate was added while stirring, and the mixture was stirred at 2.5°C for 1.2 hours. After the reaction was completed, the mixture was allowed to stand at 2.5°C for 10.5 hours, and the supernatant was collected by centrifugation at 8000 r / min for 20 minutes. The supernatant was filtered using a 500Da molecular sieve ultrafiltration membrane to collect the polypeptide to obtain sea cucumber peptide; 3.5 parts by weight of sea cucumber peptide and 0.012 parts by weight of maleimide-PEG2-succinimide ester were dispersed in 50 parts by weight of a first phosphate buffer at pH 7.4, and stirred at 24° C. in a nitrogen atmosphere for 3.2 hours. After the reaction, the mixture was ultrafiltered to terminate the reaction and remove unreacted small molecules to obtain an intermediate; 7.5 parts by weight of the intermediate and 0.055 parts by weight of 1,2-dipalmitoyl-sn-glycero-3-phosphothioethanol were dispersed in 50 parts by weight of a first phosphate buffer at pH 7.4, and stirred at 24° C. for 9 hours in a nitrogen atmosphere. After the reaction, the dialyzate was dialyzed for 30 hours using a 3500 Da dialysis bag to obtain a dialysate, which was sterilized by a 0.22 μm filter membrane and freeze-dried to obtain a modified sea cucumber peptide; 1.6 parts by weight of modified sea cucumber peptide, 14 parts by weight of soybean lecithin, and 3.4 parts by weight of cholesterol were dispersed in 500 parts by weight of ethanol, transferred to an eggplant-shaped flask, heated to 43° C., and rotary evaporated to obtain a lipid film; 500 parts by weight of a first phosphate buffer solution with a pH of 7.4 preheated to 46° C. was added, and the lipid film fell off when the eggplant-shaped flask was rotated, and the solution was allowed to swell at 46° C. for 2.5 hours. After the swelling was completed, the solution was homogenized by ultrasonication to obtain a liposome suspension; 8.5 parts by weight of chitosan oligosaccharide are dispersed in 500 parts by weight of a second phosphate buffer solution with a pH of 5.5 to obtain a chitosan oligosaccharide solution; 10 parts by weight of a liposome suspension and 11 parts by weight of the chitosan oligosaccharide solution are mixed, stirred and reacted at 24°C for 3.2 hours. After the reaction is completed, the sea cucumber peptide complex is obtained by dialyzing and freeze-drying.

[0031] Preparation Example 3: The preparation method of the sea cucumber peptide complex comprises the following steps: Fresh sea cucumbers were washed with clean water, their internal organs were removed, and their body walls were taken out and cleaned with distilled water. The body walls were pulverized in a chopper for 4 minutes to obtain sea cucumber pieces. 10 parts by weight of the sea cucumber pieces were mixed with 90 parts by weight of a first phosphate buffer solution with a pH of 7.4, and the mixture was homogenized at 3°C ​​for 12 minutes to obtain a homogenate. 0.024 parts by weight of neutral protease was added to the homogenate, the pH was adjusted to 7.0, the temperature was raised to 47° C., and the mixture was stirred for 5 hours to perform a primary enzymolysis. After the primary enzymolysis was completed, the mixture was heated to 92° C. and inactivated for 12 minutes to obtain a primary enzymolysis solution. The primary enzymolysis solution was heated to 52° C., 0.044 parts by weight of flavor protease and 0.017 parts by weight of aminopeptidase were added, the pH was adjusted to 7.4, the mixture was stirred for 10 hours to perform a secondary enzymolysis. After the secondary enzymolysis was completed, the mixture was heated to 92° C. and inactivated for 12 minutes to obtain a secondary enzymolysis solution. After 100 parts by weight of the secondary enzymatic hydrolysate was cooled to 3°C, 24 parts by weight of ammonium sulfate was added while stirring, and the mixture was stirred at 3°C ​​for 1.4 hours. After the reaction was completed, the mixture was allowed to stand at 3°C ​​for 11 hours, and the supernatant was collected by centrifugation at 8000 r / min for 20 minutes. The supernatant was filtered using a 600Da molecular sieve ultrafiltration membrane to collect the polypeptide to obtain sea cucumber peptide; 4 parts by weight of sea cucumber peptide and 0.014 parts by weight of maleimide-PEG2-succinimide ester were dispersed in 50 parts by weight of a first phosphate buffer at pH 7.4, and stirred at 24° C. in a nitrogen atmosphere for 3.4 hours. After the reaction, ultrafiltration was performed to terminate the reaction and remove unreacted small molecules to obtain an intermediate; 8 parts by weight of the intermediate and 0.06 parts by weight of 1,2-dipalmitoyl-sn-glycero-3-phosphothioethanol were dispersed in 50 parts by weight of a first phosphate buffer at pH 7.4, and stirred at 24° C. for 10 hours in a nitrogen atmosphere. After the reaction, the dialyzate was dialyzed for 36 hours using a 3500 Da dialysis bag to obtain a dialysate, which was sterilized by a 0.22 μm filter membrane and freeze-dried to obtain a modified sea cucumber peptide; 1.7 parts by weight of modified sea cucumber peptide, 16 parts by weight of soybean lecithin, and 3.7 parts by weight of cholesterol were dispersed in 500 parts by weight of ethanol, transferred to an eggplant-shaped flask, heated to 46° C., and rotary evaporated to obtain a lipid film; 500 parts by weight of a first phosphate buffer solution with a pH of 7.4 preheated to 47° C. was added, and the lipid film fell off when the eggplant-shaped flask was rotated, and the solution was allowed to swell at 47° C. for 3 hours. After the swelling was completed, the solution was homogenized by ultrasonication to obtain a liposome suspension; 9 parts by weight of chitosan oligosaccharide are dispersed in 500 parts by weight of a second phosphate buffer solution with a pH of 5.5 to obtain a chitosan oligosaccharide solution; 10 parts by weight of a liposome suspension and 12 parts by weight of the chitosan oligosaccharide solution are mixed, stirred and reacted at 24°C for 3.4 hours, and after the reaction is completed, dialyzed and freeze-dried to obtain a sea cucumber peptide complex.

[0032] Preparation Example 4: The preparation method of the sea cucumber peptide complex comprises the following steps: Fresh sea cucumbers were washed with clean water, their internal organs were removed, and their body walls were taken out and cleaned with distilled water. The body walls were pulverized in a chopper for 4.5 minutes to obtain sea cucumber pieces. 10 parts by weight of the sea cucumber pieces were mixed with 95 parts by weight of a first phosphate buffer solution with a pH of 7.4, and the mixture was homogenized at 3.5°C for 13 minutes to obtain a homogenate. 0.027 parts by weight of neutral protease was added to the homogenate, the pH was adjusted to 7.1, the temperature was raised to 48° C., and the mixture was stirred for 5.5 hours to perform a primary enzymatic hydrolysis. After the primary enzymatic hydrolysis was completed, the mixture was heated to 93° C. and inactivated for 14 minutes to obtain a primary enzymatic hydrolyzate. The primary enzymatic hydrolyzate was heated to 54° C., 0.047 parts by weight of flavor protease and 0.018 parts by weight of aminopeptidase were added to the mixture, the pH was adjusted to 7.5, the mixture was stirred for 11 hours to perform a secondary enzymatic hydrolysis, and after the secondary enzymatic hydrolysis was completed, the mixture was heated to 94° C. and inactivated for 13 minutes to obtain a secondary enzymatic hydrolyzate. After 100 parts by weight of the secondary enzymatic hydrolysate was cooled to 3.5°C, 27 parts by weight of ammonium sulfate was added while stirring, and the mixture was stirred at 3°C ​​for 1.7 hours. After the reaction was completed, the mixture was allowed to stand at 3°C ​​for 11.5 hours, and the supernatant was collected by centrifugation at 8000 r / min for 20 minutes. The supernatant was filtered using a 700Da molecular sieve ultrafiltration membrane to collect the polypeptide to obtain sea cucumber peptide; 4.5 parts by weight of sea cucumber peptide and 0.017 parts by weight of maleimide-PEG2-succinimide ester were dispersed in 50 parts by weight of a first phosphate buffer at pH 7.4, and stirred at 25° C. in a nitrogen atmosphere for 3.7 hours. After the reaction, the mixture was ultrafiltered to terminate the reaction and remove unreacted small molecules to obtain an intermediate; 8.5 parts by weight of the intermediate and 0.065 parts by weight of 1,2-dipalmitoyl-sn-glycero-3-phosphothioethanol were dispersed in 50 parts by weight of a first phosphate buffer at pH 7.4, and stirred at 25° C. for 11 hours in a nitrogen atmosphere. After the reaction, the dialyzate was dialyzed with a 3500Da dialysis bag for 42 hours to obtain a dialysate, which was sterilized with a 0.22μm filter membrane and freeze-dried to obtain a modified sea cucumber peptide; 1.8 parts by weight of modified sea cucumber peptide, 18 parts by weight of soybean lecithin, and 4.2 parts by weight of cholesterol were dispersed in 500 parts by weight of ethanol, transferred to an eggplant-shaped flask, heated to 48° C., and rotary evaporated to obtain a lipid film; 500 parts by weight of a first phosphate buffer solution with a pH of 7.4 preheated to 47° C. was added, and the lipid film fell off when the eggplant-shaped flask was rotated, and the solution was allowed to swell at 47° C. for 3.5 hours. After the swelling was completed, the solution was homogenized by ultrasonication to obtain a liposome suspension; 9.5 parts by weight of chitosan oligosaccharide are dispersed in 500 parts by weight of a second phosphate buffer solution with a pH of 5.5 to obtain a chitosan oligosaccharide solution; 10 parts by weight of a liposome suspension and 13 parts by weight of the chitosan oligosaccharide solution are mixed, stirred and reacted at 25°C for 3.7 hours. After the reaction is completed, the sea cucumber peptide complex is obtained by dialyzing and freeze-drying.

[0033] Preparation Example 5: The preparation method of the sea cucumber peptide complex comprises the following steps: Fresh sea cucumbers were washed with clean water, their internal organs were removed, and their body walls were taken out and cleaned with distilled water. The body walls were pulverized in a chopper for 5 minutes to obtain sea cucumber pieces. 10 parts by weight of the sea cucumber pieces were mixed with 100 parts by weight of a first phosphate buffer having a pH of 7.4, and the mixture was homogenized at 4°C for 15 minutes to obtain a homogenate. 0.03 parts by weight of neutral protease was added to the homogenate, the pH was adjusted to 7.2, the temperature was raised to 50° C., and the mixture was stirred for 6 hours to perform a primary enzymolysis. After the primary enzymolysis was completed, the mixture was heated to 95° C. and inactivated for 15 minutes to obtain a primary enzymolysis solution. The primary enzymolysis solution was heated to 55° C., 0.05 parts by weight of flavor protease and 0.02 parts by weight of aminopeptidase were added, the pH was adjusted to 7.6, the mixture was stirred for 12 hours to perform a secondary enzymolysis. After the secondary enzymolysis was completed, the mixture was heated to 95° C. and inactivated for 15 minutes to obtain a secondary enzymolysis solution. After 100 parts by weight of the secondary enzymatic hydrolysate was cooled to 4°C, 30 parts by weight of ammonium sulfate was added while stirring, and the mixture was stirred at 4°C for 2 hours. After the reaction was completed, the mixture was allowed to stand at 4°C for 12 hours, and the supernatant was collected by centrifugation at 8000 r / min for 20 minutes. The supernatant was filtered using a 1000Da molecular sieve ultrafiltration membrane to collect the polypeptide to obtain sea cucumber peptide; 5 parts by weight of sea cucumber peptide and 0.02 parts by weight of maleimide-PEG2-succinimide ester were dispersed in 50 parts by weight of a first phosphate buffer having a pH of 7.4, and stirred at 25° C. in a nitrogen atmosphere for 4 hours. After the reaction, ultrafiltration was performed to terminate the reaction and remove unreacted small molecules to obtain an intermediate; 9 parts by weight of the intermediate and 0.07 parts by weight of 1,2-dipalmitoyl-sn-glycero-3-phosphothioethanol were dispersed in 50 parts by weight of a first phosphate buffer at pH 7.4, and stirred at 25° C. for 12 hours in a nitrogen atmosphere. After the reaction, the dialyzate was dialyzed with a 3500Da dialysis bag for 48 hours to obtain a dialysate, which was sterilized with a 0.22μm filter membrane and freeze-dried to obtain a modified sea cucumber peptide; 2 parts by weight of modified sea cucumber peptide, 20 parts by weight of soybean lecithin, and 4.5 parts by weight of cholesterol were dispersed in 500 parts by weight of ethanol, transferred to an eggplant-shaped flask, heated to 50°C, and rotary evaporated to obtain a lipid film; 500 parts by weight of a first phosphate buffer solution with a pH of 7.4 preheated to 50°C was added, and the lipid film fell off when the eggplant-shaped flask was rotated, and the solution was allowed to swell at 50°C for 4 hours. After the swelling was completed, the solution was homogenized by ultrasonication to obtain a liposome suspension; 10 parts by weight of chitosan oligosaccharide are dispersed in 500 parts by weight of a second phosphate buffer solution with a pH of 5.5 to obtain a chitosan oligosaccharide solution; 10 parts by weight of a liposome suspension and 15 parts by weight of the chitosan oligosaccharide solution are mixed, stirred and reacted at 25° C. for 4 hours, and after the reaction is completed, dialyzed and freeze-dried to obtain a sea cucumber peptide complex.

[0034] Preparation Example 6: The preparation method of the sea cucumber peptide complex comprises the following steps: Fresh sea cucumbers were washed with clean water, their internal organs were removed, and their body walls were taken out and cleaned with distilled water. The body walls were pulverized in a chopper for 5 minutes to obtain sea cucumber pieces. 10 parts by weight of the sea cucumber pieces were mixed with 100 parts by weight of a first phosphate buffer having a pH of 7.4, and the mixture was homogenized at 4°C for 15 minutes to obtain a homogenate. 0.03 parts by weight of neutral protease was added to the homogenate, the pH was adjusted to 7.2, the temperature was raised to 50° C., and the mixture was stirred for 6 hours to perform a primary enzymolysis. After the primary enzymolysis was completed, the mixture was heated to 95° C. and inactivated for 15 minutes to obtain a primary enzymolysis solution. The primary enzymolysis solution was heated to 55° C., 0.05 parts by weight of flavor protease and 0.02 parts by weight of aminopeptidase were added, the pH was adjusted to 7.6, the mixture was stirred for 12 hours to perform a secondary enzymolysis. After the secondary enzymolysis was completed, the mixture was heated to 95° C. and inactivated for 15 minutes to obtain a secondary enzymolysis solution. After 100 parts by weight of the secondary enzymatic hydrolysate was cooled to 4°C, 30 parts by weight of ammonium sulfate was added while stirring, and the mixture was stirred at 4°C for 2 hours. After the reaction was completed, the mixture was allowed to stand at 4°C for 12 hours, and the supernatant was collected by centrifugation at 8000 r / min for 20 minutes. The supernatant was filtered using a 1000Da molecular sieve ultrafiltration membrane to collect the polypeptide to obtain sea cucumber peptide; 2 parts by weight of sea cucumber peptide, 20 parts by weight of soybean lecithin and 4.5 parts by weight of cholesterol are dispersed in 500 parts by weight of ethanol, transferred to an eggplant-shaped flask, heated to 50°C and rotary evaporated to obtain a lipid film; 500 parts by weight of a first phosphate buffer solution with a pH of 7.4 preheated to 50°C is added, the lipid film falls off when the eggplant-shaped flask is rotated, and the solution is allowed to swell at 50°C for 4 hours. After the swelling is completed, the solution is homogenized by ultrasonication to obtain a liposome suspension; 10 parts by weight of chitosan oligosaccharide are dispersed in 500 parts by weight of a second phosphate buffer solution with a pH of 5.5 to obtain a chitosan oligosaccharide solution; 10 parts by weight of a liposome suspension and 15 parts by weight of the chitosan oligosaccharide solution are mixed, stirred and reacted at 25° C. for 4 hours, and after the reaction is completed, dialyzed and freeze-dried to obtain a sea cucumber peptide complex.

[0035] Preparation Example 7: The preparation method of the sea cucumber peptide complex comprises the following steps: The 1,2-dipalmitoyl-sn-glycero-3-phosphothioethanol in Preparation Example 5 was replaced with distearoylphosphatidylethanolamine-polyethylene glycol-thiol, and the other operations were consistent with Preparation Example 5.

[0036] Preparation Example 8: The preparation method of the sea cucumber peptide complex comprises the following steps: Fresh sea cucumbers were washed with clean water, their internal organs were removed, and their body walls were taken out and cleaned with distilled water. The body walls were pulverized in a chopper for 5 minutes to obtain sea cucumber pieces. 10 parts by weight of the sea cucumber pieces were mixed with 100 parts by weight of a first phosphate buffer having a pH of 7.4, and the mixture was homogenized at 4°C for 15 minutes to obtain a homogenate. 0.03 parts by weight of neutral protease was added to the homogenate, the pH was adjusted to 7.2, the temperature was raised to 50° C., and the mixture was stirred for 6 hours to perform a primary enzymolysis. After the primary enzymolysis was completed, the mixture was heated to 95° C. and inactivated for 15 minutes to obtain a primary enzymolysis solution. The primary enzymolysis solution was heated to 55° C., 0.05 parts by weight of flavor protease and 0.02 parts by weight of aminopeptidase were added, the pH was adjusted to 7.6, the mixture was stirred for 12 hours to perform a secondary enzymolysis. After the secondary enzymolysis was completed, the mixture was heated to 95° C. and inactivated for 15 minutes to obtain a secondary enzymolysis solution. After 100 parts by weight of the secondary enzymatic hydrolysate was cooled to 4°C, 30 parts by weight of ammonium sulfate was added while stirring, and the mixture was stirred at 4°C for 2 hours. After the reaction was completed, the mixture was allowed to stand at 4°C for 12 hours, and the supernatant was collected by centrifugation at 8000 r / min for 20 minutes. The supernatant was filtered using a 1000Da molecular sieve ultrafiltration membrane to collect the polypeptide to obtain sea cucumber peptide; 5 parts by weight of sea cucumber peptide and 0.02 parts by weight of maleimide-PEG2-succinimide ester were dispersed in 50 parts by weight of a first phosphate buffer having a pH of 7.4, and stirred at 25° C. in a nitrogen atmosphere for 4 hours. After the reaction, ultrafiltration was performed to terminate the reaction and remove unreacted small molecules to obtain an intermediate; 9 parts by weight of the intermediate and 0.07 parts by weight of 1,2-dipalmitoyl-sn-glycero-3-phosphothioethanol were dispersed in 50 parts by weight of a first phosphate buffer at pH 7.4, and stirred at 25° C. for 12 hours in a nitrogen atmosphere. After the reaction, the dialyzate was dialyzed with a 3500Da dialysis bag for 48 hours to obtain a dialysate, which was sterilized with a 0.22μm filter membrane and freeze-dried to obtain a modified sea cucumber peptide; 2 parts by weight of modified sea cucumber peptide, 20 parts by weight of soybean lecithin and 4.5 parts by weight of cholesterol are dispersed in 500 parts by weight of ethanol, transferred to an eggplant-shaped flask, heated to 50°C and rotary evaporated to obtain a lipid film; 100 parts by weight of a first phosphate buffer solution with a pH of 7.4 preheated to 50°C is added, the lipid film falls off when the eggplant-shaped flask is rotated, and the mixture is allowed to swell at 50°C for 4 hours. After the swelling is completed, the mixture is homogenized by ultrasonic treatment to obtain a sea cucumber peptide complex.

[0037] Preparation Example 9: The preparation method of sea cucumber peptide comprises the following steps: Fresh sea cucumbers were washed with clean water, their internal organs were removed, and their body walls were taken out and cleaned with distilled water. The body walls were pulverized in a chopper for 5 minutes to obtain sea cucumber pieces. 10 parts by weight of the sea cucumber pieces were mixed with 100 parts by weight of a first phosphate buffer having a pH of 7.4, and the mixture was homogenized at 4°C for 15 minutes to obtain a homogenate. 0.03 parts by weight of neutral protease was added to the homogenate, the pH was adjusted to 7.2, the temperature was raised to 50° C., and the mixture was stirred for 6 hours to perform a primary enzymolysis. After the primary enzymolysis was completed, the mixture was heated to 95° C. and inactivated for 15 minutes to obtain a primary enzymolysis solution. The primary enzymolysis solution was heated to 55° C., 0.05 parts by weight of flavor protease and 0.02 parts by weight of aminopeptidase were added, the pH was adjusted to 7.6, the mixture was stirred for 12 hours to perform a secondary enzymolysis. After the secondary enzymolysis was completed, the mixture was heated to 95° C. and inactivated for 15 minutes to obtain a secondary enzymolysis solution. After 100 parts by weight of the secondary enzymatic hydrolysate was cooled to 4°C, 30 parts by weight of ammonium sulfate was added while stirring, and the mixture was stirred at 4°C for 2 hours. After the reaction was completed, the mixture was allowed to stand at 4°C for 12 hours, and the supernatant was collected by centrifugation at 8000 r / min for 20 minutes. The supernatant was filtered using a 1000Da molecular sieve ultrafiltration membrane to collect the polypeptide to obtain sea cucumber peptide.

[0038] Preparation Example 10: The preparation method of fermented yacon extract comprises the following steps: The yacon was washed and sliced ​​to obtain yacon pulp crumbs. 10 parts by weight of yacon pulp crumbs were soaked and extracted at 50°C for 1.5 hours with 80% by mass ethanol according to a material-liquid ratio of 1:10. The extract was collected by filtration. The extract was concentrated under reduced pressure to recover ethanol until the ethanol was no longer evaporated to obtain an extract. The extract was dispersed and diluted with purified water to 15 times the weight of the extract, and 0.1 parts by weight of fermentation lactobacillus was added. The temperature was adjusted to 37°C for fermentation for 72 hours. After the fermentation was completed, the temperature was raised to 95°C for insulation treatment for 10 minutes and then cooled to 23°C. The extract was collected by ultrafiltration using a 3000Da ultrafiltration membrane, and then freeze-dried and pulverized to obtain the fermented yacon extract.

[0039] Preparation Example 11: The preparation method of fermented yacon extract comprises the following steps: The yacon was washed and sliced ​​to obtain yacon pulp crumbs. 10 parts by weight of yacon pulp crumbs were soaked and extracted at 52°C for 1.7 hours with 80% by mass ethanol according to a material-liquid ratio of 1:11. The extract was collected by filtration. The extract was concentrated under reduced pressure to recover ethanol until the ethanol was no longer evaporated to obtain an extract. The extract was dispersed and diluted with purified water to 16 times the weight of the extract, and 0.2 parts by weight of plant lactobacillus was added. The temperature was adjusted to 38°C for fermentation for 78 hours. After the fermentation was completed, the temperature was raised to 96°C for insulation treatment for 11 minutes and then cooled to 24°C. The extract was collected by ultrafiltration using a 3000Da ultrafiltration membrane, and then freeze-dried and pulverized to obtain the fermented yacon extract.

[0040] Preparation Example 12: The preparation method of fermented yacon extract comprises the following steps: The yacon was washed and sliced ​​to obtain yacon pulp crumbs. 10 parts by weight of yacon pulp crumbs were soaked and extracted in 80% by mass ethanol at a solid-liquid ratio of 1:12 at 54°C for 1.9 hours. The extract was collected by filtration. The extract was concentrated under reduced pressure to recover ethanol until the ethanol was no longer evaporated to obtain an extract. The extract was dispersed and diluted with purified water to 17 times the weight of the extract. 0.3 parts by weight of fermented lactobacillus was added. The temperature was adjusted to 39°C for fermentation for 84 hours. After the fermentation was completed, the temperature was raised to 97°C for insulation treatment for 12 minutes and then cooled to 24°C. The extract was collected by ultrafiltration using a 3000Da ultrafiltration membrane. The extract was then freeze-dried and pulverized to obtain the fermented yacon extract.

[0041] Preparation Example 13: The preparation method of fermented yacon extract comprises the following steps: The yacon was washed and sliced ​​to obtain yacon pulp crumbs. 10 parts by weight of yacon pulp crumbs were soaked and extracted at 57°C for 2.3 hours with 80% by mass ethanol according to a material-liquid ratio of 1:13. The extract was collected by filtration. The extract was concentrated under reduced pressure to recover ethanol until the ethanol was no longer evaporated to obtain an extract. The extract was dispersed and diluted with purified water to 19 times the weight of the extract, and 0.4 parts by weight of Lactobacillus plantarum was added. The temperature was adjusted to 39°C for fermentation for 90 hours. After the fermentation was completed, the temperature was raised to 98°C for insulation treatment for 14 minutes and then cooled to 25°C. The extract was collected by ultrafiltration using a 3000Da ultrafiltration membrane, and then freeze-dried and pulverized to obtain the fermented yacon extract.

[0042] Preparation Example 14: The preparation method of fermented yacon extract comprises the following steps: The yacon was washed and sliced ​​to obtain yacon pulp crumbs. 10 parts by weight of yacon pulp crumbs were soaked and extracted at 60°C for 2.5 hours with 80% by mass ethanol according to a material-liquid ratio of 1:15. The extract was collected by filtration. The extract was concentrated under reduced pressure to recover ethanol until the ethanol was no longer evaporated to obtain an extract. The extract was dispersed and diluted with purified water to 20 times the weight of the extract, and 0.5 parts by weight of fermentation lactobacillus was added. The temperature was adjusted to 40°C for fermentation for 96 hours. After the fermentation was completed, the temperature was raised to 100°C for insulation treatment for 15 minutes and then cooled to 25°C. The extract was collected by ultrafiltration using a 3000Da ultrafiltration membrane, and then freeze-dried and pulverized to obtain the fermented yacon extract.

[0043] Preparation Example 15: The preparation method of yacon extract comprises the following steps: Wash and slice the yacon to obtain yacon pulp minced meat. Soak 10 parts by weight of yacon in 80% ethanol at a solid-liquid ratio of 1:15 at 60°C for 2.5 hours. Collect the extract by filtration, and concentrate the extract under reduced pressure to recover ethanol until no more ethanol is evaporated to obtain the yacon extract.

[0044] Example 1: A method for preparing an anti-aging composition containing sea cucumber peptides comprises the following steps: 3 parts by weight of the sea cucumber peptide complex prepared in Preparation Example 1, 1 part by weight of the fermented yacon extract prepared in Preparation Example 10, 1 part by weight of grape seed extract, and 0.5 part by weight of clam peptide were mixed for 10 minutes to obtain a mixture; 65 parts by weight of purified water were heated to 85° C., 0.6 parts by weight of xanthan gum and 0.3 parts by weight of sodium carboxymethyl cellulose were added, and stirred for 20 minutes to obtain a mixed system A; Mixed system A was cooled to 50° C., 0.2 parts by weight of sodium hyaluronate, 2 parts by weight of butylene glycol, 0.5 parts by weight of hydroxyethyl urea, 0.8 parts by weight of steareth-21 and 0.3 parts by weight of glyceryl glucoside were added, and stirred for 10 minutes to obtain mixed system B; The mixed system B was cooled to 40° C., the above mixture was added, and the pH was adjusted to 5.5 with lactic acid. The mixture was stirred and mixed for 20 minutes to prepare an anti-aging composition.

[0045] Example 2: A method for preparing an anti-aging composition containing sea cucumber peptides comprises the following steps: 3.5 parts by weight of the sea cucumber peptide complex prepared in Preparation Example 2, 1.2 parts by weight of the fermented yacon extract prepared in Preparation Example 11, 1.5 parts by weight of grape seed extract, and 0.7 parts by weight of clam peptide were mixed for 11 minutes to obtain a mixture; 70 parts by weight of purified water were heated to 86° C., 0.8 parts by weight of xanthan gum and 0.5 parts by weight of sodium carboxymethyl cellulose were added, and stirred for 22 minutes to obtain a mixed system A; Mixed system A was cooled to 51° C., 0.25 parts by weight of sodium hyaluronate, 2.5 parts by weight of butylene glycol, 0.6 parts by weight of hydroxyethyl urea, 0.9 parts by weight of steareth-21 and 0.5 parts by weight of glyceryl glucoside were added, and stirred for 11 minutes to obtain mixed system B; The mixed system B was cooled to 41° C., the above mixture was added, and the pH was adjusted to 5.7 with lactic acid. The mixture was stirred and mixed for 22 minutes to prepare an anti-aging composition.

[0046] Example 3: A method for preparing an anti-aging composition containing sea cucumber peptides comprises the following steps: 4 parts by weight of the sea cucumber peptide complex prepared in Preparation Example 3, 1.4 parts by weight of the fermented yacon extract prepared in Preparation Example 12, 2 parts by weight of grape seed extract, and 0.9 parts by weight of clam peptide were mixed for 13 minutes to obtain a mixture; 75 parts by weight of purified water were heated to 87° C., 0.9 parts by weight of xanthan gum and 0.7 parts by weight of sodium carboxymethyl cellulose were added, and stirred for 24 minutes to obtain a mixed system A; Mixed system A was cooled to 52° C., 0.3 parts by weight of sodium hyaluronate, 3 parts by weight of butylene glycol, 0.8 parts by weight of hydroxyethyl urea, 1.1 parts by weight of steareth-21 and 0.7 parts by weight of glyceryl glucoside were added, and stirred for 12 minutes to obtain mixed system B; The mixed system B was cooled to 42° C., the above mixture was added, and the pH was adjusted to 5.8 with lactic acid. The mixture was stirred and mixed for 24 minutes to prepare an anti-aging composition.

[0047] Example 4: A method for preparing an anti-aging composition containing sea cucumber peptides comprises the following steps: 4.5 parts by weight of the sea cucumber peptide complex prepared in Preparation Example 4, 1.8 parts by weight of the fermented yacon extract prepared in Preparation Example 13, 2.5 parts by weight of grape seed extract, and 1.2 parts by weight of clam peptide were mixed for 14 minutes to obtain a mixture; 80 parts by weight of purified water were heated to 88° C., 1.1 parts by weight of xanthan gum and 0.9 parts by weight of sodium carboxymethyl cellulose were added, and stirred for 28 minutes to obtain a mixed system A; Mixed system A was cooled to 54° C., 0.4 parts by weight of sodium hyaluronate, 3.5 parts by weight of butylene glycol, 0.9 parts by weight of hydroxyethyl urea, 1.3 parts by weight of steareth-21 and 0.8 parts by weight of glyceryl glucoside were added, and stirred for 14 minutes to obtain mixed system B; The mixed system B was cooled to 44° C., the above mixture was added, and the pH was adjusted to 5.9 with lactic acid. The mixture was stirred and mixed for 28 minutes to prepare an anti-aging composition.

[0048] Example 5: A method for preparing an anti-aging composition containing sea cucumber peptides comprises the following steps: 5 parts by weight of the sea cucumber peptide complex prepared in Preparation Example 5, 2 parts by weight of the fermented yacon extract prepared in Preparation Example 14, 3 parts by weight of grape seed extract, and 1.5 parts by weight of clam peptide were mixed for 15 minutes to obtain a mixture; 85 parts by weight of purified water were heated to 90° C., 1.2 parts by weight of xanthan gum and 1 part by weight of sodium carboxymethyl cellulose were added, and stirred for 30 minutes to obtain a mixed system A; Mixed system A was cooled to 55° C., and 0.5 parts by weight of sodium hyaluronate, 4 parts by weight of butylene glycol, 1 part by weight of hydroxyethyl urea, 1.5 parts by weight of steareth-21, and 1 part by weight of glyceryl glucoside were added and stirred for 15 minutes to obtain mixed system B; The mixed system B was cooled to 45° C., the above mixture was added, and the pH was adjusted to 6.0 with lactic acid. The mixture was stirred and mixed for 30 minutes to prepare an anti-aging composition.

[0049] Comparative Example 1: A method for preparing an anti-aging composition containing sea cucumber peptides comprises the following steps: The sea cucumber peptide complex prepared in Preparation Example 5 in Example 5 was replaced by the sea cucumber peptide complex prepared in Preparation Example 6, and other operations were consistent with Example 5.

[0050] Comparative Example 2: A method for preparing an anti-aging composition containing sea cucumber peptides comprises the following steps: The sea cucumber peptide complex prepared in Preparation Example 5 in Example 5 was replaced by the sea cucumber peptide complex prepared in Preparation Example 7, and other operations were consistent with Example 5.

[0051] Comparative Example 3: A method for preparing an anti-aging composition containing sea cucumber peptides comprises the following steps: The sea cucumber peptide complex prepared in Preparation Example 5 in Example 5 was replaced by the sea cucumber peptide complex prepared in Preparation Example 8, and other operations were consistent with Example 5.

[0052] Comparative Example 4: A method for preparing an anti-aging composition containing sea cucumber peptides comprises the following steps: The sea cucumber peptide complex prepared in Preparation Example 5 in Example 5 was replaced with the sea cucumber peptide prepared in Preparation Example 9, and other operations were consistent with Example 5.

[0053] Comparative Example 5: A method for preparing an anti-aging composition containing sea cucumber peptides comprises the following steps: The fermented yacon extract prepared in Preparation Example 14 in Example 5 was replaced by the yacon extract prepared in Preparation Example 15, and the other operations were consistent with Example 5.

[0054] Comparative Example 6: A method for preparing an anti-aging composition containing sea cucumber peptides comprises the following steps: The fermented yacon extract obtained in Preparation Example 14 of Example 5 was removed, and other operations were consistent with Example 5.

[0055] Skin irritation test: Testing Method: 220 volunteers, each group of 20, were selected to test the anti-aging compositions prepared in Examples 1-5 and Comparative Examples 1-6. Each participant used the inner sides of both arms as control tests. 0.5 mL of the anti-aging compositions prepared in Examples 1-5 and Comparative Examples 1-6 were applied to the inner side of the left hand twice daily, while the right hand remained untreated as a control. Skin reactions were recorded 24 and 72 hours later.

[0056] The results show that after 24 hours and 72 hours of applying the anti-aging compositions prepared in Examples 1 to 5 of the present invention and Comparative Examples 1 to 6, no allergic phenomena such as skin redness or erythema were observed, indicating that the anti-aging compositions prepared in the present invention are safe for use in cosmetics without any adverse reactions.

[0057] Anti-aging effect test: The anti-aging compositions prepared in Examples 1 to 5 and Comparative Examples 1 to 6 were tested for their hydroxyl radical scavenging effects and inflammation anti-aging properties. The testing method was as follows: 10 μL of a 1.25 mg / mL salicylic acid solution and a 1.5 mg / mL ferrous sulfate solution were added to an ELISA plate, followed by 150 μL of pure water, 50 μL of the anti-aging composition prepared in the present invention, and finally 10 μL of 0.03% mass concentration of hydrogen peroxide. After reacting for 10 minutes, the absorbance was measured using a microplate reader and compared with the blank control sample (50 μL of the anti-aging composition was replaced with 50 μL of pure water). The scavenging rates of the anti-aging compositions prepared in Examples 1 to 5 and Comparative Examples 1 to 6 of the present invention for strong free radicals were calculated. The test results are shown in Table 1.

[0058] Table 1. Hydroxyl radical scavenging rate Moisturizing test: Test environment: temperature 25±0.5℃, humidity 30±0.5%; 44 volunteers were selected, half male and half female, and divided into 11 groups, with 2 males and 2 females in each group. Each group used the anti-aging composition prepared in Examples 1 to 5 and Comparative Examples 1 to 6, 2 g of which was evenly applied to the forearm. The hydration content of the skin stratum corneum before and after use was measured using an instrument. The test results are shown in Table 2.

[0059] Table 2. Hydration content of the skin stratum corneum The data in Tables 1 and 2 demonstrate that the anti-aging compositions prepared in Examples 1-5 of the present invention exhibit excellent anti-aging and long-lasting moisturizing properties. As shown in Table 1, the anti-aging compositions prepared in Examples 1-5 of the present invention effectively scavenge strong free radicals, exhibit excellent anti-aging effects, and maintain a healthy skin condition. As shown in Table 2, the anti-aging compositions prepared in Examples 1-5 of the present invention maintain good moisturizing properties even after 10 hours of application. The decline in anti-aging and moisturizing properties of Comparative Example 1 may be due to the fact that the sea cucumber peptide has not been modified, the utilization rate is low, the anti-aging and moisturizing effects are poor, and the performance of the anti-aging composition is affected; the reason for the decline in anti-aging and moisturizing properties of Comparative Example 2 may be that distearoylphosphatidylethanolamine-polyethylene glycol-thiol is used to modify the sea cucumber peptide, which makes the molecular weight of the modified sea cucumber peptide too large, affecting its utilization; the reason for the decline in performance of Comparative Example 3 may be the lack of chitosan oligosaccharides, which reduces the residence time and affects the performance of the anti-aging composition; the reason for the decline in performance of Comparative Example 4 may be that the sea cucumber peptide has not been modified and encapsulated, and its utilization and residence time on the skin surface cannot be improved, thereby affecting the performance of the anti-aging composition; the reason for the decline in performance of Comparative Example 5 may be that the fermented yacon extract has not been fermented, and its biological activity cannot be enhanced, thereby affecting the performance of the anti-aging composition; the reason for the decline in performance of Comparative Example 6 may be that after the fermented yacon extract is removed, the components that can provide moisturizing properties are greatly reduced, thereby affecting the performance of the anti-aging composition.

[0060] The embodiments described above provide a detailed description of the technical solutions and beneficial effects of the present invention. It should be understood that the above descriptions are merely specific embodiments of the present invention and are not intended to limit the present invention. Various changes and modifications may be made to the present invention without departing from the spirit and scope of the present invention, and such changes and modifications fall within the scope of the present invention as claimed.

Claims

1. A method for preparing an anti-aging composition containing sea cucumber peptide, characterized in that: The preparation method comprises the following steps: The sea cucumber peptide complex, fermented yacon extract, grape seed extract, and clam peptide were mixed at 23-25° C. for 10-15 minutes to obtain a mixture; Xanthan gum and sodium carboxymethyl cellulose were dispersed in purified water at 85-90°C and stirred for 20-30 minutes to obtain a mixed system A; Mixing system A, sodium hyaluronate, butylene glycol, hydroxyethyl urea, steareth-21 and glyceryl glucoside at 50-55° C. for 10-15 minutes to obtain mixing system B; The mixed system B and the mixture are mixed at 40-45° C. for 20-30 minutes to prepare the anti-aging composition.

2. The method for preparing an anti-aging composition containing sea cucumber peptide according to claim 1, wherein: The preparation method of the sea cucumber peptide complex comprises the following steps: The chopped sea cucumber and the first phosphate buffer solution were mixed at a weight ratio of 1:8-10 and homogenized at 2-4°C for 10-15 minutes to obtain a homogenate solution; Neutral protease is added to the homogenate to perform enzymatic hydrolysis to obtain a primary enzymatic hydrolyzate; Flavor protease and aminopeptidase are added to the primary enzymatic hydrolysis solution for secondary enzymatic hydrolysis to obtain a secondary enzymatic hydrolysis solution; The secondary enzymatic hydrolysis solution and ammonium sulfate are mixed in a weight ratio of 10:2-3 and then allowed to stand, and the supernatant is collected to obtain sea cucumber peptide; Sea cucumber peptide and maleimide-PEG2-succinimide ester are reacted at a weight ratio of 3-5:0.01-0.02 at 23-25°C for 3-4 hours to obtain an intermediate; The intermediate and 1,2-dipalmitoyl-sn-glycero-3-phosphothioethanol are reacted at a weight ratio of 7-9:0.05-0.07 at 23-25° C. for 8-12 hours to obtain a modified sea cucumber peptide; Modified sea cucumber peptide, soybean lecithin and cholesterol are mixed in a weight ratio of 1.5-2:12-20:3-4.5 and then subjected to rotary evaporation to obtain a lipid film; First, phosphate buffer is added to the lipid film and subjected to swelling treatment at 45-50° C. for 2-4 hours to obtain a liposome suspension; chitosan oligosaccharide is dispersed in the second phosphate buffer at a material-liquid ratio of 8-10:50 to obtain a chitosan oligosaccharide solution; The liposome suspension and the chitosan oligosaccharide solution are reacted at a weight ratio of 10:10-15 at 23-25° C. for 3-4 hours to prepare the sea cucumber peptide complex.

3. The method for preparing an anti-aging composition containing sea cucumber peptide according to claim 2, wherein: The amount of the neutral protease used is 0.2% to 0.3% of the weight of the crushed sea cucumber.

4. The method for preparing an anti-aging composition containing sea cucumber peptide as claimed in claim 2, wherein: The amount of the flavor protease used is 0.4% to 0.5% of the weight of the chopped sea cucumber.

5. The method for preparing an anti-aging composition containing sea cucumber peptide as claimed in claim 2, characterized in that: The dosage of the aminopeptidase is 0.15% to 0.2% of the weight of the crushed sea cucumber.

6. The method for preparing an anti-aging composition containing sea cucumber peptide according to claim 1, wherein: The preparation method of the fermented yacon extract comprises the following steps: The yacon pulp minced meat is soaked in 80% ethanol by mass at a material-liquid ratio of 1:10-15 at 50-60°C for 1.5-2.5 hours, filtered and concentrated in sequence to obtain an extract, the extract is diluted with purified water to obtain a dilution, and then lactic acid bacteria is added for fermentation to obtain the fermented yacon extract.

7. The method for preparing an anti-aging composition containing sea cucumber peptide according to claim 6, wherein: The lactic acid bacteria include Lactobacillus fermentum or Lactobacillus plantarum.

8. The method for preparing an anti-aging composition containing sea cucumber peptide according to claim 6, wherein: The dosage of the lactic acid bacteria is 1% to 5% of the weight of the yacon pulp.

9. The method for preparing an anti-aging composition containing sea cucumber peptide according to claim 1, wherein: The weight ratio of the sea cucumber peptide complex, fermented yacon extract, grape seed extract, clam peptide, xanthan gum, sodium carboxymethyl cellulose, sodium hyaluronate, butylene glycol, hydroxyethyl urea, steareth-21, glyceryl glucoside and purified water is 3-5:1-2:1-3:0.5-1.5:0.6-1.2:0.3-1:0.2-0.5:2-4:0.5-1:0.8-1.5:0.3-1:65-85.

10. An anti-aging composition containing sea cucumber peptide, characterized in that: The anti-aging composition containing sea cucumber peptide is prepared by the preparation method of any one of claims 1 to 9.

Images