A composition containing an anti-wrinkle composite peptide and a preparation method and application thereof

Abstract

The application belongs to the technical field of skin care products, and particularly relates to a composition containing anti-wrinkle composite peptides, a preparation method and application thereof. The composition is composed of the following raw materials in mass percentage: acetyl hexapeptide-8 0.09-0.11%, chlorella extract 1.2-1.8%, arginine / lysine polypeptide 0.003-0.005%, dipeptide diaminobutyryl benzyl amide diacetate 0.09-0.11%, ginsenoside 0.20-0.25%, octisalate 0.70-0.75%, 1,2-hexanediol 0.5-1%, PEG-40 hydrogenated castor oil 1.02-1.14%, PPG-26-butanediol polyether-26 1.53-1.71%, butanediol 24-26%, and the balance being water. The performance of the composition is detected, and the results show that the composition has excellent anti-wrinkle effect.

Description

Technical Field

[0001] This invention belongs to the field of skin care technology, specifically relating to a composition containing anti-wrinkle complex peptides, its preparation method, and its application. Background Technology

[0002] Wrinkles are a natural phenomenon of skin aging. They are formed when the skin is affected by external environmental factors, resulting in the formation of free radicals. These free radicals damage collagen and other active substances within normal cell membranes, oxidizing cells and causing fine lines and wrinkles. The gradual appearance of wrinkles is a sign of natural aging, but it is also influenced by various factors such as genetics, age, living environment, lifestyle, and skincare. With societal development, people's awareness of skincare is gradually increasing, and they are paying more attention to the anti-wrinkle effects of skincare products.

[0003] Chlorella is a ubiquitous single-celled green algae belonging to the Chlorella genus of the Chlorophyta phylum. It is a spherical, single-celled freshwater algae. Studies have found that chlorella extract is rich in polysaccharides, polypeptides, proteins, and other cell-active substances. These active substances help maintain skin elasticity and radiance, reducing fine lines and wrinkles caused by dehydration. Chinese patent CN113476374A discloses an anti-wrinkle cosmetic and its preparation method, revealing that chlorella extract can help promote collagen and elastin synthesis in the skin, playing a repairing and anti-aging role. Furthermore, the collagen in chlorella can help reduce the depth of wrinkles, promote skin renewal after free radical and sun exposure, and regenerate damaged tissue, restoring loose skin to a youthful state, thereby removing wrinkles.

[0004] Existing technologies have disclosed various methods for preparing Chlorella extracts. Chinese Patent CN113637710A discloses a method for preparing Chlorella extract and its application. Chinese Patent CN110801413A discloses a method for preparing Chlorella fermentation extract and its preparation. However, in practice, it has been found that Chlorella extracts prepared using these two methods either have low protein content or low polysaccharide content. Therefore, it is necessary to improve the preparation process of Chlorella extracts to comprehensively increase the protein and polysaccharide content, thereby accelerating its application in the skincare field. Summary of the Invention

[0005] The first objective of this invention is to provide a composition containing an anti-wrinkle complex peptide, which has excellent anti-wrinkle effects.

[0006] A second objective of this invention is to provide a method for preparing a composition containing an anti-wrinkle complex peptide, which has simple steps.

[0007] The third objective of this invention is to provide an application of a composition containing anti-wrinkle complex peptides, which has broad application prospects.

[0008] To achieve the above objectives, the technical solution adopted by the present invention is as follows: A composition containing an anti-wrinkle complex peptide comprises the following raw materials in the indicated weight percentages: acetyl hexapeptide-8 0.09-0.11%, Chlorella extract 1.2-1.8%, arginine / lysine polypeptide 0.003-0.005%, dipeptide diaminobutyryl benzylamide diacetate 0.09-0.11%, ginsenoside 0.20-0.25%, caprylyl glycol 0.70-0.75%, 1,2-hexanediol 0.5-1%, PEG-40 hydrogenated castor oil 1.02-1.14%, PPG-26-butanol polyether-26 1.53-1.71%, butylene glycol 24-26%, and the balance being water.

[0009] Preferably, the preparation process of the Chlorella extract is as follows: (1) Under sterile conditions, Chlorella was inoculated into a modified culture medium and cultured. After a period of time, glucose was added and cultured for a longer period of time. After the culture was completed, the culture medium was collected. The modified culture medium consisted of: calcium chloride 0.2-0.5 g / L, ferrous sulfate 15-20 mg / L, magnesium sulfate 0.2-0.3 g / L, sodium chloride 1-2 g / L, dipotassium hydrogen phosphate 0.3-0.5 g / L, zinc sulfate 0.2-0.5 mg / L, sodium chloride 1-2 g / L, dipotassium hydrogen phosphate 0.3-0.5 g / L, zinc sulfate 0.2-0.5 mg / L, eugenol 0.1-0.3 mg / L, and dipotassium glycyrrhizate 0.1-0.5 mg / L. (2) The culture medium collected in step (1) is filtered and dried to obtain Chlorella powder; (3) Add the Chlorella powder from step (2) to ethanol, heat and extract, collect the extract, filter, concentrate and freeze dry the extract to obtain Chlorella extract.

[0010] Preferably, the concentration of glucose in the modified culture medium in step (1) is 5-10 g / L.

[0011] Preferably, the inoculum size of Chlorella in the modified culture medium in step (1) is 1-8 × 10⁻⁶. 5 The number of cells / mL is 5-6 days, and the culture time is 4-5 days.

[0012] Preferably, the conditions for cultivation and continued cultivation in step (1) are: temperature of 20-25℃ and light intensity of 4000-5000lx.

[0013] Preferably, the filtration in step (2) is performed using a hollow fiber microfiltration membrane; the pore size of the microfiltration membrane is 0.1-0.3 μm.

[0014] Preferably, the heating extraction temperature in step (3) is 50-60℃ and the time is 2-3h.

[0015] The preparation method of the above-mentioned composition containing anti-wrinkle complex peptides includes the following steps: According to the stated mass percentage, acetyl hexapeptide-8, arginine / lysine polypeptide, dipeptide diaminobutyryl benzylamide diacetate are stirred and mixed evenly. Then, chlorella extract, ginsenosides, caprylyl glycol, 1,2-hexanediol, PEG-40 hydrogenated castor oil, PPG-26-butanol polyether-26, butylene glycol, and water are added and stirred until evenly mixed to obtain the final product.

[0016] In the application of the above-mentioned composition containing anti-wrinkle complex peptides, the composition containing anti-wrinkle complex peptides has a mass percentage of 1-10% based on the total mass of the skin care product.

[0017] Preferably, the skin care product further includes the following raw materials in weight percentages: 1-2% carboxymethyl cellulose, 0.5-1% cetyl alcohol, 0.01-0.05% p-hydroxyacetophenone, with the balance being water.

[0018] Compared with the prior art, the main advantages of the present invention are as follows: 1. In the preparation process of Chlorella extract, this invention innovatively adds eugenol and dipotassium glycyrrhizate to the culture medium. The polysaccharide and protein contents of the Chlorella extract obtained after cultivation in the above-mentioned modified culture medium were detected. The results showed that the synergistic effect of eugenol and dipotassium glycyrrhizate could increase the protein and polysaccharide contents of the Chlorella extract, with the protein content reaching 32.59% and the polysaccharide content reaching 5.73%.

[0019] 2. The composition of the present invention has excellent anti-wrinkle effects. After using skin care products containing the composition of the present invention for 28 days, the number, area, and depth of wrinkles on the skin surface of volunteers were significantly improved, with the number of wrinkles reduced by more than 32.52%, the area of ​​wrinkles reduced by more than 42.17%, and the depth of wrinkles reduced by more than 33.48%. Detailed Implementation

[0020] The technical solution of the present invention will be further described below with reference to specific embodiments. However, those skilled in the art should understand that the following embodiments are only for illustrating the present invention and should not be regarded as limiting the present invention. Specific conditions not specified in the embodiments are performed according to conventional conditions or conditions recommended by the manufacturer. Unless otherwise specified, the reagents or instruments used are all conventional products obtained through commercial channels.

[0021] 1. Example Example 1 A composition containing an anti-wrinkle complex peptide comprises the following raw materials in the indicated weight percentages: acetyl hexapeptide-8 0.10%, Chlorella extract 1.3%, arginine / lysine polypeptide 0.004%, dipeptide diaminobutyryl benzylamide diacetate 0.10%, ginsenoside 0.24%, caprylyl glycol 0.73%, 1,2-hexanediol 0.8%, PEG-40 hydrogenated castor oil 1.10%, PPG-26-butanol polyether-26 1.62%, butylene glycol 25%, and the balance being water.

[0022] The specific preparation process of Chlorella extract is as follows: (1) Under aseptic conditions, at 4×10 5 Chlorella was inoculated into a modified medium at an inoculum size of 1 / mL and cultured at 22℃ and 4500 lx for 5.5 days. Glucose was then added, and the culture was continued at 22℃ and 4500 lx for another 4.5 days. The culture medium was collected after the culture was complete. The modified medium consisted of: 0.4 g / L calcium chloride, 17 mg / L ferrous sulfate, 0.25 g / L magnesium sulfate, 1.5 g / L sodium chloride, 0.4 g / L dipotassium hydrogen phosphate, 0.3 mg / L zinc sulfate, 0.2 mg / L eugenol, and 0.3 mg / L dipotassium glycyrrhizate. The added glucose concentration in the modified medium was 7 g / L.

[0023] (2) The culture medium collected in step (1) is filtered through a hollow fiber microfiltration membrane with a pore size of 0.2 μm, the retained material is collected, and dried to obtain Chlorella powder.

[0024] (3) According to the ratio of Chlorella powder to ethanol 1g:10mL, add Chlorella powder from step (2) to 70% ethanol, stir evenly, heat and extract at 55℃ for 2.5h, collect the extract, filter the extract to obtain the supernatant, place the supernatant in a rotary evaporator, concentrate under reduced pressure at 60℃ and freeze dry to obtain Chlorella extract.

[0025] The preparation method of the above-mentioned composition containing anti-wrinkle complex peptides is as follows: According to the above-mentioned mass percentages, acetyl hexapeptide-8, arginine / lysine peptide, dipeptide diaminobutyryl benzylamide diacetate are stirred and mixed evenly. Then, chlorella extract, ginsenosides, caprylyl glycol, 1,2-hexanediol, PEG-40 hydrogenated castor oil, PPG-26-butanol polyether-26, butylene glycol, and water are added and stirred until evenly mixed to obtain the final product.

[0026] Example 2 A composition containing an anti-wrinkle complex peptide comprises the following raw materials in the indicated weight percentages: acetyl hexapeptide-8 0.11%, Chlorella extract 1.8%, arginine / lysine polypeptide 0.005%, dipeptide diaminobutyryl benzylamide diacetate 0.11%, ginsenoside 0.25%, caprylyl glycol 0.75%, 1,2-hexanediol 1%, PEG-40 hydrogenated castor oil 1.14%, PPG-26-butanol polyether-26 1.71%, butylene glycol 26%, and the balance being water.

[0027] The specific preparation process of Chlorella extract is as follows: (1) Under aseptic conditions, at 8×10 5 Chlorella was inoculated into a modified medium at an inoculum size of 10 cells / mL and cultured for 6 days at 20°C and 4000 lx light intensity. Glucose was then added, and the culture was continued for another 5 days at 20°C and 4000 lx light intensity. The culture medium was collected after the culture was complete. The modified medium consisted of: 0.5 g / L calcium chloride, 20 mg / L ferrous sulfate, 0.3 g / L magnesium sulfate, 2 g / L sodium chloride, 0.5 g / L dipotassium hydrogen phosphate, 0.5 mg / L zinc sulfate, 0.3 mg / L eugenol, and 0.5 mg / L dipotassium glycyrrhizate. The added glucose concentration in the modified medium was 10 g / L.

[0028] (2) The culture medium collected in step (1) is filtered through a hollow fiber microfiltration membrane with a pore size of 0.1 μm, the retained material is collected, and dried to obtain Chlorella powder.

[0029] (3) According to the ratio of Chlorella powder to ethanol 1g:10mL, add Chlorella powder from step (2) to 70% ethanol, stir evenly, heat at 60℃ for 2h, collect the extract, filter the extract, obtain the supernatant, place the supernatant in a rotary evaporator, concentrate under reduced pressure at 60℃ and freeze dry to obtain Chlorella extract.

[0030] The preparation method of the above-mentioned composition containing anti-wrinkle complex peptides is as follows: According to the above-mentioned mass percentages, acetyl hexapeptide-8, arginine / lysine peptide, dipeptide diaminobutyryl benzylamide diacetate are stirred and mixed evenly. Then, chlorella extract, ginsenosides, caprylyl glycol, 1,2-hexanediol, PEG-40 hydrogenated castor oil, PPG-26-butanol polyether-26, butylene glycol, and water are added and stirred until evenly mixed to obtain the final product.

[0031] Example 3 A composition containing an anti-wrinkle complex peptide comprises the following raw materials in the indicated weight percentages: acetyl hexapeptide-8 0.09%, Chlorella extract 1.2%, arginine / lysine polypeptide 0.003%, dipeptide diaminobutyryl benzylamide diacetate 0.09%, ginsenoside 0.20%, caprylyl glycol 0.70%, 1,2-hexanediol 0.5%, PEG-40 hydrogenated castor oil 1.02%, PPG-26-butanol polyether-26 1.53%, butylene glycol 24%, and the balance being water.

[0032] The specific preparation process of Chlorella extract is as follows: (1) Under aseptic conditions, at 1×10 5 Chlorella was inoculated into a modified medium at an inoculum size of 1 / mL and cultured for 5 days at 25℃ and 5000 lx light intensity. Glucose was then added, and the culture was continued for another 4 days at 25℃ and 5000 lx light intensity. The culture medium was collected after the culture was complete. The modified medium used in the culture process consisted of: 0.2 g / L calcium chloride, 15 mg / L ferrous sulfate, 0.2 g / L magnesium sulfate, 1 g / L sodium chloride, 0.3 g / L dipotassium hydrogen phosphate, 0.2 mg / L zinc sulfate, 0.1 mg / L eugenol, and 0.1 mg / L dipotassium glycyrrhizate. The concentration of added glucose in the modified medium was 5 g / L.

[0033] (2) The culture medium collected in step (1) is filtered through a hollow fiber microfiltration membrane with a pore size of 0.3 μm, the retained material is collected, and dried to obtain Chlorella powder.

[0034] (3) According to the ratio of Chlorella powder to ethanol 1g:10mL, add Chlorella powder from step (2) to 70% ethanol, stir evenly, heat at 50℃ for 3h, collect the extract, filter the extract, obtain the supernatant, place the supernatant in a rotary evaporator, concentrate under reduced pressure at 60℃ and freeze dry to obtain Chlorella extract.

[0035] The preparation method of the above-mentioned composition containing anti-wrinkle complex peptides is as follows: According to the above-mentioned mass percentages, acetyl hexapeptide-8, arginine / lysine peptide, dipeptide diaminobutyryl benzylamide diacetate are stirred and mixed evenly. Then, chlorella extract, ginsenosides, caprylyl glycol, 1,2-hexanediol, PEG-40 hydrogenated castor oil, PPG-26-butanol polyether-26, butylene glycol, and water are added and stirred until evenly mixed to obtain the final product.

[0036] 2. Comparative Example Comparative Example 1 The difference between Comparative Example 1 and Example 1 is that: The composition of the modified culture medium is different. The modified culture medium of Comparative Example 1 consists of: calcium chloride 0.4 g / L, ferrous sulfate 17 mg / L, magnesium sulfate 0.25 g / L, sodium chloride 1.5 g / L, dipotassium hydrogen phosphate 0.4 g / L, zinc sulfate 0.3 mg / L, dipotassium glycyrrhizate 0.3 mg / L, and the rest is the same as in Example 1.

[0037] Comparative Example 2 The difference between Comparative Example 2 and Example 1 is that: The composition of the modified culture medium is different. The modified culture medium of Comparative Example 2 consists of: calcium chloride 0.4 g / L, ferrous sulfate 17 mg / L, magnesium sulfate 0.25 g / L, sodium chloride 1.5 g / L, dipotassium hydrogen phosphate 0.4 g / L, zinc sulfate 0.3 mg / L, dipotassium glycyrrhizate 0.5 mg / L, and the rest is the same as in Example 1.

[0038] Comparative Example 3 The difference between Comparative Example 3 and Example 1 is that: The composition of the modified culture medium is different. The modified culture medium of Comparative Example 3 consists of: calcium chloride 0.4 g / L, ferrous sulfate 17 mg / L, magnesium sulfate 0.25 g / L, sodium chloride 1.5 g / L, dipotassium hydrogen phosphate 0.4 g / L, zinc sulfate 0.3 mg / L, eugenol 0.2 mg / L, and the rest is the same as in Example 1.

[0039] Comparative Example 4 The difference between Comparative Example 4 and Example 1 is that: The composition of the modified culture medium is different. The modified culture medium of Comparative Example 4 consists of: calcium chloride 0.4 g / L, ferrous sulfate 17 mg / L, magnesium sulfate 0.25 g / L, sodium chloride 1.5 g / L, dipotassium hydrogen phosphate 0.4 g / L, zinc sulfate 0.3 mg / L, eugenol 0.5 mg / L, and the rest is the same as in Example 1.

[0040] Comparative Example 5 The difference between Comparative Example 5 and Example 1 is that: The modified culture media had different compositions. The modified culture medium for Comparative Example 5 consisted of: calcium chloride 0.4 g / L, ferrous sulfate 17 mg / L, magnesium sulfate 0.25 g / L, sodium chloride 1.5 g / L, dipotassium hydrogen phosphate 0.4 g / L, and zinc sulfate 0.3 mg / L.

[0041] 3. Test Case The protein and polysaccharide contents in the Chlorella extracts obtained in Examples 1-3 and Comparative Examples 1-5 were detected, and the specific procedures are as follows: Take 0.002g of the Chlorella extract obtained in Examples 1-3 and Comparative Examples 1-5 and prepare it into a 200μg / mL test sample solution.

[0042] Protein content determination: Follow the instructions of the protein quantification kit. Add 10 μL of the sample to be tested and 250 μL of working solution to a 96-well plate and mix well. Replace the sample with 10 μL of 524 μg / mL protein standard solution in the standard wells and 10 μL of water in the blank wells. Incubate at 37℃ for 30 min. Use an ELISA reader to detect the absorbance of each group of samples at 562 nm wavelength. Detect each group three times and take the average value. The total protein concentration (μg / mL) = [(A sample - A blank) / (A standard - A blank)] × standard concentration × sample dilution factor. The protein content in Chlorella extract is shown in Table 1. Determination of polysaccharide content: Take 0.5 mL of the sample solution to be tested, and immediately add 0.5 mL of 5% phenol solution and 2.5 mL of concentrated sulfuric acid to each sample solution. Mix well, incubate in a constant temperature water bath at 100℃ for 10 min, cool to room temperature, and then measure the absorbance of the sample at a wavelength of 486 nm. Each group is tested three times, and the average value is taken. The results of polysaccharide content in Chlorella extract are shown in Table 1.

[0043] As can be seen from Table 1, the Chlorella extracts obtained in Examples 1-3 have high protein and polysaccharide content, with a protein content of 32.59% and a polysaccharide content of 5.73%.

[0044] Compared to Examples 1-3, the protein and polysaccharide contents of the Chlorella extracts obtained in Comparative Examples 1-5 were reduced to varying degrees. Comparative Example 5 had the lowest protein and polysaccharide contents, with a protein content of only 17.34% and a polysaccharide content of only 0.95%. In the preparation of the Chlorella extracts, eugenol was not added to the modified culture medium in Comparative Example 1; eugenol was not added to the modified culture medium in Comparative Example 2, but the amount of dipotassium glycyrrhizate was increased; dipotassium glycyrrhizate was not added to the modified culture medium in Comparative Example 3; dipotassium glycyrrhizate was not added to the modified culture medium in Comparative Example 4, but the amount of eugenol was increased; and neither dipotassium glycyrrhizate nor eugenol was added to the modified culture medium in Comparative Example 5. The adjustments to the components of the modified culture medium all resulted in a decrease in the protein and polysaccharide contents of the Chlorella extracts, indicating that the synergistic effect of dipotassium glycyrrhizate and eugenol can increase the protein and polysaccharide contents of the Chlorella extracts.

[0045] The following describes the application effects of the compositions containing anti-wrinkle complex peptides in Examples 1-3 and Comparative Examples 1-5 of the present invention.

[0046] A skincare product comprising the following ingredients in percentages by weight: The composition contains 8% of the above-mentioned anti-wrinkle complex peptides, 1.5% carboxymethyl cellulose, 0.7% cetyl alcohol, 0.03% p-hydroxyacetophenone, and the balance is water.

[0047] 4. Application Examples Application Example 1 The safety of skincare products containing the anti-wrinkle complex peptides of Examples 1-3 and Comparative Examples 1-5 was tested, and the specific process is as follows: Volunteers: aged 18-45, 160 in total, randomly divided into 8 groups, with a male-to-female ratio of 1:1 in each group; Testing Procedure: 0.02g of skincare products containing anti-wrinkle complex peptides from Examples 1-3 and Comparative Examples 1-5 were added and placed in a patch applicator. The patch applicator was placed on the flexor side of the subject's forearm, and the palm was used to gently press it to ensure it was evenly adhered to the skin surface for 24 hours. After removing the patch applicator, the skin reaction was observed 30 minutes after the indentation disappeared. If the result was negative, the skin reaction was observed again at 24 hours and 48 hours after the patch test. The results of the patch test were judged according to the skin reaction grading criteria in Table 2. The results are shown in Table 3.

[0048] As shown in Table 3, the skin care products provided by this invention are gentle and non-irritating to the skin and safe to use.

[0049] Application Example 2 The anti-wrinkle effect of skincare products containing anti-wrinkle complex peptides from Examples 1-3 and Comparative Examples 1-5 was tested according to T / ZHCA 006-2019 "Test Method for Anti-wrinkle Efficacy of Cosmetics". The procedure is as follows: Volunteers: A total of 160 healthy female subjects aged 18-45 years were randomly divided into 8 groups.

[0050] Test environment: temperature 22℃, humidity 50%, avoid direct sunlight.

[0051] Experimental Procedure: All volunteers entered the experimental environment 30 minutes before the start of the experiment. Before the experiment began, the number, area, and depth of wrinkles on the face of each volunteer were measured using the Antera 3D Skin Analyzer. After the experiment began, volunteers applied skincare products containing the anti-wrinkle complex peptides from Examples 1-3 and Comparative Examples 1-5 to their faces twice daily, morning and evening. After using the skincare products for 28 days, the number, area, and depth of wrinkles in each volunteer group were measured again using the Antera 3D Skin Analyzer. The reduction in the number, area, and depth of wrinkles in each group was recorded, and the data for each group was the average of the test results for that group. The results are shown in Table 4.

[0052] As shown in Table 4, the composition of the present invention has excellent anti-wrinkle effects. After using skin care products containing the composition of the present invention for 28 days, the number, area, and depth of wrinkles on the volunteers' skin surface were significantly improved, with the number of wrinkles decreasing by more than 32.52%, the area of ​​wrinkles decreasing by more than 42.17%, and the depth of wrinkles decreasing by more than 33.48%.

[0053] Compared to Examples 1-3, the skincare products containing the anti-wrinkle complex peptides in Comparative Examples 1-5 showed poorer anti-wrinkle effects. The only difference between Comparative Examples 1-5 and the examples was the modification of the culture medium composition during the preparation of the Chlorella extract, which altered the polysaccharide and protein content in the Chlorella extract. Therefore, analysis based on the results of polysaccharide and protein content in the Chlorella extract shows that an increase in polysaccharide and protein content significantly promotes the anti-wrinkle effect of the skincare products.

[0054] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them. The basic principles and main features of the present invention have been described above with specific implementation schemes. Based on the present invention, some modifications or substitutions can be made, but these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of protection claimed by the present invention.

Claims

1. A composition containing an anti-wrinkle complex peptide, characterized in that, It is composed of the following raw materials in the indicated weight percentages: Acetyl hexapeptide-8 0.09-0.11%, Chlorella extract 1.2-1.8%, Arginine / Lysine polypeptide 0.003-0.005%, Dipeptide diaminobutyryl benzylamide diacetate 0.09-0.11%, Ginsenoside 0.20-0.25%, Caprylyl glycol 0.70-0.75%, 1,2-Hexanediol 0.5-1%, PEG-40 hydrogenated castor oil 1.02-1.14%, PPG-26-butanol polyether-26 1.53-1.71%, Butylene glycol 24-26%, and the balance being water; The preparation process of the Chlorella extract is as follows: (1) Under sterile conditions, Chlorella was inoculated into a modified culture medium and cultured. After a period of time, glucose was added and cultured for a longer period of time. After the culture was completed, the culture medium was collected. The modified culture medium consisted of: calcium chloride 0.2-0.5 g / L, ferrous sulfate 15-20 mg / L, magnesium sulfate 0.2-0.3 g / L, sodium chloride 1-2 g / L, dipotassium hydrogen phosphate 0.3-0.5 g / L, zinc sulfate 0.2-0.5 mg / L, eugenol 0.1-0.3 mg / L, and dipotassium glycyrrhizate 0.1-0.5 mg / L. (2) The culture medium collected in step (1) is filtered and dried to obtain Chlorella powder; (3) Add the Chlorella powder from step (2) to ethanol, heat and extract, collect the extract, filter, concentrate and freeze dry the extract to obtain Chlorella extract.

2. The composition containing anti-wrinkle complex peptides as described in claim 1, characterized in that, The concentration of glucose in the modified culture medium in step (1) is 5-10 g / L.

3. The composition containing anti-wrinkle complex peptides as described in claim 1, characterized in that, The inoculum size of Chlorella in the modified culture medium in step (1) is 1-8 × 10⁻⁸. 5 The number of cells / mL is 5-6 days, and the culture time is 4-5 days.

4. The composition containing anti-wrinkle complex peptides as described in claim 1, characterized in that, The conditions for cultivation and continued cultivation in step (1) are: temperature of 20-25℃ and light intensity of 4000-5000lx.

5. The composition containing anti-wrinkle complex peptides as described in claim 1, characterized in that, The filtration in step (2) is performed using a hollow fiber microfiltration membrane; the pore size of the microfiltration membrane is 0.1-0.3 μm.

6. The composition containing anti-wrinkle complex peptides as described in claim 1, characterized in that, The heating extraction in step (3) is performed at a temperature of 50-60℃ for 2-3 hours.

7. The method for preparing the composition containing anti-wrinkle complex peptides as described in claim 1, characterized in that, Includes the following steps: According to the stated mass percentage, acetyl hexapeptide-8, arginine / lysine polypeptide, dipeptide diaminobutyryl benzylamide diacetate are stirred and mixed evenly. Then, chlorella extract, ginsenosides, caprylyl glycol, 1,2-hexanediol, PEG-40 hydrogenated castor oil, PPG-26-butanol polyether-26, butylene glycol, and water are added and stirred until evenly mixed to obtain the final product.

8. The application of the composition containing anti-wrinkle complex peptides as described in claim 1, characterized in that, It is used in the preparation of skin care products with anti-wrinkle effects.

9. The application of the composition containing anti-wrinkle complex peptides as described in claim 8, characterized in that, The composition containing anti-wrinkle complex peptides comprises 1-10% by weight of the total weight of the skin care product; the skin care product also comprises the following raw materials by weight percentage: 1-2% carboxymethyl cellulose, 0.5-1% cetyl alcohol, 0.01-0.05% p-hydroxyacetophenone, and the balance being water.

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