阅读说明：本技术 一种促进皮肤多维防御和修复的组合物及其制备方法 (Composition for promoting skin multi-dimensional defense and repair and preparation method thereof ) 是由 杜丽丽 于 2021-10-13 设计创作，主要内容包括：本发明涉及护肤品技术领域,尤其涉及一种促进皮肤多维防御和修复的组合物及其制备方法。组合物的有效组分包括：肌肽0.01～20份,丝氨酸0.01～10份,谷氨酸0.01～10份,乙酰羟脯氨酸0.01～10份,棕榈酰三肽-5 0.001～1份,水解胶原0.01～15份,维生素B12 0.01～10份,二肽二氨基丁酰苄基酰胺二乙酸盐0.001～1份,精氨酸/赖氨酸多肽0.001～1份,植醇0.01～15份,异麦芽0.01～15份。本发明的组合物经过冷冻干燥处理后,可以保证有效组分的活性,大大延长保存时间。本发明的组合物对于皮肤的修复和抗老化功效明显,具有抗氧化、祛黄、亮肤、紧致、抗皱、保湿、促进细胞再生、提高皮肤弹性、降低皮肤粗糙度、修复皮肤屏障、均匀肤色等多种功效。(The invention relates to the technical field of skin care products, in particular to a composition for promoting skin multi-dimensional defense and repair and a preparation method thereof. The effective components of the composition comprise: 0.01-20 parts of carnosine, 0.01-10 parts of serine, 0.01-10 parts of glutamic acid, 0.01-10 parts of acetylhydroxyproline, 0. 50.001-1 parts of palmitoyl tripeptide, 0.01-15 parts of hydrolyzed collagen, 120.01-10 parts of vitamin B, 0.001-1 part of dipeptide diaminobutyrylbenzyl amide diacetate, 0.001-1 part of arginine/lysine polypeptide, 0.01-15 parts of phytol and 0.01-15 parts of isomalt. After the composition is subjected to freeze drying treatment, the activity of effective components can be ensured, and the storage time is greatly prolonged. The composition disclosed by the invention has obvious effects of repairing skin and resisting aging, and has multiple effects of resisting oxidation, removing yellow, brightening skin, tightening skin, resisting wrinkles, moisturizing, promoting cell regeneration, improving skin elasticity, reducing skin roughness, repairing skin barriers, homogenizing skin color and the like.)

1. The composition for promoting skin multi-dimensional defense and repair is characterized by comprising the following effective components in parts by weight: 0.01-20 parts of carnosine, 0.01-10 parts of serine, 0.01-10 parts of glutamic acid, 0.01-10 parts of acetylhydroxyproline, 0. 50.001-1 parts of palmitoyl tripeptide, 0.01-15 parts of hydrolyzed collagen, 120.01-10 parts of vitamin B, 0.001-1 part of dipeptide diaminobutyrylbenzyl amide diacetate, 0.001-1 part of arginine/lysine polypeptide, 0.01-15 parts of phytol and 0.01-15 parts of isomalt.

2. A composition according to claim 1, wherein: the composition comprises the following effective components in parts by weight: 3-14 parts of carnosine, 1-7 parts of serine, 1-7 parts of glutamic acid, 1-7 parts of acetylhydroxyproline, 0.8-0.8 part of palmitoyl tripeptide-50.05, 0.5-10 parts of hydrolyzed collagen, 121-7 parts of vitamin B, 0.05-0.8 part of dipeptide diaminobutyrylbenzyl amide diacetate, 0.05-0.8 part of arginine/lysine polypeptide, 1.5-10 parts of phytol and 1.5-10 parts of isomalt.

3. A composition according to claim 1, wherein: the composition comprises the following effective components in parts by weight: 5.5-9 parts of carnosine, 3-5 parts of serine, 3-5 parts of glutamic acid, 3-5 parts of acetylhydroxyproline, 50.2-0.6 part of palmitoyl tripeptide, 3-6 parts of hydrolyzed collagen, 123-5 parts of vitamin B, 0.2-0.6 part of dipeptide diaminobutyrylbenzyl amide diacetate, 0.2-0.6 part of arginine/lysine polypeptide, 4-7 parts of phytol and 4-7 parts of isomalt.

4. A composition according to claim 1, wherein: the palmitoyl tripeptide-5 is 10000U, the dipeptide diaminobutyrylbenzylamide diacetate is 60000U, and the arginine/lysine polypeptide is 80000U.

5. A method of preparing a composition according to claim 1, wherein: comprises the following steps of (a) carrying out,

the method comprises the following steps: weighing the effective components in proportion, placing the effective components in a beaker, and uniformly stirring to obtain a mixture A;

step two: placing the mixture A in a vacuum freeze dryer, and carrying out freeze drying under the conditions that the freezing temperature is-50 to-80 ℃ and the pressure is-100 to-70 Pa, wherein the drying time is 5 to 20 hours;

step three: and freeze-drying to obtain a powdery finished product.

6. A process for preparing a composition according to claim 5, wherein: the power of the vacuum freeze dryer is more than 300 kw, and the rotating speed is more than 200 r/min.

7. A process for preparing a composition according to claim 5, wherein: the drying time is 10-16 hours.

8. A process for preparing a composition according to claim 5, wherein: the water content of the prepared powdery finished product is less than 5%.

[ technical field ] A method for producing a semiconductor device

The invention relates to the technical field of skin care products, in particular to a composition for promoting skin multi-dimensional defense and repair and a preparation method thereof.

[ background of the invention ]

The skin aging is caused by many factors, mainly including physiological and natural aging and aging caused by external factors such as photoaging, glycation, oxidation, inflammation, air pollution, malnutrition, specific diseases, stress, and the like. Most cells in the body are constantly renewed, and damage is repaired through regeneration, which is a process that cells are constantly self-repaired. The essence of skin aging is that the rate of cellular damage exceeds the rate of cellular repair. With the development of biotechnology, the search for high-activity, safe and non-irritant cosmetic raw materials becomes a target of the consistent pursuit of the daily chemical industry.

When cells are damaged, the active peptide firstly stimulates the rapid repair of cell membranes and plays an important role in the cell cycle and the apoptosis process. When repairing skin cells, the skin repairing agent can not only quickly promote the growth of epidermal cells, but also promote the growth of collagen and fibrous tissues of subcutaneous connective tissues, thereby enabling the skin to be quickly healed without scars. The cell active peptide is used as an additive applied in cosmetics, and the molecular weight of each functional segment is within 3000Da, so that the polypeptide combination can be directly absorbed by skin, and the cell repair result is achieved. The research shows that the active peptide is produced in epidermal cells after being damaged by ultraviolet irradiation, which indicates that the epidermal cells carry out self-repair and a large amount of active peptide is involved. The skin care products sold in the market at present basically take the main role of maintaining the epidermis, and cannot really play the role of repairing the skin.

[ summary of the invention ]

Aiming at the defects, the invention provides a composition for promoting skin multi-dimensional defense and repair and a preparation method thereof. The skin repairing composition is prepared by mixing various amino acids, bioactive peptides, vitamin B12, moss I and other effective components, and freeze-drying. The composition is convenient to use, safe, free of irritation, good in absorption and quick in effect, and can achieve the effects of moisturizing, repairing, resisting aging and the like.

In order to achieve the purpose, the invention adopts the following technical scheme:

the composition for promoting skin multi-dimensional defense and repair is characterized by comprising the following effective components in parts by weight: 0.01-20 parts of carnosine, 0.01-10 parts of serine, 0.01-10 parts of glutamic acid, 0.01-10 parts of acetylhydroxyproline, 0. 50.001-1 parts of palmitoyl tripeptide, 0.01-15 parts of hydrolyzed collagen, 120.01-10 parts of vitamin B, 0.001-1 part of dipeptide diaminobutyrylbenzyl amide diacetate, 0.001-1 part of arginine/lysine polypeptide, 0.01-15 parts of phytol and 0.01-15 parts of isomalt.

1. As a further improvement of the invention, the composition comprises the following effective components in parts by weight: 3-14 parts of carnosine, 1-7 parts of serine, 1-7 parts of glutamic acid, 1-7 parts of acetylhydroxyproline, 0.8-0.8 part of palmitoyl tripeptide-50.05, 0.5-10 parts of hydrolyzed collagen, 121-7 parts of vitamin B, 0.05-0.8 part of dipeptide diaminobutyrylbenzyl amide diacetate, 0.05-0.8 part of arginine/lysine polypeptide, 1.5-10 parts of phytol and 1.5-10 parts of isomalt.

2. As a further improvement of the invention, the composition comprises the following effective components in parts by weight: 5.5-9 parts of carnosine, 3-5 parts of serine, 3-5 parts of glutamic acid, 3-5 parts of acetylhydroxyproline, 50.2-0.6 part of palmitoyl tripeptide, 3-6 parts of hydrolyzed collagen, 123-5 parts of vitamin B, 0.2-0.6 part of dipeptide diaminobutyrylbenzyl amide diacetate, 0.2-0.6 part of arginine/lysine polypeptide, 4-7 parts of phytol and 4-7 parts of isomalt.

As a further improvement of the invention, the palmitoyl tripeptide-5 is 10000U, the dipeptide diaminobutyrylbenzylamide diacetate is 60000U, and the arginine/lysine polypeptide is 80000U.

A method for preparing a composition for promoting multi-dimensional defense and repair of skin, which is characterized by comprising the following steps: comprises the following steps of (a) carrying out,

the method comprises the following steps: weighing the effective components in proportion, placing the effective components in a beaker, and uniformly stirring to obtain a mixture A;

step two: placing the mixture A in a vacuum freeze dryer, and carrying out freeze drying under the conditions that the freezing temperature is-50 to-80 ℃ and the pressure is-100 to-70 Pa, wherein the drying time is 5 to 20 hours;

step three: and freeze-drying to obtain a powdery finished product.

As a further improvement of the invention, the power of the vacuum freeze dryer is more than 300 kw, and the rotating speed is more than 200 r/min.

As a further improvement of the invention, the drying time is 10-16 hours.

As a further development of the invention, the finished powdered product obtained has a moisture content of < 5%.

In the composition for promoting the skin multi-dimensional defense and repair, the functions of the effective components are introduced as follows:

carnosine is a crystalline solid composed of two amino acids, beta-alanine and L-histidine. The research shows that: carnosine has a strong antioxidant capacity and is capable of stably protecting cell membranes, particularly as a water-soluble radical scavenger which prevents cell membrane peroxidation. Carnosine has been shown to bring the DNA damage signals generated by multiple replications of cells into proximity with fibroblasts, protecting the cells from oxidation by acetaldehyde, beta-amyloid peptide chains, while inhibiting sugar aging and protein cross-linking.

Serine is a neutral aliphatic hydroxyl-containing amino acid, and is a non-essential amino acid. Serine can be used as skin nutrition additive, and is one of amino acids with strong moisture retention capacity because of the existence of hydroxyl in serine. Is commonly used as skin moistening agent, can be made into liposome with phospholipid component, and is compatible with other active components to achieve better moisturizing effect.

Glutamic acid is an acidic amino acid, which is present in large amounts in cereal proteins and also in large amounts in animal brains. Glutamate plays an important role in protein metabolism in organisms, and is involved in many important chemical reactions in animals, plants, and microorganisms. Can be used for treating wrinkle by applying to skin.

Acetylhydroxyproline is one of imino acids, one of the main components of collagen tissue, and is a specific amino acid in collagen, and accounts for about 13% of the total amount of collagen amino acids. Collagen is the most abundant protein in the body, and accounts for about 1/3 total body protein. The acetylhydroxyproline is an amino acid which is specific to skin collagen, the content of the collagen is generally represented by 7.46 times of the acetylhydroxyproline, and the reduction of the content of the acetylhydroxyproline in the cortex can be used as an index of the skin aging degree. The skin is exposed to ultraviolet light or intense light, and the amount of L-acetylhydroxyproline in proteins in the stratum corneum changes the most, and the reduction is proportional to the degree of wrinkles and aging produced by the skin. The acetylhydroxyproline as exogenous nutrient additive has skin moistening, antiaging, and sunscreen effects. The acetylhydroxyproline has effect in inhibiting melanin production, and can be used as whitening auxiliary agent.

Palmitoyl tripeptide-5 is the earliest and most widely used cosmetic small molecule peptide, and belongs to the signal class peptide. The research shows that: such peptides promote collagen synthesis and extracellular matrix protein synthesis by increasing stromal cell activity. The palmitoyl tripeptide-5 is used as a skin conditioner and has the effects of supplementing collagen for skin and reducing skin wrinkles. Meanwhile, the skin care product has the characteristics of enhancing cell activity, increasing the water content of the skin and enhancing the skin luster.

The hydrolyzed collagen is a high-efficiency moisture-keeping component, has the functions of activating skin and inducing tissue regeneration, and is a main adhesive substance in the hoof tissue of a human body. The skin moisturizing cream has the advantages of large molecular weight and better moisturizing capability than polyhydric alcohols, is mainly used for supporting cells, and is particularly suitable for performing basic moisturizing on skin after the skin is exfoliated and when the skin is in a fragile stage. The collagen has good permeability and moisture-keeping ability, and can be used for removing wrinkle and conditioning rough skin. The collagen and the hydrolysate thereof have strong compatibility, can be combined with a plurality of functional components to improve the efficacy of the collagen and the hydrolysate thereof, and can be used in hair care products to effectively care hair.

Vitamin B12 is a polycyclic compound containing cobalt in 3 valency. Vitamin B12 has excellent effect of skin regeneration, and is an important component for promoting metabolism of human body, and cell regeneration and hematopoiesis are less. The collagen has a promoting effect on collagen, and has a wrinkle removing effect when applied to the skin.

The phytol and isomalt can be replaced by moss I of Switzerland Baile, and the moss I contains phytol, isomalt and water as main ingredients. Moss No. one can be used as a cell channel key: by promoting the information communication among skin cells, keratin generation in the keratinocytes is promoted, the water retention capacity of the cells is improved, and meanwhile, by increasing the cohesion among the keratinocytes, the diffusion of water and other components among the keratinocytes is controlled, the skin thickness is increased, the skin barrier function is improved, and the water loss is reduced. The specific effects are as follows: 1. maintaining the vitality of cell nucleus, keeping the cell healthy and resisting aging; 2. strengthening the skin barrier; 3. preventing the stress reaction of the skin under adverse environment, and improving the adaptability of the skin to different environments; 4. improving wrinkle depth; 5. the skin color is uniform.

Dipeptide diaminobutyrylbenzylamide diacetate is a tripeptide simulating the activity of snake venom toxin, can organize the nerve conduction of acetylcholine, achieves the effect of relaxing muscles, and has the characteristics of quick response, lasting effect and complete reversibility. Can reduce the generation of wrinkles by inhibiting the contraction of muscles, and has the effects of smoothing the skin and rapidly removing wrinkles.

Arginine/lysine polypeptide is a highly folded peptide with strong penetration and high stability. This component helps stimulate the synthesis and accumulation of structural components of the extracellular matrix, and also helps to inhibit disordered collagen cross-linking and the release of metalloproteinase 1 (MMP-1); promoting fibroblast regeneration and stimulating the synthesis of collagen. When applied to skin, the product has good compatibility with skin, is not easy to be allergic, has effect in recovering skin elasticity, and has antiaging and wrinkle resisting effects. Meanwhile, the eye cream has low irritation to skin and eyes.

According to the characteristics of the raw material components, the components are divided into signal peptides, cell channel switches and repair combination peptides according to the functions of the components in the skin repair process.

Wherein the content of the first and second substances,

signal peptide: dipeptide diaminobutyrylbenzylamide diacetate, arginine/lysine polypeptide

Cell channel switching: first moss silk

Repairing the combined peptide: carnosine, serine, glutamic acid, acetyl hydroxyproline, palmitoyl tripeptide-5, hydrolyzed collagen, and vitamin B12

The process of these components in repairing cells in senescent cells is as follows:

1. the signal peptide firstly identifies the aging cells and sends out signals;

2. the cell channel switch opens the cell channel, so that the later repairing component can enter the aging cell through the cell channel as soon as possible;

3. after the active ingredients of the repair combination peptide enter the senescent cells, the repair procedure is initiated, which is initiated in a very short time.

Eventually, as the senescent cells are repaired, the skin tissue is gradually repaired.

Compared with the prior art, the invention has the following beneficial effects:

1. the composition is prepared by compounding and combining more than 10 effective components such as various amino acids, peptides with signal transmission effect, carnosine with antioxidation effect, hydrolyzed collagen with the effects of activating skin and inducing tissue regeneration, vitamin B12 with the skin regeneration effect and the like according to a certain proportion. The composition has obvious effects of repairing skin and resisting aging, and has multiple effects of resisting oxidation, removing yellow, brightening skin, tightening skin, resisting wrinkle, keeping moisture, promoting cell regeneration, improving skin elasticity, reducing skin roughness, repairing skin barrier, and homogenizing skin color.

2. The composition of the invention can ensure the activity of the effective components after freeze drying treatment, and greatly prolong the storage time.

3. The composition of the invention is added into skin care products, and has the advantages of quick effect, safety and no irritation.

[ detailed description ] embodiments

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments. It is to be understood that the described embodiments are merely exemplary of the invention, and not restrictive of the full scope of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

In the following examples, the suppliers of the raw materials used are as follows.

Name of article	Specification of	Suppliers of goods
			Carnosine		Purchased by Hubei Liansheng bioengineering GmbH
Serine		Shandong Siyang Biotech Co., Ltd
			Glutamic acid		Shandong Siyang Biotech Co., Ltd
Acetyl hydroxyproline		Shanghai blue beauty cosmetics Co Ltd
			Palmitoyl tripeptide-5	10000U	Hangzhou Weitai bio Pharmaceutical Co.,Ltd.
Hydrolyzed collagen		Walt Laisi Biotech, Lanzhou
			Vitamin B12		Imperial Dusmann of the Netherlands
Dipeptide Diaminobutyrylbenzylamide diacetate	60000U	Chengdu Yunshi chemical Co., Ltd
			Arginine/lysine polypeptides	80000U	SHANGHAI T&W MEDICAL SCIENCE AND TECHNOLOGY Co.,Ltd.
First moss silk		Switzerland Baile Co

Example 1

A composition for promoting skin multi-dimensional defense and repair comprises the following effective components in parts by weight: 9 parts of carnosine, 3.5 parts of serine, 3.5 parts of glutamic acid, 3.5 parts of acetylhydroxyproline, 50.6 parts of palmitoyl tripeptide, 4 parts of hydrolyzed collagen, 124 parts of vitamin B, 0.6 part of dipeptide diaminobutyrylbenzyl amide diacetate, 0.3 part of arginine/lysine polypeptide and 4 parts of lichen I.

The preparation method of the composition specifically comprises the following steps:

the method comprises the following steps: weighing the effective components in proportion, placing the effective components in a beaker, and uniformly stirring to obtain a mixture A;

step two: placing the mixture A in a vacuum freeze dryer, wherein the power of the vacuum freezer is 400kw, the rotating speed is 300 r/min, and freeze drying is carried out under the conditions that the freezing temperature is-60 ℃ to-70 ℃ and the pressure is-80 ℃ to-70 Pa, and the drying time is 12 to 16 hours;

step three: freeze drying to obtain powder product; the water content of the finished product is 4.6 percent through detection.

Example 2

A composition for promoting skin multi-dimensional defense and repair comprises the following effective components in parts by weight: 5.5 parts of carnosine, 5 parts of serine, 5 parts of glutamic acid, 5 parts of acetylhydroxyproline, 50.2 parts of palmitoyl tripeptide, 6 parts of hydrolyzed collagen, 125 parts of vitamin B, 0.2 part of dipeptide diaminobutyrylbenzylamide diacetate, 0.6 part of arginine/lysine polypeptide and 6 parts of lichen silk I.

The preparation method of the composition specifically comprises the following steps:

the method comprises the following steps: weighing the effective components in proportion, placing the effective components in a beaker, and uniformly stirring to obtain a mixture A;

step two: placing the mixture A in a vacuum freeze dryer, wherein the power of the vacuum freezer is 400kw, the rotating speed is 300 r/min, and freeze drying is carried out under the conditions that the freezing temperature is-70 ℃ to-80 ℃ and the pressure is-100 Pa to-90 Pa, and the drying time is 10 to 14 hours;

step three: freeze drying to obtain powder product; the water content of the finished product is 4.3 percent through detection.

Examples 3 to 8

The preparation method of the composition for promoting skin multi-dimensional defense and repair of examples 3 to 8 is the same as that of example 2, and the weight ratio of each effective component is shown in table 1.

TABLE 1 weight ratio table of each effective component in examples 3 to 8

Example 9 human safety test

The finished products of the embodiments 1 to 8 are mixed with other auxiliary materials to prepare face cream, the face cream is subjected to safety test, and a skin sealing patch test is adopted according to a seventh human body safety inspection method of technical standards for cosmetic safety (2015 edition), and the specific test steps are as follows: selecting 50 test subjects meeting the test requirements, selecting a spot tester meeting the test requirements, taking 20mg of face cream containing the composition of the embodiment 1-8 as a test object, and adding the face cream into the spot tester; the patch test device with the test substance is applied to the flexed side of the forearm of the subject with a low-sensitivity adhesive tape, and then the patch is applied to the skin uniformly by pressing with the palm for 24 hours, and then the test substance is wiped off. Skin reactions were observed at 0.5, 24 and 48 hours after removal of the plaque tester, respectively, and the observations were recorded. From the test results, it can be seen that: after the skin reactions are observed for 0.5 hour, 24 hours and 48 hours respectively, no skin adverse reactions such as erythema, infiltration, edema, pimple, herpes and the like of the subject are found, and the composition has the characteristics of safety and no stimulation after being applied to the skin of a human body.

Example 10 efficacy evaluation test

a skin condition evaluation test

180 subjects with wrinkles or spots on their faces were selected, and 20 subjects were grouped together. The face cream containing the compositions of examples 1 to 8 was used as a test substance, and the test substance was applied to the cleaned face of the subject once a day in the morning and at night, with 30 days as a cycle.

Table 2 test results for wrinkles

The test results show that: the cream containing the composition of the invention has an average effective rate of 97.5% for wrinkles after 2 cycles of use, and 70% of the effective rate has obvious improvement. The cream of the comparative example had an average effective rate for wrinkles of 35% after 2 cycles of use. By adding the composition of the invention, wrinkles are obviously improved.

Table 3 test results for uniform skin color

Table 4 test results for skin roughness

The test results show that: the average effective rate of the face cream containing the composition of the invention on uniform skin color after 2 cycles of use is 100%, wherein 70% of the effective rate is obviously improved; the average improvement in skin roughness was 95%, with a significant improvement of 70%.

The skin of the subject area was measured colorimetrically using a KONICA MINOLTA model CR-100 colorimeter, the results of which are shown in the following table.

Table 5 skin color test results

From the above test results, it can be seen that: after 2 cycles of the cream containing the composition, the skin chromaticity L value of a tested area is obviously improved, and the b value is obviously reduced. The larger the L value is, the more white the skin color is; the larger the b value, the more yellow the skin color. The cream containing the composition has obvious effects of brightening the skin and removing yellow.

By adding the composition disclosed by the invention, wrinkles, uniform skin color, skin roughness, yellow removal and brightening are obviously improved.

b skin elasticity and collagen fiber level test

45 subjects with healthy skin were selected and divided into 9 groups, and skin care products with the same function were not used for the week before the test. After the face of the subject was cleaned, the skin of the face of the subject was tested for elasticity and collagen fiber levels after waiting for 30 minutes without rubbing any skin care product, and the test data was recorded. The face cream containing the composition of examples 1-8 was applied to the face of the subject once in the morning and once in the evening for 90 days, and the blank control was applied with the same amount of distilled water. The skin elasticity and collagen fiber level of the face of the subject were tested on days 1, 5, 10, 15, 20, and 30, respectively, and the test data were recorded.

The apparatus used in this test was a skin analysis system model CBS-1800U, manufactured by CBS corporation, Taiwan.

Table 6 facial skin elasticity test results

Table 7 facial skin collagen fiber level test results

The test result shows that the skin elasticity increasing rate is more than 50 percent and the collagen fiber water level is obviously improved after the skin care product containing the composition is used for 30 days.

c moisture test

B, testing the skin moisture content of the face of the subject in the skin elasticity and collagen fiber level test respectively before smearing, 30 minutes after smearing and 10 hours after smearing, and recording the test data. The apparatus used in this test was a skin moisture tester model CM825, manufactured by CK, Germany.

Table 8 facial skin moisture test results

The test results show that the skin care product containing the composition can improve the moisture content of the skin and reduce the loss of moisture.

Evaluation of cell regeneration promoting Effect

The effect of the composition of the present invention on the cell regeneration promoting function was evaluated by a cell scratch test method. The specific test method is as follows:

1. firstly, a marker pen is used on the back of a 6-hole plate, transverse lines are uniformly drawn by a ruler, a straight line is drawn every 1cm and transversely penetrates through the through holes, and each hole penetrates through at least 5 lines.

2. About 5X 10 additions to each well⁵Individual cells were allowed to confluent overnight for adherence.

3. The next day, cell scratches were made perpendicular to the well plate and line with a 200 μ L pipette tip.

4. The medium was aspirated and the cells were washed 3 times with sterile phosphate buffer to remove the scratched cells and smooth the scratched boundaries.

5. A composition of the present example was placed in each well as a test group; DMEM medium was placed as a blank control. At this point, the record is 0 hour, and 37 degrees 5% CO is added₂And an incubator for performing cultivation. Samples were taken at 0, 6, 12, 24 hours and photographed. After the Image was photographed by using Image J software, data analysis was performed to evaluate the effect of promoting cell regeneration by the wound area.

TABLE 9 determination of relative wound area

From the above test results, it was found that the composition of the present invention has an effect of promoting cell regeneration compared to the control group under the same experimental conditions.

e evaluation of Effect of Oxidation resistance

The effect evaluation of the antioxidant capacity is carried out by adopting an improved DPPH method, and the specific test method is as follows:

1. preparation of sample solution: weighing the mixture 1g of the invention in the embodiment, mixing the mixture with 20mL of 80% ethanol, shaking and extracting the mixture for 8 hours at 25 ℃, centrifuging the mixture for 15 minutes at 3500r/min, taking supernatant, placing the supernatant in a brown bottle, and storing the supernatant at 4 ℃ for later use.

2. The determination method comprises the following steps: 2mL of 0.2mmol/LDPPH ethanol solution and 2mL of sample solution are added into the reaction tube and mixed uniformly. The reaction was carried out in the dark at room temperature for 30 minutes in the dark with the exclusion of light, and the absorbance value was read at 517nm, which was the sample set. Meanwhile, absorbance was measured after mixing with 2mL of a buffer solution of LDPPH +2mL, which was a control group.

3. Calculation of the rate of removal of DPPH stable free radicals: clearance (%) = [ 1-sample group/control group ] × 100%.

The determination and calculation result shows that the clearance rate of DPPH free radicals of the sample is 58.23%, and the result shows that the antioxidant is good.

Evaluation of skin Barrier repair Effect

The skin barrier repairing effect evaluation is carried out by adopting a skin moisture permeation test, and the specific test method comprises the following steps:

selecting 40 subjects, using a test group consisting of a test area of 9cm × 8cm in front of the left and right upper arms of the subject and a 20% ethanol aqueous solution containing the example of the present invention applied to the test area of the upper arm on one side; the test area on the other side of the upper arm was coated with a 20% ethanol aqueous solution as a blank control. TEWL was measured before and after coating, respectively, once every day, in 30 days as a cycle.

TABLE 10 results of TEWL values

From the above test results, it is clear that TEWL tends to increase significantly after the examples of the present invention are used. The test results show that the composition of the invention has the function of repairing the skin barrier.

In summary, the active ingredients such as polypeptides and amino acids contained in the composition of the present invention improve cell activity, and promote collagen production and cell regeneration, thereby increasing skin elasticity, collagen fiber level, and skin moisture content. The composition has obvious effects of repairing skin and resisting aging, and has multiple effects of resisting oxidation, removing yellow, brightening skin, tightening skin, resisting wrinkle, keeping moisture, promoting cell regeneration, improving skin elasticity, reducing skin roughness, repairing skin barrier, and homogenizing skin color.