TWI778915B - Manufacturing method of cosmetic composition - Google Patents

Abstract

An object of the present invention is to provide a cosmetic composition containing a collagen-rich placenta extract and having excellent functionality and a method for producing the cosmetic composition. The solution is that a lotion system to which the cosmetic composition of the present invention is applied has a weight ratio of pure water: 30-95%, collagen-containing placenta extract: 0.05-0.8% (v /v), polyol: 0.1~50%, ester oil: appropriate amount (0~5%, sometimes not blended), surfactant: appropriate amount, citric acid: appropriate amount, carboxyethylene polymer: appropriate amount (lower limit: 0.1%) ), sodium hydroxide: appropriate amount, plant extracts: appropriate amount (lower limit: 0.1%) preservatives: appropriate amount of composition. Furthermore, the lotion system shown here contains epidermal growth factor (EGF) at a concentration of 0.0000075 to 0.00012% (w/v) based on the total amount.

Description

Manufacturing method of cosmetic composition

The present invention relates to a method for producing a cosmetic composition. More specifically, it relates to a method for producing a cosmetic composition containing collagen-rich placental extract and having excellent functions.

Traditionally, cosmetic materials such as lotions, lotions, and gels are often given a whitening effect for improving spots and freckles caused by skin blackening or pigmentation, and a moisturizing effect or antioxidant effect for preventing spots or sagging. , and a variety of raw materials are mixed into cosmetics.

In addition, the skin is often exposed to the outside world, resulting in aging phenomena such as wrinkles, sagging, dullness, and pigmentation with age. In particular, it is believed that collagen, which accounts for more than 90% of the dermal matrix, has a great influence on morphological changes such as wrinkles and sagging.

The amount of this collagen decreases with age, and the reduction of collagen leads to incomplete formation of the dermal structure, which becomes one of the main reasons for skin aging, wrinkles and sagging.

In addition, as a cause of the decrease in the amount of collagen, fibroblasts in the dermis deteriorate the ability to produce collagen, and as a result, wrinkles and sagging of the skin occur.

Therefore, some people try to find materials for promoting collagen production from natural products. So far, it has been reported that in addition to plant extracts or their components, various substances also have the effect of promoting collagen production.

Furthermore, collagen itself or hydrolyzed collagen is known to have a fibroblast growth effect or a collagen-producing effect.

Moreover, besides the raw material which has the effect|action of promoting collagen production, the placenta extract (placenta extract) is attracting attention as a raw material which imparts a whitening effect and many useful functions to cosmetics.

This placenta extract has the effect of regulating endocrine, strengthening the liver and detoxification, activating the immune system, acting on the autonomic nervous system, promoting blood circulation and hematopoiesis. In addition, in terms of beauty, it also has many functions, such as improving dry skin caused by anti-inflammatory effect, whitening effect caused by antioxidant effect, promoting metabolism, and rejuvenating skin.

Moreover, in the cosmetics, in order to improve the whitening effect etc. which the placenta extract has, various researches are carried out in the industry, for example, the cosmetic composition containing the placenta extract described in Patent Document 1 has been proposed.

In the cosmetic composition described in this patent document 1, the raw material which encapsulated the placenta extract and the coix seed in liposomes is used. [Prior Art Literature] [Patent Literature]

[Patent Document 1] Japanese Patent Laid-Open No. 2010-180193

[The problem to be solved by the invention]

However, among the conventional cosmetics using placenta extract as a raw material, such as the cosmetic composition disclosed in Patent Document 1, the content of collagen contained in the placenta extract itself is small, and the collagen produced by the placenta extract itself is relatively low. Further improvement is expected in the effect of fibroblast growth and collagen production.

The present invention has been developed in view of the above points, and aims to provide a method for producing a cosmetic composition containing a collagen-rich placenta extract and having excellent functionality. [Means of Solving Problems]

In order to achieve the above object, the cosmetic composition of the present invention is constituted to contain placenta extract and epidermal growth factor (EGF), and in the aforementioned placenta extract, the total nitrogen concentration (w/v) per unit of the placenta extract is The hydroxyproline concentration (w/v) is above 0.47.

Here, the hydroxyproline concentration (w/v) per unit of total nitrogen concentration (w/v) in the placental extract permeated through the placenta extract is 0.47 or more, and the placenta extract used as the raw material is rich in collagen. First of all, hydroxyproline is the main component of collagen, and it is a unique amino acid in collagen, so the amount of hydroxyproline is an indicator of collagen content. Therefore, the higher the concentration (w/v) of hydroxyproline per unit of total nitrogen concentration (w/v) in the placenta extract, the higher the collagen concentration in the placenta extract can be judged. In addition, the hydroxyproline concentration (w/v) per unit of total nitrogen concentration (w/v) of the placenta extract is above 0.47, which means that the hydroxyproline concentration in the placenta extract is higher than that in the conventional commercially available placenta extract. The concentration contains sufficient collagen. As a result, the fibroblast growth effect and collagen production effect of collagen in the placenta extract can be enhanced.

In addition, by containing the placenta extract and epidermal growth factor (EGF), the fibroblast growth effect in the cosmetic composition can be increased synergistically compared to the composition of the individual components.

Also, when the concentration of the permeated placenta extract is in the range of 0.025v/v% to 1.0v/v%, when the concentration of epidermal growth factor (EGF) is in the range of 0.00000375w/v% to 0.00015w/v% , can further improve the growth effect of fibroblasts in cosmetic compositions. In addition, coloring of the cosmetic can be suppressed.

On the other hand, when the concentration of the placenta extract was less than 0.025 v/v%, the fibroblast growth effect in the cosmetic composition was improved, but the degree of improvement was small. In addition, when the concentration of the placenta extract exceeds 1.0 v/v%, the color of the placenta extract itself is easy to appear in the cosmetic, and it is not easy to be used as a raw material when a transparent or white cosmetic is required for the color of the cosmetic application. .

On the other hand, when the concentration of epidermal growth factor (EGF) was less than 0.00000375w/v%, the fibroblast growth effect in the cosmetic composition was improved, but the degree of improvement was small. In addition, when the concentration of epidermal growth factor (EGF) exceeds 0.00015w/v%, although the effect of fibroblast growth in the cosmetic composition is improved to some extent, compared with the lower concentration, the degree of improvement has reached the maximum. , the cost of epidermal growth factor (EGF) used increases and is not good.

In addition, when the concentration of the placenta extract is in the range of 0.2 v/v% to 0.8 v/v%, the fibroblast growth effect in the cosmetic composition can be more significantly improved.

Also, when the placenta extract is obtained from horse placenta, it can be used as a placenta extract that can be used as a raw material for cosmetic compositions.

Furthermore, in order to achieve the above-mentioned object, the method for producing a cosmetic composition of the present invention includes a placenta extract production step and a mixing step, wherein the placenta extract production step includes a hot water extraction step for thawing the thawed horses. The placenta or porcine placenta is mixed with the blood exuded during the thawing, and treated with hot water; and the digestion reaction step, which is the extraction process of the above-mentioned hot water extraction step. temperature, using papain for enzymatic digestion; the mixing step is the placental extract obtained in the placental extract manufacturing step, and its hydroxyproline concentration (w/v) per unit of total nitrogen concentration (w/v) Placental extract above 0.47 was mixed with epidermal growth factor (EGF).

Here, in the hot water extraction step, by mixing the thawed horse placenta or porcine placenta with the blood exuded during thawing, and treating with hot water, water-soluble collagen can be efficiently extracted from the equine placenta or the porcine placenta. protein composition. That is, between the enzyme extract and the residue generated in the subsequent digestion reaction step, the collagen component contained in the enzyme extract can be increased, thereby improving the digestion efficiency. In addition, the hot water referred to here includes, for example, high-temperature water of about 80°C.

In addition, in the digestion reaction step, by using papain for enzymatic digestion of the extracted product after the hot water extraction step at a temperature in the range of 60°C to 70°C, the extraction treatment after the hot water extraction step can be performed. The substance is dissolved by the enzyme reaction and divided into the enzyme extract and the residue. In addition, an enzyme extract containing a water-soluble collagen component can be obtained. In addition, since papain is a heat-resistant enzyme, it can be stably supplied to the enzyme reaction while maintaining a high temperature for the extracted product treated at high temperature in the hot-water extraction step of the previous step.

In addition, in the mixing step, the placenta extract obtained in the placental extract production step, and the hydroxyproline concentration (w/v) per unit total nitrogen concentration (w/v) of the placenta extract is 0.47 or more. The extract is mixed with epidermal growth factor (EGF) to produce a cosmetic composition containing collagen-rich placenta extract and epidermal growth factor (EGF).

Furthermore, in order to achieve the above object, the cosmetic composition of the present invention is constituted to contain a placenta extract, and in the aforementioned placenta extract, the hydroxyproline concentration per unit of total nitrogen concentration (w/v) of the placenta extract is (w/v) is 0.47 or more, and the concentration of the placenta extract is in the range of 0.025v/v% to 0.8v/v%.

Here, the hydroxyproline concentration (w/v) per unit of total nitrogen concentration (w/v) in the placental extract permeated through the placenta extract is 0.47 or more, and the placenta extract used as the raw material is rich in collagen. The hydroxyproline concentration (w/v) per unit of total nitrogen concentration (w/v) of the placenta extract is above 0.47, which means that the concentration of hydroxyproline in the placenta extract is higher than that in the conventional commercially available placenta extract. , contains a sufficient amount of collagen. As a result, the fibroblast growth effect and collagen production effect of collagen in the placenta extract can be enhanced.

In addition, when the concentration of the placenta extract is in the range of 0.025v/v% to 0.8v/v%, the growth effect of fibroblasts in the cosmetic composition can be improved. In addition, coloring of the cosmetic can be suppressed.

In addition, the term "concentration of total nitrogen per unit of placenta extract (w/v)" in this specification refers to the concentration (% (w/v) of the total nitrogen concentration of placenta extract based on 1% (w/v) v)). [Effect of invention]

The method for producing a cosmetic composition of the present invention can provide a cosmetic composition containing collagen-rich placental extract and having excellent functions.

[Form of implementing the invention]

Hereinafter, the composition of a lotion, which is an example of the cosmetic composition to which the present invention is applied, will be described.

The lotion system shown here has a weight ratio of pure water: 30~95%, collagen-containing placenta extract: 0.05~0.8% (v/v), and polyol: 0.1~50% based on the total amount. , Ester oil: appropriate amount (0~5%, sometimes unmixed), surfactant: appropriate amount, citric acid: appropriate amount, carboxyethylene polymer: appropriate amount (lower limit 0.1%), sodium hydroxide: appropriate amount, plant extract : Appropriate amount (minimum: 0.1%) Preservative: Composition in appropriate amount. Furthermore, the lotion system shown here contains epidermal growth factor (EGF) at a concentration of 0.0000075 to 0.00012% (w/v) based on the total amount.

In addition, the placenta extract containing collagen refers to the "placenta extract" in the claim of this case. In addition, the term "collagen-containing placenta extract" is used below.

In addition, the composition of the lotion shown here is only an example of a cosmetic composition, and other components or different blending amounts can be appropriately set. For example, in addition to the above, a chelating agent (a chelating agent), astringent, bactericide, skin activator (vitamins, amino acid and amino acid derivative), anti-inflammatory agent, Whitening agents (arbutin, tranexamic acid, vitamin C derivatives, etc.). In addition, flavors or colorants can also be blended as required.

Pure water is the base of the lotion, and the total amount of the lotion containing the above ingredients is adjusted to 100%.

Collagen-containing placenta extract is a characteristic ingredient, which can impart fibroblast growth effect, collagen production effect, collagenase activity inhibitory effect, antioxidant effect, anti-inflammatory effect, metabolism promotion and skin rejuvenation effect to lotion. , or ingredients that enhance these preferred effects. In addition, it is a product obtained by processing the horse placenta by the production method described later.

Further, the collagen-containing placenta extract contains hydroxyproline having a hydroxyproline concentration of 0.47% (w/v) per unit of total nitrogen concentration of the placenta extract (w/v). Hydroxyproline is an indicator of the concentration of collagen in the placenta extract. The concentration of hydroxyproline in the collagen-containing placenta extract in this composition is higher than that in the conventional commercially available placenta extract. Concentration of acid, containing sufficient amount of collagen.

Epidermal growth factor (EGF) is a characteristic ingredient, and it is an ingredient that enhances the growth effect of fibroblasts in lotion. Epidermal growth factor (EGF), by combining with collagen-containing placenta extract, can synergistically enhance the growth of fibroblasts.

Polyols are humectants. As the polyhydric alcohol, for example, glycerol, 1,3-butanediol, 1,2-pentanediol and the like can be used.

Ester oils are softeners (softeners). In the above composition, ester oil is used, but other than that, for example, vegetable oil can also be used.

Surfactants are emulsifiers or cosolvents for oily components. As the surfactant, for example, lecithin and lecithin derivatives, glycerol fatty acid ester, polyoxyethylene sorbitan fatty acid ester, polyoxyethylene hardened castor oil, and the like can be used. Moreover, in this invention, the kind of the emulsifier of an oily component or a cosolvent is not limited to these, Other surfactants can also be used.

Citric acid is a buffer (pH adjuster). In the above composition, citric acid is used, but other than that, for example, sodium citrate can also be used.

Carboxy vinyl polymer is a tackifier, which imparts moderate viscosity to lotion. In the above-mentioned composition, a carboxyvinyl polymer is used, but other than that, for example, an acrylic thickener, polyvinyl alcohol, xanthan gum, carrageenan, agar, and gelatin can also be used.

Sodium hydroxide is a neutralizing agent. In the above-mentioned composition, sodium hydroxide is used, but other than this, potassium hydroxide, for example, can also be used.

A plant extract is an ingredient that imparts effective desired functions to the skin in a lotion. In addition, the preservative is an ingredient that imparts antiseptic power to the lotion.

Here, as a plant extract, the following are mentioned, for example. Coix Seed Extract, Royal Jelly Extract, Tripterygium Root Leaf Extract, Bletilla striata Root Extract, Placenta Extract, Scutellariae Extract, Jifeng Leaf Extract, Soybean Placenta Extract, Hybrid Rose Extract, Moon Peach Leaf Extract, Pilocarpus Solidago extract, yeast solute extract, creatine rhizoma peel extract, Korean ginseng root extract, peony root extract, grapevine extract.

Here, the blending amount of the collagen-containing placenta extract is 0.05 to 0.8% based on the total amount of the lotion. Here, it is not always necessary to set the blending amount of the collagen-containing placenta extract to 0.05 to 0.8% based on the total amount of the lotion. However, in order to further enhance the fibroblast growth effect of the lotion, the blending amount of the collagen-containing placenta extract is preferably set to 0.025~1.0% based on the total amount of the lotion. In addition, in order to further enhance the fibroblast growth effect of the lotion and at the same time inhibit the color of the collagen-containing placenta extract from appearing in the lotion, the blending amount of the collagen-containing placenta extract is preferably set to It is 0.2~0.8% based on the total amount of lotion.

In addition, the blending amount of epidermal growth factor (EGF) is a concentration of 0.0000075 to 0.00012% (w/v) based on the total amount of the lotion. Here, it is not necessarily necessary to set the blending amount of epidermal growth factor (EGF) to a concentration of 0.0000075 to 0.00012% (w/v) based on the total amount of the lotion. However, in order to further enhance the fibroblast growth effect of the lotion, and at the same time prevent the effect from increasing to the maximum, the blending amount of epidermal growth factor (EGF) is preferably set to be 0.00000375~0.00015 based on the total amount of the lotion. %(w/v) of the concentration range.

Next, an example of the manufacturing method of the cosmetic composition to which the present invention is applied will be described. Here, the above-mentioned lotion is used as an example for description.

First, the content of producing a collagen-containing placenta extract from horse placenta will be explained. The cryopreserved equine placenta was thawed (thawed). An amount of exudate (blood generated during thawing) equal to the fleshy part of the thawed horse placenta was measured, and the fleshy part was crushed (crushed and supplied with raw materials).

Moreover, the disintegrated product of the horse placenta and the exudate were mixed, and hot water extraction was performed at 80 degreeC for about 22 hours (hot water extraction). Thereby, the water-soluble collagen component can be efficiently extracted from the horse placenta. That is, between the enzyme extract and the residue generated in the following digestion reaction steps, the collagen component contained in the enzyme extract can be increased, thereby improving the digestion efficiency. In addition, this step belongs to the hot water extraction step in the claim of this case.

Next, the processed product was extracted with hot water, and a digestion reaction was performed at 60 to 70° C. for 3 hours using 0.2% papain (digestion reaction). Thereby, the extracted processed material can be dissolved by the enzyme reaction, and it can be divided into an enzyme extract and a residue. In addition, an enzyme extract containing a water-soluble collagen component can be obtained. In addition, this step belongs to the digestion reaction step in the claim of this case.

Next, the enzyme extract was treated with 0.8% activated carbon for 10 minutes. In addition, after that, heat treatment was performed at 90° C. for 30 minutes. Further, if necessary, centrifugation was performed at 8000 rpm for 5 minutes to separate into supernatant liquid and sediment.

In addition, the centrifuged supernatant was subjected to vacuum filtration using a filter aid (No. 2 filter paper). Furthermore, the supernatant liquid after vacuum filtration was subjected to clarification filtration.

In addition, 1.6% of 2-phenoxyethanol was added to the filtrate obtained by clear filtration, followed by stirring. Furthermore, sterilization filtration was performed using a 0.22-micrometer filter. Following the steps so far, a collagen-containing placenta extract can be produced from horse placenta.

In addition, the above-described method is merely an example, and various conditions and methods can be appropriately changed.

In addition, in the step of performing the hot water extraction, it is not necessarily necessary to perform the hot water extraction at 80° C. for about 22 hours, and the temperature and time of the water can be appropriately changed. However, in order to sufficiently and efficiently extract water-soluble collagen components from the horse placenta, it is preferable to perform hot water extraction at 80°C for about 22 hours.

In addition, in the step of carrying out the digestion reaction, it is not necessarily necessary to use 0.2% papain to carry out the digestion reaction at 60-70° C. for 3 hours, and heat-resistant digestive enzymes other than papain may also be used. In addition, the temperature and time of the digestion reaction can be appropriately changed. However, in terms of obtaining an enzyme extract rich in water-soluble collagen components from the processed material extracted with hot water, it is preferable to use 0.2% papain to carry out the digestion reaction at 60-70° C. for 3 hours.

Moreover, as an example of the manufacturing method of the cosmetic composition of this invention, the collagen-containing placenta extract obtained above is mixed with various components containing epidermal growth factor (EGF), and a lotion is manufactured.

Here, the method for mixing the collagen-containing placenta extract and the various components containing epidermal growth factor (EGF) can be a known method for producing cosmetics.

As described above, the cosmetic composition to which the present invention is applied can be produced by producing a collagen-containing placenta extract by the above-mentioned method, and mixing the collagen-containing placenta extract with various raw materials containing epidermal growth factor (EGF) to produce . In addition, the manufacturing method and conditions shown here are merely an example, and it is a matter of course that they can be appropriately set and changed according to the types of raw material components and additives.

In addition, the types of cosmetic compositions to which the present invention is applied are not limited to the above-mentioned lotions, and can be applied to, for example, emulsion/cream-based cosmetics, gels, cleansers (shampoo), facial cleansers (for face wash), and the like . In addition, as the production method, a conventional method of mixing the produced collagen-containing placenta extract and the raw materials of various cosmetic materials containing epidermal growth factor (EGF) according to the type of the cosmetic materials can be adopted.

As described above, applying the cosmetic composition of the present invention can provide a cosmetic composition containing collagen-rich placenta extract and having excellent functions. Moreover, the manufacturing method of the cosmetic composition to which this invention is applied is a method which can provide the cosmetic composition which contains the collagen-rich placenta extract and is excellent in function. [Example]

Embodiments of the present invention will be described below.

First, the following evaluations were performed on the collagen-containing placenta extract contained in the cosmetic composition of the present invention.

For the collagen-containing placenta extract produced by the above-mentioned production method of the collagen-containing placenta extract and two commercially available placenta extracts A and B (both obtained from horse placenta), the hydroxyproline Determination of acid content.

In addition, placenta extract A and placenta extract B are prepared by repeatedly freezing and immersing the horse placenta as raw materials at low temperature, disintegrating the cell tissue, extracting its protoplasm, deproteinizing and ultrafiltration. In addition, the preparation method of the placenta extract A and the placenta extract B can adopt a known method. For example, in addition to thawing, the steps of enzyme extraction can also be combined.

(1) Determination of hydroxyproline content Hydrochloric acid was added to each sample solution of the collagen-containing placenta extract, placenta extract A, and placenta extract B so that the final concentration was 6M, and hydrolysis was performed at 110° C. for 24 hours. For each of the hydrolyzed samples, amino acid analysis was performed by a post-column method labeled with o-phthalaldehyde. In addition, the total nitrogen concentration in the sample was previously measured, and the concentration (% (w/v)) of hydroxyproline per unit total nitrogen concentration of the sample was compared at 1% (w/v). Table 1 shows the hydroxyproline content in each sample.

[Table 1] Placenta Extract with Collagen Placenta Extract A Placenta Extract B Hydroxyproline concentration (%(w/v)) per unit of total nitrogen concentration 1% (w/v) 0.470 0.272 0.063

As shown in Table 1, it is apparent that the collagen-containing placenta extract contained in the cosmetic composition of the present invention contains more hydroxyproline than the commercially available placenta extract A and placenta extract B. That is, it is apparent that the collagen-containing placenta extract contains more collagen than the placenta extract A and the placenta extract B. In addition, based on horse placentas with different raw material sources, a collagen-containing placenta extract was prepared in the same way by the production method of a collagen-containing placenta extract, and the concentration of 1% (w/v) of total nitrogen per unit of the sample was measured. The concentration of hydroxyproline showed a result of 0.59 (% (w/v)).

(2) Fibroblast growth activity test Next, a fibroblast growth activity test was performed for each sample of the collagen-containing placenta extract, the placenta extract A, and the placenta extract B. Here, fibroblasts refer to cells that are located in the dermis of the skin and that can produce collagen. This fibroblast growth-promoting effect (reproductive activity) was evaluated as a sample having an excellent collagen production-promoting effect.

(Test method) Dermal fibroblasts from human neonates were seeded in a 96-well plate at 2×10 ⁴ cells/well, containing 0.5% Fetal Bovine Serum (FBS). DMEM medium (Dulbecco's Modified Eagle Medium) overnight. Placenta extract, placenta extract A and placenta extract B containing collagen were respectively replaced with the medium supplemented with the concentration of total nitrogen, and cultured in a 37°C, 5% CO ₂ incubator for 24 hours. The cells were washed with Phosphate-buffered saline (PBS), and DMEM containing 10% of a commercially available Cell Counting Kit (Cell Counting Kit-8, manufactured by Dojin Chemical Research Institute Co., Ltd.) was added. , and incubated at 37°C in a 5% CO ₂ incubator for 4 hours. After incubation, the absorbance at 450 nm was measured. In addition, the measurement results are shown as relative values when the reproductive activity when the collagen-containing placenta extract, placenta extract A, and placenta extract B are not added is taken as 100%. In addition, the cell growth activity was calculated by the calculation formula shown in the following formula 1. (Formula 1) Cell proliferation activity (%)={(absorbance at 450 nm of the sample-added group)/(absorbance of the sample-free group at 450 nm)}×100 The results of the fibroblast growth activity test are shown in Table 2.

[Table 2] Placenta Extract with Collagen Placenta Extract A Placenta Extract B control group Reproductive activity (%) 119.3 102.7 106.2 100.0

As shown in Table 2, the collagen-containing placenta extract contained in the cosmetic composition of the present invention showed the result that the growth activity of fibroblasts was significantly improved compared with the placenta extract A and the placenta extract B in the market.

Next, Examples and Comparative Examples of the cosmetic composition produced by applying the method for producing the cosmetic composition of the present invention were prepared, and the evaluation of the fibroblast growth activity test was performed. (3) Raw material components of the sample First, the raw material components were added so that the compositions shown in Tables 3 to 6 were obtained, and the samples of Examples 1 to 15 and Comparative Examples 1 to 9 were prepared. In addition, the numerical values of the components shown in Tables 3 to 6 below are the volume ratio (%(v/v)) based on the total amount for the collagen-containing placenta extract; for the epidermal growth factor (EGF) , it represents the weight-to-volume ratio (%(w/v)).

[table 3] Example 1 Example 2 Example 3 Example 4 Example 5 Example 6 Comparative Example 7 Collagen-containing placenta extract (%) 0.8 0.4 0.2 0.1 0.05 0.025 - Reproductive activity (%) 119.3 118.5 119.9 109.5 108.5 103.5 100.0

[Table 4] Comparative Example 2 Comparative Example 3 Comparative Example 4 Comparative Example 5 Comparative Example 6 Comparative Example 7 Comparative Example 8 EGF Concentration (%(w/v)) 0.00012 0.00006 0.00003 0.000015 0.0000075 0.00000375 - Reproductive activity (%) 114.6 121.0 122.1 129.5 124.5 124.7 100.0

[table 5] Example 7 Example 8 Example 9 Example 10 Example 11 Collagen-containing placenta extract (%) 2.0 1.0 0.8 0.4 0.2 EGF Concentration (%(w/v)) 0.0003 0.00015 0.00012 0.00006 0.00003 Reproductive activity (%) 131.0 132.0 137.4 133.2 137.3

[Table 6] Example 12 Example 13 Example 14 Example 15 Comparative Example 9 Collagen-containing placenta extract (%) 0.1 0.05 0.025 0.0125 - EGF Concentration (%(w/v)) 0.000015 0.0000075 0.00000375 0.000001875 - Reproductive activity (%) 129.8 127.7 129.0 118.0 100.0

In addition, the fibroblast growth activity test was carried out as follows. (Test method) Skin fibroblasts from human neonates were seeded in 96-well plates at 2×10 ⁴ cells/well, and cultured overnight in DMEM medium containing 0.5% fetal bovine serum (FBS). . The collagen-containing placenta extract, epidermal growth factor (EGF), and the mixture of collagen-containing placental extract and epidermal growth factor (EGF) were replaced with the medium supplemented with the amounts shown in Tables 3 to 5, respectively. , cultured for 24 hours in a 37°C, 5% CO ₂ incubator. The cells were washed with phosphate-buffered saline (PBS), added with DMEM containing 10% of a commercially available Cell Counting Kit (Dojin Chemical Laboratory Co., Ltd.: Cell Counting Kit-8), and incubated at 37° C. Incubate for 4 hours in a 5% CO ₂ incubator. After incubation, the absorbance at 450 nm was measured. In addition, the measurement results are based on the reproduction activity when the collagen-containing placenta extract, epidermal growth factor (EGF) and the mixture of the collagen-containing placental extract and epidermal growth factor (EGF) are not added as 100 Relative value in %. In addition, the cell growth activity was calculated by the calculation formula shown in the following formula 1. (Formula 1) Cell proliferation activity (%)={(the absorbance at 450 nm of the sample-added group)/(the absorbance at 450 nm of the sample without the addition of the sample)}×100 In addition, Comparative Examples 1, 8 and 9 showed that no collagen-containing group was added. Proliferative activity of protein-containing placental extract, epidermal growth factor (EGF), and a mixture of collagen-containing placental extract and epidermal growth factor (EGF).

The results of the fibroblast growth activity test are shown in Figures 1 to 3 and Tables 3 to 6. In addition, Fig. 1 graphs the numerical values shown in Table 3, Fig. 2 graphs the numerical values shown in Table 4, and Fig. 3 graphs the numerical values shown in Tables 5 and 6.

Regarding Examples 1 to 6 and Examples 7 to 15, it was shown that the growth activity of fibroblasts was improved. Especially in Examples 7 to 14, compared with Comparative Examples 2 to 4, 6, and 7, the results also showed that the growth activity of fibroblasts was significantly improved.

In addition, an example of the various cosmetics manufactured by the manufacturing method of the cosmetic composition to which this invention is applied is shown below.

(Lotion/Cream) [Table 7] constituents Representative raw materials Addition (%) water pure water 30~95 Humectant (polyol) Glycerol, 1,3-butanediol, 1,2-pentanediol, etc. Appropriate amount (0.1~50%) Placenta Extract with Collagen 0.05~0.8%(v/v) Epidermal Growth Factor (EGF) 0.00012~0.0000075%(w/v) Oil (softener, softener) Higher alcohols, fatty acids, ester oils, vegetable oils, silicone oils, waxes, etc. Appropriate amount (3~50%) Emulsifier Surfactant Moderate Buffer (pH adjuster) Citric Acid, Sodium Citrate Moderate Tackifier Carboxy vinyl polymer, acrylic tackifier, polyvinyl alcohol, xanthan gum, carrageenan, agar, gelatin, etc. Appropriate amount (lower limit 0.1%) Neutralizer Sodium Hydroxide, Potassium Hydroxide Moderate plant extract Appropriate amount (lower limit 0.1%) preservative Moderate

(gel) [Table 8] constituents Representative raw materials Addition (%) water pure water 30~95 Humectant (polyol) Glycerol, 1,3-butanediol, 1,2-pentanediol, etc. Appropriate amount (0.1~50%) Placenta Extract with Collagen 0.05~0.8%(v/v) Epidermal Growth Factor (EGF) 0.00012~0.0000075%(w/v) Oil (softener, softener) Higher alcohols, fatty acids, ester oils, vegetable oils, silicone oils, waxes, etc. Appropriate amount (1~50%) Emulsifier Surfactant Moderate Buffer (pH adjuster) Citric Acid, Sodium Citrate Moderate Tackifier Carboxy vinyl polymer, acrylic tackifier, polyvinyl alcohol, xanthan gum, carrageenan, agar, gelatin, etc. Appropriate amount (lower limit 0.1%) Neutralizer Sodium Hydroxide, Potassium Hydroxide Moderate plant extract Appropriate amount (lower limit 0.1%) preservative Moderate preservative Moderate

(Detergent (Shampoo)) [Table 9] constituents Representative raw materials Addition (%) water pure water 30~95 Humectant (polyol) Glycerol, 1,3-butanediol, 1,2-pentanediol, etc. Appropriate amount (0.1~50%) Placenta Extract with Collagen 0.05~0.8%(v/v) Epidermal Growth Factor (EGF) 0.00012~0.0000075%(w/v) detergent Surfactant 5~50% Buffer (pH adjuster) Citric Acid, Sodium Citrate Moderate plant extract Appropriate amount (lower limit 0.1%) preservative Moderate

(Facial Cleanser (Face Wash)) [Table 10] constituents Representative raw materials Addition (%) water pure water 10~50 Humectant (polyol) Glycerol, 1,3-butanediol, 1,2-pentanediol, etc. Appropriate amount (1~50%) Placenta Extract with Collagen 0.05~0.8%(v/v) Epidermal Growth Factor (EGF) 0.00012~0.0000075%(w/v) fatty acid Stearic acid, palmitic acid, myristic acid, lauric acid 5~50% Buffer (pH adjuster) Calcium hydroxide Appropriate amount (with neutralization degree) plant extract Appropriate amount (lower limit 0.1%) preservative Moderate

In addition, in the above-mentioned various cosmetic materials, other components or different blending amounts can be appropriately set. For example, in addition to the above, a chelating agent (a chelating agent), astringent, bactericide, skin activator (vitamins, amino acid and amino acid derivative), anti-inflammatory agent, Whitening agents (arbutin, tranexamic acid, vitamin C derivatives, etc.). In addition, flavors or colorants can also be blended as required.

Fig. 1 is a graph showing the results of a fibroblast growth activity test in placental extracts. Fig. 2 is a graph showing the results of a fibroblast growth activity test in epidermal growth factor (EGF). [ Fig. 3] Fig. 3 is a graph showing the results of a fibroblast growth activity test in a mixture of placental extract and epidermal growth factor (EGF).

Claims (4)

A method for producing a cosmetic composition, comprising a step of producing a placenta extract and a step of mixing, wherein, The placenta extract manufacturing steps include: The hot-water extraction step is to mix the thawed horse placenta or porcine placenta with the tissue fluid such as blood exuded during thawing, and heat the mixture to perform hot-water extraction treatment; and The digestion reaction step is to use papain for enzymatic digestion of the extracted product after the hot water extraction step at a temperature in the range of 60°C to 70°C; The mixing step is the placenta extract obtained in the placenta extract manufacturing step, and the placenta extract and epidermis of which the hydroxyproline concentration (w/v) per unit total nitrogen concentration (w/v) is 0.47 or more Cell Growth Factor (EGF) mix. The manufacturing method of the cosmetic composition of claim 1, wherein, The concentration of the aforementioned placenta extract is in the range of 0.025v/v%~1.0v/v%, The concentration of the aforementioned epidermal growth factor (EGF) is in the range of 0.00000375 w/v% to 0.00015 w/v%. The manufacturing method of the cosmetic composition of claim 2, wherein, The concentration of the aforementioned placenta extract is in the range of 0.05v/v% to 0.8v/v%. The method for producing a cosmetic composition according to claim 1, 2 or 3, wherein, The aforementioned placenta extract is obtained from horse placenta.

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