KR102316110B1 - Functional cosmetic composition comprising complex extract of walnut shell and persimmon calyx as effective ingredient, and manufacturing method thereof - Google Patents

Abstract

A functional cosmetic composition including a complex extract of walnut shells and persimmon calyxes as an active ingredient of the present invention has excellent wrinkle or elasticity improvement effects, and further has anti-inflammatory, skin moisturizing, sebum reduction, keratin reduction or antioxidant effects.

Description

Functional cosmetic composition containing complex extract of walnut shell and persimmon as an active ingredient, and manufacturing method thereof

The present invention relates to a functional cosmetic composition comprising a complex extract of walnut shells and persimmons.

NO is known to have various functions such as blood pressure regulation, platelet aggregation, and signal transduction in vivo. In particular, pro-inflammatory cytokines or LPS act on immune cells to increase NO production by the action of inducible nitric oxide synthase (iNOS) inducible from L-arginine. Excessive NO production is associated with chronic inflammation, and many studies have been conducted to develop selective NO biosynthesis inhibitors to control chronic inflammation through the regulation of iNOS activity (Sharma et al., 2007; Korhonen et al., 2005).

On the other hand, aging of skin cells closely related to skin wrinkles can be classified into natural aging that occurs according to aging and aging caused by external environment. Natural aging is not easy to control because genetic factors are involved, and the majority of studies are to control aging by controlling environmental factors such as UV rays, oxidation, and drying.

The environmental factors mainly cause wrinkles by destroying or denaturing collagen protein, elastin protein, etc. included in the dermal layer. In particular, UV rays oxidize skin cells (fibroblasts) and cause a decrease in the production and destruction of elastin. will bring In addition, the active oxygen mass-produced by the above factors may act as a cause of aging and wrinkles in the skin by causing protein destruction, sugar oxidation, genetic modification, etc. in skin cells by oxidation.

There have been attempts to manufacture cosmetics by adding extracts extracted from various plants, fruits, and herbal medicines, as various side effects and problems due to skin damage have emerged due to the use of cosmetics containing a large amount of chemical ingredients. In particular, it was required to search for a material that can be safely used in the human body while retaining functions such as anti-oxidation and anti-wrinkle.

Vegetable raw materials, such as fruits, may be candidates for meeting these needs due to their low cytotoxicity while containing various substances useful for physiological activity. However, in order to extract a desired active ingredient in a high content from the fruit, a huge amount of raw materials are required, so there is a problem of an increase in the cost of the product, and there is also a problem of consumption of food resources and environmental pollution due to the generation of by-products.

Most of the skins and fruit stalks generated as by-products after processing vegetable raw materials are disposed of as garbage, and it is time to discuss and take measures for environmentally friendly disposal or recycling.

Accordingly, the need for the development of cosmetic raw materials capable of securing a high content of the desired active ingredient, without consuming resources, and increasing the cost or reducing environmental pollution increases.

It is an object of the present invention to provide a cosmetic composition that inhibits elastase and exhibits the efficacy of preventing or improving wrinkles.

Another object of the present invention is to provide a cosmetic composition exhibiting anti-inflammatory efficacy by inhibiting NO production.

Another object of the present invention is to provide a cosmetic composition having no cytotoxicity and less skin irritation.

Another object of the present invention is to provide a cosmetic composition having an increased content of natural phenolic compounds.

Another object of the present invention is to provide a method for preparing the cosmetic composition.

According to one aspect of the present invention, it is possible to present a cosmetic composition containing a complex extract of walnut shells and persimmons as an active ingredient and exhibiting an effect of improving skin wrinkles or improving elasticity.

The cosmetic composition may further have an anti-inflammatory effect and a skin moisturizing effect.

Specifically, the mixing ratio of the walnut shell extract and the persimmon extract of the complex extract may be 1: 0.5 to 1: 2.

The complex extract may be contained in an amount of 0.001% to 60% by weight based on the total weight of the cosmetic composition.

The cosmetic composition is not limited thereto, but may be in the form of a lotion, serum, essence, lotion, cream, powder, foundation, pack, patch, mask sheet, gel ointment, spray or detergent.

In addition, according to another aspect of the present invention, it is possible to present a method for producing a cosmetic composition having the effect of improving skin wrinkles or improving elasticity, containing the above-described complex extract of walnut shell and persimmon as an active ingredient.

The method may include a step of preparing a complex extract by mixing the extract of walnut shells and extracts of persimmons, or by mixing and then extracting the pulverized dried walnut hulls and pulverized dried persimmons. At this time, the blending ratio of the walnut shell extract and the persimmon stalk extract among the complex extracts or the blending ratio of the dried walnut shell pulverized product and the dried persimmon stalk extract may be 1: 0.5 to 1: 2, and specifically may be 1:1.

In the method for preparing the cosmetic composition, 1 to 100 times the weight of a solvent is added to the dried raw material pulverized of walnut shells and persimmons, and then extracted by hot water extraction at a temperature of 10 to 120° C. for 30 minutes to 360 hours. , performing one or more of ultrasonic extraction, cold extraction, and percolation.

In addition, the manufacturing method of the cosmetic composition is after adding 0.001% to 60% by weight of the complex extract to the general composition for preparing a cosmetic composition, and then, according to a conventional method, a lotion, serum, essence, lotion, cream, powder, foundation, pack, It may include formulating into a formulation such as a patch, a mask sheet, a gel ointment, a spray or a cleaning agent.

The cosmetic composition containing the complex extract of walnut shells and persimmons of the present invention as an active ingredient has excellent skin wrinkle improvement or elasticity improvement effect, and further possesses the functionality of anti-inflammatory effect and skin moisturizing effect.

1 is a graph showing the cytotoxicity test results for Preparation Examples and Comparative Preparation Examples of the present invention.
2 is a graph showing the anti-inflammatory test results for Preparation Examples and Comparative Preparation Examples of the present invention.
3 is a graph showing the evaluation results of elastase inhibitory activity for Preparation Examples and Comparative Preparation Examples of the present invention.
4 is a graph showing the skin improvement evaluation results of the retention rate of moisture for Examples and Comparative Examples of the present invention.

The complex extract of walnut shell and persimmon of the present invention has anti-wrinkle effect and anti-inflammatory effect, and improves and maintains skin moisture of the cosmetic composition containing it through increasing the content of natural phenolic compounds, reduces the amount of sebum, each It is possible to realize the effect of reducing the mass.

Walnut (Juglans sinensis) is a fruit of a deciduous broad-leaved tree belonging to the genus Walnut and Walnut. There are more than 10 species of walnuts around the world, including the Persian walnut (J. sinensis).

The walnut fruit contains alpha-linolenic acid (ALA), an unsaturated fatty acid omega-3, along with various phenolic components. However, the functions and effects of walnut shell extract have not been actively studied.

Persimmon stalk is the calyx of the fruit of Disopyros kaki L. Persimmons are known to be effective in relieving vomiting and coughing. The main components of persimmons are succinic acid, quercetin, campferol, neodiospyrin, oleanolin, ursolic, betulin and daucosterol. ) and so on.

In the present invention, the walnut shell extract or persimmon extract is not particularly limited, and specifically, it may be used in various forms, such as an extract, a powder obtained by drying the extract, and a concentrate obtained by concentrating and filtering the extract.

The extract may be in the form of, for example, a tinc, a concentrate, an extract, a liquid extract, etc. containing alcohol, purified water, oil, etc. as a leaching solvent.

The detailed extraction method of the walnut shell or persimmon extract will be dealt with in more detail later in the present application.

Specifically, the mixing ratio of the walnut shell extract and the persimmon extract of the complex extract may be 1: 0.5 to 1: 2.

It is possible to implement improved skin moisturizing ability along with improved NO production inhibition and elastase inhibitory activity within the above compounding ratio range.

In one embodiment, the blending ratio of the walnut shell extract and the persimmon extract of the complex extract may be measured based on the solid weight of the extract. At this time, the solid weight of the extract may be a solid obtained by freeze-drying after extracting by adding a solvent to the dried walnut shell pulverized product and dried persimmon stalk or a mixture thereof.

In another embodiment, the blending ratio of the walnut shell extract and the persimmon extract in the complex extract may be measured based on the solids weight of the raw material. At this time, the weight of the solids may be the solids for the dried walnut shell pulverized and dried persimmons pulverized or a mixture thereof before adding the extraction solvent.

The complex extract may be contained in an amount of 0.001 wt% to 60 wt%, specifically 1 wt% to 55 wt%, more specifically 20 wt% to 55 wt%, based on the total weight of the cosmetic composition. Expected functionality may be expressed within the above range.

The cosmetic composition may be in any one formulation of lotion, serum, essence, lotion, cream, powder, foundation, pack, patch, mask sheet, gel ointment, spray, and detergent, but is not limited thereto.

The cosmetic composition of the present invention may further include a composition for preparing a cosmetic together with the complex extract.

The composition for cosmetic preparation acts together with the complex extract to realize a cosmetic effect according to a desired cosmetic use and formulation, and is not particularly limited as long as it is a component used as a general cosmetic subject or excipient.

Specifically, as the composition for preparing the cosmetic, a solvent, a fat and oil component, a moisturizer, an emollient, a surfactant, an organic and inorganic pigment, an organic powder, an ultraviolet absorber, a preservative, a disinfectant, an antioxidant, a plant extract, a pH adjuster, a plant oil , a pigment, a fragrance, a blood circulation promoter, a cooling agent, an inhibitor of sweat secretion, etc. can be mixed and used.

For example, the cosmetic composition is purified water, alcohol, glycerin, myristic acid, butylene glycol, potassium hydroxide. , polyethylene glycol, stearate, stearic acid, behenyl alcohol, microcrystalline wax, phenoxyethanol, ethylhexylglycerin ( ethylhexylglycerin), tocopheryl acetate, 1,2-hexanediol, caprylyl glycol, hydroxyethylene cellulose, hydroxyethyl cellulose ethyl cellulose), xanthan gum, collagen, sodium citrate, citrate, magnesium ascorbate, allantoin, panthenol, betaine ( betaine), Cetearyl Alcohol, Sunflower oil, Cetearyl Olivate, Sorbitan Olivate, Glyceryl Stearate and fragrance ) may include one or more of.

Another aspect of the present invention relates to a method for preparing a functional cosmetic composition comprising the above-described complex extract of walnut shell and persimmon as an active ingredient. The method includes mixing an extract of walnut hulls and an extract of persimmon, or mixing and then extracting a pulverized dried walnut hull and pulverized dried persimmon to prepare a complex extract.

In the manufacturing method of the cosmetic composition of the present invention, the process of preparing the extract of walnut shell, extract of persimmon or complex extract is not particularly limited and may be obtained through various methods known in the art to which the present invention pertains.

For example, in the process of preparing the extract of walnut shell, extract of persimmon or complex extract, 1 fold to 100 fold weight of a solvent is added to the dry raw material pulverized material of each extraction target, and then a temperature range of 10 to 120 ° C. It may include performing at least one of hot water extraction, reflux cooling extraction, ultrasonic extraction, cold extraction extraction, and percolation for 30 minutes to 360 hours at .

Each of the extraction targets means a mixture of walnut shells and persimmons in the case of walnut shell extracts, persimmons extracts, and complex extracts in the case of walnut shell extracts.

The solvent is not particularly limited as long as it can be used as an extraction solvent for the cosmetic composition, and specifically, it may be water, an organic solvent, or a mixed solvent thereof.

The water may be alkaline water, and the organic solvent may be an anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms such as ethanol, propanol, methanol, n-butanol; polar organic solvents such as acetone and 1,3-butylene glycol; non-polar organic solvents such as ether, hexane, benzene, chloroform, and ethyl acetate; vegetable organic solvents such as soybean oil and sesame oil; and the like. The organic solvent may be used alone or as a mixture of two or more. In this case, when a mixed solvent is used, the mixing composition and ratio may be appropriately mixed according to the purpose.

In one embodiment, as a solvent used in the extraction, one or more lower alcohols of ethanol, methanol, n-butanol, and propanol; 1,3-butylene glycol, at least one polar organic solvent of acetone; and a mixed solvent including alkaline water. In this case, the extraction efficiency of the active ingredient in the preparation of the extract of the walnut shell, the extract of persimmon or complex extract can be improved.

In another embodiment, as a solvent used for the extraction, one or more lower alcohols of ethanol, methanol, n-butanol, and propanol; and a mixed solvent including water. In this case, the extraction balance for both the walnut shell and the persimmon stalk is excellent, and the extraction efficiency of the active ingredient in the preparation of the complex extract can be improved.

The amount of the solvent added during the extraction is not particularly limited, but may be 1 to 100 times the weight of the dry raw material pulverized for each extraction target, specifically 2 to 80 times, and more specifically 5 to 50 times. Within the above range, it is possible to more efficiently increase the time required for extraction and the content of the extracted active ingredient.

The extraction temperature is not particularly limited, but may be performed in a temperature range of 10 to 120 °C, for example, 50 to 120 °C or 10 to 40 °C. Within the above range, it is possible to more efficiently increase the time required for extraction and the content of the extracted active ingredient.

The extraction time is not particularly limited, but may be performed for 30 minutes to 360 hours, for example, 30 minutes to 72 hours, 30 minutes to 24 hours, or 168 hours to 360 hours. Within the above range, it is possible to more efficiently increase the time required for extraction and the content of the extracted active ingredient.

The extraction method is not particularly limited, but may include performing any one or more of hot water extraction, reflux cooling extraction, ultrasonic extraction, cold extraction, and percolation. Within the above range, it is possible to more efficiently increase the time required for extraction and the content of the extracted active ingredient.

The number of times of extraction during the extraction is not particularly limited, but may be performed one or more times for each extract, and specifically may be repeated 1 to 5 times. Within the above range, it is possible to more efficiently increase the time required for extraction and the content of the extracted active ingredient.

In one embodiment, the extraction may be extraction by heating and reflux cooling at 50 to 120° C. for 30 minutes to 24 hours after each extraction target and extraction solvent are added to an extractor equipped with a reflux cooler. In this case, the heating and reflux cooling extraction may be a method in which each target material to which a solvent is added is heated to reflux and then cooled and extracted. In this case, the extraction efficiency can be further improved while minimizing the amount of solvent loss due to heating.

In another embodiment, the extraction may be performed by cold extraction by immersion at 10 to 40° C. for 1 to 15 days after adding an extraction solvent to each extraction target. In this case, the manufacturing cost can be reduced by omitting the equipment required for heating and reducing the amount of energy used.

In another embodiment, the extraction may be performed for 1 to 200 hours at room temperature after adding an extraction solvent to each extraction target. In this case, the content of the active ingredient may be further increased.

In another embodiment, the extraction may be performed by adding an extraction solvent to each extraction target and then boiling at 50 to 120° C. for 30 minutes to 72 hours. In this case, the content of the active ingredient can be increased more efficiently.

In addition, each of the extracts may include an extract material extracted through a conventional purification process as well as an extract material extracted by the above-described extraction solvent. For example, the active fraction obtained through various additional purification methods, such as separation using an ultrafiltration membrane having a constant molecular weight cut-off value, may also be included in the extract of the present invention.

To prepare the complex extract in the cosmetic composition preparation method of one embodiment, after preparing each of the walnut shell extract and the persimmon extract, they may be mixed in a weight ratio of 1: 0.5 to 1: 2.

Preparing the complex extract in the cosmetic composition preparation method of another embodiment is to mix the dried walnut shell pulverized product and the dried persimmon stalk pulverized product in a weight ratio of 1: 0.5 to 1: 2, and then add an extraction solvent to extract. .

The manufacturing method of the cosmetic composition is after adding 0.001% to 60% by weight of the complex extract to a general cosmetic composition, lotion, serum, essence, lotion, cream, powder, foundation, pack, patch, mask sheet, gel It may further include formulating in any one formulation of an ointment, a spray, and a cleaning agent. The composition for cosmetic preparation acts together with the complex extract to realize a cosmetic effect according to the intended use and formulation of makeup, and is not particularly limited as long as it is a component used in general cosmetic subjects, excipients, etc., and specific examples are described above. Follow the same content as the description.

In addition, the formulation method is not particularly limited, and may be obtained through various methods known in the art to which the present invention pertains.

Hereinafter, the configuration and operation of the present invention will be described in more detail through preferred embodiments of the present invention. However, this is presented as a preferred example of the present invention and cannot be construed as limiting the present invention for any reason.

Content not described herein will be omitted because it can be technically inferred sufficiently by those skilled in the art to which the present invention pertains.

Preparation Example 1: Preparation of a complex extract of walnut shells and persimmons (weight ratio 1:1)

The shell was separated from the walnut fruit, and then completely dried and pulverized. After separating the stems from the persimmon fruit, they were completely dried and pulverized.

After mixing 15 g of the dried walnut shell pulverized product obtained as described above and 15 g of dried persimmon stalk obtained above, 20 times 70% ethanol based on the total weight of the mixture was added, and the complex extract was extracted by heating at 80° C. for 2 hours. prepared. After repeating the above process once more, the filtrate obtained by filtration through a 0.45 μm filter was concentrated under reduced pressure and freeze-dried.

Preparation Example 2: Preparation of a complex extract of walnut shells and persimmons (weight ratio 2:1)

After separating the hull from the walnut fruit, it was completely dried and pulverized. After separating the stem from the persimmon fruit, it was completely dried and ground.

After mixing 20 g of the pulverized dried walnut shells and 10 g of pulverized dried persimmons obtained as described above, 20 times 70% ethanol based on the total weight of the mixture was added, and the complex extract was extracted by heating at 80° C. for 2 hours. prepared. After repeating the above process once more, the filtrate obtained by filtration through a 0.45 μm filter was concentrated under reduced pressure and freeze-dried.

Preparation Example 3: Preparation of a complex extract (weight ratio 1:2) of walnut shells and persimmons

After separating the hull from the walnut fruit, it was completely dried and pulverized. After separating the stem from the persimmon fruit, it was completely dried and ground.

After mixing 10 g of the pulverized dried walnut shells and 20 g of pulverized dried persimmons obtained as above, 20 times 70% ethanol based on the total weight of the mixture was added, and the complex extract was extracted by heating at 80° C. for 2 hours. prepared. After repeating the above process once more, the filtrate obtained by filtration through a 0.45 μm filter was concentrated under reduced pressure and freeze-dried.

Comparative Preparation Example 1: Preparation of walnut shell extract

After separating the hull from the walnut fruit, it was completely dried and pulverized. 20 times 70% ethanol based on the total weight was added to 30 g of the dried walnut shell pulverized product obtained as described above, and a complex extract was prepared by heat extraction at 80° C. for 2 hours. After repeating the above process once more, the filtrate obtained by filtration through a 0.45 μm filter was concentrated under reduced pressure and freeze-dried.

Comparative Preparation Example 2: Preparation of persimmon extract

The stems were isolated and obtained from the persimmon fruit, and then completely dried and pulverized. 20 times 70% ethanol based on the total weight was added to 30 g of the dried persimmon pulverized product obtained as described above, and a complex extract was prepared by heat extraction at 80° C. for 2 hours. After repeating the above process once more, the filtrate obtained by filtration through a 0.45 μm filter was concentrated under reduced pressure and freeze-dried.

Comparative Preparation Example 3: Preparation of walnut egg extract

The skin was removed from the walnut fruit and the kernels were isolated and then completely dried and pulverized. 20 times 70% ethanol based on the total weight was added to 30 g of the dried walnut kernel pulverized product obtained as described above, and a complex extract was prepared by heat extraction at 80° C. for 2 hours. After repeating the above process once more, the filtrate obtained by filtration through a 0.45 μm filter was concentrated under reduced pressure and freeze-dried.

Comparative Preparation Example 4: Preparation of persimmon peel extract

The peel was separated from the persimmon fruit, and then completely dried and pulverized. 20 times 70% ethanol based on the total weight was added to 30 g of the dried persimmon peel pulverized product obtained as described above, and a complex extract was prepared by heat extraction at 80° C. for 2 hours. After repeating the above process once more, the filtrate obtained by filtration through a 0.45 μm filter was concentrated under reduced pressure and freeze-dried.

Example 1: Preparation of lotion formulation containing complex extract

30% by weight of the complex extract prepared in Preparation Example 1, 62.65% by weight of purified water, 5% by weight of butylene glycol, 1.5% by weight of 1,2-hexanediol, 0.5% by weight of allantoin, 0.05% by weight of panthenol and 0.3% by weight of betaine % was added to a stirrer, and then mixed to prepare a cosmetic composition of a lotion formulation.

Example 2: Preparation of serum formulation containing complex extract

50% by weight of the complex extract prepared in Preparation Example 1, 39.7% by weight of purified water, 7% by weight of butylene glycol, 1.5% by weight of 1,2-hexanediol, 1% by weight of glycerin, 0.5% by weight of betaine and 0.3% by weight of xanthan gum After the weight % was added to a stirrer, a cosmetic composition of a serum formulation was prepared by mixing.

Example 3: Preparation of cream formulation containing complex extract

55% by weight of the complex extract prepared in Preparation Example 1, 26.6% by weight of purified water, 3% by weight of glycerin, 6% by weight of butylene glycol, 0.5% by weight of betaine, 1.5% by weight of hydroxyethyl cellulose, 0.8% by weight of cetearyl alcohol %, sunflower oil 1.8% by weight, cetearyl olivate 0.75% by weight, sorbitan olivate 0.75% by weight, glyceryl stearate 2.5% by weight, ethylhexyl glycerin 0.5% by weight, and xanthan gum 0.3% by weight were added to a stirrer Then, by mixing, a cosmetic composition of a cream formulation was prepared.

Comparative Example 1

A cosmetic composition of a lotion formulation was prepared in the same manner as in Example 1, except that the complex extract prepared in Preparation Example 1 was replaced with the same amount of purified water.

Comparative Example 2

A cosmetic composition of a serum formulation was prepared in the same manner as in Example 2, except that the complex extract prepared in Preparation Example 1 was replaced with the same amount of purified water.

Comparative Example 3

A cosmetic composition of a cream formulation was prepared in the same manner as in Example 3, except that the complex extract prepared in Preparation Example 1 was replaced with the same amount of purified water.

Test Example 1: Toxicity evaluation test

DMSO was added to the extract samples according to Preparation Examples and Comparative Preparation Examples and diluted to the concentrations shown in Table 1 below, and the degree of cytotoxicity according to the concentration was evaluated.

Using DMEM medium (Dulbecco's Modified Eagle Medium) containing 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin, the mouse macrophage cell line RAW 264.7 was equally counted and dispensed at ^{1Х10 4 cells/ml in a 96-well plate.} After that, it was incubated for 24 hours at 37° C. and 5% carbon dioxide.

After mixing the samples according to Preparation Examples and Comparative Preparation Examples to the cultured cells with the medium for each concentration, 1 mL of each was added to each well and cultured at 37° C. and 5% carbon dioxide conditions for 24 hours. After incubation for 24 hours, only the supernatant of each well was taken separately, WST-1 assay solution (ez-cytox) was added to each well, reacted for 2 hours in an incubator, and absorbance was measured at 450 nm with an ELISA reader, and the following formula 1 to calculate the cell viability, and the results are shown in Table 1 below.

[Formula 1]

Cell viability (%) = [(absorbance of sample treated group / absorbance of untreated group) Х 100]

density
(μg/ml) Preparation Example 1 manufacturing example
2 Preparation 3 Comparative Preparation Example 1 Comparative Preparation Example 2 Comparative Preparation Example 3 Comparative Preparation Example 4 0 100.00 100.00
100.00
100.00 100.00
100.00
100.00
3.125 102.20 104.36 103.09 98.85 100.66 97.31 100.64 6.25 107.81 104.41 103.71 92.25 97.80 89.33 94.92 12.5 107.02 104.98 103.34 96.71 100.04 91.12 96.91 25 102.01 103.86 102.00 94.59 98.75 90.10 92.67 50 104.72 100.70 99.40 90.67 98.53 92.78 95.75 100 100.07 101.64 80.48 93.19 52.80 87.16 93.28

As can be seen in Table 1 and FIG. 1, Preparation Examples 1 to 3 of the complex extract of the present invention showed that the cell viability exceeded 100 at most concentrations tested, confirming that it can be safely used.

Test Example 2: Anti-inflammatory activity evaluation

DMSO was added to the extract samples according to Preparation Examples and Comparative Preparation Examples and diluted to the concentrations shown in Table 1 below, and the degree of cytotoxicity according to the concentration was evaluated.

Using DMEM medium (Dulbecco's Modified Eagle Medium) containing 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin, the mouse macrophage cell line RAW 264.7 was equally counted and dispensed at ^{1Υ10 4 cells/ml in a 96-well plate.} After incubation for 24 hours at 37 ℃, 5% carbon dioxide conditions.

After mixing the samples according to Preparation Examples and Comparative Preparation Examples to the cultured cells with a medium for each concentration, 1 mL of each was added to each well and cultured at 37° C. and 5% carbon dioxide condition for 24 hours.

At this time, LPS (Lipopolysaccharide), an inflammatory factor that expresses NO, was treated together at a concentration of 1 ug/ml and cultured at 37° C. and 5% carbon dioxide condition for 24 hours. Then, after taking only the culture solution from each well separately, 100 μl of the culture solution was respectively taken in a 96-well plate and 50 μl of Griess reagent A (N-1-Naphthylethylenediamine (NEDHC)) and 50 μl of Griess reagent B (Sulfanilamide) using NO assay kit. were added and reacted for 10 minutes, respectively, and absorbance was measured at 540 nm using an ELISA plate reader. The results are shown in Table 2 as a percentage based on 1 mM nitrite standard solution.

density
(μg/ml) Preparation Example 1 Preparation 2 Preparation 3 Comparative Preparation Example 1 Comparative Preparation Example 2 Comparative Preparation Example 3 Comparative Preparation Example 4 0 0.00 0.00 0.00 0.00 0.00 0.00 0.00 3.125 12.38 7.49 -0.86 4.32 1.15 -7.78 -8.06 6.25 8.35 5.18 12.10 3.46 2.59 -1.44 -3.74 12.5 14.69 14.69 10.94 11.52 10.08 -9.50 -4.90 25 24.48 26.79 20.45 15.26 18.15 2.02 -2.02 50 32.83 31.11 31.11 32.26 30.53 2.30 -4.03 100 45.22 39.75 33.00 46.95 52.42 8.64 5.18

As a result, as can be seen in Table 2 and Figure 2, the composite extract of the present invention showed the greatest NO production inhibitory effect when the two extracts of walnut shell and persimmon were mixed at a weight ratio of 1:1.

Test Example 3: Wrinkle improvement or elasticity improvement evaluation

DMSO was added to each sample of the extracts according to the Preparation Examples and Comparative Preparation Examples and diluted to the concentrations shown in Table 3 below, and the elastase inhibitory activity according to the concentrations was evaluated.

N-succinyl-(Ala)3-p-nitroanilide 1.6 mM concentration of N-succinyl-(Ala)3-p-nitroanilide dissolved in 10 mM Tris-HCl buffer (pH 8.0) by concentration of samples according to Preparation Examples and Comparative Preparation Examples and Elastase derived from porcine pancreas type IV 0.4 U/ml was mixed and reacted at 22° C. for 5 minutes, and then placed in a 96-well plate and absorbance was measured at 410 nm.

At this time, the sample without the sample was used as a control group and calculated using Equation 2 below, and the results are shown in Table 3 below. At this time, oleanolic acid was used as a positive control, and when the elastase inhibitory activity was 50%, the concentration of oleanolic acid was 9.37 μg/ml .

[Formula 2]

Elastase inhibitory activity (%) = 100 - [(absorbance of the group treated with the sample / absorbance of the group not treated with the sample) Х 100]

density
(μg/ml) Preparation Example 1 Preparation 2 Preparation 3 Comparative Preparation Example 1 Comparative Preparation Example 2 Comparative Preparation Example 3 Comparative Preparation Example 4 0 0.00 0.00 0.00 0.00 0.00 0.00 0.00 1.5625 17.03 9.18 13.95 5.3 9.01 4.91 -0.55 3.125 27.94 16.52 22.74 5.97 15.36 7.93 -1.11 6.25 34.83 21.26 35.52 12.09 20.68 21.71 4.52 12.5 54.40 42.13 50.06 17.76 37.88 48.75 -0.55 25 68.19 65.02 67.76 24.1 57.95 63.57 7.20 50 77.83 78.36 82.30 38.73 77.82 76.20 16.97 100 86.56 83.96 87.22 50 88.67 83.40 28.41 IC50 10.60 17.77 11.48 95.20 20.06 17.61 -

As can be seen in Table 3 and Figure 3, the complex extract of the present invention shows a large synergistic effect of elastase inhibitory activity compared to the case of using walnut shell extract or persimmon extract alone in all compounding ratios, in particular, 1 The highest activity was shown at a mixing ratio of :1. The above results mean the expression of wrinkle improvement or elasticity improvement effect when the complex extract of the present invention is applied to the skin.

Test Example 4: Measurement of the content of phenolic compounds

It is known that plant-derived natural phenolic compounds act on the skin, and have skin improvement effects by promoting cell regeneration by skin cell physiological activity, improving antioxidant capacity, reducing sebum amount, and reducing dead skin mass.

DMSO was added to each sample of the extracts according to Preparation Examples and Comparative Preparation Examples to measure the contents of phenols and flavonoids belonging to phenolic compounds, respectively.

For the total phenol content, 100 μl of Folin-Ciocalteu reagent was added to 100 μl of the sample, and after stabilization at room temperature for 5 minutes, 100 μl of 10% sodium carbonate was added, reacted at 30° C. for 1 hour, and absorbance was measured at 760 nm. The total phenol content of each sample was calculated from the standard calibration curve obtained by preparing gallic acid as a standard by concentration.

To measure the total flavonoid content, 1 ml of diethylene glycol was added to 100 μl of the sample, mixed well, 100 μl of 1N soduum hydroxide was added, reacted at 37° C. for 1 hour, and absorbance was measured at 420 nm. The total phenol content of each sample was calculated from the standard calibration curve obtained by preparing naringin for each concentration as a standard material.

Table 4 shows the content of the phenolic compound compared to each reference compound.

density Preparation Example 1 Preparation Example 2 Preparation 3 comparison
Preparation Example 1 comparison
Preparation 2 comparison
Preparation 3 comparison
Preparation 4 Gallic acid vs.
Total phenol content
(mg gallic acid/g) 42.434 40.486 41.239 35.919 39.534 27.814 3.906 Naringin contrast
Total tannin content
(mg·naringin/g) 35.211 34.947 33.576 27.954 32.998 15.976 3.087

As can be seen in Table 4, it was confirmed that Preparation Examples 1 to 3 of the composite extract of the present invention contained a large amount of phenolic compounds compared to Comparative Preparation Examples of the single extract. Among the preparation examples, Preparation Example 1 showed the highest phenolic compound and tannin content. From this, the complex extract of the present invention can have beneficial effects in improving skin conditions, such as increasing antioxidant capacity and lowering the amount of sebum and dead skin cells.

Test Example 5: Evaluation of skin improvement

When the serum of Example 2 containing the complex extract of the present invention and the serum of Comparative Example 2 without it were applied to the skin, the degree of change in skin moisture content, the degree of change in sebum amount, and the degree of change in the amount of dead skin cells were evaluated, and the feeling of use was evaluated. and sensory evaluation for efficacy. Each moisture level, sebum amount, and dead skin mass were evaluated using a wireless portable skin diagnostic device (product name: dpViso, manufacturer: CHOWIS) as values calculated according to the measurement index built into the device.

(1) Evaluation of the degree of change in moisture content

20 testers in their 20s and 30s were allowed to use the serum of Example 2 and the serum of Comparative Example 2 in the same amount in the morning and evening for 2 weeks, and then on the last day for 15 hours, the change in the moisture content of the skin was measured at the following time intervals. The average values of the measured values are shown in Table 5 and FIG. 4 .

hr 0 3 6 9 12 15 Example 2 100 63 55 54 49 48 Comparative Example 2 100 50 40 38 39 39

As a result, the serum containing the complex extract of the present invention maintained a significantly higher moisturizing rate for the entire period compared to the serum without it.

(2) sensory evaluation

20 testers in their 20s and 30s were asked to use the serum of Example 2 and the serum of Comparative Example 2 in the same amount, once in the morning and once in the evening for 10 days, respectively, and then applied according to the 5-point scale method, Sensory evaluation was performed on the degree of improvement and stimulation. The results are shown in Table 6 below by averaging the total scores.

Example 2 Comparative Example 2 spreadability 4.5 4.2 absorptivity 4.4 4.2 moisturizing power 3.8 3.6 degree of improvement 3.5 3.2 irritation 0 0

As shown in Table 6, it was confirmed that the serum containing the complex extract of the present invention was superior to the serum containing the same as compared to the serum.

Although the embodiments of the present invention have been described above, the present invention is not limited to the above embodiments, but may be implemented in various different forms, and those of ordinary skill in the art to which the present invention pertains will appreciate the technical spirit of the present invention. However, it will be understood that the invention may be embodied in other specific forms without changing essential features. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not restrictive.

Claims (8)

A cosmetic composition having a wrinkle improvement or elasticity improvement effect containing a complex extract of walnut shell and persimmon as an active ingredient, wherein the compounding weight ratio of the walnut shell extract and the persimmon extract of the complex extract is 1:1.
The cosmetic composition according to claim 1, wherein the composition further has at least one of an anti-inflammatory effect and a skin moisturizing effect.
delete The cosmetic composition according to claim 1, wherein the complex extract is contained in an amount of 0.001 wt% to 60.0 wt% with respect to the total weight of the cosmetic composition.
The cosmetic composition according to claim 1, wherein the cosmetic composition is one of lotion, serum, essence, lotion, cream, powder, foundation, pack, patch, mask sheet, gel ointment, spray, and cleaning agent.
walnut hull extract and persimmon extract in a weight ratio of 1:1, or by mixing the dried walnut hull extract and dried persimmon stalk extract in a weight ratio of 1:1 and then extracting to prepare a complex extract, A method of manufacturing a cosmetic composition having an effect of improving wrinkles or improving elasticity.
delete 7. The method of claim 6, wherein after adding a solvent of 1 to 100 times the weight of the dry raw material pulverized to be extracted, heat extraction, reflux cooling extraction, ultrasonic extraction, chilled extraction at a temperature range of 10 to 120° C. for 30 minutes to 360 hours. A method for producing a cosmetic composition, comprising performing at least one of extraction and percolation.

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