KR102107193B1 - Cosmetic composition comprising extract of calendula arvensis fermented by aureobasidium pullulans - Google Patents

Abstract

The present invention relates to a cosmetic composition containing a calendula black yeast fermentation broth fermented with calendula using a black yeast mycelium, specifically, to a cosmetic composition containing a calendula black yeast fermentation broth obtained with high yield of flavonoids among active ingredients. . According to the present invention, the antioxidant effect, skin moisturizing, skin barrier improvement effect is excellent.

Description

Cosmetic composition containing fermentation broth of black marigold {COSMETIC COMPOSITION COMPRISING EXTRACT OF CALENDULA ARVENSIS FERMENTED BY AUREOBASIDIUM PULLULANS}

The present invention relates to a cosmetic composition containing a calendula black yeast fermentation broth fermented with a black yeast mycelium, and more particularly, to a cosmetic composition excellent in antioxidant, skin moisturizing, and skin barrier improvement effects.

The goal of cosmetics is to maintain beautiful and healthy skin, and to delay the aging of naturally occurring skin with age. In general, aging is due to oxidative damage of biomaterials by free radicals. In other words, highly reactive free radicals such as free radicals, such as hydrogen peroxide, hydroxy radicals, and superoxide radicals, peroxidize the lipids that make up the cell membrane, destroying the cell membrane, oxidizing proteins, and attacking nucleic acids to cause oxidative damage to biomaterials Destroy the cells and eventually kill the cells. The most significant skin characteristics due to aging include wrinkles, decreased elasticity, and dull skin. In addition, in addition to wrinkles and reduced elasticity, after the 40s, the face color is often unclear and gray and dark. Particularly, in the case of young women, as they experience childbirth, blood bleeding occurs, and blood circulation disorders occur, resulting in a decrease in metabolic ability and skin roughness and dullness. When the metabolic ability decreases and the skin becomes rough, the amount of moisture is also easily reduced. In general, the amount of moisture in the skin is about 70% in the dermal layer, but decreases toward the epidermal layer, resulting in about 10-30% in the keratinocyte layer. It is known that in the keratinocyte layer, a substance having a hydrophilic water retention ability called natural moisturizing factor (NMF) exists and plays an important role in moisturizing the skin. In the normal keratinocyte layer, when moisture is maintained at about 10 to 30%, the skin is smooth and soft, and the body protects normally.

Therefore, it is very important to properly maintain the moisture content of the keratinocyte layer of the skin to prevent skin aging, but other natural moisturizing factors such as sodium pyrrolidone carboxylate (PCA-Na), sodium lactate, and urea are electrolytic. This strong, there is a problem that impairs the emulsion stability of the cosmetic. Amino acids, collagen, and elastin also have moisturizing abilities, but have limited moisturizing abilities. The ability to retain moisture in the stratum corneum can be controlled by a natural moisturizing factor (NMF) composed of sebum components, amino acids, lactic acid, urea and inorganic salts. In addition, the development of substances with high moisturizing effects is one of the important research tasks in cosmetics. In addition, substances known to have antioxidant activity include ascorbic acid, dl-α-tocopherol, beta-carotene, dibutylhydroxytoluene (BHT), etc. Since these materials are used as synthetic products, there is an urgent need for the development of new antioxidant cosmetics that can be applied to the skin more safely and usefully because they can cause skin safety problems when used for a long time or when used in large quantities.

Therefore, research on improving skin aging and moisturization is expected to continue, and cosmetics using natural products are being developed to solve these skin problems and reduce skin irritation caused by various chemicals. Natural ingredients have fewer side effects on the skin, and recently, as the consumer's response to cosmetics using natural ingredients has increased, the development value of cosmetics has increased.

To this end, studies are being actively conducted to search for the skin physiological activity of various natural plant components and to develop substances having excellent safety and excellent skin beauty effects.

Korean Patent Publication No. 10-2013-0026955 discloses a skin protective cosmetic composition, while using a natural herbal complex composition comprising distilled extracts of Eoseongcho, Angelica, wormwood, gold silver, green tea, gold purple, bellflower and black bean pulverized, have.

In addition, Korean Patent Registration No. 10-1114141 discloses a cosmetic composition that contains a daisy, a marigold, a safflower, a geranium, and a pansy flower extract together, which has a high antioxidant effect and an excellent skin texture improvement and elasticity promoting effect.

The object of the present invention is to reduce the risk of problems with existing cosmetic cosmetics, increase the content of useful active ingredients present in trace amounts in natural materials, and to provide a cosmetic composition having various physiologically active effects such as moisturizing skin and elasticity of the skin. Is to provide.

In order to achieve the object of the present invention, in the present invention, the marigold extract, which is a natural extract, is fermented with a black yeast mycelium, and contains a marigold black yeast fermentation solution with an increased flavonoid content as an active ingredient, for antioxidant, moisturizing, or skin. Provided is a cosmetic composition for improving barriers.

Preferably, the calendula black yeast fermentation broth can be obtained by fermenting the calendula extract with the black yeast for 1 to 7 days while maintaining pH 5 to 7 under a temperature condition of 20 to 40 ° C.

Also preferably, the marigold extract is selected from the group consisting of purified water, methanol, ethanol, glycerin, ethyl acetate, butylene glycol, propylene glycol, dichloromethane, chloroform, ethyl ether, butylene glycol, hexane, and mixtures thereof. It can be extracted using one or more solvents.

In addition, preferably, the calendula black yeast fermentation broth may be contained in an amount of 0.0001 to 30.0% by weight relative to the total weight of the cosmetic composition.

Also preferably, the cosmetic composition is a group consisting of solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation and spray It may have any one formulation selected from.

The cosmetic composition according to the present invention uses a natural extract, is harmless to the human body, has excellent safety, and contains anti-oxidation, moisturizing, and skin barrier improvement effects by containing the calendula black yeast fermentation broth with an increased flavonoid content.

All technical terms used in the present invention, unless defined otherwise, have the following definitions and conform to the meaning as commonly understood by those skilled in the art in the relevant field of the present invention. In addition, although a preferred method or sample is described herein, similar or equivalent ones are included in the scope of the present invention.

The term “about” means 30, 25, 20, 15, 10, 9, 8, 7, 6, 5, for reference amount, level, value, number, frequency, percent, dimension, size, amount, weight or length It means a quantity, level, value, number, frequency, percentage, dimension, size, quantity, weight or length that varies by 4, 3, 2 or 1%.

In the present invention, the terms “comprise” and “comprising”, unless the context requires otherwise, include the presented steps or components, or groups of steps or components, but any other steps or components, or steps Or it should be understood that it implies that the group of components is not excluded.

The present invention relates to a cosmetic composition containing an extract produced by fermenting marigold extract with a black yeast mycelium. More specifically, it provides a cosmetic composition containing the calendula black yeast fermentation extract having an increased component content of flavonoids by fermenting the marigold with a black yeast mycelium as an active ingredient.

Calendula ( Cendula arvensis ) is an asteraceae plant, grows up to 20-50 cm in height, and is a pale yellow to orange flower. Marigold has long been used as a medicine, tea, and spice, and its stability has been proven, and it is widely used to treat various diseases. In the old days, it was used as a substitute for antibiotics and hemostatic agents.

The marigold extract of the present invention is obtained by extracting from various organs or parts (eg leaves, branches, flowers, roots, stems, etc.) of the marigold tree, preferably an extract obtained from an outpost or a flower, most preferably a flower Means an extract obtained from

On the other hand, the marigold extract of the present invention can be prepared using conventional solvents, under conditions of temperature and pressure, according to conventional methods known in the art. That is, the extract of marigold is water, anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms, acetone, ethyl acetate, butyl acetate and 1,3-butylene glycol, propylene glycol, dichloromethane, chloroform, ethyl ether, butylene glycol, After extraction using a solvent selected from the group consisting of hexane and mixtures thereof, or after extraction of marigolds using the solvent, it may be prepared by mixing, but preferably, the solvent is preferably alcohol, more preferably 70% It is good to use ethanol. And the suitable amount of the extraction solvent is 1 to 15 times the dry weight of the marigold, more preferably 5 to 10 times, and most preferably 7 times.

On the other hand, the marigold extract of the present invention may be not only an extract by the above-described extraction method, but also an extract that has undergone a conventional purification process. Obtained through various purification methods additionally performed, such as separation using an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatography (made for separation according to size, charge, hydrophobicity or affinity) It is interpreted that the fraction is also included in the marigold extract of the present invention. In addition, the marigold extract of the present invention may be prepared in powder form by additional processes such as distillation under reduced pressure and freeze drying or spray drying.

Black yeast ( Aureobasidium pullulans ) is a microorganism used to make beer, wine, etc., and is a group of single-celled organisms that do not have hyphae and have no photosynthetic ability or motility, although they are molds or mushrooms. It is a microorganism that humans have used for food since 5,000 years ago, and the yeast itself is used as a cheap source of fat or protein. It contains a rich group of vitamin B and some contain vitamin D. In addition, black yeast is a microorganism that produces beta-glucan (β-Glucan). Black yeast beta-glucan (β-glucan) contains a large amount of components known to have physiological activities, such as anti-cancer and insulin secretion suppression.

And the calendula black yeast fermentation broth of the present invention means that the produced calendula extract is fermented with the black yeast ( Aureobasidium pullulans ) for 1 to 7 days while maintaining pH 5 to 7 under a temperature condition of 20 to 40 ° C.

According to a preferred embodiment of the present invention, the content of the calendula black yeast fermentation liquid added to the cosmetic composition is 0.0001 to 30.0% by weight, preferably 0.001 to 25.0% by weight, most preferably 0.1 to 0.1% based on the total weight of the cosmetic composition. 10.0% by weight. When the content of the calendula black yeast fermentation solution is less than 0.0001% by weight, the antioxidant, moisturizing, and skin barrier improvement effects are not exhibited, and when it exceeds 30% by weight, the degree of increase in the effect of increasing the content is minimal, and the safety and stability of the formulation There is a problem and it is not economical.

The cosmetic composition of the present invention can be prepared in any formulation conventionally prepared in the art, for example, solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing , Oil, powder foundation, emulsion foundation, wax foundation and spray, etc., and more specifically, flexible lotion, nutrition lotion, nutrition cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, It can be prepared in the form of cleansing water, packs, sprays or powders. In addition, the components included in the cosmetic composition of the present invention include ingredients commonly used in cosmetic compositions in addition to the active ingredients as active ingredients, such as stabilizers, solubilizers, vitamins, pigments and fragrances, and conventional auxiliaries. It includes.

When the formulation of the present invention is a paste, cream, or gel, animal oil, vegetable oil, wax, paraffin, starch, trakant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide, etc. may be used as a carrier component. You can.

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder can be used as a carrier component, and in the case of a spray, additionally chlorofluorohydrocarbon, propane / Propellant such as butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer or emulsifier is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid ester.

When the formulation of the present invention is a suspension, liquid diluents such as water, ethanol or propylene glycol as carrier components, suspensions such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters, microcrystals Sex cellulose, aluminum metahydroxide, bentonite, agar or trakant, etc. can be used.

When the formulation of the present invention is a surfactant-containing cleansing, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivatives, methyltaurate, sarcosinate, fatty acid amide as a carrier component Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters and the like can be used.

When the cosmetic composition of the present invention is a soap, a surfactant-containing cleansing agent, or a surfactant-free cleansing formulation, it may be wiped, peeled off, or washed with water after being applied to the skin. As a specific example, the soap is liquid soap, powdered soap, solid soap and oil soap, and the surfactant-containing cleansing formulation is a cleansing foam, cleansing water, a cleansing towel and a cleansing pack, and the surfactant-free cleansing formulation is a cleansing cream , Cleansing lotions, cleansing water and cleansing gels, but are not limited thereto.

Hereinafter, the present invention will be described in more detail through manufacturing examples. These manufacturing examples are only for explaining the present invention in more detail, and according to the gist of the present invention, the scope of the present invention is not limited by these manufacturing examples to those skilled in the art to which the present invention pertains. It will be obvious.

< Manufacturing example  1> Preparation of marigold solvent extract

The marigold was washed with purified water, dried, and then pulverized, then finely sieved using a 100-mesh sieve. After adding reflux 3 times for 5 hours by adding 70% (V / V) ethanol to 100 g / L of the calendula pulverized product, and then cooling it, filter with Whatman # 3 filter paper and concentrate under reduced pressure at 50 ° C or less. And lyophilized. The lyophilized product was dissolved to 2% (W / V) using 50% glycerin to prepare a marigold solvent extract.

< Manufacturing example  2> Preparation of marigold fermentation extract

As a result of fermentation of calendula and extract, the calendula was screened as a sample, washed sufficiently, and then finely pulverized to pass through a 300 mesh sieve. This was added to the culture solution of the genus Saccharomyces (Saccharomyces cerevisia KCTC 7904) strain to be 20 g / L. Glucose 2% (W / V) was added as a carbon source, and peptone 0.5% (W / V) was additionally added to culture. Cultivation was carried out using a 5L fermenter, and maintained at pH 5-7 at 37 ° C, preferably at pH 6, and cultured for 1 to 7 days, preferably 5 days. After the culture, the culture medium was centrifuged to remove the culture bacteria first.

After the cultured bacteria were first removed, the marigold fermentation product was added with ethanol so as to be a final 70% (V / V) ethanol aqueous solution, refluxed 3 times for 5 hours, cooled, and then filtered with Whatman # 2 filter paper. . The filtered extract was transferred to a concentration tank to concentrate the filtered extract under reduced pressure and freeze-dried. This powder was dissolved to 2% (W / V) using 50% glycerin to prepare a marigold fermentation extract.

< Manufacturing example  3> Marigold Black yeast  Preparation of fermentation broth

To 5 g of the marigold solvent extract prepared in Preparation Example 1, glucose 2% (W / V) was added as a carbon source, and peptone 0.5% (W / V) was further added to sterilize. Here, 20g / L of black yeast ( Aureobasidium pullulans ) strain culture was inoculated. The culture was maintained for 7 days at 37 ° C and pH 6 using a 5L fermenter. After the culture, the culture solution was centrifuged to remove the culture bacteria first, and sterilized to prevent further culture (121 ° C, 15 minutes, 1.5 atmosphere).

After fermentation, the calendula fermentation product from which the cultured bacteria were first removed is added to ethanol to make a final 70% (V / V) ethanol aqueous solution, refluxed 3 times for 5 hours, cooled, and filtered with Whatman # 3 filter paper Did. When the filtration was completed, the extract obtained in the extraction step was transferred to a concentration tank and concentrated under reduced pressure or freeze-dried at 50 ° C or lower. The freeze-dried product was dissolved to 2% (W / V) using 50% glycerin to prepare a calendula black yeast fermentation broth.

< Manufacturing example  4> Marigold Black yeast  Preparation of fermentation broth

A marigold solvent extract was prepared in the same manner as in Preparation Example 1, except that purified water was used instead of 70% (V / V) ethanol. To 5 g of the marigold solvent extract, 2% (W / V) of glucose was added as a carbon source, and 0.5% (W / V) of peptone was further added to sterilize. Here, 20g / L of black yeast ( Aureobasidium pullulans ) strain culture was inoculated. The culture was maintained for 7 days at 37 ° C and pH 6 using a 5L fermenter. After the culture, the culture solution was centrifuged, and the culture bacteria were first removed and sterilized (121 ° C, 15 minutes, and 1.5 atmospheres) to prevent further culture.

After fermentation, purified water was added to the marigold fermentation product after the culture was first removed, reflux was extracted three times for 5 hours, and then cooled, followed by filtration with Whatman # 3 filter paper. When the filtration was completed, the extract obtained in the extraction step was transferred to a concentration tank and concentrated under reduced pressure or freeze-dried at 50 ° C or lower. The freeze-dried product was dissolved to 2% (W / V) using 50% glycerin to prepare a calendula black yeast fermentation broth.

< Manufacturing example  5> Marigold Black yeast  Preparation of fermentation broth

A marigold solvent extract was prepared in the same manner as in Preparation Example 1, except that 50% (V / V) ethyl acetate was used instead of 70% (V / V) ethanol. To 5 g of the marigold solvent extract, 2% (W / V) of glucose was added as a carbon source, and 0.5% (W / V) of peptone was further added to sterilize. Here, 20g / L of black yeast ( Aureobasidium pullulans ) strain culture was inoculated. The culture was maintained for 7 days at 37 ° C and pH 6 using a 5L fermenter. After the culture, the culture solution was centrifuged, and the culture bacteria were first removed and sterilized (121 ° C, 15 minutes, and 1.5 atmospheres) to prevent further culture.

After fermentation, the marigold fermentation product from which the cultured bacteria were first removed is refluxed 3 times for 5 hours by adding ethyl acetate so as to be a final 50% (V / V) ethyl acetate aqueous solution, cooled and then cooled, whatman # 3 filter paper Filtered with. When the filtration was completed, the extract obtained in the extraction step was transferred to a concentration tank and concentrated under reduced pressure or freeze-dried at 50 ° C or lower. The freeze-dried product was dissolved to 2% (W / V) using 50% glycerin to prepare a calendula black yeast fermentation broth.

< Formulation example  1 to 3>

Formulation examples of the cream in the cosmetic composition containing the calendula black yeast fermentation solution (Preparation Examples 3 to 5) are shown in Table 1 below.

Ingredient (unit: weight%) Formulation Example 1 Formulation Example 2 Formulation Example 3 Calendula black yeast fermentation broth (Production Example 3) 5.0 - - Calendula black yeast fermentation broth (Production Example 4) - 5.0 - Calendula black yeast fermentation broth (Production Example 5) - - 5.0 glycerin 8.0 8.0 8.0 Sodium hyaluronate 3.0 3.0 3.0 Polysorbate 60 1.5 1.5 1.5 Mineral oil 3.0 3.0 3.0 Squalane 5.0 5.0 5.0 Triethanolamine 1.0 1.0 1.0 Lipophilic monostearin glycerin 2.0 2.0 2.0 Cetearyl alcohol 2.2 2.2 2.2 Stearic Acid 1.5 1.5 1.5 Sorbitan stearate 0.6 0.6 0.6 Sodium magnesium silicate 0.1 0.1 0.1 Betaine 3.0 3.0 3.0 antiseptic a very small amount a very small amount a very small amount incense a very small amount a very small amount a very small amount Coloring a very small amount a very small amount a very small amount Purified water Balance Balance Balance system 100 100 100

< Comparative Formulation Example  1>

Comparative formulation example 1 of the cream containing no marigold fermentation broth was shown in Table 2 below.

Ingredient (unit: weight%) Comparative Formulation Example 1 glycerin 8.0 Sodium hyaluronate 3.0 Polysorbate 60 1.5 Mineral oil 3.0 Squalane 5.0 Triethanolamine 1.0 Lipophilic monostearin glycerin 2.0 Cetearyl alcohol 2.2 Stearic Acid 1.5 Sorbitan stearate 0.6 Sodium magnesium silicate 0.1 Betaine 3.0 antiseptic a very small amount incense a very small amount Coloring a very small amount Purified water Balance system 100

< Comparative Formulation Example  2>

Comparative formulation example 2 of the cream in the cosmetic composition containing the marigold fermentation extract (Production Example 2) is shown in Table 3 below.

Ingredient (unit: weight%) Comparative Formulation Example 2 Marigold fermentation extract (Production Example 2) 5.0 Sodium hyaluronate 8.0 Polysorbate 60 3.0 Mineral oil 1.5 Squalane 3.0 Triethanolamine 5.0 Lipophilic monostearin glycerin 1.0 Cetearyl alcohol 2.0 Stearic Acid 2.2 Sorbitan stearate 1.5 Sodium magnesium silicate 0.6 Betaine 0.1 antiseptic 3.0 incense a very small amount Coloring a very small amount Purified water Balance system 100

<Comparison Formulation example  3>

Comparative formulation example 2 of the cream in the cosmetic composition containing the marigold extract (Preparation example 1) is shown in Table 4 below.

ingredient Content (unit:% by weight) Marigold extract (Production Example 1) 5.0 Sodium hyaluronate 8.0 Polysorbate 60 3.0 Mineral oil 1.5 Squalane 3.0 Triethanolamine 5.0 Lipophilic monostearin glycerin 1.0 Cetearyl alcohol 2.0 Stearic Acid 2.2 Sorbitan stearate 1.5 Sodium magnesium silicate 0.1 Betaine 3.0 antiseptic a very small amount incense a very small amount Coloring a very small amount Purified water Balance system 100

< Experimental example  1> Active ingredient of marigold ( Triterpenoid  Measurement of content)

The content of the material of each phenolic compound of the marigolds of Preparation Examples 1 to 3 was analyzed using HPLC as shown in Table 5.

Time (min) water Acetonitrile 0 80 20 5 80 20 25 72 28 35 67 33 40 59 41 45 20 80 47 20 80 50 80 20

As for the analysis column, the optipack C18, the column temperature 30 ° C, and the UV detector absorbance wavelength were analyzed at 203 nm, and the results are shown in Table 6 below.

Preparation Example 1 Preparation Example 2 Preparation Example 3 Preparation Example 4 Preparation Example 5 Flavonoids (% content) 3.21 3.42 5.85 3.75 4.21

As a result of measuring the active ingredient content of the calendula of Preparation Examples 1 to 5, the flavonoid components of the calendula black yeast fermentation broth (Preparation Examples 3 and 5) extracted by fermenting the calendula of the present invention using black yeast mycelium have different extraction conditions (Preparation Example) Compared to 1 to 2), it was confirmed that it increased significantly.

< Experimental example  2> Antioxidant effect measurement

The antioxidant activity of the extracts prepared in Preparation Examples 1 to 3 of the present invention was measured by a thiobarbituric acid method (TBA assay) using a lipid peroxidation system by a Fenton reaction. That is, 10 μl of ethyl linoleate, 0.2% sodium dodecyl sulfoxide (SDS), 10 μmol of ferrous chloride, and 2 μmol of hydrogen peroxide were prepared in Preparation Examples 1 to 3 in 5 ml of 25 mM Trizma hydrochloride / 0.75 mM potassium chloride buffer. After adding dimethylsulfoxide (DMSO) test solution of one extract and incubating at 55 ° C. for 16 hours, the reaction was stopped by adding 50 μl of 4% dibutylhydroxytoluene (BHT). 0.3 ml of this solution was placed in a test tube, and 3.0 ml of 0.05 N hydrochloric acid and 0.67% thiobarbituric acid (TBA) solution were added and boiled for 30 minutes. After cooling the reaction solution, 4.0 ml of 85% butanol was added, and after centrifugation, the absorbance of the butanol layer was measured at 535 nm, and the antioxidant effect of each extract was calculated by the following equation (1). Table 7 shows the results.

Test substance Antioxidative effect(%) Experiment concentration (10㎍ / ml) Experimental concentration (50㎍ / ml) Experimental concentration (100㎍ / ml) Preparation Example 1 25 52 58 Preparation Example 2 30 58 64 Preparation Example 3 46 72 88 Preparation Example 4 33 60 65 Preparation Example 5 38 66 74

Antioxidant effects of the calendula extracts of Preparation Examples 1 to 5 showed mostly high activity, and in particular, it can be seen that the antioxidant effect of Preparation Example 3 according to the present invention is very excellent.

In addition, the calendula extract of Preparation Examples 1 to 5 and the existing antioxidants ascorbic acid (ascorbic acid), D-alpha-tocopherol (dl-α-tocopherol), beta-carotene (β-carotene) and dibutyl hydroxyl toluene (BHT) The antioxidant activity of the was measured in the same manner as in Experimental Example 2 to compare IC ₅₀ , which is a 50% inhibitory concentration. The results are shown in Table 8 below.

Test substance IC ₅₀ (10 μg / ml) Preparation Example 1 67.3 Preparation Example 2 51.5 Preparation Example 3 12.8 Preparation Example 4 45.2 Preparation Example 5 29.7 Ascorbic acid 21.90 Dl-a-tocopherol 3.36 Beta-carotene (β-carotene) 29.8 Dibutyl hydroxyl toluene (BHT) <1

The marigold extracts of Preparation Examples 1 to 5 showed high activity comparable to other antioxidants, and it can be seen that Preparation Example 3 particularly exhibits high activity. In particular, it can be seen that these marigold extracts have significant antioxidant activity despite being a natural mixture.

< Experimental example  3> Marigold Black yeast  Evaluation of the effect of fermentation broth on improving skin moisturization

The anti-wrinkle effect of the calendula black yeast fermentation broth cream (Formulation Example 1) and Comparative Formulation Examples 1 to 3 was evaluated as follows. 25 people in their 20s to 40s who do not have skin disorders are divided into two groups of 20 people per group, and the nutrition cream of the creams of the above formulation examples 1 and comparative formulations 1 to 3 for each group is faced twice a day for 1 month and It was made to apply to the forearm. Before starting application, skin conductivity was measured using a corneometer (CM820 courage Khazaka electronic GmbH, Germany) at constant temperature and humidity conditions (24 ° C, humidity 40%), and used as a basic value.After 1, 2, and 4 weeks The rate of increase in skin conductivity (%) was measured to evaluate the rate of increase. The results are shown in Table 9 below.

Experimental product 1 week later After two weeks 3 weeks later Formulation Example 1 45 50 54 Formulation Example 2 33 35 40 Formulation Example 3 35 38 42 Comparative Formulation Example 1 17 20 25 Comparative Formulation Example 2 31 34 37 Comparative Formulation Example 3 19 26 32

As shown in Table 9, in the case of the cream containing the marigold calendula fermentation broth (Formulation Examples 1 to 3), the skin conductivity increase rate was very excellent compared to Comparative Formulation Examples 1 to 3. In general, since the skin conductivity is proportional to the amount of moisture in the skin, the above results confirm that the cosmetic composition containing the calendula black yeast fermentation liquid maintains the skin moisture content higher than that of the other cosmetic composition. Skin irritation was not observed in most of the subjects who applied the cosmetic to the skin.

< Experimental example  4> Marigold Black yeast  Evaluation of the effect of fermentation broth on improving the skin barrier

In order to investigate the effect of improving the skin barrier of the cosmetic composition containing the calendula black yeast fermentation broth of the present invention, an experiment was conducted as follows. This experimental example was evaluated by performing a comparative experiment using Tewameterr (TM300, Courage and Khazaka Electronic Co., Germany) for the transdermal water loss (TEWL) for the cosmetic of formulation example 1 for humans. For 40 women in their 20s and 40s, apply each sample to the right and left faces, and apply the percutaneous moisture loss of 1 hour, 2 hours, 4 hours, and 6 hours before and after applying the area 1 cm below the right side 3 cm from the eye area. Tewameterr (TM300, Courage and Khazaka Electronic Co., Germany) was measured 3 times to evaluate the percutaneous moisture loss using the average value. The results are shown in Table 10 below.

division 1 hours 2 hours 4 hours 6 hours Formulation Example 1 14.9 13.9 12.1 8.9 Formulation Example 2 16.8 15.7 13.9 11.1 Formulation Example 3 16.2 15.4 13.2 10.7 Comparative Formulation Example 1 17.4 15.7 13.8 11.4 Comparative Formulation Example 2 19.9 17.5 15.8 14.6

As shown in Table 10 above, as a result of measuring the amount of transdermal moisture loss, it was found that the amount of transdermal moisture loss in the facial skin of the experimenter who applied the cream containing the calendula black yeast fermentation liquid had a better effect on improving the skin barrier. Could know.

Since the specific parts of the present invention have been described in detail above, it is clear that for those skilled in the art, this specific technology is only a preferred embodiment, and the scope of the present invention is not limited thereto. Therefore, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

Claims (7)

The calendula black yeast fermentation broth obtained by fermenting it with black yeast for 1 to 7 days while maintaining the pH 5 to 7 under the temperature condition of 20 to 40 ° C is contained in an amount of 0.0001 to 30.0% by weight relative to the total weight of the cosmetic composition. Antioxidant cosmetic composition. The calendula black yeast fermentation broth obtained by fermenting it with black yeast for 1 to 7 days while maintaining the pH 5 to 7 under the temperature condition of 20 to 40 ° C is contained in an amount of 0.0001 to 30.0% by weight relative to the total weight of the cosmetic composition. A cosmetic composition for moisturizing the skin. The calendula black yeast fermentation broth obtained by fermenting it with black yeast for 1 to 7 days while maintaining the pH 5 to 7 under the temperature condition of 20 to 40 ° C is contained in an amount of 0.0001 to 30.0% by weight relative to the total weight of the cosmetic composition. Cosmetic composition for improving skin barrier. The method according to any one of claims 1 to 3,
The marigold extract is purified water, methanol, ethanol, glycerin, ethyl acetate, butylene glycol, propylene glycol, dichloromethane, chloroform, ethyl ether, butylene glycol, hexane and one or more solvents selected from the group consisting of mixtures thereof Extracted using, cosmetic composition. The method according to any one of claims 1 to 3,
The cosmetic composition is any one selected from the group consisting of solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation and spray The cosmetic composition having a formulation of. delete delete