KR102308756B1 - Cosmetic Composition Containing Mixture Extracts of Citrus Unshiu Peel Extract and Andrographis Paniculata As Active Ingrediend - Google Patents

Abstract

The present invention relates to a cosmetic composition containing a mixed extract of Citrus unshiu peel and Andrographis paniculata. More specifically, the present invention relates to a cosmetic composition comprising a mixed extract of Citrus unshiu peel and Andrographis paniculata, which is highly effective in enhancing the moisture of the skin, alleviating skin irritation, and improving wrinkles in the skin. The cosmetic composition of the present invention can be extensively used for the purpose of protecting the skin while alleviating various skin troubles caused by the environmental pollution, stress, and the like.

Description

Cosmetic Composition Containing Mixture Extracts of Citrus Unshiu Peel Extract and Andrographis Paniculata As Active Ingrediend}

The present invention relates to a cosmetic composition containing a mixed extract of tangerine peel and Calmeg as an active ingredient, and more particularly, to a cosmetic composition having effects of improving skin wrinkles, moisturizing the skin, and alleviating skin irritation.

Human skin experiences various skin troubles such as dullness, sensitivity, and deterioration of moisture retention due to environmental changes, internal and external stress, and the like. In particular, the skin characteristics that appear the most due to aging are dryness of the skin and loss of skin elasticity, the generation of wrinkles increases, and the skin becomes dull. In addition, it is easily sensitive to weak external stimuli and it takes a long time to recover.

Therefore, many efforts are being made to solve these skin troubles, and as a representative method, natural plant ingredients and stimulants are used to solve the problem, and the cosmetic industry uses these methods a lot. Cosmetics keep the skin beautiful and healthy, and are mainly used to relieve skin troubles and dry skin caused by aging. Recent cosmetic market trends are trying to develop materials using fermented products generated by fermentation along with basic characteristics of natural products by fermenting natural products in addition to using extracts of natural fruits.

Korean Patent No. 10-1814498 discloses a cosmetic composition containing a mixed extract of eucalyptus leaves and red shamrock leaves to have skin protection effects such as skin moisturizing enhancement and skin irritation relief.

The present invention has excellent moisturizing ability than the prior art among several natural plant extracts, and while searching for a substance that helps to alleviate skin troubles, a mixed extract of tangerine peel and Calmeg has a moisturizing effect, skin wrinkle improvement effect and skin stress. It was confirmed that it can help to alleviate skin irritation caused by, and thus completed the present invention.

Accordingly, it is an object of the present invention to provide a cosmetic composition containing a mixed extract of tangerine peel and Calmeg to improve various skin troubles, improve skin wrinkles, and moisturize the skin.

The above object is achieved by a cosmetic composition for improving skin wrinkles comprising a mixed extract of tangerine peel and Calmeg as an active ingredient.

In addition, it provides a cosmetic composition for moisturizing the skin comprising a mixed extract of tangerine peel and Calmeg as an active ingredient.

In addition, it provides a cosmetic composition for alleviating skin irritation comprising a mixed extract of tangerine peel and Calmeg as an active ingredient.

Preferably, the mixed extract contains 0.0001 to 30% by weight based on the total weight of the cosmetic.

Preferably, the mixed extract may be extracted by an extraction method selected from the group consisting of a solvent extraction method, a fermentation extraction method, an ultra-high pressure treatment extraction method, and an ultra-high pressure extraction method after fermentation.

The mixed extract of tangerine peel and Calmeg of the present invention has an excellent skin irritation alleviation effect, and excellent anti-wrinkle effect due to the excellent effect of inhibiting MMP-1 production. In addition, the composition of the present invention containing the mixed extract of tangerine peel and the mixed extract of Calmeg has excellent skin moisturizing effect. Therefore, the cosmetic composition of the present invention can be used extensively as a cosmetic that protects the skin and improves various skin troubles caused by environmental pollution, stress, and the like.

All technical terms used in the present invention, unless otherwise defined, have the following definitions and have the meanings as commonly understood by one of ordinary skill in the art in the relevant field of the present invention. In addition, although preferred methods and samples are described herein, similar or equivalent ones are also included in the scope of the present invention. The contents of all publications herein incorporated by reference are incorporated herein by reference. The term about means 30, 25, 20, 15, 10, 9, 8, 7, 6, 5, 4, means an amount, level, value, number, frequency, percentage, dimension, size, amount, weight or length varying by 3, 2 or 1%.

Throughout this specification, unless the context requires otherwise, the terms include and comprising includes a given step or component, or group of steps or components, but any other step or component, or step or component. It should be understood to imply that a group of these is not excluded.

The present invention relates to a cosmetic composition containing a mixed extract of tangerine peel and Calmeg, and the cosmetic composition according to the present invention is excellent in skin moisturizing enhancement, skin irritation relief and skin wrinkle improvement effects.

Citrus unshiu is an evergreen tree belonging to the genus Unhyang and Citrus. It is native to Jeju Island in Korea and is known to be distributed in Japan, China, Central and South America, and the Black Sea. The fruit of the tangerine tree is not only edible by itself, but also consumes more than 20% of processed products such as juice and canned food, and has abundant juice, unique flavor and high functional ingredients compared to other fruits. For example, tangerines contain essential oils such as d-limonene, linalool, linalylacetate, and flavonoids hesperidin, naringin, and porcirin. ), nobiletin, alkaloid synephrine, pectin, ascorbic acid, beta-cryptoxanthin, coumarin, etc. They are known to have digestive stimulation, obesity, cough, increase blood pressure, anti-allergy, and antibacterial action.

Calmeg (Andrographis paniculata) is an annual plant of the family Acanthaceae that grows mainly in shady, moist areas throughout India, especially in southern fields and forests. The height is 0.3-1.0 meters, the stem is dark green, 2-6mm thick, the leaves are 8cm long and 2.5cm wide, willow leaf-shaped, pointed, and the flowers have purple spots on white or light pink petals.

The mixed extract of tangerine peel and Calmeg used as an active ingredient in the composition of the present invention refers to an extract obtained by mixing extracts extracted from each of tangerine peel and Calmeg, or an extract extracted after mixing tangerine peel and Calmeg.

In addition, the mixed extract of tangerine peel and Calmeg may be contained in an amount of preferably 0.0001 to 30% by weight, more preferably 0.001 to 20% by weight, and most preferably 0.01 to 10% by weight based on the total weight of the cosmetic composition. % may be included. When the content of the extract is less than 0.0001% by weight, the skin improvement effect does not appear, and when it exceeds 30% by weight, the degree of increase in the effect of increasing the content is insignificant, there is a problem in safety and stability of the formulation, and it is economical also not desirable

In the present invention, the mixed extract may include tangerine peel and Calmeg in a weight ratio of 1 to 3:1 to 3, preferably in a weight ratio of 1:1.

Among the extraction methods of tangerine peel and calmeg of the present invention, pulverized extraction refers to pulverizing tangerine peel and calmeg with a fine grinder and filtering the tangerine peel and calmeg through a sieve. At this time, it is preferable to use a pulverized product filtered through a sieve of 300 mesh or less.

In addition, solvent extraction in the extraction method of tangerine peel and Calmeg of the present invention may be prepared using a conventional solvent according to a conventional method known in the art. Typical solvents include, for example, water, anhydrous or hydrous lower alcohols having 1 to 4 carbon atoms (e.g., methanol, ethanol, propanol and butanol), propylene glycol, 1.3-butylene glycol, glycerin, acetone, diethyl ether, ethyl It refers to extraction with a solvent selected from the group consisting of acetate, butyl acetate, dichloromethane, chloroform, hexane, and mixtures thereof.

In addition, the fermentation extraction in the extraction method of mandarin orange peel and Calmeg of the present invention includes an extract obtained by fermenting mandarin orange peel and Calmeg. At this time, the fermentation extraction is performed by fermenting orange peel and Calmeg with a bacteria selected from the group consisting of yeast, lactic acid bacteria, Bacillus genus and mixtures thereof, maintaining pH 5 to 7 under a temperature condition of 20 to 40 ° C., and fermenting for 1 to 7 days. It means that the extract is carried out, preferably, the bacterium is Saccharomyces cerevisisae, and most preferably, it is Casaromyces cerevisiae KCTC 7904.

In addition, in the extraction method of the mixed extract of mandarin orange peel and Calmeg of the present invention, ultra-high pressure extraction includes an extract obtained by ultra-high pressure treatment of mandarin orange peel and Calmeg. At this time, ultra-high pressure treatment means processing for 1 to 24 hours at a temperature of 40 to 90 ℃ at a pressure of 100 to 3000 MPa, preferably at a pressure of 500 to 1000 MPa at a temperature of 40 to 60 ℃ and 12 to 24 time treatment, and most preferably means treatment at a pressure of 1000 MPa at a temperature of 50° C. and for 24 hours.

In addition, when the mixed extract of tangerine peel and Calmeg of the present invention is extracted by mixing and applying the above-described extraction method, it is possible to obtain an extract more effective in improving skin wrinkles, improving skin moisture, and improving skin troubles. For example, the mixed extract of the present invention is obtained by fermenting the pulverized product of finely pulverized orange peel and Calmeg with bacteria selected from the group consisting of yeast, lactic acid, Bacillus and other mixtures, and then at a pressure of 100 to 3000 MPa. It also includes the ultra-high pressure treatment extract after fermentation of mandarin peel and Calmeg obtained by treating the step of ultra-high pressure extraction at a temperature of 40 to 90° C. for 1 to 24 hours.

In addition, the mixed extract of tangerine peel and Calmeg of the present invention is subjected to filtration, organic solvent reflux extraction, reduced pressure concentration, freeze-drying, etc. to control the concentration of the extract extracted by the above extraction method and increase the concentration of active ingredients. may be It is obvious in the art that the extracts obtained by filtering, purifying and concentrating the extract by the above-described extraction method are also included in the present invention.

Meanwhile, the mixed extract of tangerine peel and Calmeg of the present invention may be extracted using other extraction methods as well as the above-described extraction method.

According to a preferred embodiment of the present invention, the mixed extract of tangerine peel and Calmeg is prepared by the following method. First, the tangerine peel and Calmeg are thoroughly washed and then pulverized, and then finely made using 300 mesh. It is added to the culture medium of the saccharomyces genus (Saccharomyces cerevisiae KCTC 7904) strain of the tangerine peel and Calmeg so that this is 100g/L. Here, 1-4% of glucose (preferably 1-2%) as a carbon source and 0.1-1% (preferably 0.2-0.5%) of peptone as a nitrogen source were added and cultured. The culture was cultured for 1 to 7 days while maintaining the pH at 5-7 under a temperature condition of 20 to 40 °C using a 5L fermenter. After culturing, the culture medium was centrifuged to remove the culture medium. The culture solution of tangerine peel and Calmeg from which the culture has been removed is extracted by ultra-high pressure treatment at a pressure of 100 to 3000 MPa at a temperature of 40 to 90° C. for 1 to 24 hours, and preferably 40 to 60° C. at a pressure of 500 to 1000 MPa. It is to be treated at a temperature of 12 to 24 hours, most preferably at a pressure of 1000 MPa at a temperature of 50 ° C. for 24 hours. Mandarin peel and Calmeg treated with ultra-high pressure are extracted with reflux three times for 5 hours by adding 70% (V/V) aqueous ethanol solution, cooled, and filtered with Whatman #2 filter paper. When the filtration is completed, in the present invention, the extract obtained in the extraction step is transferred to a concentration tank, concentrated under reduced pressure, and freeze-dried. Concentration under reduced pressure was performed using a rotary vacuum concentrator, and the concentration condition was to put 700 ml of extract in a 1,000 ml round flask and concentrated under reduced pressure for 2 hours while rotating at 50 to 70 ° C and 600 to 1200 rpm at a pressure of 650 mmHg to 750 mmHg. The concentrated solution concentrated under reduced pressure was subjected to pre-freezing, the temperature was lowered to -40°C, maintained at -40°C for 2 to 4 hours, then dropped to -80°C through trap freezing, and the vacuum was maintained at 10-30 mTorr. Reheat to -40°C over 120 minutes. After that, maintain at -20°C for 240 minutes, at 0°C for 40 minutes, at 15°C for 340 minutes, at 30°C for 130 minutes, and then at 1300 minutes until completely dry.

Ingredients included in the cosmetic composition of the present invention include ingredients commonly used in cosmetic compositions in addition to the active ingredient as an active ingredient, for example, conventional adjuvants such as antioxidants, stabilizers, solubilizers, vitamins, pigments and fragrances; and a carrier.

The cosmetic composition of the present invention may be prepared in any formulation conventionally prepared in the art, for example, a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing agent , oil, powder foundation, emulsion foundation, wax foundation, spray, etc., but is not limited thereto. More specifically, it may be prepared in the form of a flexible lotion (skin), a nourishing lotion (milk lotion), a nourishing cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray, or a powder. .

When the cosmetic composition of the present invention is a soap, surfactant-containing cleansing or surfactant-free cleansing formulation, it may be applied to the skin and then wiped off, removed, or washed off with water. As a specific example, the soap is liquid soap, powder soap, solid soap, and oil soap, the surfactant-containing cleansing formulation is a cleansing foam, cleansing water, cleansing towel, and cleansing pack, and the surfactant-free cleansing formulation is a cleansing cream , cleansing lotion, cleansing water, and cleansing gel, but not limited thereto.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention in more detail, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention. .

Example 1: Preparation of solvent extracts of orange peel and Calmeg

500g of tangerine peel and 500g of Calmeg were selected as samples, washed thoroughly, and then pulverized and passed through a 300-mesh sieve to make the pulverized product fine. 70% (V/V) aqueous ethanol solution was added thereto, extracted under reflux 3 times for 5 hours, cooled and filtered with Whatman #2 filter paper. The filtered extract was concentrated under reduced pressure and then freeze-dried. This powder was dissolved to 2% (W/V) using 50% glycerin to prepare a solvent extract of orange peel and Calmeg.

Example 2: Preparation of fermented extract of Tangerine Peel and Calmeg

500g of tangerine peel and 500g of Calmeg were selected as samples, washed thoroughly, and the pulverized product was made fine so that it could pass through a 300-mesh sieve. Tangerine peel and Calmeg were added to the culture medium of the Saccharomyces genus (Saccharomyces cerevisiae KCTC 7904) strain to be 10g/L in these. Here, 1 to 4% of glucose is used as a carbon source, preferably 2% (W/V) is added, and 0.1 to 1% of peptone is added, preferably 0.5% (W/V) is additionally added and cultured. did. The culture was carried out using a 5L fermenter, at 37°C, preferably at pH 6 while maintaining at pH 5-7, and preferably for 1 to 7 days, preferably for 5 days. After culturing, the culture medium was centrifuged to remove the culture medium.

The fermented product from which the culture was first removed was refluxed and extracted three times for 5 hours by adding 70% (V/V) aqueous ethanol solution, followed by cooling, and then filtered with Whatman #2 filter paper. The filtered extract was transferred to a concentration tank, and the filtered extract was concentrated under reduced pressure and freeze-dried. This powder was dissolved to 2% (W/V) using 50% glycerin to prepare a fermented extract of orange peel and Calmeg.

Example 3: Preparation of ultra-high pressure extract of tangerine peel and Calmeg

500g of orange peel and 500g of Calmeg are selected as samples, washed thoroughly, and then finely pulverized so that they can pass through a 300-mesh sieve. cooled to

The ultra-high pressure-treated mixed extract was refluxed and extracted three times for 5 hours by adding 70% (V/V) aqueous ethanol solution, followed by cooling, followed by filtration with Whatman #2 filter paper. When the filtration is completed, in the present invention, the extract obtained in the extraction step is transferred to a concentration tank, the filtered extract is concentrated under reduced pressure, and freeze-dried. The freeze-dried product was dissolved to 2% (W/V) using 50% glycerin to prepare an ultra-high pressure extract of tangerine peel and Calmeg.

Example 4: Preparation of fermented ultra-high pressure extract of Tangerine Peel and Calmeg

500g of tangerine peel and 500g of Calmeg were selected as samples, washed thoroughly, and the pulverized product was made fine so that it could pass through a 300-mesh sieve. This pulverized product was added to the culture medium of the Saccharomyces genus (Saccharomyces cerevisiae KCTC 7904) strain at 10 g/L. Here, 1 to 4% of glucose is used as a carbon source, preferably 2% (W/V) is added, and 0.1 to 1% of peptone is added, and preferably 0.5% (W/V) is additionally added and cultured. did. The culture was carried out using a 5L fermenter at 37°C, preferably at pH 6 while maintaining at pH 5-7, and culturing for 1 to 7 days, preferably for 5 days. After culturing the microorganisms, the culture medium was centrifuged to remove the first culture.

The fermented product of tangerine peel and Calmeg from which the culture was first removed was heated to 50° C. under a pressure of 1000 MPa using an ultra-high pressure processor, followed by ultra-high pressure treatment for 24 hours, and then cooled to room temperature. The ultra-high pressure-treated fermented tangerine peel and Calmeg was added with 70% (V/V) aqueous ethanol solution, extracted under reflux three times for 5 hours, cooled, and filtered with Whatman #2 filter paper. Upon completion of filtration, in the present invention, the extract obtained in the extraction step was transferred to a concentration tank, concentrated under reduced pressure at 50° C. or less, and then freeze-dried. Concentration under reduced pressure was concentrated using a rotary decompression concentrator. As for the concentration condition, 700 ml of extract was placed in a 1,000 ml round flask and concentrated under reduced pressure for 2 hours while rotating at 50 to 70 ° C and 600 to 1200 rpm at a pressure of 650 mmHg to 750 mmHg. did. The concentrated solution concentrated under reduced pressure was subjected to pre-freezing, the temperature was lowered to -40°C, maintained at -40°C for 2 to 4 hours, then dropped to -80°C through trap freezing, and the vacuum was maintained at 10-30 mTorr. Reheat to -40°C over 120 minutes. After that, the temperature was maintained at -20°C for 240 minutes, at 0°C for 40 minutes, at 15°C for 340 minutes, at 30°C for 130 minutes, and maintained at 1300 minutes until completely dried. The freeze-dried product was dissolved to 2% using 50% glycerin to prepare a fermented ultra-high pressure extract of orange peel and Calmeg.

Comparative Example 1: Preparation of Tangerine Peel Solvent Extract

1000 g of tangerine peel was selected as a sample, and a tangerine peel solvent extract was prepared according to the method of Example 1.

Comparative Example 2: Preparation of Calmeg Solvent Extract

1000 g of Calmeg was selected as a sample, and according to the method of Example 1, a solvent extract of a thorn bamboo stem was prepared.

<Preparation Example 1> Preparation of cosmetics

Cosmetics containing the mixed extract of tangerine peel and Calmeg of the present invention were prepared as shown in Table 1 below. Formulation Examples 1 to 4 are, respectively, the solvent extract of Tangerine peel and Calmeg of Example 1, the ferment extract of Tangerine peel and Calmeg of Example 2, the ultra-high pressure extract of Tangerine peel and Calmeg of Example 3, and Example 4 It contains a fermented ultra-high pressure extract of mandarin orange peel and Calmeg as an active ingredient, and Comparative Formulation Example 1 is a cosmetic formulation in which no orange peel and Calmeg are added. And Comparative Formulation Examples 2 to 3 are formulation examples containing Comparative Examples 2 to 3 obtained by solvent-extracting each of Tangerine Peel and Calmeg.

Ingredients (unit: weight %) Formulation Example 1 Formulation Example 2 Formulation Example 3 Formulation Example 4 Comparative Formulation Example 1 Comparative Formulation Example 2 Comparative Formulation Example 3 Glyceryl Stearate SE 2.0 2.0 2.0 2.0 2.0 2.0 2.0 stearyl alcohol 1.5 1.5 1.5 1.5 1.5 1.5 1.5 lanolin 1.5 1.5 1.5 1.5 1.5 1.5 1.5 Polysorbate 60 1.3 1.3 1.3 1.3 1.3 1.3 1.3 Sorbitan Stearate 0.5 0.5 0.5 0.5 0.5 0.5 0.5 hydrogenated vegetable oil 1.0 1.0 1.0 1.0 1.0 1.0 1.0 mineral oil 5.0 5.0 5.0 5.0 5.0 5.0 5.0 squalane 3.0 3.0 3.0 3.0 3.0 3.0 3.0 trioctanoin 2.0 2.0 2.0 2.0 2.0 2.0 2.0 dimethicone 1.0 1.0 1.0 1.0 1.0 1.0 1.0 tocopherol acetate 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Carboxyvinyl Polymer 0.14 0.14 0.14 0.14 0.14 0.14 0.14 glycerin 6.0 6.0 6.0 6.0 6.0 6.0 6.0 1.3-Butylene Glycol 2.0 2.0 2.0 2.0 2.0 2.0 2.0 Sodium Hyaluronate 1.0 1.0 1.0 1.0 1.0 1.0 1.0 triethanolamine 0.12 0.12 0.12 0.12 0.12 0.12 0.12 methylparaben 0.2 0.2 0.2 0.2 0.2 0.2 0.2 incense 0.02 0.02 0.02 0.02 0.02 0.02 0.02 Example 1 3 - - - - - - Example 2 - 3 - - - - - Example 3 - - 3 - - - - Example 4 - - - 3 - - - Comparative Example 1 - - - - - 3 - Comparative Example 2 - - - - - - 3 Comparative Example 3 - - - - - - Distilled water remaining amount remaining amount remaining amount remaining amount remaining amount remaining amount remaining amount Sum 100 100 100 100 100 100 100

<Experimental Example 1> Measurement of skin moisturizing effect

The cosmetic formulations of Formulation Examples 1 to 4 and Comparative Formulation Examples 1 to 4 were used to compare the skin moisturizing effect.

The experiment was divided into two groups of 20 people per group for 25 people in their 20s and 40s without skin diseases, and the creams of Formulation Examples 1 to 4 and Comparative Formulation Examples 1 to 3 were applied to the face and face for 1 month in each group twice daily. It was applied to the forearm. Before the start of application, the skin conductivity was measured in a corneometer (CM820 courage Khazaka electronic GmbH, Germany) under constant temperature and humidity conditions (24℃, 40% humidity) and used as the default value, and the skin conductivity after 1 week, 2 weeks, and 4 weeks was measured and the rate of increase was evaluated. The results are shown in Table 2 below.

sample 1 week later 2 weeks later after 4 weeks Formulation Example 1 32 31 31 Formulation Example 2 40 38 40 Formulation Example 3 35 35 39 Formulation Example 4 44 43 45 Comparative Formulation Example 1 14 13 18 Comparative Formulation Example 2 23 17 23 Comparative Formulation Example 3 24 19 23 Comparative Formulation Example 4 22 18 21

As can be seen from the results in Table 2, Formulation Examples 1 to 4 containing the mixed extract of tangerine peel and Calmeg of the present invention Comparative Formulation Example 1 which did not contain the mixed extract of Tangerine peel and Calmeg at all The skin conductivity increase rate was excellent compared to Comparative Formulation Examples 2 to 3 containing extracts of tangerine peel and Calmeg alone.

In particular, formulations containing extracts subjected to fermentation or ultra-high pressure extraction (Formulation Examples 2 to 4) had better skin conductivity than the mixed extracts obtained by solvent extraction (Formulation Example 1), and extracts subjected to ultra-high pressure extraction after fermentation Formulation Example 4 contained had the best skin conductivity.

In addition, in general, since skin conductivity is proportional to the amount of skin moisture, from the above results, Formulation Examples 1 to 4 containing the mixed extract of Tangerine peel and Calmeg of the present invention did not include the mixed extract of Tangerine peel and Calmeg. It can be seen that the skin moisture content is maintained high compared to Formulation Example 1 and Comparative Formulation Examples 2 to 3 containing the extracts of tangerine peel and Calmeg alone.

From these results, it can be seen that the mixed extract of tangerine peel and Calmeg is effective in enhancing skin moisture. In particular, it can be seen that the extract obtained by fermenting tangerine peel and Calmeg at ultra-high pressure has the most excellent effect.

This is presumed to be because many active ingredients including polyphenols and isoflavones among the components of the mixed extract of mandarin orange peel and Calmeg were transformed or concentrated into more effective active ingredients through fermentation and ultra-high pressure extraction steps.

<Experimental Example 2> Skin irritation alleviation effect test by lactic acid

In order to find out the skin irritation alleviation effect of the mixed extract of mandarin orange peel and Calmeg of the present invention, a skin irritation alleviation effect test by lactic acid was conducted, and the test method is as follows.

Human skin cells, fibroblasts (Korea Cell Line Bank, Korea) were cultured in T-75 flasks (Falcon, USA) until they grew to about 80%. This was again transferred to a 96-well plate (Falcon, USA) at 3×10 ⁴ cells/well and cultured for 24 hours. After culturing, it was checked whether the cells were completely attached and well grown through a microscope, and the experiment was conducted using lactic acid as a skin cell stimulant. At this time, 0.2% of lactic acid was used. That is, 200 μl of DMEM (Sigma, USA) medium containing 0.2% lactic acid was added to each 96-well, and the mixed extracts of mandarin peel and Calmeg prepared in Examples 1 to 4 were each added at 1.0% (v/ v) and added. At this time, lactic acid and mixed extracts of tangerine peel and Calmeg were not added as a control group, and those with only lactic acid added were used as a control group. 12 hours after addition of the test substance, in order to compare the cell viability, MTT (Sigma, USA) solution (3mg/ml) was added and the cell viability was measured at 570 nm using an ELISA READER (Molecular Devices, USA). and the cell viability (%) was calculated according to Equation 1 below, and the experimental results are shown in Table 3 below.

Test substance (concentration %) Cell viability (%) control 100 Extract of Example 1 (1%) 99.8 Extract of Example 2 (1%) 99.8 Extract of Example 3 (1%) 99.8 Extract of Example 4 (1%) 99.9 Comparative group (lactic acid 0.2%) 0.0 Lactic acid (0.2%) + extract of Example 1 (1%) 70.8 Lactic acid (0.2%) + extract of Example 2 (1%) 75.4 Lactic acid (0.2%) + extract of Example 3 (1%) 78.9 Lactic acid (0.2%) + extract of Example 4 (1%) 81.3 Lactic acid (0.2%) + extract of Comparative Example 1 (1%) 38.6 Lactic acid (0.2%) + extract of Comparative Example 2 (1%) 42.6 Lactic acid (0.2%) + extract of Comparative Example 3 (1%) 38.6

As can be seen from the results in Table 3, the addition of the mixed extract of tangerine peel and Calmeg showed a mitigating effect on cell generation inhibition by lactic acid treatment. Compared with Comparative Examples 1 to 3, when the mixed extract was added, it was confirmed that the effect was improved. In addition, each extract had no effect on cell viability on its own, indicating no cytotoxicity.

<Experimental Example 3> MMP-1 production inhibitory effect - wrinkle improvement effect

Human normal skin cells, fibroblasts (Korea Cell Line Bank, Korea) were inoculated into a 48-well microplate (Nunc, Denmark) at 1×10 ⁶ cells per well, DMEM medium (Sigma, USA) and 37°C conditions. After culturing for 24 hours in a serum-free DMEM medium added to a final concentration of 1.0% of the mixed extracts of Examples 1 and 4, and as a control, in a serum-free DMEM medium that does not contain the mixed extract of tangerine peel and Calmeg 48 Incubated for additional hours. After incubation, the supernatant of each well was collected and the amount of newly synthesized MMP-1 (ng/ml) was measured using an MMP-1 assay kit (Amersham, USA), and MMP-1 was generated according to Equation 2 below. The inhibition rate was calculated, and the results are shown in Table 4. In this case, TGF-β (10 ng/ml, Roche, USA) was used as a positive control for inhibition of MMP-1 production.

Test substance (concentration %) MMP-1 production inhibition rate (%) TGF-β (10 ng/ml) 75.1 Extract of Example 1 (1%) 78.8 Extract of Example 2 (1%) 83.1 Extract of Example 3 (1%) 83.4 Extract of Example 4 (1%) 84.9 Extract of Comparative Example 1 (1%) 36.9 Extract of Comparative Example 2 (1%) 57.8 Extract of Comparative Example 3 (1%) 54.3

As shown in the results of Table 4, for the inhibition of MMP-1 production, the group treated with the mixed extract of Tangerine peel and Calmeg of Examples 1 to 4 at a concentration of 1.0% was treated with TGF-β as a positive control. Compared to the group and Comparative Examples 1 to 3, it showed a better inhibition rate of MMP-1 production. Here, MMP-1 is a collagen-degrading enzyme, and is a protein whose expression is increased by stress caused by various external stimuli (ultraviolet rays) and the like. When exposed to ultraviolet rays, the synthesis of the skin cells increases, and it dissolves and decomposes the matrix protein of the skin. The inhibition of the production of this MMP-1 enzyme means that it is effective in improving skin wrinkles. Therefore, the fact that the mixed extract of tangerine peel and Calmeg reduces the synthesis of this protein means that it is effective in resolving skin troubles caused by stress, etc. and improving wrinkles.

Hereinafter, based on the results of the above experimental examples, a cosmetic composition comprising a mixed extract of tangerine peel and Calmeg of the present invention is formulated and presented. However, it is not intended to limit the composition of the present invention to the following formulation examples.

Prescription Example 1: Softening lotion

Prescription examples of softening lotion among cosmetics containing mixed extracts of tangerine peel and Calmeg are as follows.

ingredient content(%) Mixed extract of Tangerine Peel and Calmeg 3.0 glycerin 3.0 1.3-Butylene Glycol 2.0 allantoin 0.1 DL-Panthenol 2.0 E.D.T.A-2NA 0.02 Benzophenone-9 0.05 Sodium Hyaluronate 5.0 Nicole HCO 60 0.4 methyl paraben 0.2 incense 0.01 Distilled water remaining amount Sum 100

Prescription Example 2: Nutrient lotion

Among the cosmetics containing the mixed extract of tangerine peel and Calmeg, the prescription example of the nutrient lotion is as follows.

ingredient Content (unit: wt%) Mixed extract of Tangerine Peel and Calmeg 5.0 Glyceryl Stearate SE 1.5 stearyl alcohol 2.5 lanolin 1.5 Polysorbate 60 1.3 Sorbitan Stearate 0.5 hydrogenated vegetable oil 1.0 mineral oil 4.0 squalane 3.0 trioctanoin 2.0 dimethicone 1.0 tocopherol acetate 0.5 Carboxyvinyl Polymer 0.12 glycerin 5.0 1.3-Butylene Glycol 3.0 Sodium Hyaluronate 1.0 triethanolamine 0.12 methylparaben 0.2 incense 0.02 Distilled water remaining amount Sum 100

Prescription Example 3: Nourishing Cream

Among the cosmetics containing the mixed extract of tangerine peel and Calmeg, the prescription example of the nourishing cream is as follows.

ingredient Content (unit: wt%) Mixed extract of Tangerine Peel and Calmeg 5.0 lipophilic monostearate glycerin 1.0 cetearyl alcohol 2.2 stearic acid 2.5 beeswax 1.0 Polysorbate 60 2.5 Sorbitan Stearate 0.5 hydrogenated vegetable oil 1.0 squalane 2.0 mineral oil 3.0 trioctanoin 8.0 dimethicone 2.0 Sodium Magnesium Silicate 0.1 glycerin 5.0 betaine 3.0 triethanolamine 1.0 Sodium Hyaluronate 3.0 methylparaben 0.1 incense 0.1 Distilled water remaining amount Sum 100

As described above in detail a specific part of the present invention, for those of ordinary skill in the art, this specific description is only a preferred embodiment, and it is clear that the scope of the present invention is not limited thereto. Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

Claims (5)

A cosmetic composition for improving skin wrinkles comprising a mixed extract of tangerine peel and Calmeg as an active ingredient. A cosmetic composition for moisturizing the skin comprising a mixed extract of tangerine peel and Calmeg as an active ingredient. A cosmetic composition for alleviating skin irritation comprising a mixed extract of tangerine peel and Calmeg as an active ingredient. The cosmetic composition according to any one of claims 1 to 3, wherein the mixed extract is contained in an amount of 0.0001 to 30% by weight based on the total weight of the cosmetic. The cosmetic composition according to any one of claims 1 to 3, wherein the mixed extract is extracted by an extraction method selected from the group consisting of a solvent extraction method, a fermentation extraction method, an ultra-high pressure treatment extraction method, and an ultra-high pressure treatment extraction method after fermentation.