KR20050018321A - Extracts from mixture of herbal medicine which have excellent effect, It's manufacturing process and usage - Google Patents

Abstract

PURPOSE: Provided are the extract from the mixture of herbal medicines which has excellent skin whitening effect and a manufacturing process and usage thereof. The extract inhibits melanogenesis, lipid peroxides, scavenges free radicals, and proliferates cells. CONSTITUTION: The extract from the mixture of herbal medicines having excellent skin whitening effect is manufactured by the steps of: mixing Clematidis Radix, Prunellae Spica (Herba) and Trichosanthis Radix and heating the mixture with water and organic solvent, followed by stirring and filtration of the extract; adding water or organic solvent to the remnant to give extract; and mixing the above extracts, followed by centrifugation using ethanol, and concentration under reduced pressure to give powdery extract.

Description

Extract of botanical herb with excellent skin whitening effect, preparation method thereof and use thereof {Extracts from mixture of herbal medicine which have excellent effect, It's manufacturing process and usage}

The present invention relates to a medicinal herb extract excellent in skin whitening efficacy, a method for producing the same, and a use thereof, more specifically, by mixing and extracting the gastric glands, haejucho and cheonhwa in appropriate ratios, inhibiting melanogenesis, inhibiting lipid peroxide, and free It is excellent in radical scavenging and cell proliferation, and particularly relates to a medicinal herb extract having excellent skin whitening effect by thinning the deposited melanin pigment, a preparation method thereof, and a skin whitening cosmetic composition containing the same.

Human skin color is determined by hemoglobin (hemoglobin), bilirubin (bile pigment), and melanin (skin pigment), and the most important factor in determining the color of skin is melanin, which is determined by the amount, nature and distribution of melanin. do. Melanin pigment is converted into dopa (DOPA, dihydroxy phenylalanine) and dopachrome by tyrosinase (YPA) contained in the inside of melanocytes in the basement layer of the epidermis, tyrosine, a type of amino acid normally present in the human body After oxidation and condensation, it finally becomes a dark brown pigment called melanin, a polymer.

Melanin in the epidermal layer of the skin acts as a sunscreen and protects the skin organs under the dermis and also protects the skin from external harmful factors such as trapping free radicals and free radicals generated in the skin. To perform.

However, abnormal pigmentation of skin pigments such as blemishes, freckles, and senile plaques is caused by abnormally activated melanin production in melanoma cells, resulting in excessive melanin production and deposition. Such hyperpigmentation can be a serious mental burden from the point of view of skin beauty and can interfere with normal social activities. In addition, Asian women have preferred white and clean skin like white jade, making it an important standard of beauty.

Therefore, the development of a whitening agent for the treatment of abnormal skin pigmentation and satisfying the beauty desire has been actively made. For example, substances that inhibit tyrosinase enzyme activity such as kojic acid, arbutin, hydroquinone, vitamin-C (L-Ascorbic acid), derivatives thereof, and various plant extracts are used. Has been.

However, kojic acid inhibits enzymatic activity by adsorbing copper ions present in the active site of tyrosinase, while there is a problem of instability and skin side effects when blended into cosmetics, and vitamin-C and its derivatives are cosmetic raw materials Hydroquinone has excellent skin whitening effect, but hydroquinone is highly irritating to the skin such as allergic properties, toxicity to melanocytes and permanent depigmentation to the skin. Arbutin is a derivative in which glucopyranoside is combined with hydroquinone and has little side effects when using hydroquinone and inhibits the synthesis of melanin pigment without toxicity to the human body, increasing melanin pigmentation. Applicability has been suggested as a treatment for skin diseases, but Has the disadvantage of being partially degraded by cattle,

In recent years, many studies have been conducted on extracting herbal ingredients of plants and using the extracts as whitening cosmetics. Whitening cosmetics using the whitening skin component [Japanese Patent Publication No. 55-44375] and whitening cosmetics using the components of licorice [0007] Whitening cosmetics using a number of herbal ingredients such as red ginseng, green tea, bilberry leaf, cornus milk, etc. have been known as well as [Japanese Laid-open Patent No. 60-214721].

However, the conventional whitening cosmetics using the herbal ingredients are effective to use at a high concentration because most of the extract of the herbal medicines, there is a problem such as discoloration, discoloration during long-term storage and formulation application concentration.

Therefore, the present inventors solve the problems of the conventional single active ingredients and single drug herbal whitening efficacy drug, formulation instability, skin side effects, blocking the melanin pigmentation process of the whitening mechanism known to date (ultraviolet ray blocking, melanin biosynthesis rate determining step Inhibitors of intyrosinase enzyme activity, signal transduction (ET-1, IL-1a, IL-6) and melanin decolorization (melanin reduction, photooxidation inhibition, exfoliation and turnover promotion). As a result of the exploration and research, the whitening effect was tested on various extracts to confirm the whitening effect of whitening cosmetics containing fucobacterium herbal extracts of gastric ulcer, haejucho and cheonhwa powder, and it is a safe substance without formulation stability and skin side effects. Thus, the present invention has been completed.

Accordingly, an object of the present invention is to provide a herbal extract and a method for preparing the same, which are hypoallergenic and effective for whitening, mixing dry matter, and extracting the ingredients of the gastric ginseng, haejucho and cheonhwa at a certain ratio.

In addition, an object of the present invention is to provide a skin whitening cosmetic composition containing the above herbal extract.

The present invention is characterized by the medicinal herb extracts of the stomach, wild vinegar and cheonhwabun and its preparation method.

In addition, the skin whitening cosmetic composition containing the herbal medicine extract is characterized by another.

Referring to the present invention in more detail as follows.

Clematis chinensis Osbeck refers to the roots of the creeper and the Buttercup S. chinensis. Useful Plant Resources Research, Korea Research Institute of Chemical Technology, 1988, Dohae Hyangjeomsa Dictionary-Yeonglimsa 1990]. Flavonone glycosides such as anemonin, anemonol, sterols, sugars, glucose, lactones, amino acids, cretinins, etc. Saponin, including Flavanone glycosides, Cremontanoside A, Cremontanoside B, Cremontanoside C, and Cremontanoside S ), Etc. [Korea Useful Plant Resources Exhibition, Dohae Hyangdae Dictionary, Chinese Dictionary]. In addition, gastric glands have been widely used for diseases with arthritis, mumps, acute jaundice-type infectious liver disease, conjunctivitis, tonsillitis, etc.

Prunella vulgaris Linne (Prunella vulgaris Linne) is a fruit tree of perennial herbaceous plants, which is collected from summer, and is harvested in summer. The component of the vinegar contains Tritpenoid saponin, and its saponenin is Oleanolic acid. In addition, free oleanolic acid, ursolic acid, rutin, hyproside, cis-caffeic acid, trans-caffeic acid, vitamin B ( Vitamin B), Vitamin C, Vitamin K, Carotene, Resin, Amaroid, Tannin, Essential Oil, Alkaloid, Chlorogenic Acid, Water Soluble Salts (about 3.5%, 68% of which are potassium chloride) are known, and the presence of rosmarinic acid is also known [Korea Use Plant Resources Research Exhibition, Korea Research Institute of Chemical Technology, 1988] Ph.D., Beijing Medical University, 1991, Chinese Medicine Dictionary]. Pleurotus has been used to treat chronic boils, cramps, acute mastitis, and lymph node tuberculosis in Chinese medicine. It has been used to destroy jinga (soak in the lower abdomen or lumps of blood), remove individual parts, and treat numbness and numbness.

Cheonhwabun (Trichosanthes Kirilowii Max), also known as Fruit Root or White Pea, collects the roots of perennial vines, Hanultari and Yellow Hanultari, in autumn. Beat until the yellow layer on the outside was used as a medicine. The known components of cheonhwa flour are trichosanthin, which is a protein, and arginine, citrulline, and palmitic acid, linoleic acid, and the like, which are known as amino acids, and arginine. Recently, stenols such as bronolic acid, cucurbitacin B, and alpha-spinasterol have been identified [Korea Use Plant Resources Research, Korea Research Institute of Chemical Technology-1988, illustration] Pharmacological Ambassador-Younglimsa 1990]. Cranberries are effective in stopping boils, in various boils, bruises, skin eczema, wounds and dental mammary glands, and have been widely used for drainage, small seedling, detoxification, and antipyretic effects.

Method of producing a composite herbal extract excellent in whitening effect with the three herbal drugs are as follows.

1) mixing the yeongyeongseon, haejucho and cheonhwa after heating and stirring with water or an organic solvent of 10 to 15 times the weight of the mixed herbal medicine, primary extraction and filtration;

2) heating the organic residue with 7-12 times the weight of the mixed drug weight to the residue remaining after the filtration, and performing secondary extraction and filtration; And

3) The filtrates obtained after the first and second extraction are combined, centrifugation using ethanol is carried out to obtain a supernatant, and the mixture is concentrated under reduced pressure to obtain a powder extract.

To explain this in detail, the mixture of the gastric glands, haejucho and cheonhwa powder in a weight ratio of 1: 0.5 to 1.5: 0.5 to 2 and put water or organic solvent of 10 to 15 times the weight of the mixed herbal medicine, 80 ± 5 ℃, 6 ~ The mixture was heated, stirred for 8 hours, and extracted firstly, filtered, and then the residue was added with water or an organic solvent of 7-12 times the weight of the mixed herbal medicine, heated and extracted secondly, filtered, and then mixed with the previous filtrate. Increase the efficiency If the amount of water is too small, the stirring is difficult, solubility of the extract is lowered, the extraction efficiency is lowered, if too large, the amount of the ethanol solvent used in the next purification step is large, there is a problem that is not economical. In this case, the organic solvent is one or two or more selected from ethanol, protanol, butanol, glycerol, propylene glycol, 1,3-butylene glycol, methyl acetate, ethyl acetate, benzene, hexane, diethyl ether, dichloromethane This is preferable.

In the present invention, a method of re-extracting after primary extraction is adopted. However, when the herbal extract is mass-produced, even though it is effectively filtered, the water content of the herbal medicine itself is high, so that loss occurs. To do this.

As described above, the extract obtained by the first and second extraction is filtered, and then the impurities are purified by adding the filtrate with the same amount of ethanol. At this time, when the content of ethanol is less than the filtrate, it is uneconomical because the incorporation of impurities is relatively high and the content of the active ingredient is lowered. The ethanol fraction obtained after the Winsim separation is concentrated under reduced pressure at 55-65 ° C. to remove the solvent remaining in the sample.

The extract thus obtained is lyophilized to obtain a powdery extract, which has a complex whitening effect such as melanin inhibition, lipid peroxide suppression, free radical scavenging, and cell proliferation than the hydrothermal extracts of the three herbs. It is excellent and can be usefully applied to the whitening cosmetic composition.

In addition, the present invention includes a skin whitening cosmetic composition that satisfies the action of inhibiting melanin production, inhibiting lipid peroxidation, free radical scavenging, and cell proliferation simultaneously containing the extract powder prepared as an active ingredient.

In particular, in the present invention, the extraction of the three herbal medicines by mixing in the appropriate weight ratio of 1: 0.5 to 1.5: 0.5 to 2 without extracting the gastric ginseng, haejucho and cheonhwa, respectively, is increased between the herbal ingredients compared to the respective extracts. Due to its action, it shows excellent whitening effect. In this case, the mixing ratio of each used herbal medicine also shows the most advantageous medicinal effect. If the mixing ratio of the herbal medicine is out of the above range, tyrosinase enzyme inhibitory action, lipid peroxide inhibitory action, There is a problem that the whitening effect such as free radical scavenging action, cell proliferation action is reduced and the indicator components such as rosmarinic acid and oleanoic acid decrease.

As a result of analyzing the indicator material of the powder extract extracted from the gastric agent mixed with the gastric glands, hazelweeds and cheonhwa in a certain ratio by using liquid chromatography, rosmarinic acid, oleanolic acid ) Was found to contain. Herein, rosmarinic acid has been reported for anti-inflammatory action such as inhibition of biosynthesis of prostacyclin and metabolism of arachidonic acid and elimination of harmful free radicals produced in multinucleated leukocytes [Biochem. Pharmac, 29, pp533 to 538 and Agents Actions, 17, pp375 to 376 (1985), and oleanolic acid has been reported by several researchers to have an excellent effect on anti-inflammatory and anti-inflammatory analgesic activity [Jouranal of pharm. Phamacol, 44 (5), pp 456-458 (1992).

Rosmarinic acid and oleanolic acid are the main index components of the releasing agent obtained by the present invention. These components are synergistic effects of each other, and they inhibit the production of tyrosinase enzymes, inhibiting lipid peroxidation, and free radical scavenging. It can exert powerful medicinal effects on cell proliferation, so that a whitening effect can be expected even with a small amount of skin application. Moreover, the action | action of other active ingredients other than rosmarinic acid and oleanolic acid cannot also be excluded as an index component of the releasing agent obtained by this invention.

According to the present invention, in order to exhibit a significant effect throughout the whitening effect, it was found that the case of containing rosmarinic acid in the range of 0.01 to 0.08 wt% and oleanoic acid in the range of 0.1 to 0.8 wt% is preferable. In other words, the three herbal medicines are mixed and extracted at a constant ratio, so that the whitening effect is best when the indicator components such as rosmarinic acid and oleanolic acid are contained at a predetermined level.

In particular, when applying the herbal medicine extract of the present invention on the skin, there is less concern of side effects than other synthetic single raw materials, because it is a natural extract, in fact, the skin toxicity test results of the herbal medicine extract, there is no effect on the living body It turned out to be.

As described above, the present invention has a marked difference in the whitening effect compared to the extract obtained by extracting and extracting the herbal medicines by boiling extracts of three kinds of herbal medicines of the gastric glands, hachichos and cheonhwa in a certain ratio.

Therefore, when applying the complex herbal extracts in cosmetics, it is possible to formulate such as softening lotion, astringent lotion, lotion, essence, cream, gel, pack, soap, essence sheet, hydrogel patch or mask, the complex herbal extract is a whole It is preferable to mix | blend 0.001-20.0 weight% with respect to cosmetic composition.

Hereinafter, the present invention will be described in more detail based on the following examples, but the present invention is not limited thereto.

Example 1

Wash the dried liquor line with tap water, dry it 100g, 200 g of chopped chunks of 1-2 cm, 100 g of fruit tree part collected in late summer, add 4.5 liters of water and stir well After heating, stirring and extraction at 80 ° C. for 7 hours, the filtrates were collected, and 3 liters of water was added to the residue, followed by warm extraction for 3 hours. The same amount of ethanol was added to the mixture, followed by centrifugation to collect the supernatant and concentrated under reduced pressure until the herbal extract was dried at 60 ° C. to obtain 21 g of powder extract.

As a result of analyzing the obtained powder extract by liquid chromatography, the content of each indicator component was 0.62% of oleanolic acid and 0.08% of rosmarinic acid.

Example 2

The liquor was washed and dried in 100 g of 1.5 g of dry water, heated at 80 ° C. for 7 hours, and extracted. The filtrates were collected, and 1 liter of water was added to the residue, followed by warm extraction for 3 hours.

200 g of hwahwabun was cut into 1.0-2.0 cm, 3 liters of water was added, heated and stirred for 8 hours at 80 ° C, and extracted. The filtrates were collected, and 2 liters of water was added to the residue, followed by warm extraction for 3 hours.

100 g of fruit tree was taken, 1.5 liters of water was added, heated at 80 ° C. for 7 hours, and extracted with extraction. The filtrates were collected, and 1 liter of water was added to the residue, followed by warm extraction for 3 hours.

Each of the extracted filtrates was combined and the same amount of ethanol was added to the mixture, followed by stirring. The supernatant was collected by centrifugation and concentrated under reduced pressure until the herbal extract was dried at 60 ° C. to obtain 28 g of powder extract.

As a result of analyzing the obtained powder extract by liquid chromatography, the content of each indicator component was 0.56% of oleanoic acid and 0.06% of rosmarinic acid.

Example 3

100 g each of the commercially available gasoline, hagocho and cheonhwa were mixed in equal amounts, and 4.5 liters of water was added to the mixture and stirred well. The mixture was heated at 80 ° C for 7 hours, stirred and extracted to collect the filtrate. The mixture was extracted by heating for 3 hours and the filtrates were combined.

Equivalent amount of ethanol was added to the mixture, followed by centrifugation to collect the supernatant, and concentrated under reduced pressure until the herbal extract was dried at 60 ° C. to obtain 26 g of powder extract.

As a result of analyzing the obtained powder extract by liquid chromatography, the content of each indicator component was 0.36% of oleanoic acid and 0.04% of rosmarinic acid.

Comparative Example 1

Commercially available commercial gastric glands were extracted with water using a standard liquid solution method, filtered, and the filtrate was lyophilized to obtain a powder extract.

As a result of analyzing the obtained powder extract by liquid chromatography, the content of each indicator component was 0.09% of oleanoic acid and 0.02% of losmarinic acid.

Comparative Example 2

The commercially available Hagocho was extracted by water extraction with a standard melt solution, filtered, and the filtrate was lyophilized to obtain a powder extract.

As a result of analyzing the obtained powder extract by the method by the liquid chromatography method, the content of the indicator component was 0.12% or less of oleanoic acid or 0.01% or less of rosmarinic acid.

Comparative Example 3

Commercially purchased cranberries were extracted with water using a standard solution and filtered, and the filtrate was lyophilized to obtain a powder extract.

As a result of analyzing the obtained powder extract by liquid chromatography method, the content of each indicator component was 0.06% of oleanoic acid and 0.01% of losmarinic acid.

Comparative Example 4

The gastric glands, hachichos and cheonhwa were mixed in a weight ratio of 1: 1: 2, water extracted by standard liquid solution according to the method described above, filtered, and the filtrate was lyophilized to obtain a powder extract.

As a result of analyzing the obtained powder extract by the method by the liquid chromatography method, the content of the indicator component was 0.24% or less of oleanoic acid or 0.02% or less of rosmarinic acid.

Comparative Example 5

Mix 100 g of commercially available gasoline, 100 g of Hagocho, and 200 g of cheonhwa, add 6 liters of ethanol and stir well, heat and stir at 80 ° C for 7 hours to collect the filtrate, and add 4 liters of ethanol to the residue. The mixture was extracted by heating for about 3 hours, the filtrates were combined, and concentrated under reduced pressure until the herbal extract was dried at 60 ° C. to obtain 13 g of powder extract.

As a result of analyzing the obtained powder extract by liquid chromatography, the content of each indicator component was 0.48% of oleanoic acid and 0.03% of rosmarinic acid.

Reference Example 1

Analyzing liquid extract extract powder prepared by the same method as the present invention for the Chinese herbal medicine extract powder extract according to the present invention and the Chinese imported products of the three herbal medicines as a domestic herbal medicine, The surface composition comparison experiment was performed and the results are shown in Table 1 below.

division Oleanolic acid Los marine acid Domestic Botanical Extract 0.51% 0.08% Chinese Herbal Herbs Extract 0.59% 0.06%

Reference Example 2: Toxicity Test

A) Single dose toxicity test by intravenous administration and oral administration

Single-dose toxicity test by intravenous administration and oral administration was performed in Wistar male rats 5 weeks old. The dry powder extract obtained in Example 1 was observed intravenously at a dose of 500 mg / kg and 1000 mg / kg and orally administered at a dose of 2000 mg / kg for 14 days. In addition, no abnormalities were found in other anatomical findings compared to the control group, and thus the extract obtained in Example 1 was confirmed to be a very safe substance.

B) 3-month repeated dose toxicity test by oral administration

Sprague Dawley male rats at 5 weeks of age were tested for repeated dose toxicity for 3 months by oral administration. The extract of Example 1 was orally administered three days a day at a dose of 1000 mg / kg to observe the general condition every day, and the body weight was measured at 7 day intervals. As a result of performing qualitative, quantitative, hematological, necropsy, and long-term weight measurement of urine at the end of the administration period, the effect of the administration of the extract of Example 1 was not shown at all for all observation items and test items, and thus very safe. It was confirmed to be a substance.

Reference Example 3: Skin Safety Test

A) skin primary irritation test

Primary skin irritation tests were performed using six New Zealand white rabbits. The left and right midline of the rabbit's midline were depilated in a forward direction of about 3 × 10 cm, and four test sites were set by dividing each back and forth. 18 gauge gauge needles were made in two parts of left and right diagonal so that bleeding did not occur, and the remaining two parts were placed in the dry area. The extract of Example 1 was dissolved in butylene glycol to prepare a 20% solution, 0.5 ml of the solution was infiltrated into 2 sheets of 2.5 × 2.5 cm lint, and the blockage adhered to the right 2 parts, and the left 2 parts. After only butylene glycol was infiltrated into 2 sheets of 2.5 x 2.5 cm lint, occlusion adhered to the left 2 parts, and the skin was observed for 24, 48 and 72 hours.

As a result of the test, both the dry and damaged areas to which the extract obtained in Example 1 was applied did not have skin abnormalities such as redness and edema.

B) Continuous skin irritation test

Continuous skin irritation test was performed using a Hartley guinea pig of about 350 g in weight. The extract of Example 1 was dissolved in butylene glycol to make a 5% solution, and 0.1 ml was applied to the left abdomen of the animal once a day for four consecutive weeks (six times a week). Skin reactions such as redness and swelling of the application site were observed daily, and the skin irritation of the extract of Example 1 was evaluated based on this compared with the positive control group to which the same 2% sodium lauryl sulfate solution was applied.

As a result of the test, no skin irritation response by repeated application of the extract of Example 1 was observed.

C) Human patch test

According to the Japanese Information Service Research Council, the closed patch test was conducted on 30 healthy males, and the sample was closed for 48 hours on the back and 2 hours after removal and twice the next day (72 hours). In the optical patch test by the optical stimulus response determination method, the sample was attached to the dorsal area for 24 hours, and after 2 hours of removal, the DERMARAY UV-A light (BLB 10 light) was irradiated at a distance of 20 cm for 7 minutes. Samples were attached to the dorsal area for 2 hours and 10 minutes after removal for urticaria reaction.

As a result of the test, no skin irritation response by repeated application of the extract obtained in Example 1 was observed.

Therefore, the fucobacterium herbal extracts of the gastrointestinal tract, haejucho and cheonhwamin according to the present invention have been identified as a very safe material when applying the skin.

Experimental Example 1: Confirming the inhibitory effect of mushroom tyrosinase

In order to measure the whitening effect of the extract extracted in Examples 1 to 3 and Comparative Examples 1 to 5, it was examined whether the extract plays a role in inhibiting tyrosinase enzyme activity that acts as an important factor in melanin production. Koji acid (Kojic acid), arbutin (Arbutin), hydroquinone (Hydroquinone), vitamin-C (L-ascorbic acid), ethyl ascorbyl ether, epidermis extract In comparison with the tyrosinase enzyme activity inhibitory effect was measured.

The inhibitory activity against the tyrosinase was in vitro extracts of Examples 1 to 3 and Comparative Examples 1 to 5 and positive control groups of kojic acid, arbutin, hydroquinone, vitamin-C, ethyl ascorbyl ether, and extracts of lettuce extract, respectively. Put μL into a microplate (96 wells), add 150 μL of 0.1M phosphate buffer (pH 6.8), 25 μL of substrate 1.5 mM L-tyrosine solution, and add 16 unit / mL mushroom tyrosinase (0.05M phosphate buffer, pH 6.8). After adding 7 μl and reacting for 10 minutes at 37 ° C., the reaction was stopped for 5 minutes on ice. Then, the absorbance was measured at 475 nm, the absorption wavelength of dopachrome, the reaction product, and the tyrosinase inhibition rate. Saved. Inhibition rate for tyrosinase (%) is calculated by the following Equation 1, the results are shown in Table 2 and FIG.

A: absorbance before reaction of the inhibitor

B: Absorbance after the reaction with the inhibitor

C: Absorbance before reaction of the absence of inhibitor

D: absorbance after the reaction without the inhibitor

division % Inhibition against tyrosinase Kojisan 87.5 Hydroquinone 99.4 Arbutin 67.8 Vitamin-C 98.7 Ethyl ascorbyl ether 72.1 Lettuce extract 8.2 Example 1 Extract 84.2 Example 2 Extract 81.4 Example 3 Extract 76.9 Comparative Example 1 Extract 42.3 Comparative Example 2 Extract 45.8 Comparative Example 3 Extract 38.6 Comparative Example 4 Extract 58.0 Comparative Example 5 Extract 66.4

As shown in Table 2, the tyrosinase enzyme activity inhibition rate of the herbal medicine extract of Example 1 was found to be 84.2%, which is much better than the extracts of alveolar extract, arbutin and ethyl escorbyl ether. It has an effect. In addition, it was shown that the inhibitory activity was weaker than vitamin-C, hydroquinone and kojic acid in terms of other substances, but considering that it is not a single ingredient but a tablet extract extracted from mixed herbal medicines, it is effective as a skin whitening agent compared to conventional substances. It can be seen that is superior.

Experimental Example 2: Confirmation of the melanin production inhibitory effect using the pigment cells of the rat

In this experimental example, in order to observe the skin whitening effects of the extracts of Examples 1 to 3 and Comparative Examples 1 to 5 at the cellular level, the effect of inhibiting melanogenesis in melanocytes was confirmed by in vitro methods. Melanin black pigment is reduced by treating the extract of the present invention and the existing whitening raw materials while artificially culturing B16 melanocytes that secrete melanin black pigment, a cell strain derived from mouse, from ATCC (American Type Culture Collection). The degree was compared and evaluated.

To determine melanin biosynthesis inhibition of B16 melanocytes, B16 melanocytes were suspended in suspension containing 10% fetal bovine serum at a concentration of 5 × 10 ⁵ (cells / Dish-35mm), and then suspended cells were cultured. 10 ㎍ / ㎖, 50 ㎍ / ㎖, 100 ㎍ / ㎖ each sample was incubated at 37 ℃ in a CO ₂ incubator (5% carbon dioxide / 95% air conditions) for 3 days, and then The primary melanin production was determined by color, cells were washed with phosphate buffer, and then trypsinized to separate them from the flask. The collected cells were collected in a tube, washed with phosphate buffer, and centrifuged to evaluate the melanin secretion inhibition through the amount of melanin precipitated at the bottom of the tube. The evaluation method was evaluated by measuring the absorbance at 475 nm and calculating the number of melanin-producing cells by measuring the absorbance at 475 nm according to the treatment of nothing in the cell culture solution and the degree of secretion of the black melanin pigment when each sample was treated. .

division Concentration (µg / ml) Melanin amount (mg) Melanin / cell (pg) % Inhibition No processing (voice control) - 0.167 436 - Kojisan 10 0.120 384 11.9 50 0.056 298 31.7 100 0.032 183 58.0 Hydroquinone 10 0.074 121 72.2 50 0.028 58 86.7 100 0.017 13 97.0 Arbutin 10 0.145 394 9.6 50 0.099 246 43.6 100 0.065 119 72.7 Vitamin-C 10 0.112 272 37.6 50 0.041 101 76.8 100 0.020 24 94.5 Ethyl ascorbyl ether 10 0.128 376 13.8 50 0.084 206 52.8 100 0.047 122 72.0 Lettuce extract 10 0.135 435 0.0 50 0.163 431 1.1 100 0.152 368 15.6 Example 1 Extract 10 0.125 293 32.8 50 0.066 121 72.2 100 0.038 72 83.5 Example 2 Extract 10 0.129 298 31.7 50 0.072 126 71.7 100 0.043 84 80.7 Example 3 Extract 10 0.132 302 30.7 50 0.082 139 68.1 100 0.054 101 76.8 Comparative Example 1 Extract 100 0.098 261 40.5 Comparative Example 2 Extract 100 0.102 273 37.4 Comparative Example 3 Extract 100 0.104 296 32.1 Comparative Example 4 Extract 100 0.118 197 54.8 Comparative Example 5 Extract 100 0.056 148 66.1

As shown in Table 2, when the concentration of each sample is 100 ㎍ / ㎖ hydroquinone, vitamin-C, the extract of Example 1, Example 2, Example 3, arbutin, ethyl ascorbyl ether, koji acid, Inhibitory effect was shown in the order of the extract of lettuce extract. This is the melanin production inhibition rate of the extract of Example 1 at a concentration of 100 ㎍ / ㎖ 83.5%, which has a much better effect than koji acid, lettuce extract and arbutin is known to have excellent whitening effect.

Experimental Example 3: Confirm the skin whitening effect

A) whitening effect of animal skin

Brown guinea pigs are animals that cause pigmentation due to ultraviolet rays, allergic reactions, etc. Like humans, they have the same color of hair and skin, so if you choose a uniform hair color, you can use a wide area after removing hair. . The improvement effect of pigmentation can be verified by the method of causing a pigmentation in the following method, and apply | coating each said sample to a pigmentation site | part. Thus, substances whose effects have been recognized by this method can be usefully used as a primary in vivo screening method because they have a high probability of being effective in evaluation of human UV pigments.

Formation of pigmentation by ultraviolet (UVB) was performed by attaching a 2.5 × 2.5 cm square hole-shielding aluminum foil to the skin from which the hairs behind the brown guinea pigs were removed, followed by a SE lamp (wavelength 290 to 320 nm). After irradiating ultraviolet rays so that the amount of irradiation energy was 1350 ml / cm ² , the aluminum foil was peeled off, and each whitening sample was applied by the following method for evaluation. Pigmentation after UV irradiation appears 3-4 days later, reaching peak after about 2 weeks. It is important that the resulting color tone of the ultraviolet pigmentation spot is the same and no staining occurs, and the individual whose UV pigmentation spot is uneven is excluded. After 14 days of pigmentation after UV irradiation, each whitening sample was applied twice a day for 30 days. Each whitening sample was dissolved in a mixed solvent of butylene glycol: water: ethanol (50: 30: 20), diluted with a cotton swab, and a negative control site where a solvent was applied.

The whitening effect of the skin was measured using a Minolta color difference meter to determine the whitening effect. In order to display the color, the L * a * b colorimeter was used to measure mainly L * value as an index, and the measurement was repeated three times or more at one site, and the pigmentation part was measured evenly. The difference in skin color (ΔL *) between the start time and the completion time of application was calculated according to the following Equation 2 and is shown in the following Table 4 and FIG. 2. The whitening effect is determined by comparing the ΔL * of the sample application site and the control site. When the ΔL * value is 2 or more, the whitening effect of the deposited pigment is clear and the whitening effect is about 1.5.

division Concentration (% by weight) Whitening effect (ΔL *) No processing (voice control) - 0.72 Kojisan 0.05 0.78 0.10 0.82 Hydroquinone 0.05 1.14 0.10 1.54 Arbutin 0.05 0.89 0.10 1.02 0.50 1.58 Vitamin-C 0.05 1.59 0.10 2.07 Ethyl ascorbyl ether 0.05 0.91 0.10 1.08 0.50 1.61 Lettuce extract 0.10 0.74 5.00 1.26 Example 1 Extract 0.05 1.39 0.10 1.88 Example 2 Extract 0.05 1.21 0.10 1.84 Example 3 Extract 0.05 1.09 0.10 1.73 Comparative Example 1 Extract 0.10 1.38 Comparative Example 2 Extract 0.10 1.26 Comparative Example 3 Extract 0.10 1.34 Comparative Example 4 Extract 0.10 1.52 Comparative Example 5 Extract 0.10 1.69

As shown in Table 4, the herbal medicine extract of Example 1 of the present invention has a whitening effect of the pigment deposited at a concentration of 0.05% by weight, and a clear whitening effect at 0.1% by weight, followed by vitamin-C. Good to know. However, the kojic acid is rather pigment deposition due to the side effects such as the experimental animal erythema can know the fact that the whitening effect halved, hydroquinone, also there is half a whitening effect compared to the result of Experimental Examples 1 and 2, which in It may be due to skin side effects in vivo animal testing.

B) whitening effect on human skin

After attaching opaque tape with a hole of 1.5 cm diameter to the back of the subject, 15 healthy men were irradiated with UVB twice as much as Minimal Erythema Dose. After induction of blackening of the skin, each whitening sample was applied, and after 30 days, the degree of pigmentation of the skin was measured using a color contrast meter using a black and white contrast determination method. Each sample is an extract of Examples 1 to 3 and a positive control group, and arbutin, vitamin-C, ethyl ascorbyl ether, and lettuce extract having relatively low skin irritation were added to a mixed solvent of butylene glycol: water: ethanol (50: 30: 20). Melt and apply twice a morning and evening. The determination of the effect was obtained by evaluating the L * value representing the light and dark of the skin as in the animal test (representing the skin color 50-80 of our country), and calculating ΔL *, and is shown in Table 5 below.

division Concentration (% by weight) Whitening effect (ΔL *) No processing (voice control) - 0.78 Arbutin 0.05 0.83 0.10 1.12 0.50 1.48 Vitamin-C 0.05 1.54 0.10 1.95 0.20 2.45 Ethyl ascorbyl ether 0.05 0.91 0.10 1.08 0.50 1.53 Lettuce extract 0.10 0.84 1.00 1.38 Example 1 Extract 0.05 1.34 0.10 1.90 0.20 2.12 Example 2 Extract 0.05 1.11 0.10 1.68 0.20 2.01 Example 3 Extract 0.05 1.06 0.10 1.52 0.20 1.98

As shown in Table 5, the whitening effect of each sample is good effect of the extract of Examples 1 to 3 next to vitamin-C, the extract of Example 1 whitening effect when applied to human skin than ethyl ascorbyl ether and arbutin It was confirmed.

Experimental Example 4: Free radical scavenging effect experiment

Preradial scavenging activity was tested using the extracts of Examples 1 to 3 and Comparative Examples 1 to 5, vitamin-C, alpha-rutin, ethyl ascorbyl ether, and BHT (butylated hydroxy toluene) as positive controls. . 1 ml of a 0.1 mM DPPH (1,1-diphenyl-2-picrylhydrazyl) [sigma] solution dissolved in methanol was added to a test tube, and 0.988 ml of ethanol was added to 200 µg of each test sample. The reaction was carried out for a minute (final concentration of each sample in the reaction solution was 0.1% (v / v)). At this time, 1 ml of ethanol containing no test substance was added to the negative control group. After the reaction, the absorbance was measured at a wavelength of 516 nm with a spectrophotometery to calculate the free radical scavenging rate of each sample by the following Equation 3, and the results are shown in Table 6 below.

division Free radical scavenging rate (%) Vitamin-C 94.7 Alpha-routine 88.8 Ethyl ascorbyl ether 65.1 BHT 88.4 Lettuce extract 21.3 Example 1 Extract 89.1 Example 2 Extract 87.5 Example 3 Extract 84.9 Comparative Example 1 Extract 61.4 Comparative Example 2 Extract 59.0 Comparative Example 3 Extract 57.1 Comparative Example 4 Extract 64.5 Comparative Example 5 Extract 73.1

As shown in Table 6, the comparative radicals 1 to 3 and the extraction solvent of the extracts of Examples 1 to 3, the free radical scavenging rate of the extracts of the sole herbal extracts, were more effective than the extracts of the Comparative Examples 4 to 5, the solvents of water or ethanol alone. It was superior to the positive control vitamin-C, alpha-rutin (α-rutin) and BHT, but less effective than ethyl ascorbyl ether and lettuce extract. This shows that the free radical scavenging component of the herbal medicine extract extracts more synergistically than the active ingredient extracted from the herbal medicine alone.

Experimental Example 5: Confirmation of the inhibitory effect on lipid peroxide

Inhibition rate of lipid peroxidation induced by bioenzyme lipoxygenase was measured on the extract samples of Examples 1 to 3 and Comparative Examples 1 to 5, and vitamin-C and alpha- are well known as antioxidants as positive controls. The routine, BHT, was used.

12 ml of 50 mM sodium-phosphate buffer (pH 6.8) were placed in a test tube, where 0.1 ml of 0.2% (v / v) sample solution dissolved in 50 mM sodium-phosphate buffer solution and 6.6 mM arachidonic acid [Sigma] 0.1 Ml was added. 0.1 ml of lipoxygenase [Sigma] dissolved in 50 mM sodium-phosphate buffer was added to each test tube and reacted for 10 minutes in a 25 ° C thermostat (final concentration of the sample added to each reaction solution was 100 µg / ml). At this time, 0.1 ml of 50 mM sodium-phosphate buffer solution was added to the negative control group. After the reaction, 1 ml of 20% (v / v) trichloroacetic acid solution and 2 ml of 0.67% (v / v) thiobarbituric acid solution are added, and the mixture is boiled in boiling water for 10 minutes. After cooling, the mixture was centrifuged (1800 rpm, 3 minutes), and only the supernatant was taken, and the absorbance was measured at a wavelength of 535 nm with a photospectrometer. Inhibition rate of lipid peroxidation of each sample was calculated by the following Equation 4 and the results are shown in Table 7 below.

division Lipid peroxidation inhibition rate (%) Vitamin-C 65.7 Alpha-routine 71.2 Ethyl ascorbyl ether 24.6 BHT 66.8 Lettuce extract 18.7 Example 1 Extract 69.1 Example 2 Extract 66.3 Example 3 Extract 61.0 Comparative Example 1 Extract 34.9 Comparative Example 2 Extract 39.3 Comparative Example 3 Extract 37.1 Comparative Example 4 Extract 49.1 Comparative Example 5 Extract 60.3

As shown in Table 7, the extracts of Examples 1 to 3 were higher in inhibition of lipid peroxidation than BHT, ethyl ascorbyl ether and vitamin-C except alpha-rutin, which is a positive control. Therefore, it can be seen that the extracts of Examples 1 to 3 of the present invention have an excellent anti-aging effect of high skin lipids.

Experimental Example 6: Confirming the effect of cell proliferation

The cell proliferation effect of the extracts of Examples 1 to 3 and Comparative Examples 1 to 5 was measured.

100 μl of a 5% FBS solution was added to a 96 microwell plate, and each sample was diluted and dispersed in a DMEM medium containing 2% FBS. Using hemocytometer, fibroblasts obtained from neonatal epidermal tissue (Foreskin) tissue were dispersed in 100 μl medium at 3500 per well, incubated for 5 days, and then the supernatant was removed and washed again by adding 200 μl of 5% FBS (phosphate buffered saline). 150 μl of DMSO was added per well, followed by Vortexing and melting for 5 minutes to measure absorbance at a wavelength of 570 nm (final concentration of the sample contained in the reaction solution was 100 μg / ml). At this time, the negative control group without the sample was used as the same experimental method, and the relative absorbance was obtained, and the cell proliferation effect was compared from the difference. The results are shown in Table 8 and FIG. 3.

division Cell growth rate (%) Vitamin-C 137 Lettuce extract 102 Example 1 Extract 136 Example 2 Extract 133 Example 3 Extract 130 Comparative Example 1 Extract 121 Comparative Example 2 Extract 128 Comparative Example 3 Extract 119 Comparative Example 4 Extract 127 Comparative Example 5 Extract 128

As shown in Table 8, the herbal medicine extract of Example 1 of the present invention showed a cell proliferation effect similar to the vitamin C-positive control of the same concentration, and also has a better cell proliferation effect than the extract of Comparative Examples 1 to 3 alone. And it was found.

Formulation Example 1 and Comparative Formulation Example 1: Preparation of Facial Skin

Prescription example of a lotion in the cosmetic containing the extract of Example 1 of the present invention is as follows.

number Raw material name Prescription Example 1 Comparative Prescription 1 One 1,3-butylene glycol 4.0 4.0 2 Disodium ID 0.1 0.1 3 Fiji-Fiji 18/4 copolymer 1.0 1.0 4 Di-panthenol 0.1 0.1 5 Methylparaben 0.2 0.2 6 ethanol 8.0 8.0 7 incense Quantity Quantity 8 Sebum-26-butes-26 / sebum-40 hardening castor oil 0.5 0.5 9 Purified water Remaining amount Remaining amount 10 Example 1 Extract 0.1 -

1) After weighing the raw materials of Nos. 1, 2, and 3, most of the purified water of No. 9 was added, and stirred at room temperature to dissolve.

2) The raw materials of Nos. 4, 5, 6, 7, and 8 were weighed, stirred and dissolved, and added to 1) above.

3) After adding No. 10 and a small amount of purified water to dissolve it, the solution was added to 1), stirred properly, and filtered to prepare a lotion.

Formulation Example 2 and Comparative Formulation Example 2: Preparation of Lotion

Prescription example of a lotion in the cosmetic containing the extract of Example 1 of the present invention is as follows.

number Raw material name Prescription Example 2 Comparative Prescription 2 One Shebutter 0.5 0.5 2 Cetyl alcohol 2.0 2.0 3 Glyceryl Monostearate 1.0 1.0 4 Butylmethoxydibenzoylmethane 1.0 1.0 5 Fiji-2 Orleans 0.5 0.5 6 Polysorbate 60 1.5 1.5 7 Macadamia oil 2.0 2.0 8 Squalane 5.0 5.0 9 Dimethicone 0.3 0.3 10 Pyromethicone 2.0 2.0 11 Propylparaben 0.1 0.1 12 Ethylhexylmethoxycinnamate 1.0 1.0 13 Purified water Remaining amount Remaining amount 14 Di-panthenol 0.05 0.05 15 Methylparaben 0.2 0.2 16 Disodium ID 0.05 0.05 17 1,3-butylene glycol 8.0 8.0 18 Triethanolamine 0.1 0.1 19 Xanthan Gum 0.05 0.05 20 Carbomer 0.1 0.1 21 incense Quantity Quantity 22 Example 1 Extract 0.1 -

1) The raw materials of Nos. 1 to 12 were weighed and placed in an oil phase heating tank to be dissolved by stirring at 75 ° C.

2) The raw materials of Nos. 13 to 18 were weighed and placed in an aqueous tank to be stirred and dissolved at 75 ° C.

3) The 2) was transferred to the emulsification tank under vacuum pressure, and then 1) was transferred to emulsify at 3500 rpm for 5 minutes.

4) Weigh the raw materials of Nos. 19 to 20 to make 1% solution solution, inject into the emulsification tank under vacuum pressure, homogeneously mix at 2500 rpm, stir the pedal mixer at 25 rpm while vacuuming and defoaming, and cool to 55 ° C. It was.

5) The extract obtained in Example 1 was weighed, dissolved in purified water at room temperature, and then weighed and placed in an emulsification tank, and then homogenously dispersed and cooled.

Formulation Example 3 and Comparative Formulation Example 3: Preparation of Cream

Prescription example of the cream of the cosmetic composition containing the extract of Example 1 of the present invention is as follows.

number Raw material name Prescription Example 3 Comparative Prescription 3 One Mango Butter 1.0 1.0 2 Cetyl alcohol 2.0 2.0 3 Glyceryl Monostearate / Pig 100 Glyceryl Stearate 1.5 1.5 4 Butylmethoxydibenzoylmethane 1.0 1.0 5 Cetearyl Alcohol / Cetearyl Glucoside 1.0 1.0 6 Hydrogenated Lecithin / C _12-16 Alcohol / Palmitate 1.5 1.5 7 Macadamia oil 2.0 2.0 8 Squalane 6.0 6.0 9 Dimethicone 0.3 0.3 10 Pyromethicone 2.0 2.0 11 Propylparaben 0.1 0.1 12 Ethylhexylmethoxycinnamate 1.0 1.0 13 Purified water Remaining amount Remaining amount 14 Di-panthenol 0.05 0.05 15 Methylparaben 0.2 0.2 16 Disodium ID 0.05 0.05 17 1,3-butylene glycol 8.0 8.0 18 Triethanolamine 0.1 0.1 19 Methylcellulose 0.05 0.05 20 Carbomer 0.1 0.1 21 incense Quantity Quantity 22 Example 1 Extract 0.1 -

1) The raw materials of Nos. 1 to 12 were weighed and placed in an oil phase heating tank to be dissolved by stirring at 75 ° C.

2) The raw materials of Nos. 13 to 18 were weighed and placed in an aqueous tank to be stirred and dissolved at 75 ° C.

3) The above 2) was transferred to the emulsification tank under vacuum decompression, and then 1) was transferred to emulsify at 3500 rpm for 5 minutes.

4) Weigh the raw materials of Nos. 19 to 20 to make 1% solution solution, inject into the emulsion tank under vacuum and reduced pressure, homogeneously mix them at 2500 rpm, stir the pedal mixer at 25 rpm while vacuum and defoaming, and 55 ° C. Cooled to.

5) The extract of Example 1 was weighed and dissolved in purified water at room temperature, and then the fragrance was weighed and placed in an emulsification tank, homogeneous dispersion, and cooling was completed.

Formulation Example 4 and Comparative Example 4: Hydrogel Patch

Prescription example of the hydrogel patch of the cosmetic containing the extract of Example 1 of the present invention is as follows.

number Raw material name Prescription Example 4 Comparative Prescription 4 One Purified water Remaining amount Remaining amount 2 glycerin 15.0 15.0 3 Sodium alginate 0.5 0.5 4 Acrylate / acrylamide copolymer 1.5 1.5 5 cornstarch 0.5 0.5 6 Polyacrylic acid 2.0 2.0 7 ethanol 1.0 1.0 8 Di-panthenol 0.1 0.1 9 Methylparaben 0.2 0.2 10 Disodium ID 0.05 0.05 11 Aluminum hydroside 0.1 0.1 12 Tartaric acid 0.2 0.2 13 Sorbitan oleate 0.2 0.2 14 1,3-butylene glycol 5.0 5.0 15 incense Quantity Quantity 16 Example 1 Extract 0.1 -

1) The raw materials of Nos. 1 to 6 were weighed and homogenized and stirred at room temperature to form a viscous liquid. The raw materials of Nos. 7 to 9 were separately dissolved at room temperature and injected into the dissolving tank, and then homogenized and stirred.

2) The raw materials of Nos. 10 to 14 were heated and dissolved at 60 캜, and then injected into small amounts in a dissolution tank to form gels.

3) Using a coating machine having an applicator gap on the hydrogel composed of the above formulation, the hydrogel composition was uniformly coated at 700 g / m ² on a siliconized polyester release film at a width of 1 m × 1 m in length. Laminated with polar polyethylene nonwovens. Finally, the multi-layer laminate size was double-cut into shapes that can be attached to the eyes and the entire face, and then packaged in a pouch laminated film composed of three layers such as paper, low density polyethylene, and aluminum.

Experimental Example 7

The experimental example of the present invention was carried out to measure the whitening effect of the human body of the cosmetic composition containing the herbal medicine extract of Example 1 as an active ingredient. 15 women 25-35 years old were selected as the creams of Prescription 3 and Comparative Prescription 3, and the subjects received the cream of Comparative Example 3 on the facial left side and the cream of Prescription 3 on the facial right side. It was applied twice a day for 1 month in a row.

15 women aged 20 to 45 years were selected as hydrogel patches of Formulation Example 4 and Comparative Example 4, and those subjects received the patch of Comparative Example 4 on the lower left eye area, and the formulation of Formula 4 on the lower right eye area. The patch was applied once a day for 30 minutes, and then applied for one month continuously by a peeling method.

After 1 month, the skin color of each subject's left / right application area and left / right eye area was compared by image analyzer, and the lightest color was 5, the middle color was 3, and the darkest color was 1 The color corresponding to was evaluated by the numerical value of 0.5 unit. Table 9 shows the cream (prescription example 3) and the patch containing the extract of the present invention (prescription example 4) and the cream (comparative example 3) and the patch (comparative example 4) containing no extract of the present invention. Comparing the skin color of the left and right facial and sub-eye areas of the subject shows that the cream and patch containing the extract of the present invention have an excellent whitening effect.

Subject Number One 2 3 4 5 6 7 8 9 10 11 12 13 14 15 Prescription Example 3 2.5 3.5 3.0 3.0 3.5 4.0 3.5 3.0 2.5 2.0 4.0 2.5 3.0 2.5 3.5 Comparative Prescription 3 1.0 1.5 1.5 1.0 2.0 1.0 1.0 1.5 1.0 1.0 1.5 2.0 1.0 1.0 1.5 Prescription Example 4 3.5 4.0 3.5 3.0 3.0 2.5 3.5 4.0 3.5 3.5 2.5 3.5 4.0 3.5 3.0 Comparative Prescription 4 2.0 1.5 1.5 2.5 1.5 1.0 1.5 2.0 1.5 1.0 1.0 1.5 2.0 1.0 1.5

Experimental Example 8: Confirmation of Skin Elasticity Effect

In order to measure the effect of increasing the skin elasticity when applying the cream of Prescription Example 3 and Comparative Example 3, 20 women of 20 to 35 years old had the cream of Prescription 3 on the right side of the face and the left side of the face. The cream of Comparative Example 3 was applied twice a day in the morning and evening, and after 1 week, 2 weeks, 3 weeks, and 4 weeks, the skin elasticity of the skin was measured by using a ball Istometer. The results are shown in the following Table 10 and FIG.

division 3 on prescription Comparative Prescription 3 Week 0 1 week 2 weeks 3 weeks 4 Weeks Week 0 1 week 2 weeks 3 weeks 4 Weeks Elasticity amplitude first 1.7 3.8 5.4 6.8 7.6 1.7 3.2 4.0 4.2 4.4 second 1.6 3.3 4.3 5.4 6.2 1.5 2.3 2.6 2.9 3.1 third 0.0 2.4 2.8 4.6 5.8 0.0 0.0 1.8 2.1 2.6 fourth 0.0 0.0 1.7 3.8 4.4 0.0 0.0 0.0 0.0 0.4 medium 0.83 2.38 3.55 5.15 6.00 0.80 1.38 2.10 2.30 2.63

As shown in Table 10, it was confirmed that the cream of the formulation of Comparative Example 3 containing the herbal medicine extract according to the present invention has an improvement in skin elasticity compared to the cream of Comparative Example 3.

As described above, the herbal medicine extract according to the present invention has a tyrosinase inhibitory activity, has an action of inhibiting melanin production, has a free radical scavenging effect and a cell proliferation effect such as vitamin-C, and has a skin elasticity enhancing effect. In addition, it has been confirmed that there is a lipid peroxide inhibitory effect such as alpha-routine, and is expected to be very useful as a cosmetic.

Figure 1 is a graph showing the inhibitory effect of mushroom tyrosinase of the herbal medicine extract according to the present invention.

Figure 2 is a graph showing the whitening efficacy of the herbal medicine extract according to the present invention in the animal skin.

Figure 3 is a graph showing the cell proliferation efficacy of the herbal extract according to the present invention.

Figure 4 is a graph showing the skin elasticity effect of the herbal medicine extract according to the present invention.

Claims (10)

1) mixing the yeongyeongseon, haejucho and cheonhwa after heating and stirring with water or an organic solvent of 10 to 15 times the weight of the mixed herbal medicine, primary extraction and filtration; 2) heating the organic residue with 7-12 times the weight of the mixed drug weight to the residue remaining after the filtration, and performing secondary extraction and filtration; And 3) combining the filtrates obtained after the first and second extraction, centrifugation using ethanol to take a supernatant, and concentrated under reduced pressure to obtain a powder extract, characterized in that the preparation method of the complex herbal extract. The method according to claim 1, wherein the gastric gland, hachicho, and cheonhwa are mixed in a weight ratio of 1: 0.5 to 1.5: 0.5 to 2.0. The method of claim 1, wherein the organic solvent used in the primary and secondary extraction is ethanol, protanol, butanol, glycerol, propylene glycol, 1,3-butylene glycol, methyl acetate, ethyl acetate, benzene, hexane, di Method of producing a complex herbal extract, characterized in that one or two or more selected from ethyl ether and dichloromethane. Herbal herbal extract, characterized in that obtained by the method selected from 1-3. The method of claim 4, wherein the herbal extracts, the herbal extracts, characterized in that containing the marine acid and oleanolic acid as an indicator component. The method of claim 5, wherein the herbal medicine extract, characterized in that the content of rosmarinic acid in the range of 0.01 to 0.08% by weight. According to claim 5, the herbal medicine extract, characterized in that the content of oleanolic acid in the herbal medicine extract ranges from 0.1 to 0.8% by weight. Skin whitening cosmetic composition, characterized in that containing the herbal medicine extract of claim 4. 9. The skin whitening cosmetic composition according to claim 8, wherein the extract of the botanical herbal medicine is contained in an amount of 0.001 to 20.0% by weight based on the total cosmetic composition. The skin whitening cosmetic composition according to claim 8, wherein the cosmetic is in the form of a softening cream, astringent makeup, lotion, essence, cream, gel, pack, soap, essence sheet, hydrogel patch or mask.