KR101107614B1 - Cosmetic Composition Containing Fermented of Viola mandshurica Extract - Google Patents

Abstract

The present invention relates to a cosmetic composition comprising a viola mandshurica extract as an active ingredient, and more particularly, inhibits tyrosinase activity, which is an enzyme involved in melanin biosynthesis, and produces melanin. Inhibiting the whitening effect (whitening effect), by having an antioxidant effect of removing free-radical (anti-aging effect) by showing the anti-aging effect (violet), violet ( Viola mandshurica ) relates to a cosmetic composition comprising the fermented extract as an active ingredient.

Viola mandshurica, whitening, tyrosinase, melanin, anti-oxidant, anti-aging, free-radical, anti -bacterial), cosmetic composition

Description

Cosmetic composition containing violet fermented extract {Cosmetic Composition Containing Fermented of Viola mandshurica Extract}

The present invention relates to a cosmetic composition comprising a violet ( Viola mandshurica ) extract as an active ingredient, and more specifically, a whitening, anti-aging or antimicrobial cosmetic composition containing a violet ( Viola mandshurica ) fermentation extract as an active ingredient. It is about.

Determining the skin color of a person involves various factors such as the activity of melanocytes (melanocytes) that make melanin pigment, the distribution of blood vessels, the thickness of the skin and the presence or absence of pigments in and outside the body such as carotenoids and bilirubin. The most important factor, melanin, is produced by the cascade enzyme reaction in the melanosome of melanocytes. Melanin in the surface layer forms a cap-like structure around the nucleus to protect skin organs from ultraviolet rays and at the same time It plays an important role in protecting cellular proteins by removing free radicals generated in vivo. Melanin is influenced by physiological factors such as heredity, hormone secretion, stress, and environmental factors such as UV irradiation, and melanin produced by internal and external stress stimulation is not released to the outside through the dead skin It remains stable. The most important step in the biosynthesis of melanin from Tyrosine is the initial reaction to produce DOPA through catalysis of Tyrosinase (Hwaring and Tsukamoto, FASEB J., 5 (14): 2902- 2909, 1991) may inhibit melanin biosynthesis through inhibition of this reaction. At this time, when free radicals are generated in the skin, inflammatory reactions, or ultraviolet rays are applied, melanin production is increased and accumulated in the epidermal layer of the skin, forming pigmentation such as blemishes, freckles, and mushrooms, and promoting skin aging. It is also known to be involved in inducing skin cancer (Kim et al., J. Korean Ind. Eng. Chem., 16 (3): 348-353, 2005). In order to treat or alleviate such pigmentation, various methods of combining whitening agents such as ascorbic acid, kojic acid, arbutin, glutathione, hydroquinone, etc. have been used in cosmetics and medicines, but for skin stability problems and cosmetics Because of its limitation in use due to the formulation problem, research has been conducted to find a whitening material having tyrosinase inhibitory activity from natural extracts such as lettuce, licorice and mulberry.

Viola ( Viola mandshurica ) and violet (violet) is a perennial herb that commonly grows on sunny meadows or hillsides in the spring, and includes essential oils (methyl salicylates), glycosides, flavonoids, saponins, and pigments. And use; Jan. Ill., 1991). The pharmacological effect is effective in purulent diseases, so it shows bactericidal, antipyretic and anti-inflammatory effects on boils, lymphatic tuberculosis and skin swelling. (Chinese medicinal dictionary) Sangwon 1607-myeon; > 756, p. 769 in the lower volume. Detoxification, anti-inflammatory, swelling, branch, top soil, diuretic, etc. It is used for jaundice, hepatitis, species, etc. It is also used as a fragrance.

Conventional techniques related to violet extract include extracting anticancer active substance from violet (Korean Patent Registration No. 10-0612087), cosmetic composition for acne prevention and treatment (Korean Patent Publication No. 10-2004-0056079), herbal composition ( Republic of Korea Patent Application Publication No. 10-2007-0051260), the effect of violet extract to protect nerve cells from oxidative stress by Glutamate (Lee, Mi-ra, Korean Journal of Applied Biochemistry, 2008), antioxidant activity and α-Amylase of violet extract and the inhibitory activity (this treasure me, Journal of Korea Food and nutrition, 2008) for α-Glucosidase and magnetic basement (紫花地下) Comparative study of the kinds of effects - violets in the anti-inflammatory antimicrobial (andeokgyun and bakseonggyun me, prime for Society , Vol . 6, No. 1, p77, 1991).

However, in the above research reports, there is no disclosure about the whitening effect, anti-aging effect or antibacterial effect of the violet fermentation extract according to the present invention.

Accordingly, the present inventors have made efforts to develop a natural extract excellent in whitening effect while having less skin side effects in terms of stability, violet fermentation extract inhibits tyrosinase activity, an enzyme involved in melanin biosynthesis, and melanin ( It inhibits melanin production and shows whitening effect, and it has anti-aging effect by removing free radicals. It shows anti-aging effect and antibacterial effect. This invention was completed.

An object of the present invention to provide a cosmetic composition for whitening, anti-aging or antimicrobial containing violet ( Viola mandshurica ) fermented extract as an active ingredient.

In order to achieve the above object, the present invention is added to 50 to 200 parts by weight of sugar to 100 parts by weight of violet ( Viola mandshurica ) and extracted by fermentation and maturation for 10 to 30 days for 150 to 200 days violet ( Viola mandshurica ) fermentation It provides a cosmetic composition for whitening, anti-aging or antimicrobial containing the extract as an active ingredient.

Viola mandshurica fermentation extract according to the present invention inhibits tyrosinase activity, an enzyme involved in melanin biosynthesis, inhibits melanin production, and exhibits a whitening effect, free radical ( Since it has an antioxidant effect of removing free-radical), it can be usefully used as a cosmetic composition for whitening, anti-aging or antibacterial.

In the present invention, violet ( Viola mandshurica ) fermented extract inhibits tyrosinase activity, which plays an important role in melanin pigment biosynthesis, thereby inhibiting melanin production and exhibits a skin whitening effect. By removing the free radicals (Free-radical) that causes the has a skin aging prevention effect, it may be characterized by having an antibacterial effect.

Human skin color is determined by the amount of melanin, carotene and hemoglobin, of which melanin is the most decisive factor. Melanin is synthesized in melanocytes (melanocytes), which are pigment cells in the basal layer of the skin (epidermis), and are transferred to peripheral keratinocytes (Keratinocytes) to represent human skin color. An abnormally low production of melanin causes skin lesions such as vitiligo. Overproduction, on the other hand, forms one of the main concerns of middle-aged women, blemishes and freckles, and is also closely related to skin cancer (Melanoma).

The whitening action is described in more detail as follows. Melanin is produced by tyrosine (Enzyme and non-enzymatic oxidation) in cells called melanocytes in the basal layer of the skin epidermis and is transferred to keratinocytes that make up the epidermis. Important enzymes involved in melanin biosynthesis include tyrosinase, tyrosinase-related protein-1 (TRP-1), and dopachrome tautomerase (DCT, TRP-2). Two enzymes besides tyrosinase have been recently developed and recently attracted attention, but the enzyme that plays a decisive role in the synthesis of melanin is tyrosinase. The first step in the biosynthesis of melanin is caused by tyrosinase, which is tyrosine hydroxylase activity that converts tyrosine to dopa and dopaquinone (Dopa). ) Has both dopa oxidase activity. In the process of melanin synthesis in melanocytes, first, tyrosine is used as a substrate, and tyrosinase produces dopachrome, and dopachrome produces dopa-quinone. The enzyme reaction produces melanin.

Recently, a new theory of melanin formation following the expression of signal transduction agents in skin cells has been proposed. That is, when the skin is exposed to ultraviolet light, interlukin-α, which causes inflammation, is expressed from keratinocytes in the epidermis, and this interleukin-alpha is keratinocyte endothelin. Promotes expression. This series of processes increases the formation of melanocytes and activates melanocytes. The melanin is formed by tyrosine tyrosine in the melanoma in the melanocytes thus formed.

The living body acquires energy through respiration using oxygen as an electron acceptor, and oxygen is absolutely necessary for life support. However, in the air, oxygen, which is a stable molecular state, is composed of superoxide radicals (O2-) and hydrogen peroxide (H2O2) due to physicochemical and environmental factors such as enzymes, chemicals, pollutants, photochemical reactions, and stress. It is converted into highly reactive oxygen species (ROS) and excessive free radicals such as singlet oxygen (1O2) and has a double side effect that causes fatal toxicity to the living body. Here, free radicals refer to substances with unpaired electrons generated during oxidative metabolism, and are characterized by very strong reactivity and very short lifespan.

In addition, since free radicals are very unstable in the state of molecules or atoms, they take away electrons from other materials nearby or pass their electrons to other materials. May cause functional damage. In addition, it is known as a cause of various diseases as well as various diseases by attacking and destroying enzymes or nucleic acids of the biological system or inducing mutations of chromosomes (Janssen YM et al., Lab Invest, 69: 261, 1993). .

In particular, since the skin is always in contact with oxygen and exposed to ultraviolet rays, lipid peroxides generated as a result of lipid peroxidation reactions by free radicals and free radicals may cause skin aging. Therefore, the antioxidant material having active oxygen and free radical scavenging activity can be used as an anti-aging cosmetic material which is very useful for skin.

In the present invention, violet ( Viola mandshurica ) extract may be used in the form of fermented extract.

The present invention adds 50 to 200 parts by weight of saccharides to 100 parts by weight of violet ( Viola mandshurica ) and extracts by fermentation ripening at 10 ~ 30 ℃ for 150 to 200 days and then contains the extract ( Viola mandshurica ) fermentation extract as an active ingredient It relates to a cosmetic composition for whitening, anti-aging or antibacterial.

In the present invention, with respect to 100 parts by weight of the violet (Viola mandshurica), it can be characterized in that 50 to 200 parts by weight of sugar is added, if less than the above range or exceeds the above range violet fermentation extract of an appropriate concentration Cannot be obtained.

In the present invention, the extraction temperature and the extraction time may be characterized in that it is carried out for 10 to 30 days at 150 ~ 200 days, if less than the range or exceeds the range is not made smooth extraction or the final extract Component deformation and the like.

In the present invention, the sugar may be selected from the group consisting of sugar, fructose, glucose, lactose, malt, syrup, dextrin, and oligosaccharide. In the present invention, the results of cell viability confirmation, tyrosinase activity inhibitory effect, melanogenesis inhibitory effect and DPPH radical scavenging effect confirmation experiment were carried out using violet methanol extract as a comparative example for violet fermentation extract. It was confirmed that the violet fermentation extract is excellent in whitening and anti-aging activity while having a lower cytotoxicity than the violet methanol extract extracted using a conventional organic solvent.

The cosmetic composition may be characterized in that it contains 0.001 to 90 parts by weight, preferably 2 to 10 parts by weight of the violet (Viola mandshurica) extract based on 100 parts by weight of the composition. If the amount is less than 0.001 parts by weight, the whitening effect or antioxidant effect is greatly reduced, and if it exceeds 90 parts by weight, a slight problem occurs on the skin.

In the present invention, the cosmetic composition may be selected from the group consisting of lotion, milky lotion, cream, gel, essence (essence) and pack cosmetics.

"Cosmetic water" is generally a clear liquid cosmetic applied to the skin surface to keep the skin clean and healthy. The basic function of the lotion is to supply moisture or moisturizing ingredients to the stratum corneum of the skin, and the lotion also has the function of softening the skin. Dissolves and stabilizes substances that are difficult to dissolve in water to make the appearance transparent. Transparent or semi-transparent using microemulsion or lipid nanosphere, and contains several percent oil in water (O / W type: Oil in Water Type), an opaque lotion emulsified in a water in oil (W / O type) or a water in silicon (W / S) type, and a mixture of a water-soluble polymer. In the present invention, well-known "cosmetics" can be appropriately used depending on the purpose thereof.

The lotion according to the present invention is purified water for supplying moisture to the stratum corneum by dissolving water-soluble components such as ion-exchanged water in addition to the violet fermentation extract; alcohol for dissolving and sterilizing useful components such as ethanol and propanol; and glycerin; polyethylene glycol Moisturizing for stratum corneum (PEG: polyethylene glycol), hyaluronic acid

Emulents for improving moisture retention and usability, such as ester oils and vegetable oils (emolients), solubilizers for solubilizing raw materials such as polyoxyethylene oleyl alcohol ether; citric acid; Buffer for adjusting the pH of products such as lactic acid, lactic acid and amino acids; flavors for adding flavors such as vanillin, orange, milk, geraniol (rose) and linalool (lemon) Antiseptics for inhibiting decay by inhibiting microorganisms such as methylparaben and phenoxy ethanol; colorants for coloring; anti-fading agents for preventing discoloration and discoloration such as metal ion blocking agents and ultraviolet absorbers; and astringents, bactericides, vitalizers It may contain a mixture of two or more drugs, such as anti-inflammatory agents or whitening agents.

The lotion according to the present invention may include about 0.001 to 90 parts by weight, preferably 0.01 to 10 parts by weight, and more preferably about 0.1 to 5 parts by weight of the violet fermented extract when the total amount of the lotion is 100 parts by weight. have.

"Emulsion" is an intermediate between a lotion and a cream and is generally a fluid emulsion. Latex is a cosmetic mainly used to supply moisture, moisturizer, oil, etc. to the skin in order to maintain the skin's moisture balance, moisturizing and flexibility. The component contained in the milky milk is similar to the ingredient included in the cream described later, but the amount of the solid oil and lead is less than that of the cream because the milky milk is fluid. In this invention, a well-known "milk" can be used suitably according to the use.

The emulsion according to the present invention may include about 0.001 to 90 parts by weight, preferably 0.01 to 10 parts by weight, and more preferably about 0.1 to 5 parts by weight of the violet fermented extract when the total amount of the emulsion is 100 parts by weight. have.

"Cream" is a type of emulsion in which one side of a liquid that does not mix with each other, such as water and oil, is dispersed in a stable state in the other dispersion medium. In the present invention, a known "cream" can be appropriately used depending on the use thereof.

The cream contains, for example, the following ingredients: That is, the aqueous phase components included in the cream include purified water such as ion-exchanged water; alcohols for dissolving and sterilizing useful components such as ethanol and propanol; and moisturizing agents for moisturizing the stratum corneum such as glycerin, PEG and hyaluronic acid; Mucus, such as a seed, pectin, a cellulose derivative, can be illustrated. Oil components included in the cream include hydrocarbons such as squalene, liquid paraffin, petrolatum and solid paraffin; oils such as olive oil, almond oil, cacao fat and castor oil; fatty acids such as stearic acid, oleic acid and palmitic acid, cetanol and stearyl alcohol Higher alcohols such as IPM (isopropyl myristate), glycerin triester, pentaerythritol tetraester, and the like; silicone oils such as polysiloxanes; Although the quantity of each component contained in a cream changes with the kind and use greatly, in this invention, a well-known thing may be used suitably.

The cream according to the present invention may include about 0.001 to 90 parts by weight, preferably 0.01 to 10 parts by weight, more preferably about 0.1 to 5 parts by weight of the violet fermented extract, when the total amount of the cream is 100 parts by weight. have.

A "gel" is a cosmetic that is uniform in appearance and transparent to translucent, such as a gel or a sol. In this invention, a well-known "gel" can be used suitably according to the use. The gel according to the present invention may include 0.001 to 90 parts by weight, preferably 0.01 to 10 parts by weight, and more preferably about 0.1 to 5 parts by weight of the violet fermentation extract when the total amount of the gel is 100 parts by weight. have.

"Essence" (essence) is basically a component such as the lotion, emulsion and cream described above, it can be exemplified to include an emulsion, a solubilizer, a moisturizer, water and other drugs. In the present invention, a known cosmetic liquid can be used. Emulsions included in the essence include natural oils such as olive oil, camellia oil, sesame oil, sunflower oil, sweet almond oil and jojoba oil; diglycerin esters or triglycerin esters of fatty acids such as capric acid, myristic acid, oleic acid and isostearic acid. Etc. can be illustrated. These emulsions function primarily as emollients. Solubilizers included in the cosmetic liquid include propylene glycol fatty acid esters such as glycerin fatty acid esters, polyglycerol fatty acid esters, sorbitan fatty acid esters, and monostearic acid propylene glycol, glycerin, diglycerine, ethylene glycol, propylene glycol, 1,3-butylene Polyhydric alcohols, such as glycol, etc. can be illustrated. On the other hand, the solubilizer may be exemplified, such as glycerin or diglycerin. Distilled water and deionized water can be illustrated as water contained in cosmetic liquid. Other pharmaceutical agents include fungicides, preservatives, vitamins, natural extracts from plants, colorants, flavors and the like.

The quantity of each component which comprises a cosmetic liquid can use a well-known component amount. For example, when the total amount of the cosmetic liquid is 100 parts by weight, the violet fermentation extract may contain 0.001 to 90 parts by weight, preferably 0.01 to 10 parts by weight, more preferably about 0.1 to 5 parts by weight.

A "pack cosmetic" is applied to the target area for the purpose of moisturizing the skin, promoting blood circulation, and purifying it. Pack cosmetics generally include a jelly, a paste, a powder, a creamy liquid, and the like, and may be applied to the face to form a film and then peeled off, wiped off after application or washed off after application. have. Powdered materials are used by dissolving or suspending uniformly in water or the like during use. Moreover, the pack cosmetics which impregnated cosmetics with the nonwoven fabric and collagen sheet of shapes, such as an eye mask, and the pack cosmetics etc. which are immersed in cosmetics at the time of using the said nonwoven fabrics etc. may be sufficient.

Pack cosmetics according to the present invention, when the total amount of the pack cosmetics to 100 parts by weight, 0.001 to 20 parts by weight, preferably 0.01 to 10 parts by weight, more preferably 0.1 to 5 parts by weight, most of the violet fermented extract Preferably it may comprise about 0.2 to 2 parts by weight. The other composition may be a known composition used for a pack cosmetic, and may be prepared according to a known production method.

However, without being limited to the above description, the above cosmetics may be used according to the method in which each is commonly used.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention, it will be apparent to those skilled in the art that the scope of the present invention is not to be construed as limited by these examples.

Comparative Example 1: Preparation of Violet Methanol Extract

1 kg of Viola mandshurica cultivated on a farm in Haman- gun , Gyeongnam, washed, drained and chopped , lyophilized and crushed, and then cooled at 20 ° C for 10 hours with 10 times the weight of the dried sample. Extraction and concentration under reduced pressure gave violet methanol extract.

In the present invention, the violet fermented extract of the violet methanol extract obtained in the above to confirm that the cytotoxicity is lower than the extract extracted with a conventional organic solvent and excellent in anti-aging activity, Experimental Example 1 to 4 Was performed.

Example 1 Preparation of Violet Fermented Extract

8 kg of Viola mandshurica cultivated on a farm in Haman- gun , Gyeongnam were washed, drained and sliced. Then, 4 kg of brown sugar was added and sealed, fermented and aged at about 18 ° C. for 6 months to obtain about 3 L of fermented extract ( Yield 37.5%).

Experimental Example 1: Effect of violet fermented extract on cell survival

B16F10 mouse melanin in a 96-well plate containing DMEM culture medium (Dulbecco's modified Eagle's medium, Gibco-BRL, USA) containing 10% fetal bovaine serum (FBS) Cells (KCLB, Seoul, Korea) were added at 1 × 10 ⁴ per well and incubated for 24 hours in a 37 ° C., 5% CO ₂ incubator. The plate was centrifuged, the supernatant of the medium was discarded, washed with PBS (phosphate buffer solution), and then serum-free medium (Arbutin, Sigma, USA: 20 μl, (1 μg / ml), (1 μg). / ML) 20µl, (100µg / ml) 20µl}, Violet Fermentation Extract of Example 1 1x (50mg / ml) 20µl, (5mg / ml) 20µl, (0.5mg / ml) 20µl And 1 ml (50 mg / ml) 20 ml, (5 mg / ml) 20 ml, (0.5 mg / ml) 20 ml of violet methanol extract, and 2 ml of 10% fetal bovine serum containing DMEM culture medium for melanocyte stimulating hormone. (a-melanocyte stimulating hormomone; a-MSH, Sigma, USA , 200 nM) was added to three wells for each experimental group, followed by incubation for 24 hours in a 37 ° C., 5% CO ₂ incubator. At this time, the control group was to add a-MSH without adding violet fermented extract or arbutin.

The plate was centrifuged and the supernatant of the medium was discarded and 10 μl of MTT solution was added with fresh DMEM culture medium containing 10% fetal bovine serum. After 4 hours of incubation, the plate was centrifuged, the supernatant of the medium was discarded, washed with PBS, 100 μl of DMSO (demethyl sulfoxide) was added to dissolve formazan and absorbance at 570 nm using an ELISA reader. The cell viability was calculated by the measurement. The data were performed three independent experiments each, and the results were expressed as mean ± SEM. The significance test was confirmed by ANOVA according to the biological statistical standard (P <0.001).

As a result of the MTT assay, as shown in Figure 1, the arbutin treatment group, a chemical whitening agent, showed a cell survival rate of 75-93% compared to the non-treatment group without treatment of the test solution, and the violet methanol extract treatment group compared to the no treatment group. While cell viability of 76-93% was shown, the experimental group treated with violet fermented extract showed a survival rate of 94% or more.

Experimental Example 2: Inhibitory Effect of Violet Fermentation Extract on Tyrosinase Activity

In order to confirm the whitening effect, tyrosinase activity, which plays an important role in melanin pigment biosynthesis, was tested in the following manner. B16F10 melanocytes (1 × 10 ⁵ ) were plated on 60 mm plates in 10% fetal bovine serum (FBS; fetal bovaine serum, Gibco-BRL, USA) and DMEM culture medium (Dulbecco's modified Eagle's medium, Gibco-) with 1% antibiotics. BRL, USA), and incubated for 24 hours at 37 ℃, 5% CO ₂ incubator. The plate was centrifuged, the supernatant of the medium was discarded, washed with PBS, and then serum-free medium (Arbutin, Sigma, USA: 20 μl, (10 μg / ml), 20 μl 20 μl of (100 μg / ml)}, 20 μl of the stock solution of violet fermentation extract of Example 1 (50 mg / ml), 20 μl of (5 mg / ml), 20 μl of (0.5 mg / ml) and violet methanol extract stock solution 1 μl (50 mg / ml) 20 μl, (5 mg / ml) 20 μl, (0.5 mg / ml) 20 μl and 2 ml of 10% fetal bovine serum containing DMEM culture medium and a-melanocyte stimulating hormomone a-MSH, Sigma, USA , 200 nM) was added to each well and treated in three wells, followed by incubation for 24 hours in a 37 ° C., 5% CO ₂ incubator. At this time, the control group was to add a-MSH without adding violet fermented extract or arbutin. The plate was centrifuged, the supernatant of the medium was discarded, washed twice with PBS, the cells were recovered with a scraper, the PBS was simply removed by a centrifuge and suspended in 1 ml of PBS. Dispense 0.5 ml into two 1.5 ml tubes and centrifuge at 3,000 rpm for 5 minutes to discard supernatant and obtain cell pellet. One is for measuring tyrosinase activity and the other is for measuring melanin pigment content. Used to.

First, tyrosinase activity measurement was performed as follows. 55 μl of lysis buffer (1% Triton X-100, 10 mM sodium phosphate pH 7.0, 0.1 mM PMSF) was added to the cell pellet, which was left on ice for 30 minutes and centrifuged at 5,000 rpm for 5 minutes. . 50 μl of supernatant was added to a 96-well plate, 25 μl of 10 mM tyrosinase solution and 50 mM sodium phosphate buffer were added to a total volume of 200 μl at 37 ° C. After reacting for 30 minutes, the absorbance was measured at 475 nm using an ELISA reader. Next, 200 μl of 1N NaOH containing 10% DMSO was added to the cell pellet for measuring the content of melanin pigment, and left at 80 ° C. for 1 hour to obtain intracellular melanin. This amount 100 μl was placed in a 96-well plate and the absorbance was measured at 405 nm using an ELISA reader, and then the melanin inhibition rate was compared with the control.

As a result, as shown in FIG. 2, it was confirmed that the tyrosinase activity was more suppressed in the violet fermentation extract treatment group than the arbutin treatment group or violet methanol extract treatment group.

Experimental Example 3: Inhibitory Effect of Violet Fermentation Extract on Melanogenesis

Intracellular melanogenesis inhibition experiments were cultured cells in the same manner as the measurement of tyrosinase activity. Intracellular melanin quantification was performed as follows. 200 μl of 1N NaOH containing 10% DMSO was added to the cell pellet prepared in Experimental Example 2, and left at 80 ° C. for 1 hour. After dissolving the melanin in the cells, 100 μl of this solution was placed in a 96-well plate. After measuring the absorbance at 490 nm using the melanin production inhibition rate was compared with the control to measure the whitening effect.

As a result, as shown in Figure 3, the violet fermentation extract treatment group significantly inhibited melanin production compared to the untreated group, the activity was more inhibited than violet methanol extract treatment group, similar to albumin chemical whitening agent It was confirmed that the melanin production inhibitory effect.

Experimental Example 4: DPPH radical scavenging effect of violet fermented extract

Antioxidant activity was tested using the Blois method to measure the free-radical scavenging effect of the sample using 1,1-diphenyl-2-picrylhydrazyl (DPPH, Aldrich, USA) as follows. . Samples were added to 0.1 mM DPPH methanol solution, mixed well, and reacted at room temperature for 10 minutes. Absorbance was measured at 517 nm using an ELISA reader to measure free radical scavenging effect. As a control, violet methanol extract (MVS) was added instead of violet fermentation extract (FVS), and the antioxidant activity of the test solution was compared with that of the synthetic antioxidant BHA (butylated hydroxyanisole).

As a result, as shown in Figure 4, violet fermentation extract was confirmed that there is no difference in antioxidant capacity when using 50ul compared to the synthetic antioxidant BHA (0.05%).

     In Experimental Examples 1 to 4 according to the present invention, the violet fermentation extract has a tyrosinase inhibitory effect and a melanin inhibitory effect to assay the whitening effect, compared with violet methanol extract or arbutin, which is a synthetic whitening agent, but with a chemical whitening agent. In contrast, it was confirmed that there is little cytotoxicity. In addition, it also has an antioxidant function that eliminates free radicals, so it can be seen that it also has an anti-aging effect.

Experimental Example 5: Antibacterial Activity of Violet Fermented Extract

In order to examine the antimicrobial activity of the violet fermentation extract was experimented by the following method using a disc diffusion method (disc diffusion). B. subtilis 168 and E. coli incubated in LB medium for 24 hours were diluted in 0.75% NaCl, and then prepared by adjusting the OD value to a concentration of 0.2 or more in 600 nm spectrophotometer. B. subtilis 168 and E. coli prepared above were evenly plated with a cotton swab on the surface of plate solid LB medium. The fermented extract obtained in Example 1 was placed on a sterilized tinfoil paper and placed on a sterilized tweezers with dry heat and sterilized 8 mm in diameter, and 10 μl, 100 μl, 200 μl, and 300 μl, respectively. 50 mg / mL) was absorbed into the filter paper to prepare an antimicrobial paper disc. The antimicrobial disc was picked up with sterile tweezers and placed on a medium on which bacteria were smeared, incubated at 37 ° C. for 24 hours, and then the growth of each strain around the disc was examined. As a control group, the experimental group was not absorbed violet fermentation extract was prepared under the same conditions as above.

 As a result, it was found that clear inhibitory zone (inhibitory zone, clear zone) occurred at all concentrations of 10 μl, 100 μl, 200 μl, and 300 μl (50 mg / mL), respectively. It was confirmed that high. Since the experiment is to qualitatively check the presence or absence of antibacterial activity of the violet fermentation extract, measuring the size of the transparent low-ring ring was omitted.

Therefore, when combining the above results, it can be seen that the violet fermentation extract of the present invention can be usefully used as a cosmetic composition for whitening, anti-aging or antibacterial.

Hereinafter, an example of a cosmetic formulation using a whitening effect, anti-aging effect or antibacterial effect of violet fermentation extract is intended to explain in detail, not intended to limit the present invention.

Formulation Example 1: Lotion

100 mg of violet fermented extract of Example 1; Glycerin 250 mg; 1.5 mg of 1,3-butylene glycol PEG 1500 allantoin DL-panthenol; EDTA 1 mg; 2 mg of benzophenone-9; 250 mg sodium hyaluronate; Ethanol 500 mg; Octicdodeces-16 polysorbate-20 10 mg; Preservatives, flavors, pigments 2 mg; And the distilled water was mixed with the appropriate amount to formulate the lotion.

Formulation Example 2: Cream

75 mg of violet fermented extract of Example 1; Lipophilic monostearineglycerin 100 mg; Stearyl alcohol 110 mg; 50 mg of stearic acid beeswax; Polysorbate-60 75 mg; Sorbitan stearate 30 mg; 50 mg of hardened vegetable oil; Squalane 150 mg; Mineral oil trioctanoyl 250 mg; Dimethicone 50 mg; Sodium magnesium silicate 5 mg; Glycerin 250 mg; Betaine 150 mg; 200 mg triethanolamine sodium hyaluronate; Preservatives, fragrances, pigments 2 mg; And a distilled water was mixed to formulate a cream according to a conventional method.

Formulation Example 3: Beauty Soap

Violet Fermentation Extract of Example 1 5g, wild green tea powder 2g, cactus powder 2g, pine needle powder 1g, palm oil 90g, natural fresh Rose floral flavors were mixed and formulated cosmetic soap according to a conventional method.

Although the composition ratio is a composition that is relatively suitable for preference cosmetics in a preferred embodiment, the composition ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, and use.

The specific parts of the present invention have been described in detail above, and it is apparent to those skilled in the art that such specific descriptions are merely preferred embodiments, and thus the scope of the present invention is not limited thereto. something to do. Thus, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

1 is a graph showing the results of confirming the cell activity change by the violet extract treatment in B16F10 cells using the MTT assay (FVS: violet fermentation extract of Example 1; MVS: violet methanol extract of Comparative Example 1).

Figure 2 is a graph showing the result of comparing the whitening effect with the sample untreated group and arbutin treated group by measuring the tyrosinase activity by violet extract treatment in B16F10 cells (FVS: violet fermentation of Example 1 Extract: MVS: violet methanol extract of Comparative Example 1.

Figure 3 is a graph showing the results of comparing the melanin (melanin) production amount by the violet extract treatment in B16F10 cells and the sample untreated group and arbutin treated group (FVS: violet fermentation extract of Example 1; MVS: of Comparative Example 1 Violet methanol extract).

4 is a graph showing the free radical removal ability of the violet extract as a DPPH scavenging effect (FVS: violet fermentation extract of Example 1; MVS: violet methanol extract of Comparative Example 1).

Figure 5 is a photograph showing the antibacterial test of violet fermentation extract of Example 1, (A) is the result for B. subtilis 168, (B) is the result for E. coli .

Claims (6)

Whitening cosmetics containing 50-200 parts by weight of saccharides in 100 parts by weight of Viola mandshurica , fermented and matured at 10-30 ° C. for 150-200 days, and extracted as fermented extract of Viola mandshurica as an active ingredient. Composition. Anti- aging cosmetics containing 50-200 parts by weight of saccharides in 100 parts by weight of violet ( Viola mandshurica ), fermented and matured at 10-30 ° C. for 150-200 days, and then extracted as fermented extract of Viola mandshurica as an active ingredient. Composition. 50 to 200 parts by weight of sugars are added to 100 parts by weight of violets ( Viola mandshurica ), and then fermented and matured at 10 to 30 ° C for 150 to 200 days, followed by extraction of fermented extract of Viola mandshurica as an active ingredient. Composition. The cosmetic composition according to any one of claims 1 to 3, wherein the sugar is selected from the group consisting of sugar, fructose, glucose, lactose, malt, syrup, dextrin, and oligosaccharide. The cosmetic composition according to any one of claims 1 to 3, wherein the cosmetic composition contains 0.001 to 90 parts by weight of the Viola mandshurica fermented extract, based on 100 parts by weight of the composition. The cosmetic composition according to any one of claims 1 to 3, wherein the cosmetic composition is selected from the group consisting of lotion, milky lotion, cream, gel, essence (essence) and pack cosmetics.

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