KR20130019186A - Skin care preparations for whitening, anxi-oxidation or anti-biotics with fermented extract of lycium chinensis miller - Google Patents

Abstract

PURPOSE: A cosmetic composition containing a fermented Lycium Chinense Miller extract and a method for preparing the same are provided to ensure skin whitening function and antioxidation against reactive oxygen and to strong antibacterial activity against Staphylococcus aureus. CONSTITUTION: A functional cosmetic composition with skin whitening, antioxidative, and antibacterial effects contains a fermented Lycium Chinense Miller extract. A method for preparing the extract comprises: a step of pulverizing Lycium Chinense Miller; a step of inoculating Lactobacillus sp. and fermenting at 25-35 deg. C. for 72-96 hours; a step of centrifuging the fermented Lycium Chinense Miller and removing medium ingredients and other floating matters; a step of completely removing remaining Lactobacillus sp.; and a step of freeze-drying.

Description

Skin care preparations for whitening, anxi-oxidation or anti-biotics with fermented extract of Lycium Chinensis Miller}

Fermented goji berry to obtain extracts with different molecular size or composition from existing ingredients to enhance the effect of cosmetic effects (whitening, antioxidant, antibacterial, etc.)

Lycium Chinensis Miller (Lycium Chinensis Miller) is a deciduous shrub belonging to the branch family, native to Asia Minor, and our country is mainly produced by Chungyang-gun, Chungnam, and Jindo-gun, Jeonnam. Goji berries are the fruit of the Goji berries (Lycii fructus), the roots of the phalanges of phalanges, leaves are Gujiyeob, young shoots are said to be pure grass (Lee SR 1983). According to the agreement, the wolfberry is effective in nourishing, tonic, blood, and jigal (Her J 1999). According to the herbaceous tree, the wolfberry is not toxic, and the root canal is hardened and does not get hot and cold. It is good for diabetes and neurological paralysis, which promotes lung and kidney function and improves vision (Lee SJ 1987). Wolfberry contains various functional ingredients such as betaine, cholin, physalien, rutin, β-sitosterol and zeazanthin (Lee BY et al. 1995), antioxidant, antimicrobial and anticancer effects, immune enhancing effect, liver function improvement, blood cholesterol lowering It is known to exhibit various physiological activities such as effects (Joo IS et al. 1997, Chung HK et al. 2004, Kang KI et al., 2006).

The present invention relates to a functional cosmetic composition comprising a fermentation extract of goji berry having whitening, antioxidant and antibacterial activity, and more particularly, the composition has a function of reducing melanin and an antioxidant effect of eliminating free radicals. The present invention relates to a functional cosmetic composition having an antimicrobial effect against Staphylococcus aureus, which secretes toxins that act as superantigens that cause skin problems.

The skin color of the human body is generally determined by melanin, redox hemoglobin, carotene and melanoids, and the most influential one is melanin. Melanin is made from melanocytes and serves to protect the skin from harmful free radicals generated from sunlight, especially ultraviolet light. After melanin is formed, the original skin color is restored by skin metabolism in healthy skin, but in aging skin, melanin pigment does not fall off the skin with dead keratin and is deposited on the skin as it becomes blemishes and blemishes. Therefore, to maintain skin health, it is necessary to suppress the synthesis of excessive melanin.

Melanin is formed through a waveguide, an intermediate metabolite of dopachrome, by an enzyme called tyrosinase as a substrate in skin cells. Such a method of inhibiting the synthesis of melanin may include preventing or reducing exposure to ultraviolet rays by using a cosmetic containing a suitable sunscreen, inhibiting melanin synthesis by inhibiting tyrosinase activity, or melanocyte synthetic melanocytes. The method of inhibiting the cell activity of the, and the like. However, currently used substances such as albutin, kojic acid (5-hydroxy-2-hydormethyl-v-pyrone), and hydroquinone (Hydroquinone) have low inhibitory activity, instability of the substance itself, and toxicity to skin. Is holding. Therefore, there is a need for a natural inhibitor that is not toxic to skin and stable and has a high melanin inhibitory effect. Recently, with increasing interest in the usefulness of natural materials, traditional herbal ingredients for skin beauty have been used.

Free radicals, which are free radicals, are biochemical mechanisms that naturally occur during aerobic energy metabolism in the cells of all aerobic organisms that breathe oxygen, and have a system that can fundamentally protect cells from damage caused by free radicals. . However, the production of free radicals beyond the self-defense ability by the influence of the external environment promotes the oxidation of organelles, and the DNA is modified to cause various diseases.

Oxygen, which is breathed by living things, is the most stable form when the energy level is low and there are two electrons in the outer orbital. This form is called triplet oxygen. When triplet oxygen absorbs a large amount of energy by thermal energy, light energy, chemical energy, etc., the outermost orbital is removed by a reduction reaction, and the outermost orbital is converted into singlet oxygen which is completely empty. Is called active oxygen.

Free radicals require electrons to fill because the outermost orbitals that are not paired with electrons are removed, and they can easily react with the materials in which they exist and speed up the oxidation reaction very quickly.

As a common example, in skin cells exposed to ultraviolet rays, strong light energy converts triplet oxygen into singlet oxygen, a harmful oxygen, and rapidly oxidizes the cells, causing rapid aging of the skin. (Yun Sun, Kwak Ho Kyung , Functional Food Science , Life Science , 2006, pp95 -101)

SOD (Superoxide dismutase), Catalase, Glutathione Peroxydase, Vitamin E (Vitamin E), etc. are present as catalysts for mechanisms to prevent the oxidation of organs from free radicals in the body. As antioxidant enzymes age, their defense against free radicals gradually decreases, resulting in various diseases and rapid aging. Therefore, there is a strong demand for the development of natural antioxidants with strong antioxidant power without harm to the human body.

In addition to skin aging caused by free radicals, skin problems that appear in various forms can be fatal for members of the modern society who are pursuing external beauty.The cause of allergic problems in the form of allergy is mainly on the skin surface. Many are parasitic microorganisms. Toxins released by Staphylococcus aureus among various microorganisms act as superantigens, causing atopic dermatitis problems on the skin surface. (MR Kim , SY Yang , TY Han , JJ Seo , Effects of Emulsion Containing the Specific Antibodies against Staphylococcus aureus and Staphylococcus Toxins on Atopic Dermatitis , Journal of Society Cosmetics Scientists of Korea , v32 n4 , pp233 -239 )

Chemical antimicrobials applied to the surface of the skin cause strong skin irritation irrespective of effects, so it is burdensome to use it in the state with various troubles of skin diseases, so it is necessary to develop natural antibacterial agent without skin irritation, especially atopic skin There is a strong demand for the development of natural antibacterial agents against Staphylococcus aureus.

Meanwhile, goji berry fermented extract contains crude protein 18.1%, crude fiber 11.2%, free amino acid 12.39 mg%, free sugar 6.2%, tannin 633.1 mg% and organic acid 1877.5mg%, which contains a large amount of ingredients that have a unique taste. It has antimicrobial activity and is used as a main component of natural antibacterial agents.

In addition, the fermentation of these natural materials tends to increase the antioxidant activity as well as the antibacterial activity as well as the content of polyphenols as a whole, and the experimental results of increasing the antioxidant activity of garlic-fermented black garlic have been proved. (EK Jang , JH Seo , SB Lee , Physiological Activity and Antioxidative Effects of Aged Black Garlic Extract , Korean J. Food Sci . Technology , v40 n4 , pp443-448 )

Ginger's antimicrobial activity has already been suggested through Oriental medicine books such as Dongbobogam, Herbal Wood, etc. In particular, ginger and shogaol have strong antimicrobial activity, and free radical activity. In addition, the effect of inhibiting the DNA damage of the cells by oxygen has been reported, which demonstrates the antioxidant activity. (Park Jong-chul, Natural Products of Functional Foods , Hyoil Publishing Co., 2002, pp83 -86)

The functionality of the flavonoids of the buttercups, the hyperoside and the persicarin, has been demonstrated many times in the field of health functional foods and contributes greatly to the antioxidant activity that is characteristic of the flavonoids. (Bakjongcheol of functional foods Natural Science, published by the effective date, 2002, pp37 -42)

The present inventors fermented and extract goji (leaf, fruit, root) while screening natural products with excellent whitening, antimicrobial and antioxidant activity that can be applied as a cosmetic material, functionality having excellent whitening, antibacterial and antioxidant effects It was found that it can be applied as a cosmetic composition, to complete the present invention based on this.

       Fermented goji berry with Bacillus, Aspergillus and Kefir to produce ingredients useful for skin care.

Goji berries are fermented with Bacillus, Aspergillus and Kefir. It is better known by the name Tibetan mushrooms, and is commonly used as a fermentation agent for milk fermentation products for colon health and obesity prevention in homes. Tibetan mushroom is the name is the name attached to that Tibetan local monks consumed by mixing milk to cache peer (kefir) for health care, not actual Mushrooms inde lumps of lactic acid bacteria (Lactobacillus) and yeast (Yeast) united like a small mushroom Looks like.

Since most of the components are Lactobacillus , fermentation of general foods is possible in addition to dairy fermentation. Since most fermentation cosmetics use fermentation methods using Bacillus , most of the fermentation cosmetics are used for fermentation. Fermented raw material manufacturing is expected to be able to differentiate from other companies.

As described above through the above Examples and Experimental Examples, the cosmetic composition of the present invention exhibited not only an excellent whitening effect, an antioxidant effect, but also an antimicrobial activity, such as antioxidant and atopic dermatitis, which are the basis of skin aging. It may be usefully used as a cosmetic composition and cosmetic additive for preventing or improving trouble.

In order to achieve the above object, the present invention provides a functional cosmetic composition which essentially contains fermented extract fermented with goji berry and lactic acid bacteria.

Specifically, the composition of the present invention is characterized by consisting of fermented extract extracted through the centrifugation and filtering process after inoculating and fermenting lactic acid bacteria ( Lactobacillus sp ) on the leaves, berries and roots.

The present invention is inoculated with lactic acid bacteria ( Lactobacillus sp ) by crushing gojija, fermented for 72-96 hours at 25 ~ 35 ℃ and centrifuged to remove the media components and other suspended matter, and then passed through the filter again 2㎛ remaining lactic acid bacteria Can be completely removed to form a liquid fermentation extract, or lyophilized to a powder state.

The cosmetic composition may be included in an amount of 0.001% to 100% by weight based on the total weight of the cosmetic.

Hereinafter, the present invention will be described in detail.

In order to manufacture fermentation extract of wolfberry used in the present invention,

Specifically, the wolfberry is prepared and washed as it is, and then sterilized by autoclaving to remove various bacteria, dried sufficiently in the shade, crushed small, and mixed in a basic medium at a constant concentration (about 10 ⁵ to 10 ⁷ cells / mL). After inoculating Lactobacillus sp on the medium, it is fermented in shaker incubator for 72-96 hours at 27 ℃ ~ 35 ℃.

After centrifugation, only the supernatant except for the precipitate was taken and again passed through a 2㎛ filter to obtain a final fermentation extract from which the lactic acid bacteria contained in the supernatant were completely removed.

The present invention provides an antioxidant cosmetic composition for skin whitening and aging prevention, wherein the amount of fermented extract has 0.01 to 30% by weight of the total cosmetic composition, preferably 0.01 to 5.0% by weight. Provided are antimicrobial cosmetic compositions useful for improvement and treatment.

The formulation of the cosmetic composition comprising the active ingredient of the fermentation extract of the wolfberry of the present invention as described above is not particularly limited, and may be appropriately selected according to the purpose. For example, the cosmetic composition of the present invention may be prepared in a formulation such as flexible cosmetics, nourishing cosmetics, nutrition cream, massage cream, essence, pack, emulsion, oil.

The flexible longevity may be prepared to contain 1 to 15% by weight of polyhydric alcohols and 0.01 to 5% by weight of a surfactant in addition to the fermentation extract of the present invention. In addition, nutrient cosmetics, nutrition cream and massage cream may be prepared to contain 1 to 20% by weight of oils and 2 to 15% by weight of waxes in addition to the fermentation extract of the present invention, the essence polyhydric alcohols 3 in addition to the fermentation extract of the present invention It can be prepared by containing ˜30% by weight.

The cosmetic composition of the present invention requires oils, surfactants, moisturizers, lower alcohols, thickeners, chelating agents, pigments, preservatives, fragrances, purified water, etc., which are formulated to general basic cosmetics in addition to the components described above in each formulation. It can mix | blend suitably according to the use.

Hereinafter, to present a preferred embodiment to help the understanding of the present invention,

Example  1. Preparation of wolfberry fermented extract

Chungcheongsan Cheongyang-si, Chungcheongsan prepared by washing, high-pressure sterilization to remove various bacteria, dried in the shade and then crushed small, mixed with a constant concentration (about 10 ⁵ ~ 10 ⁷ cells / mL) to the basal medium and this medium Lactobacillus sp ) after inoculation and incubated in shake incubator for 72-96 hours at 27 ℃ ~ 35 ℃.

The fermentation broth was centrifuged at 10,000 x g for 30 minutes to take only the supernatant except for the precipitate, and again passed through a 2㎛ filter to completely remove the lactic acid bacteria that may be included in the supernatant to obtain a final fermentation extract.

Example  2. Manufacturing of Soft Cosmetics

Ingredients content(%)   Fermentation extract of Example 1 10.0   ethanol 5.0   glycerin 2.0   1,3-butylene glycol 5.0   Sodium hyaluronate 2.0   Disodium ID 0.01   Preservative, coloring, fragrance Proper amount   Purified water Balance

Example  3. Manufacture of nutritional cream

Ingredients content(%)   Fermentation extract of Example 1 5.0   glycerin 3.0   1,3-butylene glycol 5.0   Cetyl alcohol 3.0   Sorbitan stearate 1.0   Stearic acid 1.0   Polysorbate 60 2.0   Triethanolamine 0.3   Grapeseed Oil 3.0   Preservative, coloring, fragrance Proper amount   Purified water Balance

Example  4. Emulsion  Produce

Ingredients content(%)   Fermentation extract of Example 1 5.0   glycerin 3.0   1,3-butylene glycol 5.0   Cetyl alcohol 2.0   Sorbitan stearate 1.0   Stearic acid 1.0   Polysorbate 60 2.0   Triethanolamine 0.3   Olive oil 2.0   Preservative, coloring, fragrance Proper amount   Purified water Balance

Example  5. Manufacture of Essence

Ingredients content(%)   Fermentation extract of Example 1 7.0   glycerin 5.0   1,3-butylene glycol 7.0   ethanol 4.0   Polyoxyethylene alcohol ether 1.0   Sodium hyaluronate 5.0   Grapeseed Oil 2.0   Camellia oil 1.0   Preservative, coloring, fragrance Proper amount   Purified water Balance

Experimental Example  1. Whitening effect experiment of fermented extract of the present invention

In order to confirm the whitening effect of the fermentation extract of Example 1 used in the present invention, a mouse-derived B-16 melanoma (B16 BL6) cell line (KCLB 80006) was purchased and 10% FCS (<52> fetal calf serum) was obtained. After inoculation into the included DMEM (Dulbemlo's modified Eagle's medium) medium was incubated for 24 hours at 37 ℃, CO2 incubator (CO2 concentration 5%). 28.8 μl of the extract prepared in Example 2 (2) was treated with the cultured melanoma cell culture solution and cultured again for 5 days under the same conditions. Thereafter, the cultures were washed with PBS (phosphate buffered in saline) and treated with 0.05% trypsin and 0.53 mM EDTA solution to separate adsorbed cells, followed by centrifugation for 5 minutes to obtain only cells. After the cell number was measured and the cell number of each experiment was constant, centrifugation was carried out again to obtain precipitated melanin, and treated with 1N NaOH to dissolve the melanin, and the absorbance was measured at 405 nm. At this time, using the cells not treated with the extract as a control, the melanin synthesis inhibitory effect of each extract was measured compared to the amount of melanin in the control.

Melanin quantification was used by modifying the method of Hosoi et al. Cells were dispensed at 3 × 10 5 cells / well in 6 well plates, the samples were treated and incubated in a 37 ° C. CO 2 incubator for 48 hours. Cells were collected, cell number was measured, precipitated by centrifugation at 1200 rpm for 5 minutes, and then dissolved in 1 ml homogenization buffer (50 mM Sodiumphosphate pH 6.5, 1% Triton X-100, 2 mM PMSF). 200 μl of 1 N NaOH (10% DMSO) was added to the pellet, and the O.D405 value was measured at 405 nm after vortex. The melanin produced in each well was calculated using a standard calibration curve obtained with melanin standard (Sigma, USA). Melanin was compared with melanin production in 104 cells.

Intracellular tyrosinase activity measurement

Intracellular tyrosinase activity was measured using Pawelk and Pomerantz15,16). The cells were dispensed into 5 × 105 cells / well in 6 well plates and incubated for one day before processing the samples. After 24 hours, cells were collected and lysed, and then 0.1Msodium phosphate buffer (pH 6.8) added with 0.2% L-DOPA was added thereto, incubated at 37 ° C for 2 hours, and absorbance was measured at 490 nm.

To investigate the inhibitory effect of tyrosinase activity of the compositions, 10 mM L-DOPA (L-dihydroxyphenylalanine, in 0.1M Sodium Phosphate buffer, pH 6.8) dissolved in 0.1M sodium phosphate buffer (pH 6.8), a substrate of tyrosinase 0.2 mL of the extract of the compositions prepared in Example 1 was added to 0.4 mL and mixed well at room temperature. 0.4 mL of tyrosinase (100 units / mL) was added to the mixed solution and reacted at 25 ° C. for 15 minutes. Thereafter, the reaction was stopped under refrigeration conditions and the absorbance was measured at 475 nm. In the control group, distilled water, which is an extraction solvent, was added instead of the extract. Tyrosinase inhibitory effects were calculated by the formula:

              % Inhibition of tyrosinase activity = 1- (absorbance of extract / absorbance of control) × 100

Experimental Example  2. Antioxidant Effect Test of Fermented Extract of the Present Invention

In order to confirm the antioxidant efficacy of the fermentation extract of Example 1 used in the present invention, a relatively stable free radical and maximum absorbance at 517 nm in the presence of radical state, 1,1-diphenyl- having the characteristic of losing absorbance when erased 2-pycrylhydrazyl (DPPH) was used.

DPPH purchased from Sigma Co., Ltd. was dissolved in ethanol to a concentration of 0.15 mM, and 0.5 ml was added to a quartz cell for UV-Spectrophotometer, and 0.5 ml of the fermentation extract of Example 1 was added thereto and left for 30 minutes. After sufficiently allowing the reaction to proceed, absorbance was measured at 517 nm with a UV-Spectrophotometer. In this case, methyl alcohol was used as a control group, and a control group containing vitamin C was also measured to compare the efficacy of the fermentation extract, and the fermentation extract when the absorbance of the treated fermentation extract became half the absorbance of the control group. The concentration of is expressed in IC ₅₀ (inhibitory concentration 50) value is shown in Table 5 below.

Table 5

division IC ₅₀ Methyl alcohol 0.0032% Vitamin C 0.0006% Fermented Extract 0.0009%

As shown in Table 5, the wolfberry fermented extract used in the cosmetic composition of the present invention can be said to be an antioxidant that can eliminate very strong free radicals.

Experimental Example  3. Functionality of the present invention Cosmetics  Antioxidant Effect of the Composition

Using the cosmetic composition of Example 2 containing goji berry fermentation extract as a sample with reference to the result obtained in Experimental Example 1 was carried out an experiment on the antioxidant efficacy in the same manner as Experimental Example 1, vitamin C and vitamin E as a control , Methyl alcohol was used.

The IC ₅₀ values shown here are shown in Table 6 below.

Table 6


division IC ₅₀ Methyl alcohol 0.0033% Vitamin E 0.0014% Vitamin C 0.0005% Example 2 Cosmetic Composition 0.0015%

Experimental Example  4. Antimicrobial Effect Experiment of Fermented Extract of the Present Invention

In order to verify the antimicrobial activity of the fermentation extract of Example 1 against Staphylococcus aureus, a minimal inhibition concentration (MIC) measurement method was performed.

After fermentation extract of Example 1 was added to BHI medium, which is Staphylococcus aureus culture medium, at a predetermined concentration interval from 0.01% by weight to 0.5% by weight, and then solidified. Inoculation was followed by incubation for 24 hours in a fixed incubator at 37 ℃ to observe the degree of bacterial growth.

In addition, to compare the degree of actual antimicrobial activity, triclosan (triclosan) and salicylic acid (salicylic acid), which are frequently used in antibacterial cosmetics due to the wide antimicrobial range as a control group was added to the medium to evaluate the efficacy.

TABLE 7


Sample concentration 0.01 0.05 0.10 0.15 0.20 0.25 0.30 0.35 0.40 0.50 Example 1 + + + + - - - - - - Triclosan + + - - - - - - - - Salicylic acid + + + - - - - - - -

          (+; No antimicrobial activity,-; antimicrobial activity)

As shown in Table 7, the goji berry fermentation extract showed an antimicrobial activity against Staphylococcus aureus at 0.20% by weight, and showed similar levels compared to triclosan or salicylic acid, which exhibited antimicrobial activity at 0.10% and 0.15% by weight, respectively. have.

Experimental Example  5 Functionality of the Invention Cosmetics  Antimicrobial Effect Test of Composition

Referring to the results obtained in Experimental Example 1, using the cosmetic composition of Example 2 containing the wolfberry fermentation extract as a sample was carried out antimicrobial activity experiment in the same manner as in Experimental Example 3.

Table 8
Sample concentration 0.01 0.05 0.10 0.15 0.20 0.25 0.30 0.35 0.40 0.50 Example 2 + + + + + + - - - -

 (+; No antibacterial activity,-; antibacterial activity)

As shown in Table 8, the antimicrobial activity was shown at 0.30% by weight, and the concentration of the antimicrobial activity was slightly higher than that of the fermentation extract as a sample, but the difference was not significant.

Claims (2)

      Functional cosmetic composition with whitening, antioxidant effect and antibacterial effect extracted by fermenting goji berry with lactic acid bacteria The method of claim 1, wherein the cosmetic is specified to be a lotion, cream, emulsion, essence, pack, oil