KR20130049980A - Fermented mulberry tree's extract and its cosmetic usage - Google Patents

Abstract

PURPOSE: A fermented extract of Morus alba L. by-products, and a cosmetic material containing the same are provided to increase useful ingredients for skin and to enhance moisturizing effect and elasticity. CONSTITUTION: A method for fermenting Morus alba L. by-products comprises: a step of mixing Morus alba L. leaves, fruits, branches, and barks with water, inoculating with lactic acid bacteria and yeast and fermenting; and a step of maturing the fermented liquid. The lactic acid bacteria includes one or more selected from the group consisting of Lactobacillus sakei, Lactobacillus brevis, Lactobacillus plantarum, Lactococcus lactis, Bacillus coagulance, and a mixture thereof. A cosmetic composition containing 0.001-80wt% of the fermented extract of Morus alba L. by-products.

Description

Fermented mulberry tree by-products and skin cosmetics containing same {Fermented mulberry tree's extract and its cosmetic usage}

The present invention relates to a method for preparing a mulberry by-product fermentation extract and a cosmetic composition comprising the same as an active ingredient, to a mulberry by-product fermentation and a moisturizing, elasticity, whitening, cosmetic composition for improving skin wrinkles containing the same as an active ingredient.

Fermentation refers to a process in which microorganisms decompose or synthesize organic matter using their own enzymes. Representative fermentation microorganisms include Bacillus subtilis, lactobacillus, koji, yeast, mold, and acetic acid.

Recently, as the interest in fermented foods and cosmetics using natural materials has increased in the cosmetics field, researches on whitening, skin wrinkle improvement, antioxidant, moisture, elasticity, etc. are active.

The fermentation broth extract does not use any other solvents and, through fermentation and maturation, converts sugar components contained in natural materials into alcohols, and is effective in flavonoids, vitamin C, vitamin P, amino acids, catechins, etc. It is possible to increase the extraction efficiency of the components. In addition, sometimes irritating or toxic natural materials can be used as a method of stabilizing natural materials, reducing toxicity or converting them into stable derivatives through fermentation extraction, and can obtain much more cosmetic effects. In addition, by the fermentation process such as lactic acid bacteria and yeast, it is possible to maximize the efficacy of natural materials in the process of conversion to low molecular weight materials that are easily absorbed, from the unstable or inactive form to the active form, to enhance moisturizing and elasticity, whitening, skin Wrinkle improvement effect can be maximized.

Mulberry is a genus of rosewood mulberry family. Most of the species are native to Asia. It is genetically close to Broussonetia. Mulberry is a fast-growing water when it is young, but grows slowly, and is less than 10-15m tall. Fruits ripen, dark purple to black, can be eaten, and some fruits are very sweet. Among the by-products of Mulberry Nammu, the root bark is called baekbaekpi, the fruit is mulberry, and the leaves are called sangji.

Sangbaikpi ( Cortex Mori : mulberry tree root bark) is the root bark of Morus bombycis koidzumi, Morus alba Linne , or other similar plants belonging to the Moraceae family.

Audi is a mulberry fruit, which contains glucose, fructose, calcium, iron, potassium, magnesium, zinc, vitamins A, B1 and C. Audi contains 4 to 5 times more iron than other fruits, 2 times more calcium than strawberries, 2 times more potassium than apples, and magnesium and zinc. Neohesperidine is a powerful antioxidant that is known to prevent skin lipids and oxidation, strengthening skin tissue and increasing skin elasticity. AHA (alpha hydroxy acid) is always used to control abnormal skin It keeps your skin clean. It refers to a substance that enhances or inhibits the function of a living body in living life, and is a substance that plays a role in correcting abnormal conditions caused by a lack or excessive secretion of a substance involved in function control in a living body. Can be defined The creation of bioactive substances is of great importance for a better and healthy life of the living body. The creation of new bioactive substances can be obtained from natural products, such as plants and animals, extracted and purified from microorganisms and metabolites of animal and plant cell lines, or by chemical synthesis.

At present, it is used as a by-product related to mulberry, mulberry leaves are used for slumber, and the fruit, Audi, is a fruit that is sold for some food. Thus, although the extract by using the overall by-products of the mulberry tree has been extended to the cosmetics field as a cosmetic food extract, there is no study on the fermentation extract of the mulberry by-products that can maximize the active ingredient through the fermentation process.

Accordingly, the present invention relates to a cosmetic composition for exploring physiologically active substances possessed by the mulberry by-product extract, and to enhance the content of the active ingredient of the mulberry by-product extract through the fermentation process, by-product of the mulberry by-product fermentation and moisturizing, elasticity To develop a cosmetic composition for improving skin whitening, wrinkles.

The present invention relates to a method for preparing a mulberry by-product fermentation extract and a cosmetic composition comprising the same as an active ingredient, the fermentation by the fermentation process through lactic acid bacteria and yeast and moisturizing, elasticity, whitening, skin wrinkles containing it as an active ingredient It is to provide a cosmetic composition for.

In order to achieve the above object, the present invention is a cosmetic having a mulberry by-product fermentation extract through the fermentation process and moisturizing, elasticity, whitening, skin wrinkle improvement activity containing the mulberry by-product fermentation extract.

Mulberry by-products used in the present invention are (1) mixing the dried mulberry by-products with water and fermenting by inoculating alone or mixed inoculation with lactic acid bacteria and yeast; (2) aging the fermentation broth; Fermentation broth obtained from the fermentation broth, or concentrates obtained by partially or totally depressurizing and lyophilizing the fermentation broth, or chemicals having a main effect in the fermentation broth and the concentrate themselves.

Mulberry by-products were washed with the same amount, and dried at room temperature and then ground to prepare a fine powder. The dried mulberry by-products are dried and used to maintain the nutritional and functional ingredients of the mulberry by-products for a long time, and are easy to handle when stored.

The mulberry by-product fermentation extract was dried at room temperature, mixed with a powder sample at a stable temperature in water, and then fermented broth was obtained through a fermentation process of lactic acid bacteria and yeast alone or mixed. After the fermentation process, it may include a low temperature aging or high temperature aging process.

Lactobacillus used for the fermentation is Lactobacillus sakei, Lactobacillus brevis, Lactobacillus brevis, Lactobacillus plantarum, Lactococcus lactis, Bacillus coagulance ) And yeast Bacillus subtilis (Bacillus subtilis) was used, the fermentation process is characterized in that one or two or more selected from the group consisting of these strains. The lactic acid bacteria were cultured to pH 4.0-5.0, yeast pH 6.5-7.5, lactic acid bacteria were incubated for 3 to 7 days at 20-37 city temperature, yeast was incubated for 24 to 72 hours at 50-60 city temperature. The cosmetic composition according to the present invention may contain, in addition to the above-mentioned extracts, other ingredients that can give a synergistic effect to the main effect without impairing the main effect of the present invention. The mulberry by-product fermentation extract may be present in the composition in an amount of 0.001 to 80% by weight based on the dry weight of the composition.

If the mulberry by-product fermentation extract is present in the form of a solution or powder in the form of nanocapsules, microcapsules, nanosomes, etc. those skilled in the art will be able to control the amount of the solution or powder in the composition according to the present invention within the concentration range . As described above, the present invention is provided to cosmetics, foods, medicines, and the like processed into a composition having a whitening and skin wrinkle improving activity having a moisturizing and elasticity improving effect.

The mulberry by-product fermentation extract of the present invention is not particularly limited in its formulation as an external preparation for skin, and it is a foundation cosmetic and foundation such as softener, milk lotion, nutrition cream, massage cream, essence, cleansing foam, cleansing water, pack or body oil It may be prepared in the form of a color cosmetic such as a lipstick, mascara or a makeup base. When the cosmetic of the present invention is used as a detergent, a face wash and a bath may be prepared. Hereinafter, the present invention will be described in detail with reference to Examples and Experimental Examples, which are only presented to aid the understanding of the present invention, but the present invention is not limited thereto.

As described above, the present invention relates to a method for preparing a mulberry by-product fermentation extract and a cosmetic composition comprising the same as an active ingredient, the mulberry by-product fermentation extract is an active ingredient including flavonoid component sugars, organic acids, vitamins and the like, such as mulberry In addition, the fermentation broth produced through the fermentation process using lactic acid bacteria and yeast is increased in the moisturizing, elasticity, whitening effect by increasing the useful substances on the skin by the conversion of the polymer compound into a low molecular compound by the enzyme produced during the fermentation process It is expected to be useful.

In addition, the cosmetic composition containing the mulberry by-product fermentation extract is excellent in antioxidant activity, the production of collagen, the inhibitory effect of MMP-1 (Matrix metalloproteinase; enzymes that promote the decomposition of collagen) exerts a skin wrinkle improvement effect.

<Example 1> Experimental preparation

Mulberry by-products have the same amount of mulberry leaves, berries, branches, roots, washed, and then dried at room temperature and ground to prepare a powder form.

<Example 2> Preparation of Mulberry By-Product Fermentation Extract

After gave good mixing was added to distilled water of 10 times the amount of dry weight of mulberry by-products, adjusting the pH to 4.0 to 5.0 to 6.5-7.5, and the pre-cultured lactic acid bacteria (Lactobacillus plantarum) and yeast (Bacillus subtilis), each 10 ⁷ - Add to 10 ⁸ CFU / mL concentration. Lactic acid bacteria were incubated for 3-7 days at 30-37 city temperature, yeast was incubated for 24 to 72 hours at 50-60 city temperature. In order to terminate the fermentation, heat was applied for 10 minutes at 80 cities to induce a loss of function of lactic acid bacteria and yeast. The precipitate was removed using a centrifugal separator and the supernatant was collected, or the filtered supernatant was concentrated under reduced pressure at 60 ° C. or lower and then lyophilized.

<Comparative example 1> Preparation of mulberry by-product general extract

Mulberry by-product general extract used as a control was obtained through the same hot water extraction process without the fermentation process. That is, 10 times the amount of dry weight of distilled water was added and mixed well, heated reflux extraction for 3 hours at 80 cities to separate the supernatant and the precipitate was extracted three times. Each extract was chilled and then filtered through a filter paper having a transmission size of 8 micrometers. The filtered extract was concentrated under reduced pressure at 60 ° C. or lower and dried completely using a vacuum dryer to obtain a mulberry by-product general extract.

Experimental Example 1 Determination of Components and Contents of Fermented Extract of Mulberry By-Product

<1-1> Total Flavonoid Content Determination

Total flavonoid content measurement was performed as follows. Add 100% of sample solution, 300ul of ethanol, 20ul of 10% aluminum nitrate solution, 20ul of 1M potassium acetate solution and 560ul of water, and stir thoroughly. After 40 minutes at room temperature, the liquid layer is measured for absorbance at 405 nm using a microplate reader (Chromate, USA). For the blank test, ethanol solution was used instead of the sample solution, and blanks were used for water instead of aluminum nitrate solution. The standard curve was prepared by diluting the Quercetin 0.05% concentration, and the total flavonoid content of the sample was calculated from the calibration curve.

Table 1

Total flavonoid content

Experimental Example 2 Antioxidant Activity

DPPH (2.2-diphenyl-1-picryl hydrezyl) radical scavenging ability was used to measure the antioxidant activity of the mulberry by-product fermentation extract.

500 uM DPPH (2.2-diphenyl-1-picryl hydrezyl) solution was dissolved in ethanol and filtered. The prepared DPPH solution and the sample solution were mixed 1: 1 and mixed vigorously, and left in a dark room for 20 minutes. After measuring the absorbance at 540 nm using a Microplate reader (Chromate, USA), DPPH radical removal rate (%) was calculated through the following equation (1). At this time, 1 mM of ascorbic acid was used as a positive control for removing radicals.

Where A is the absorbance obtained from the blank and B is the absorbance obtained from the sample

Table 2

Antioxidant Activity of Fermented Extract of Mulberry By-Product

<Experiment 3> Skin wrinkle improvement effect

<3-1> Collagen biosynthesis evaluation

A procollagen assay was performed to measure the amount of newly formed collagen in Fibroblast cells. Fibroblast, one of the cell lines involved in collagen production, was treated with Dulbecco's modified essential medium (DMEM, Welgene, Korea) containing 1% Antibiotic-Antimycotic (AA, Welgene, Korea) and 10% Fetal bovine serum (FBS, Welgene, Korea). Incubated in 37 cities, 5% CO ₂ incubator using. Fibroblast cells of 2 × 10 ⁵ concentration were inoculated into 6 well plates, incubated for 24 hours, medium was removed, and serum-free medium was treated for 24 hours. After 24 hours, the serum-free medium was removed, washed with PBS, and irradiated at 12.5 mJ at a UV 312 nm wavelength. Thereafter, the samples were treated by concentration in phenolred free DMEM medium and further incubated for 48 hours. Ascorbic acid 50 uM as a positive control, UV (+) as a negative control, and a control group as a control. After incubation for 48 hours after the extract treatment, the medium was collected. The amount of collagen was measured using a Procollagen Type-I C-Peptide (PIP) EIA kit (Takara Bio Inc. MK101). 100ul of antibody-POD conjugate solution was added to each well, and 20ul of sample or standard was added and mixed well and left for 3 hours in 37 cities. After the reaction, the contents were removed and washed four times with 400ul of PBS. After removal, 100ul of substrate solution was added and reacted for 15 minutes at room temperature. Then, 100ul of stop solution was added and mixed gently. The absorbance at 450nm was measured using a microplate reader (Chromate, USA), and a standard concentration curve was prepared. It was calculated (Table 3).

As shown in Table 3, in order to examine the anti-wrinkle activity of the extracts by UV-stimulation, the mulberry by-products yeast fermentation extracts were compared with the mulberry by-products general extract (80 cities, 3h) to compare with . When the mulberry by-product yeast fermentation extract than the mulberry by-products general extract (80 cities, 3h), it was confirmed that the amount of newly generated collagen increased.

Table 3

Collagen Biosynthesis by UV-stimulation of Mulberry By-Product Fermentation Extracts

<3-2> MMP-1 inhibition evaluation

There are various kinds of enzymes that degrade collagen, and the most well known is MMP-1 (Matrix metalloproteinase I, collagenase) that degrades collagen type I. Fibroblast, one of the cell lines involved in collagen production, was treated with Dulbecco's modified essential medium (DMEM, Welgene, Korea) containing 1% Antibiotic-Antimycotic (AA, Welgene, Korea) and 10% Fetal bovine serum (FBS, Welgene, Korea). Cultured in a 37%, 5% CO ₂ incubator. Fibroblast cells of 2 × 10 ⁵ concentration were inoculated into 6 well plates, incubated for 24 hours, medium was removed, and serum-free medium was treated for 24 hours. After 24 hours, the serum-free medium was removed, washed with PBS, and irradiated at 12.5 mJ at a UV 312 nm wavelength. Thereafter, the samples were treated by concentration in phenolred free DMEM medium and further incubated for 48 hours. Ascorbic acid 50 uM as a positive control, UV (+) as a negative control, and a control group as a control. After incubation for 48 hours after the extract treatment, the medium was collected. The collected medium was measured for absorbance at 450 nm with an ELISA reader using an MMP-1 measurement kit (RPN 2610, Amersham Bioscience, UK). Add a buffer to a well in a 96 well plate coated with an Anti-MMP1 antibody, leave it blank, and place the free sample in the medium and 100ul of the standard solution in each well. Fill wells with washing buffer and wash them three times. Add 100ul peroxide coujugate and react for 2 hours without shaking at room temperature. Fill well with washing buffer and shake for 30 minutes. After stopping the reaction with 100ul 1M H2SO4 was compared by measuring the absorbance at 450nm using a microplate reader (Chromate, USA) (Table 4).

As shown in Table 4, in order to examine the anti-wrinkle activity of the extracts by UV-stimulation, the mulberry by-products yeast fermentation extracts were compared with the mulberry by-products general extract (80 cities, 3h) for comparison . MMP-1 activity was newly inhibited when treated with the mulberry by-product yeast fermentation extract compared to the mulberry by-product general extract (80 cities, 3h), which is consistent with the collagen biosynthesis results (Table 3).

Table 4

Inhibition of MMP-1 Activity by UV-stimulation of Mulberry By-product Yeast Fermentation Extract

As shown in Table 4, in order to examine the anti-wrinkle activity of the extracts by stimulation by UV light, compared with the mulberry by-product fermentation extract and the mulberry by-product general extract (80 shown, 3h) for comparison. It was confirmed that the amount of collagen newly generated by the mulberry by-product fermentation extract than when treated with the mulberry by-product general extract (80 cities, 3h). This shows the result corresponding to the collagen biosynthesis of Table 3.

Experimental Example 4 Emulsion Base Preparation

An emulsion base was prepared comprising a cosmetic containing mulberry by-product fermentation extract. The cosmetics used in this experiment are in the form of an emulsion cosmetic composition, the composition of which is shown in the table. First, add the mulberry by-product yeast fermentation extract and other contents recorded in the table by weight, and (a) heat the phase to maintain at 90 degrees, and then purified water (30%) to 90 degrees and then slowly (a) in the water It is added and emulsified uniformly with a homomixer. Thereafter, the mixture was cooled to room temperature, and the remaining (b) purified water was added and stirred uniformly to prepare a cosmetic liquid.

Table 5

Emulsion base manufacturing

<Experiment 5> Measurement about skin elasticity effect

In order to measure the test for skin elasticity effect of the cosmetic according to the present invention, 10 healthy males and females in their 40s and 40s were included. The cosmetics prepared in Experiment 5 were used, and the subjects were treated in a constant temperature and humidity chamber (temperature 20-22 degrees, relative humidity 40-60%) at baseline, 2, 4, and 6 weeks for 1 hour after cleansing. After stabilizing the skin, measurements were taken. Skin elasticity was measured by cutometer SEM 474 (Courage + Khasaka, Germany), and the average value was measured by measuring 500 mbar sound pressure, 2 sec inhalation time, and 5 times repeated measurement.

Table 6

Skin elasticity measurement

As a result of measuring skin elasticity by using cutometer SEM 474, it was confirmed that the mulberry by-product yeast fermentation extract has about 4 times better skin elasticity than the common extract of mulberry by-product.

<Experiment 6> Skin safety test

In order to measure the skin safety test of the cosmetic according to the present invention, a conventional patch test was performed. The subjects were 10 healthy males and females in their 30s and 40s, and the cosmetics prepared in Experimental Example 4 were patched inside the lower arm of the forearm to remove the sulcus after 48 hours to observe the condition of the skin. The results are shown in Table 7 below.

Table 7

Skin safety measurement

<Experiment 7> Whitening effect test on human skin

The subjects were 10 healthy males and females in their 30s and 40s, and 1.5 ~ 2 times the minimum erythema dose (Minimal Erytherna Dose) of each subject. UV light was irradiated to a degree to induce skin blackening. After irradiation, 1% of each test substance (solvent is 1,3-butylene glycol: ethanol = 7: 3), 2% arbutin as a control, only a solvent, and one place without change of state for 10 weeks Observed. Skin color was measured on a weekly basis with a colorimeter CR2002 (Minolta, Japan). The difference between the skin color (ΔL *) at the start and completion of the coating was calculated according to Equation 3 below, and the results are shown in Table 10 below. The whitening effect is determined by comparing ΔL * between the sample application site and the control site. When the value of ΔL * is about 2, the whitening of the deposited pigment is apparent, and when it is about 1.5 or more, the whitening effect may be determined. .

Table 8

Whitening effect on skin

From the results of Table 8, it was confirmed that the mulberry by-product yeast fermentation extract according to the present invention showed an excellent whitening effect than the arbutin, a positive control as well as the mulberry by-products general extract.

Claims (9)

(1) mixing the same amount mixture of mulberry by-products of the upper leaves, mulberry, upper limbs, and lettuce leaves with water and fermenting by inoculating single or mixed lactic acid bacteria and yeast; (2) aging the fermentation broth; Fermentation method characterized in that it comprises a. According to claim 1, The lactic acid bacteria Lactobacillus sakei (Lactobacillus sakei), Lactobacillus brevis (Lactobacillus brevis), Lactobacillus plantarum, Lactococcus lactis (Lactococcus lactis), Bacillus coagulus (Bacillus) coagulance) and a fermentation method characterized in that one or more selected from the group consisting of these. The fermentation method of claim 1, wherein the yeast is Bacillus subtilis. The cosmetic composition according to claim 1, which contains 0.001 to 80% by weight based on the total weight of the mulberry by-product fermentation extract. The cosmetic composition for improving whitening and skin wrinkles according to claim 1, wherein the mulberry by-product fermentation extract is concentrated and freeze-dried after the fermentation broth or the extract is characterized by a mulberry by-product. The pH of the culture medium is adjusted to 4.0-5.0 to 6.5-7.5, lactic acid bacteria are incubated for 3 to 7 days at 20-37 city temperature and yeast is incubated for 24 to 72 hours at 50-60 city temperature Cosmetic composition for improving whitening and skin wrinkles, characterized in that. According to claim 1, wherein the cosmetic composition for moisturizing, elasticity, whitening, skin wrinkle improvement in the formulation of nanocapsules, microcapsules, nanosomes, etc. containing the mulberry by-product fermentation extract According to claim 1 or 2, wherein the formulation of the cosmetic composition is a flexible lotion, gel, water-soluble liquid, milk lotion, nutrition cream, massage cream, essence, oil-in-water (O / W) or water-in-oil (W / O) type Basic cosmetics such as emulsions, flakes or solid anhydrous products, oil dispersions in aqueous phase using globules, forms of lipid vesicles in ionic or nonionic form, ointments, cleansing foams, cleansing water, packs or body oils, It is selected from color cosmetics such as oil-based or oil-in-water makeup base, foundation, skin cover, lipstick, lip gloss, face powder, two-way cake, eye shadow, mascara, cheek color and eyebrow pencils. Moisturizing, elasticity, whitening, cosmetic composition for improving skin wrinkles. The cosmetic, food, pharmaceutical or household product according to claim 1, which is prepared from a mulberry by-product fermentation extract.