KR102465293B1 - Preparation method of fermented korean rice wine lees extract and cosmetic composition comprising the same - Google Patents

Abstract

The present invention relates to a manufacturing method of a rice wine lees fermented extract and a cosmetic composition containing the same as an active ingredient. According to the present invention, the method includes the steps of: (A) inoculating rice wine lees with Lactobacillus bacteria; (B) fermenting the rice wine lees while culturing the same in the dark for 22 to 26 hours at 25 to 30℃; (C) fermenting the same while irradiating light of 450-480 nm for 10-15 hours to the same; (D) fermenting the same while irradiating light of 540-560 nm for 10-15 hours to the same; (E) fermenting the same while removing the light source and culturing the same in the dark for 22 to 26 hours; (F) extracting a fermented product of rice wine lees with a solvent; and (G) performing concentration and drying after filtration. The rice wine lees fermented extract of the present invention, fermented and extracted by irradiation with light of a specific wavelength, exhibits an excellent skin inflammation relieving effect, a skin moisturizing effect, and an exfoliation effect.

Description

Preparation method of fermented korean rice wine lees extract and cosmetic composition comprising the same

The present invention relates to a technology for producing a sake lees fermented extract and using it as a cosmetic, and specifically, by irradiating a specific wavelength of light during the fermentation process, a sake lees fermented extract excellent in skin inflammation relief, moisturizing, and exfoliating effects is produced, and it is used as a cosmetic. It is about the technology used as

The stratum corneum of the skin is the first barrier that has a defense mechanism against external environmental stimuli and is an important factor determining the skin condition. In addition, the stratum corneum of healthy skin contains 15-20% of moisture, and when the moisture falls below 10%, it becomes dry and wrinkles increase due to decreased gloss and elasticity. In general, skin keratin formation proliferates from the lowermost layer of the epidermis, gradually rises to the outermost layer of the epidermis, and then naturally separates from the epidermal layer after a certain period of time has elapsed. However, when defects are caused by extrinsic factors such as penetration of irritants or intrinsic factors such as aging, hormones, and stress, the dead skin cells of the stratum corneum do not fall off in a normal cycle and are delayed, making the skin look rough and dull. Furthermore, as the skin barrier function is weakened by stimulation, the resistance is lowered, which leads to a decrease in the function of the epidermal double lipid membrane and increases the sensitivity to irritants, which causes skin inflammation. becomes Accordingly, the need for a skin scrub product that can be used without irritation while protecting the skin barrier is increasing.

A general scrub cosmetic removes old dead skin cells by rubbing particulate matter into the skin. However, hard particles have a limitation in that they can cause microscopic wounds by stimulating the skin.

Under this background, the present inventors studied to develop a cosmetic that can remove dead skin cells without irritating the skin. , the present invention was completed by confirming that the exfoliating effect was excellent.

(0001) Republic of Korea Patent Publication No. 10-2016-0000959 (2016.01.06) (0002) Republic of Korea Patent Publication No. 10-2012-0051180 (2012.05.22) (0003) Republic of Korea Patent Publication No. 10-2006-0062665 (2006.06.26)

An object of the present invention is to provide a method for producing a fermented sake lees fermented extract, which is extracted after irradiating light of a specific wavelength during the fermentation process to produce a fermented sake lees extract having excellent skin inflammation relief, moisturizing, and skin exfoliation effects.

Another object of the present invention is to provide a cosmetic composition containing the fermented sake lees extract as an active ingredient.

According to the present invention to achieve the above object,

(A) inoculating the Lactobacillus bacteria in sake lees; (B) fermenting while culturing in the dark for 22 to 26 hours at 25-30 ° C; (C) fermenting while irradiating light of 450-480 nm for 10-15 hours; (D) fermentation while irradiating light of 540-560 nm for 10-15 hours; (E) fermenting while removing the light source and culturing in the dark for 22 to 26 hours; (F) extracting the fermented sake lees with a solvent; And (G) there is provided a method for producing a fermentation extract of sake lees comprising the step of concentration and drying after filtration.

More preferably, the Lactobacillus is Lactobacillus plantarum (Lactobacillus plantarum).

The extraction in step (F) is water, ethanol, methanol, butanol, propanol, butylene glycol, glycerin, chloroform, ethyl acetate, dichloromethane, hexane, acetone, acetonitrile, petroleum ether and diethyl ether consisting of It consists of extraction with at least one solvent selected from

According to the present invention in order to achieve the above other object, there is provided a cosmetic composition containing 0.001 to 20% by weight of the fermented sake lees extract prepared by the above method based on the total weight of the cosmetic composition.

The cosmetic composition is characterized in that for relieving skin inflammation, for moisturizing the skin or for exfoliating the skin.

The sake lees fermented extract produced by the method of the present invention has excellent skin inflammation relief, moisturizing and skin exfoliating effects, so it can be usefully used as a skin soothing and moisturizing cosmetic. In addition, since it has an excellent effect of removing dead skin cells without skin irritation, it can be usefully used as a cosmetic for a scrub.

1 shows the skin keratin improvement effect of the cosmetic composition containing the fermented sake lees extract of the present invention.

Hereinafter, the present invention will be described in more detail.

Sake lees refer to the leftover liquor residues after brewing and squeezing liquor. In addition to starch and protein, sake lees contain a large amount of nutrients such as fiber, minerals, vitamins, alcohol and organic acids, enzymes, and yeast.

Research on sake lees has been conducted in various fields, and as a result, it has been reported that sake lees have a blood sugar control effect in diabetic patients and lower blood pressure in hypertensive patients. In addition, the antibacterial, antioxidant activity, anti-inflammatory activity, and collagen synthesis promoting effect of sake lees are also known.

The present invention uses such sake lees as a raw material, fermenting it with Lactobacillus bacteria while irradiating light of a specific wavelength, and extracting it to produce a sake lees fermented extract excellent in skin inflammation relief, skin moisturizing and skin exfoliation effects, and using it as a cosmetic. do it with

According to the present invention, (A) the step of inoculating Lactobacillus bacteria in sake lees; (B) fermenting while culturing in the dark for 22 to 26 hours at 25-30 ° C; (C) fermenting while irradiating light of 450-480 nm for 10-15 hours; (D) fermentation while irradiating light of 540-560 nm for 10-15 hours; (E) fermenting while removing the light source and culturing in the dark for 22 to 26 hours; (F) extracting the fermented sake lees with a solvent; And (G) there is provided a method for producing a fermentation extract of sake lees comprising the step of concentration and drying after filtration.

First, the sake lees are inoculated with Lactobacillus bacteria. At this time, when using Lactobacillus plantarum among Lactobacillus bacteria, it is more preferable because it exhibits more excellent skin improvement activity.

After inoculation, primary fermentation is performed while dark cultured at 25-30℃ for 22-26 hours. Then, the second fermentation is carried out while irradiating 450-480 nm light for 10-15 hours under the same temperature condition. At this time, more preferably, light of 453 nm is irradiated.

Then, the tertiary fermentation is carried out under the same temperature condition while irradiating light of 540-560 nm for 10-15 hours. More preferably, light with a wavelength of 552 nm is used. When irradiating such light, the distance from the lamp is maintained at 30-40 cm.

After the third fermentation, the light source is removed and the fourth fermentation is performed while dark cultured again for 22 to 26 hours to prepare a fermented sake lees.

Using the prepared fermented product as a solvent, preferably water, ethanol, methanol, butanol, propanol, butylene glycol, glycerin, chloroform, ethyl acetate, dichloromethane, hexane, acetone, acetonitrile, petroleum ether and diethyl extraction with at least one solvent selected from the group consisting of ethers. The extract is filtered, concentrated and dried to prepare the fermented sake lees extract of the present invention.

In the process of fermentation in this way, after dark culture for a certain period of time, 450-480 nm light and 540-560 nm light having different wavelengths are irradiated for a certain period of time in order, and the light source is removed again for dark culture and fermentation. The sake lees fermented extract produced through the process has a superior skin inflammation alleviation effect (Test Examples 2 and 3) and skin moisturizing effect (Test Example) compared to the sake lees extract extracted by not irradiating light or irradiating a specific single wavelength of light during the fermentation process. 4, 5) and skin exfoliation effect (Test Example 6) were confirmed.

The fermented sake lees extract as the active ingredient is contained in an amount of 0.001 to 20% by weight based on the total weight of the cosmetic composition.

The cosmetic composition is used for alleviating skin inflammation, moisturizing the skin, or exfoliating the skin.

The cosmetic composition of the present invention can be prepared in the form of general emulsified formulations and solubilized formulations. Cosmetics with emulsified formulations include nourishing lotions, creams, essences, and exfoliants, and solubilized cosmetics include softening lotions and feminine washes. In addition, by containing a dermatologically acceptable medium and base in addition to cosmetics, it can be prepared in the form of an adjuvant that can be applied topically and systemically commonly used in the field of dermatology.

In addition, the formulation of the cosmetic composition in the present invention is not limited, but skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nourishing lotion, exfoliant, massage cream, nourishing cream, moisture cream, hand Cream, essence, pack, soap, shampoo, treatment, cleansing foam, cleansing lotion, cleansing cream, body lotion, body cleanser, feminine cleanser, emulsion, press powder, loose powder, and eye shadow can be applied.

[Example]

Hereinafter, the present invention will be described in more detail based on the following examples and test examples. However, the following examples are only for illustrating the present invention, and the present invention is not limited by the following examples.

Comparative Example 1: Preparation of sake lees extract

Sake lees were added to purified water 10 times the weight, extracted at 80° C. for 3 hours while stirring, and filtered after cooling. And after concentration at a temperature of 50 ~ 60 ℃ using a vacuum concentrator and freeze-dried with a freeze dryer to prepare an extract powder.

Examples 1 to 5: Preparation of fermented sake lees extract

The lactic acid bacteria Lactobacillus plantarum (Lactobacillus plantarum) was inoculated to 1×10 ⁵ CFU/g of sake lees, and then dark culture fermentation was performed at 27° C. for 24 hours. Then, photo-culturing was carried out while irradiating LED light for each wavelength under the conditions shown in Table 1 below. Example 1 was fermented for 24 hours without light irradiation, Example 2 was fermented with light of 380 nm wavelength for 24 hours, Example 3 was fermented with light of 453 nm wavelength for 24 hours, Example 4 was fermented by irradiating light with a wavelength of 545 nm for 24 hours, and Example 5 was fermented by irradiating light with a wavelength of 660 nm for 24 hours.

And after irradiating light with a wavelength of 453 nm for 12 hours, fermented while irradiating light with a wavelength of 552 nm for 12 hours as Example 6, after irradiating light with a wavelength of 453 nm for 12 hours, fermented while irradiating light with a wavelength of 410 nm for 12 hours This was referred to as Example 7.

In the case of LED irradiation, the culture temperature was maintained at 27 °C and the distance from the lamp was maintained at 30 cm. After this fermentation, the LED light source was removed and dark culture fermentation was performed again at 27°C for 24 hours.

Thereafter, it was added to purified water 10 times the weight of the fermented sake lees, extracted at 80° C. for 3 hours while stirring, and filtered after cooling. And after concentration at a temperature of 50 ~ 60 ℃ using a vacuum concentrator and freeze-dried with a freeze dryer to prepare an extract powder.

division LED wavelength (nm) Irradiation time (h) Example 1 - - Example 2 380 24 Example 3 453 24 Example 4 545 24 Example 5 660 24 Example 6 453 / 552 12 /12 Example 7 453 / 410 12 /12

Test Example 1: Confirmation of skin cell protection effect

The skin cytoprotective effect was confirmed by the treatment of the sample of the lees extract prepared in Comparative Examples and Examples. Human-derived fibroblasts were cultured in a 96-well plate at a concentration of 1×10 ³ cells/well, and 0.2 ml of IMDM medium containing 10% FBS was supplied, and then cultured in a CO ₂ incubator for 24 hours. Thereafter, MTT assay was performed to determine the viability of the cells. For MTT assay, 0.1 ml of MTT solution was added to the cultured cells, followed by incubation for 4 hours, then the medium was removed and the formazan formed by adding 0.5 ml of DMSO was measured for absorbance at 570 nm using ELISA. Cell viability (%) was measured by the following formula, and the results are shown in Table 2 below.

Cell viability (%) = [(St-Bo/Bt-Bo)] × 100

Bo: Absorbance at 570 nm of wells in which only the cell culture medium was subjected to color development

Bt: Absorbance at 570 nm of wells subjected to color reaction without sample treatment

St: Absorbance at 570 nm of wells treated with samples and subjected to color reaction

division Cell viability (%) 0.1% 0.5% Comparative Example 1 99 88 Example 1 99 94 Example 2 100 97 Example 3 100 100 Example 4 100 98 Example 5 99 91 Example 6 100 100 Example 7 100 97

As can be seen from Table 2, cell viability of 90% or more was confirmed for all of the samples to which the extract of Example was applied, which had a low effect on cytotoxicity and was confirmed to be safe and relatively less likely to cause skin irritation.

Test Example 2: Inflammatory mediator COX-2 and PGE2 expression reduction effect

In order to examine the effect on the cytokine production of Human Epidermal Keratinocyte (HaCaT), 100 μl of 5×10 ⁵ cells/ml cells were put into a 96 well plate, incubated in a CO ₂ incubator for 48 hours, and then the medium was discarded, After washing the surface with PBS solution, it was replaced with a medium not added with FBS, starvated for 18 hours, and cultured for 24 hours. After culturing, the cells were stimulated by treatment with the stimulating factor SLS (10 ppm), treated with 0.1% of each sample in Examples and Comparative Examples, and cultured for 24 hours, then mRNA expression levels were investigated using RT-PCR. .

Using the mRNA expression level increased by each of the stimuli as a control, the expression reduction rate was calculated by the following formula, and the results are shown in Table 3.

mRNA expression reduction rate (%) = [1-(sample mRNA expression level / control group mRNA expression level)] × 100

division COX-2 expression reduction rate PGE2 expression reduction rate Comparative Example 1 5.9 4.4 Example 1 11.4 14.1 Example 2 20.8 18.7 Example 3 24.4 23.2 Example 4 26.2 21.0 Example 5 12.7 10.8 Example 6 28.6 24.9 Example 7 18.4 13.9 Dexamethasone 26.5 23.4

As can be seen in Table 3, the expression of inflammatory factors induced by stimulation, COX-2 and PGE2, was further reduced in the fermented sake lees fermented extract of the present invention as compared to the hot-water extract of the comparative example. The samples of Examples 3 and 4, which were fermented and extracted while irradiated with light of 453 nm wavelength and 545 nm wavelength, showed a better expression reduction effect. The sample of Example 6 showed the most excellent expression reduction effect.

Test Example 3: Inflammatory mediators NF-kB and TNF-a expression reduction effect

In order to examine the effect on the cytokine production of Human Epidermal Keratinocyte (HaCaT), 100 μl of 5×10 ⁵ cells/ml cells were put into a 96 well plate, incubated in a CO ₂ incubator for 48 hours, and then the medium was discarded, After washing the surface with PBS solution, it was replaced with a medium not added with FBS, starvated for 18 hours, and cultured for 24 hours. After incubation, the cells were stimulated by treatment with a stimulus factor SLS (10ppm), treated with 0.1% of each sample of the above examples and comparative examples, and after culturing for 24 hours, the cells were examined for mRNA expression level using RT-PCR did.

Using the mRNA expression level increased by each of the stimulatory factors as a control, the expression reduction rate was calculated by the following formula, and the results are shown in Table 4.

mRNA expression reduction rate (%) = [1-(sample mRNA expression level / control group mRNA expression level)] × 100

division NF-kB expression reduction rate TNF-a expression reduction rate Comparative Example 1 9.4 8.8 Example 1 13.8 10.8 Example 2 16.8 13.9 Example 3 19.4 18.9 Example 4 20.7 19.2 Example 5 14.2 10.2 Example 6 22.0 21.7 Example 7 17.3 14.4 Dexamethasone 20.3 21.8

As can be seen in Table 4, the expression of inflammatory factors induced by stimulation, NF-kB and TNF-a, in the fermented sake lees fermented extract of Examples of the present invention compared to the hot-water extract of Comparative Example further reduced. In particular, the sample of Example 6, which was fermented and extracted by irradiating light with wavelengths of 453 nm and 552 nm in parallel for 12 hours, showed the most excellent expression reduction effect.

Test Example 4: Hyaluronic acid production amount measurement

Hyaluronic acid production amount by the fermented sake lees fermented extract prepared in Examples of the present invention was measured. HaCaT cells were aliquoted in a 96-well plate at a concentration of 1×10 ⁵ cells/well and cultured, then replaced with a medium not added with FBS, starvated for 18 hours, and cultured for 24 hours. HaCaT cells were treated with 0.1% of each sample and cultured for 24 hours. The recovered culture solution was measured for hyaluronic acid (HA) production using an ELSIA kit. The results are shown in Table 5

division HA production (ug/ml) Example 1 30.1 Example 2 35.7 Example 3 41.3 Example 4 40.6 Example 5 28.8 Example 6 43.2 Example 7 36.0 Comparative Example 1 23.2 hyaluronic acid 44.7

As can be seen in Table 5, the sake lees fermentation extract sample of the example of the present invention showed an excellent amount of hyaluronic acid production compared to the sample of the comparative example. of the sample showed the best moisturizing effect.

Examples 8 to 9: Preparation of cream containing fermented sake lees extract

The cream containing the sake lees fermented extract of the above example was prepared according to a conventional method with the composition and content shown in Table 6 below. A cream not containing it was used as Control Example 1.

ingredient Content (wt%) Control Example 1 Example 8 Example 9 Sake lees fermented extract (Example 1) - 2.0 - Sake lees fermented extract (Example 6) - - 2.0 lipophilic monostearate glycerin 2.0 2.0 2.0 stearyl alcohol 2.2 2.2 2.2 stearic acid 1.5 1.5 1.5 beeswax 1.0 1.0 1.0 Polysorbate 60 1.5 1.5 1.5 Sorbitan Stearate 0.6 0.6 0.6 purified vegetable oil 1.0 1.0 1.0 squalane 3.0 3.0 3.0 mineral oil 5.0 5.0 5.0 trioctanoin 5.0 5.0 5.0 dimethicone 1.0 1.0 1.0 Sodium Magnesium Silicate 0.1 0.1 0.1 glycerin 5.0 5.0 5.0 betaine 3.0 3.0 3.0 triethanolamine 1.0 1.0 1.0 Sodium Hyaluronate 4.0 4.0 4.0 preservatives, fragrances, colors a very small amount a very small amount a very small amount Purified water remaining amount remaining amount remaining amount Sum 100 100 100

Test Example 5: Confirmation of skin moisturizing improvement effect

As a clinical test for skin moisturizing, skin moisturizing ability was evaluated by measuring skin conductivity using a skin moisture meter (Corneometer, Courage+Khazaka, GmbH, Germany). In order to confirm the skin moisturizing effect, the moisture content was measured before and 2 weeks after use while applying the formulations of Examples 8 and 9 and Control Example 1 twice per day, divided into 10 people each. Skin moisture was measured under constant temperature and humidity conditions maintained at a room temperature of 20 to 25 °C and a relative humidity of 40 to 55%. The results are shown in Table 7 below.

division Initial (D0) mean 2 weeks (D14) average Moisture increase (%) Control Example 1 34 37 8.8 Example 8 32 39 21.9 Example 9 33 44 33.3

As can be seen in Table 7, the formulations of Examples 8 and 9 containing the sake lees fermented extract of the present invention showed a better moisture increase, and in the formulation of Example 9 containing the sake lees fermented extract of Example 6 It showed the best skin moisturizing effect.

Test Example 6: Confirmation of skin keratin improvement effect

In order to confirm the effect of improving the skin keratin, the formulations of Examples 8 and 9 and Control Example 1 were divided into 10 people and applied twice a day (morning and evening) for 2 weeks. After 2 weeks, the degree of improvement of keratin was evaluated through image analysis.

Before the start of the experiment, the keratin of the face to be measured was measured and applied (2 times/day) for 4 weeks. The measurement result of the skin keratin state by image analysis is shown in FIG. 1 .

As can be seen in FIG. 1 , the cream of Example 9 containing the fermented sake lees extract of the present invention exhibited a more excellent keratin improvement effect.

Claims (7)

delete delete delete (A) inoculating the sake lees Lactobacillus plantarum (Lactobacillus plantarum);
(B) fermenting while culturing in the dark for 22 to 26 hours at 25-30 ° C;
(C) fermenting while irradiating light of 450-480 nm for 10-15 hours;
(D) fermentation while irradiating light of 540-560 nm for 10-15 hours;
(E) fermenting while removing the light source and culturing in the dark for 22 to 26 hours;
(F) extracting the fermented sake lees with a solvent; and
(G) A cosmetic composition containing 0.001 to 20% by weight of the fermentation extract of sake lees prepared by a method comprising the steps of concentration and drying after filtration, based on the total weight of the cosmetic composition. The cosmetic composition according to claim 4, wherein the cosmetic composition is for skin inflammation relief. The cosmetic composition according to claim 4, wherein the cosmetic composition is for moisturizing the skin. The cosmetic composition according to claim 4, wherein the cosmetic composition is for exfoliating the skin.

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