KR20160137798A - Whitening composition comprising the fermentated extract of Inula britannica as an active ingredient - Google Patents

Abstract

The present invention relates to a cosmetic composition comprising a fermented extract of Inula britannica having a skin whitening effect as an active ingredient. According to the present invention, it is confirmed through experiments that the fermented extract of Inula britannica, obtained through the solvent extraction of fermented Inula britannica which is fermented by lactobacillus, shows antioxidant and whitening effects, so the composition improves skin whitening functions.

Description

[0001] The present invention relates to a cosmetic composition comprising a fermentation starch extract as an active ingredient,

More particularly, the present invention relates to a whitening functional cosmetic composition comprising, as an active ingredient, an extract obtained by fermenting Lactobacillus sp.

Inula britannica L. is a perennial herb that lives in wetlands of Korea, Japan and China, especially in Korea. It is concentrated in Gyeonggi province and Gangwon-do mountainous area, (Kobayashi T. et al., Preventive active effects of the flowers of Inula britannica on autoimmune diabetes in C57BL / KsJ mice induced by multiple low doses of streptozotocin, Phytother Res., 16, 377-382, 2002). The flowering form of the plant is 10 ~ 15 mm in diameter, spherical or spherical, composed of several guns and arranged like a tile. Quercetin is known to have high antioxidant activity (Yang C. et al., Sesquiterpenes and other constituents from the aerial parts of inula japonica, planta Med., 69 , 662-666, 2003). The efficacy of such a laxative has been shown to be effective against sputum, cough, chick embryo, trimming, asthma chronic bronchitis and acute pleuritis. Especially in oriental medicine, it is widely used for the treatment of dyspepsia by smooth circulation of confluent (Sunwoo S. et al. , Studies on the volatile components of inulae flos, J. Korean Agric. Chem. Soc., 34, 312-317, 1991).

On the other hand, fermentation is a process in which microorganisms decompose organic substances using their own enzymes, and fermentation methods are being studied in foods, medicines, and cosmetics for the purpose of increasing the extraction rate of natural products and increasing physiological functions. In a study on this, Joe et al. Reported that the ginsenosides of ginseng were replaced with fungal Aspergillus niger ) and then increased its physiological function (Jo M. et al., Cytotoxicity of the white ginseng extract and red ginseng extract treated with partially purified β-glucosidase from Aspergillus usamii KCTC 6954, Food Sci. Biotechnol., 23, 215-219, 2014) Kim et al. Studied the antioxidant activity of ginseng flowers using Bacillus subtilis (Kim, Kyunghee et al., Antioxidant activity of ginseng flowers according to fermented microorganisms The Korean Society of Food Science and Nutrition, 42 (5), 663-669, 2013). Lactic acid bacteria in microorganisms are safe microorganisms easily found in intestines and fermented foods of humans and animals (Orrhge, K. et al., 2000, Bifidobacteria and lactobacilli in human health, Drugs Exptl. Clin . Res., 26, 95-111). It has been shown to be parasitic by adhering to intestinal epithelial cells and improving the properties of intestinal microflora, thereby stabilizing intestinal microflora, reducing the generation of decay products due to inhibition of harmful bacteria, Anticancer action, cholesterol lowering, and many other benefits to the human body.

Functional cosmetics are cosmetics prescribed by the Ministry of Health and Welfare, and are classified into products that help to whiten skin, products that help to improve the wrinkles of the skin, and products that help to protect the skin from ultraviolet rays. Recently, functional cosmetics have attracted a lot of attention from consumers who are seeking beauty and healthy skin in the current society, which is improving the living standard and aging society. In particular, whitening cosmetics are products that reduce melanin pigment, a major pigment of skin cells. That is, the melanine pigment is a substance produced by an oxidation reaction from tyrosine to tyrosinase in epidermal cells, which is known as a very stable substance. In addition, peroxides, which are physiological metabolites of human cells, are also known to be responsible for the formation of melanin pigments. These substances are also generated by intracellular enzymatic action and are highly unstable and reactive, attacking and damaging intracellular DNA, (Kim, Hye-Jung et al., 2, inhibitory effect of UV extract of ethanol extract on the skin on skin photoaging, Journal of the Korean Society of Food Science and Nutrition, 41, 1540-1545, 2012) ). Therefore, recent patent applications related to whitening cosmetics have been increasing. Especially, as consumer interest in health and environment has increased, use of natural materials rather than chemicals has been increasing. For example, natural materials such as mushrooms, which are relatively safe from the skin, such as mushrooms from mulberry, top pine, pine needles, green tea, mung bean, Whitening agents, anti-aging agents, etc., are a major part of functional cosmetics applications. Therefore, the future of the functional cosmetics industry is very positive as a high-tech futuristic industry that can create high added value by using environment-friendly natural materials and will continue to develop. In addition, the active study of functional cosmetics will be an opportunity to improve the quality of the cosmetics industry in general and further promote the development of the overall cosmetics industry.

The present invention relates to a method for purifying para-coumarinic acid and a functional cosmetic and pharmaceutical composition using the para-coumarinic acid (Korean Patent Registration No. 10-0947148-0000) , And a composition for preventing and treating whitening and skin aging which contains bokbunja seed extract as an active ingredient (Korean Patent Registration No. 10-1510092-0000). In addition, a cosmetic composition containing juniper or juniper fruit extract as an active ingredient has been filed (Korean Patent Application No. 10-2013-0049314). On the other hand, the present inventors have disclosed a patent (Korean Patent Registration No. 10-1373173) on a composition for the treatment and prevention of an inflammatory disease or an allergic disease which contains an extract of a complex herbal medicine as an active ingredient, (Korean Patent Registration No. 10-1150643) for a composition for the prevention and treatment of an inflammatory or allergic disease containing an extract of L. monocytogenes extracted with a solvent or a non-polar solvent as an active ingredient has been disclosed. Also disclosed is a patent (Korean Patent Registration No. 10-1195120) on a pharmaceutical composition for prevention and treatment of highly pathogenic avian influenza H5N1 infection containing a methanol extract of L. monocytogenes as an active ingredient. However, none of the aforementioned literatures discloses a whitening functional cosmetic composition containing a lyophilized extract fermented with lactic acid bacteria as an active ingredient.

Accordingly, it is an object of the present invention to provide a cosmetic composition containing a lyophilized extract fermented with lactic acid bacteria as an active ingredient, and a process for producing the same.

The above object of the present invention is achieved by a method for producing a fermented product, comprising the steps of: obtaining a crude fermentation product having a whitening functional effect fermented using lactobacillus bacterium Lactobacillus plantarum KCCM 11613; Recovering the supernatant by centrifugation of the supernatant obtained in the above step, filtering the supernatant, and freeze-drying the supernatant; The method according to any one of claims 1 to 3, wherein the step (b) comprises the steps of: (a) extracting ethanol from the lyophilized sample; and (c) measuring the pH, titratable acidity and viable cell count of the extract. Measuring the solids content, total polyphenol and flavonoid content from the extract; The antioxidative activity and the whitening activity of the extract were confirmed, and the cosmetic composition formulation was prepared.

The fermented starch extract according to the present invention exhibits superior antioxidative activity and whitening activity than the non-fermented starch extract. The fermented starch extract has an excellent whitening effect as well as skin aging. The fermented starch extract contains the fermented starch extract as an active ingredient There is an excellent effect of providing a cosmetic composition.

FIG. 1 shows the pH (A), total titable acidity (TTA) (B) and viable cell count (L) of the fermentation broth of L. plantarum KCCM 11613 according to Experiment 1 of the present invention (viable cells) (C).
FIG. 2 is a graph showing the measurement results of the whitening activity of the fermentation starch extract of the present invention. FIG.

The present invention provides a cosmetic composition comprising a lactic acid bacteria fermentation starch extract as an active ingredient.

Hereinafter, the present invention will be described in more detail by way of examples. However, the following examples are intended to illustrate the invention and are not intended to limit the scope of the invention.

< Example  1> Disclosure of the present invention and fermentation of lactic acid bacteria Embarkation Fermentation product  Produce

Lactobacillus plantarum ( Lactobacillus plantarum KCCM 11613), a useful lactic acid bacterium, was used in this experiment and cultivation was carried out by liquid culture. That is, the microorganism was cultured at 30 ° C for 1 day inoculation using a MRS culture medium in a 1 L Erlenmeyer flask. On the other hand, in this culture, the embryo and distilled water were extracted at 60 ° C for 24 hours at a ratio of 1:10 (v / v). 2 &lt; / RTI &gt; 300 mL of the filtrate (solid content, 16 mg / mL) obtained in the above step and 0.3 g of peptone (total 0.1%, v / v) were mixed and the pH was adjusted to 6.5. Sterilized for 15 minutes and allowed to cool. In the above step, 1 ml of 0.1% peptone solution was added to the sterilized culture so that the number of cells was 1 × 10 ⁶ / ml. In the non-cultured product (control), 1 ml of the 0.1% peptone solution was added, and the mixture was shaken at 30 rpm at 50 rpm After fermentation, the mixture was centrifuged at 12,000 rpm for 20 minutes. The supernatant was filtered with a 0.45 μm membrane filter and stored at -20 ° C. until used in the next experiment.

< Experimental Example  1> invention Embarkation Fermented  pH, total Titratable acidity  And Viable cell count  Measure

The pH of the fermented product was measured by time during fermentation, and the pH was titrated to pH 8.1 with 0.1 N NaOH solution to determine the acidity as the volume of 0.1 NaOH consumed. The number of viable cells was counted by diluting the sample taken at each fermentation time and culturing at 30 ° C for 48 hours on a MRS agar medium.

As shown in FIG. 1, the number of bacteria reached to the stopper at the end of 12 hours fermentation and showed a maximum value of 8.6 log cfu / mL. The pH value was the lowest at 4.39 in the 24 hour fermentation and the total titratable acidity And the maximum consumption of 0.1 N NaOH was 1.6 mL. There was no significant change in pH and total titratable acidity after 24 hours, but there was a tendency to decrease the bacterial number.

< Experimental Example  2> invention Embarkation Fermentation product  Determination of solids, total polyphenols and total flavonoid content

The solids content was determined by drying at 105 ℃, and the content of solids remaining after drying was measured and calculated.

The total polyphenol content was measured colorimetrically using the Folin-Denis method. After mixing 0.1 mL of the sample and 2 mL of 2% aqueous sodium carbonate solution and incubating at room temperature for 3 minutes, add 0.1 mL of 50% Folin-Ciocalteau's reagent (Sigma Chemical Co., St. Louis, MO, USA in DW) And the absorbance at 750 nm was measured. The total polyphenol contents (mg of gallic acid equivalent, mg GAE) were determined using standard curves of Gallic acid (0, 12.5, 25, 50, 100 μg / Respectively.

Total flavonoids were also assayed by aluminum nitrate assay. 100 μL of sample, 100 μL of 10% ammonium nitrate solution, 100 μL of 1 M potassium acetate solution and 4.7 mL of 80% ethanol were mixed and incubated at 25 ° C. for 40 minutes, and the absorbance was measured at 415 nm. The total flavonoid content in the sample was calculated using a standard curve using the standard reagent quercetin, and the unit was quercetin equivalent (QE) mg / g.

As shown in [Table 1], the solid content of the fermented product after fermentation for 12 hours was the highest, indicating that the yield was increased. As the fermentation time was increased, the solid content was decreased. Total polyphenol content The total polyphenol content of the control group was 6.17 mg GAE / g but the content decreased slightly after fermentation for 72 hours. The total polyphenol content of the samples was 6.03 mg GAE / g and 6.25 mg GAE / g, respectively. Total flavonoid content was increased during the fermentation process from 1.95 ± 0.02 to 2.23 ± 0.04 after 72 hours fermentation, but there was no significant increase thereafter.

Therefore, the change of total flavonoid contents was found to be the highest at 72 hours. Therefore, the fermentation conditions were determined to be 72 hours for the fermentation time of L. plantarum KCCM 11613.

As a result of measuring the solid content, total polyphenol and total flavonoid content of the fermented product of the present invention sample Fermentation time
(hr) Solids content
(mg / mL) Total polyphenol content
(mg GAE / g of solid) Total flavonoid content
(mg quercetin / g of solid) NF 0 17.45 ± 0.00 6.17 ± 0.33 1.95 + 0.02 IFLP 0
12
24
48
72 17.10 ± 0.00
17.80 ± 0.00
17.35 ± 0.00
16.20 ± 0.00
15.80 ± 0.00 6.03 ± 0.04
6.22 ± 0.19
6.07 ± 0.21
6.25 + 0.21
6.05 + 0.26 1.96 + 0.02
2.01 + 0.04
2.02 + 0.02
2.17 ± 0.03
2.23 0.04

^{NF : Non-fermented, IFLP : Inula  britannica  fermented by Lactobacillus plantarum KCCM  11613}

< Example  2> Embarkation Fermented  Solvent extract preparation

The lyophilized fermented product prepared in Example 1 and the control group were lyophilized, and the lyophilized fermented product was mixed with 20 g of the control group and 200 mL of 70% ethanol (10 times) And extracted. After extraction, the mixture was filtered with filter paper (whatman No. 1) and concentrated using a rotary vacuum concentrator (EYELA, Tokyo Rikakikai Co., Japan) to obtain an ethanol extract. The concentrate was lyophilized, pulverized and dissolved in 70% ethanol at any appropriate concentration to verify its functionality (antioxidant and whitening effect).

In the present invention, it is preferable that the fermented product of lyophilized lactic acid bacteria lyophilized is mixed with ethanol or water of 60% to 80% by weight, which is 5 to 15 times (w / v) Extraction yield decreased.

< Experimental Example  3> Fermentation of the present invention Embarkation  Extract Antioxidant power  Measure

The ferric tripyridyltrizaine (Fe ³⁺ -TPTZ) complex was reduced to ferrous tripyridyltriazine (Fe ^{2 +} -TPTZ) by a reducing agent at low pH, and the ferric reducing antioxidant power , And it is a method to evaluate the antioxidant activity by measuring the absorbance of Fe ^{2 +} when the sample reduces Fe ^{3 +} to Fe ^{2 +} . As an experimental method, preparation of FRAP reagent was performed as follows. Twenty-five mL of acetate buffer (300 mM, pH 3.6) was warmed at 37 ° C and then 5 mL of 10 mM 2,4,6-tris (2-pyridyl) -s-triazine (TPTZ) dissolved in 40 mM HCl And 2.5 mL of 20 mM ferric sulfate (FeSO ₄ ). To the 0.95 mL FRAP reagent prepared, 0.05 mL of the sample dissolved at the desired concentration was added and reacted at 37 ° C. for 30 minutes and the absorbance was measured at 593 nm. As a blank sample, 300 mM acetate buffer or solvent was used instead of the sample.

From the measured absorbance, the antioxidant power of the sample was calculated as follows.

 [Equation 1]

Antioxidant power (%) = [1- (OD _s / OD _o )] x 100

(OD _s : Absorbance of sample after X hours, OD _o : absorbance of blank sample after X hours)

As shown in [Table 2], the reducing power of ferrous ions was significantly increased with fermentation time of the fermented lyophilized extracts compared with the control group, and when the solid content of the extract after fermentation for 72 hours was 1.0 mg / mL, it was 0.59 0.73 FeSO ₄ eq mM, indicating an increase of about 23%.

The result of the antioxidant activity measurement of the fermentation liquor extract of the present invention sample Fermentation time Ferric reducing antioxidant power (FeSO4 eq mM) Solids concentration (mg / mL) One 0.5 0.1 NF 0 0.48 + 0.03 0.26 ± 0.01 ND IFLP 0
12
24
48
72 0.59 + 0.02
0.62 ± 0.01
0.61 + 0.02
0.68 ± 0.03
0.73 + 0.04 0.27 ± 0.01
0.30 + 0.04
0.29 + 0.03
0.33 + 0.02
0.34 + 0.03 ND
ND
ND
0.01 ± 0.02
0.02 0.03

^{NF : Non-fermented, IFLP ; Inula  britannica  fermented by Lactobacillus plantarum KCCM  11613,}

^{ND: Not detected.}

< Experimental Example  4> Fermentation of the present invention Embarkation  Extract Tyrosinase ( tyrosinase ) Inhibition activity measurement

Tyrosinase is the most important enzyme in the production of melanin, a black or red skin pigment. That is, this enzyme acts as a kind of DOPA oxidase which oxidizes DOPA produced from tyrosine in melanosome, which is a small intestine of skin cells, to form dopachrome, and ultimately synthesizes melanin polymer which is very stable. Therefore, in the development of the skin whitening agent, tyrosinase inhibition experiment is recognized as the first evaluation method for the usefulness of the test substance.

The inhibitory effect of tyrosinase activity was measured using the fermentation liquor extract prepared by the method of the present invention [Example 2]. That is, 880 μL of L-3,4-dihydroxyphenylalanine (DOPA) solution dissolved in 3.8 mM of 0.1 M phosphate buffer (pH 6.8) and 100 μL of the fermentation supernatant extract of the present invention were mixed. To this mixture was added 20 μL of tyrosinase (1,000 U / mL), incubated at 25 ° C for 5 minutes, and absorbance was measured at 475 nm.

The whitening activity of the sample from the measured absorbance was calculated as follows, where the blank was reacted without the lyophilized extract and the control was the non-fermented lyophilized extract.

&Quot; (2) &quot;

Tyrosinase inhibition rate (%) = [1- (absorbance of sample / absorbance of control group)] × 100

As shown in Table 3, the inhibitory effect against tyrosinase was 15.53 to 19.32% in the control group, while it was 5.4 times higher in the 72-hour fermentation line-up extract than in the control group was a clear difference could be seen in the city or inhibitory effect of the tie of Figure fleet flower extract at a concentration such as shown in Figure 2 (control) in the present invention into effect the fleet flower extract (sample).

The whitening activity measurement results of the fermentation liquor extract of the present invention Fermentation time (h)
Tyrosinase inhibition activity (%) NF IFLP 0 16.41 + - 4.16 27.91 + - 4.42 24 18.74 ± 4.02 77.77 + - 0.83 48 19.32 + - 2.41 80.20 ± 1.93 72 15.53 ± 94.1 83.61 ± 2.05

^{NF  : Non-fermented, IFLP  : Inula  britannica  fermented by Lactobacillus plantarum KCCM  11613}

The cosmetic composition having a whitening effect according to the present invention may be in the form of a water-soluble liquid, cream, lotion, gel, pack, essence, oil / water, water / Powder, and the like. The composition of each of these formulations may contain various kinds of bases and additives necessary for formulation of the formulation, and the kinds and amounts of these ingredients can be easily selected by those skilled in the art. Hereinafter, a cream and a lotion composition will be described as a typical application example of a cosmetic composition.

< Formulation Example  1> Fermentation of the present invention Embarkation  Preparation of cream containing extract

Table 4 shows examples of formulations of cream in cosmetics containing the starch extract of Example 2. Creams were prepared at the composition ratios shown in Table 4 below. After the carboxy polymer was impregnated with purified water for 30 minutes, butylene glycol and glycerin were added, and the mixture was heated and stirred at 70 ° C to prepare an aqueous layer. The oily layer was liquefied by heating to 70 DEG C while mixing and stirring stearic acid, cetyl alcohol, glyceryl monostearate, wax, liquid paraffin, caprylic / capric glyceride and squalane. The aqueous layer prepared in the stirrer at 70 ° C was added, and the oily layer was gradually added thereto. The mixture was mixed with triethanolamine to adjust the pH to about 6.5 to 7.2, followed by degassing with stirring. The degassed formulation was cooled to room temperature, and the fermented starch extract was added and stirred to prepare a cream.

The blending ratio of the cream containing the fermentation supernatant extract of the present invention Raw material Content (% by weight) The fermented starch extract (Example 2) 2 to 3 Stearic acid 1-2 Cetyl alcohol 1-2 Glyceryl monostearate 1-2 Wax 1 to 1.5 Liquid paraffin 3.5 ~ 4 Caprylic / capric triglyceride 3.5 ~ 4 Carboxyvinyl polymer 0.1 to 0.2 Butylene glycol 5 ~ 5.5 glycerin 2.5 to 3 Triethanolamine 0.3 to 0.5 Squalane 4 to 4.5 Purified water 67.8 ~ 75.1

< Formulation Example  2> Fermentation of the present invention Embarkation  Preparation of Lotions Containing Extracts

Table 5 shows formulation examples of lotion in the cosmetic containing the starch extract of Example 2. Lotions were prepared at the composition ratios shown in Table 5 below. After the carboxy polymer was impregnated with purified water for 30 minutes, ethanol and propylene glycol were added and the mixture was heated and stirred at 70 캜 to prepare an aqueous layer. The oily phase was liquefied by heating wastewater, fluid pyraphene, sorbitan stearate, protonated monostearic acid glycerin and stearic acid while stirring and mixing at 70 ° C. The aqueous layer prepared in the 70 ° C agitator was added, and the oily layer was slowly added thereto. The mixture was mixed with triethanolamine to adjust the pH to about 6.0 to 6.5, followed by degassing with stirring. The degassed formulations were cooled to room temperature and the fermented starch extract was added and stirred to prepare a lotion.

The blending ratio of the lotion containing the fermentation supernatant extract of the present invention Raw material Content (% by weight) The fermented starch extract (Example 2) 1.5 to 2 Wax 0.5 to 1 Floating pyrapin 9.5 to 10 Sorbitan stearate 0.5 to 1 Chin type glycerin monostearate 0.5 to 0.6 Stearic acid 1.5 to 2 ethanol 5 ~ 7 Propylene glycol 2.5 to 3 Carboxy polymer 0.1 to 0.2 Triethanolamine 0.2 to 0.3 Purified water 72.9-78.2

INDUSTRIAL APPLICABILITY The present invention has an effect of providing a cosmetic composition containing an extract of fermentation starch as an active ingredient and has an excellent effect that can be used as a new material for cosmetics, especially in formulations such as creams, lotions and ointments. In addition, since it provides an excellent effect of improving the whitening function by providing a cosmetic composition containing a useful ingredient contained in the fermentation starch extract, it is an extremely useful invention in the cosmetics industry.

Claims (5)

A cosmetic composition comprising a lactic acid bacterium fermentation extract of the present invention as an active ingredient.
The method according to claim 1, wherein the step of fermenting the lactic acid bacteria by inoculation comprises lactic acid bacteria inoculated and fermented to obtain a fermented product; The supernatant is recovered by centrifuging the supernatant obtained in the above step, and the supernatant is filtered and freeze-dried.
[Claim 3] The method according to claim 2, wherein the lactic acid bacteria fermentation extract is obtained by mixing 60 to 80% ethanol or water of 5 to 15 times (w / v) by weight with respect to the lyophilized lactic acid bacteria freeze- Then concentrating under reduced pressure to obtain a concentrate; And lyophilizing the concentrated liquid obtained in the above step to obtain a powdered cosmetic composition.
The cosmetic composition according to claim 1, wherein the cosmetic composition exhibits whitening function.
The cosmetic composition according to any one of claims 1 to 4, wherein the cosmetic composition is a water-soluble liquid, cream, lotion, gel, pack, essence, oil / water, w / And a heavy water (W / Si) type powder.

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