KR101280306B1 - A cosmetic composition for anti-aging containing an extract of poncirus trifoliata by bioconversion - Google Patents

Abstract

The present invention relates to an anti-aging cosmetic composition containing an extract of Poncirus trifoliata bioconverted by Ganoderma lucidum mycelium, and more specifically, extracting the extract from the fruit ; Mixing the whole culture solution of Ganoderma lucidum mycelium or Ganoderma lucidum mycelium to the extract, followed by main culture; Removing the Ganoderma lucidum mycelium after the present culture; relates to an anti-aging functional cosmetic composition comprising the filtrate obtained from the active ingredient, antioxidant effect, collagen synthesis promoting effect, collagenase (MMP- It shows the activity and expression inhibitory effect of 1) and the improvement of skin wrinkles.

Description

Anti-aging cosmetic composition containing fermented fruit extract {A COSMETIC COMPOSITION FOR ANTI-AGING CONTAINING AN EXTRACT OF PONCIRUS TRIFOLIATA BY BIOCONVERSION}

The present invention is a bioconverted cell ( Poncirus) trifoliata ) relates to an anti-aging cosmetic composition containing an extract, and more particularly, to an anti-aging functional cosmetic composition containing a filtrate obtained by fermenting the fruit extract as an active ingredient.

Human skin is constantly changing with age. Skin aging is largely divided into natural aging (or endogenous aging) and external aging (EB Doris, Cosmetics & Toiletories, 1996; 111: 31-37), and natural aging (endogenous aging) is artificial because it is influenced by genetic factors. While it is difficult to control, aging is relatively easy because it is influenced by environmental factors. Representative external aging factors include ultraviolet rays, and the most prominent external aging phenomenon is wrinkle formation (HW Daniell, Ann Intern Med, 1971; 75 (6): 873-880, GL Grove et al., J Am Acad Dermatol, 1989; 21 (3): 631-637, CE Griffiths et al., Arch Dermatol, 1992; 128: 347-351).

On the other hand, one of the photoaging mechanisms caused by ultraviolet light is via free radical pathways (D Harman, J Gerontol, 1956; 11 (3): 298-300). Free radicals are known to cause breakdown of connective tissue formation such as skin collagen, inhibition of cell membrane function, promotion of DNA mutation, modification of protein function, intercellular energy transfer, and modification of metabolic related molecules (RM Lavker and AM Kligman, J Invest Dermatol, 1988; 90: 325-330). The involvement of free radicals in aging means that antioxidants or various chemical scavengers can be used to delay the aging process.

On the other hand, the synthesis and degradation of extracellular matrix such as collagen in vivo is properly controlled, but its synthesis decreases as aging progresses, and matrix metalloproteinase (MMP), an enzyme that degrades collagen, In particular, the expression of collagenase (hereinafter referred to as 'MMP-1') and gelatinase (gelatinase (hereinafter referred to as 'MMP-2')) is promoted to reduce the elasticity of the skin and wrinkles are formed. Ultraviolet radiation and free radicals also activate these degrading enzymes.

MMPs are classified into MMP-1, MMP-2, and MMP-9 according to their substrate specificity. MMP-1 is a fibroblast type collagenase, an enzyme of epilepsy, and collagen types I, II, III, and Ⅶ as substrates. There are, Ⅷ, 젤 and gelatin, which are cut into peptides of 3/4 or 1/4 length of original collagen to facilitate the action of nonspecific enzymes (gelatinase). MMP-2 belongs to 72 kD gelatinase (A) and degrades gelatin, collagen types IV, V, VIII, VIII, elastin and fibronectin as substrates.

MMPs of neutrophils and polymorphonuclear leukocytes are primarily activated by oxidation, which first generates hydrogen peroxide in the cell and is converted to hypochlorous acid by myeloperoxidase in the presence of chlorine ions. Activate it.

Until now, the most common method was to apply sunscreen products directly to the skin to prevent aging due to ultraviolet rays.However, since 1988, retinoic acid was reported to be effective in reducing roughness and fine wrinkles of aged skin. JS Weiss et al., J Am Acad Dermatol, 1988; 19 (1): 169-175), research is being actively conducted to develop substances that inhibit or improve skin aging worldwide. Among them, derivatives of vitamins such as vitamins A, C, and E and AHA (alpha-hydroxy acid) are representative skin aging improving agents known to date (DS Rosenthal et al., J Invest Dermatol, 1990; 95: 510-515, CM Ditre et al., J Am Acad Dermatol, 1996; 34 (2): 187-195). However, they are very unstable in the natural environment, so there is a problem in use, or somewhat insufficient to show a visible effect. Therefore, there is an urgent need for the development of a new skin wrinkle improving material that is safe for the living body, scavenges free radicals, inhibits the activity and expression of collagenase (MMP-1), and collagen synthesis.

Therefore, the present inventors have studied the method to further increase the efficacy of the fruit extract excellent in antioxidant properties among the plants that grow in nature as a result of Ganoderma extract ( Ganoderma) lucidum ) When the bioconversion to mycelia, the antioxidant effect, collagenase (MMP-1) activity and expression inhibitory effect and collagen synthesis promoting effect significantly increased and completed the present invention.

Thus, the present invention is a Poncirus It is an object of the present invention to provide an anti-aging functional cosmetic composition comprising a filtrate obtained by bioconverting trifoliata ) extract into Ganoderma lucidum mycelium as an active ingredient.

In order to achieve the above object, the present invention comprises the steps of extracting the extract from the fruit; Mixing the whole culture solution of Ganoderma lucidum mycelium or Ganoderma lucidum mycelium to the extract, followed by main culture; Removing the Ganoderma lucidum mycelium after the present culture; provides an anti-aging functional cosmetic composition comprising the filtrate obtained from the active ingredient.

Hereinafter will be described in detail for the anti-aging functional cosmetic composition containing the fermented fruiting extract of the present invention.

In the case of extract extracted simply as a solvent, fruit is effective in vitro, but when it is applied to skin cosmetics, there is a problem that the effect is not satisfactory substantially.

On the other hand, the present inventors confirmed that the main component showing the efficacy effect in the fruit fruit extract is flavonoids. Plant-rich phenolic compounds such as flavonoids, tannins and anthocyanins are excellent antioxidant candidates for protecting skin damaged by UV rays (T Shibamoto, American Chem Soc, 1994; 564: 247-256), particularly Voids or derivatives thereof have been reported to have excellent antioxidant activities such as radical scavenging activity and lipid peroxidation inhibitory activity caused by ultraviolet rays (RJ Nijveldt et al., Am J Clin Nutr, 2001; 74: 418-425).

In general, hydrophobic substances are more effective for skin penetration than hydrophilic substances, and the ceramide component distributed in the stratum corneum is hydrophobic, which is more effective for interacting with hydrophobic substances than hydrophilic substances, so that the hydrophobic substance is more easily skin outermost layer. Because it can pass through.

On the other hand, most of flavonoids are glycosides in which sugars are combined with one or more molecules, and are present in nature, have a relatively high molecular weight, and have some water solubility. There is a problem that absorption is difficult.

Therefore, the present invention comprises the steps of extracting the extract from the fruit room to provide a skin anti-aging cosmetic composition that acts as an effective external skin for skin permeability is easy; Mixing the whole culture solution of Ganoderma lucidum mycelium or Ganoderma lucidum mycelium to the extract, followed by main culture; Removing the Ganoderma lucidum mycelium after the main culture; containing the filtrate obtained from the step as an active ingredient, the filtrate obtained through the above step that the total phenol content and total flavonoids content than the extract extracted from the fruit room It was confirmed in this experiment.

In addition, the present invention is characterized in that the filtrate contained as an active ingredient has excellent anti-wrinkle effect by the antioxidant effect, collagenase (MMP-1) activity and expression inhibition effect and collagen synthesis promoting effect.

The present invention also relates to an anti-aging cosmetic composition containing fermented fruiting extract, characterized in that the content of the extract is 0.001 to 30.0% by weight based on the total skin cosmetic composition. When the extract is included in less than 0.001% by weight, there is almost no skin improvement effect, when the extract is more than 30.0% by weight it is not economical because the degree of increase in the effect of increasing the content is insignificant.

In addition, the present invention relates to a cosmetic composition, characterized in that using the aqueous solution containing 40 to 95% (v / v) of ethanol as the extraction solvent during the extraction.

In addition, the present invention relates to a cosmetic composition, characterized in that the extract is concentrated under reduced pressure or lyophilized after extraction.

In addition, the present invention relates to a cosmetic composition, characterized in that the gamyeo concentrated or lyophilized extract is dissolved in 0.001 ~ 30.0% by weight in at least one solvent selected from purified water, ethanol, butylene glycol and propylene glycol.

In addition, the present invention after adjusting the pH of the culture medium at the time of the main culture to 5.0 ~ 7.0, inoculate the Ganoderma lucidum mycelium so as to 4.0 to 6.0% (v / v), the temperature 22 ~ 32 ℃, rotation speed 120 ~ 180rpm, It relates to a cosmetic composition characterized by culturing for 5 to 7 days in aeration 1.2 ~ 1.8vm conditions.

The present invention further relates to an anti-aging functional cosmetic composition containing fermented fruiting extract.

The present invention also relates to a cosmetic composition exhibiting skin irritation-releasing effect containing fermented fruiting extract.

In addition, the present invention, the cosmetic composition is a lotion, gel, water-soluble liquid, cream, essence, other oil-in-water (O / W) and water-in-oil (W / O) type base cosmetic formulation and oil-in-water or water-in-oil makeup base , Foundation, skin cover, lipstick, lip gloss, face powder, two-way cake, eye shadow, teak color and eyebrow pencil is characterized in that the cosmetic composition selected from.

The cosmetic composition containing the fermented fruiting extract of the present invention exhibits anti-aging functions such as free radical scavenging effect, collagen synthesis promoting effect, skin wrinkle improvement effect such as inhibiting activity and expression of collagenase (MMP-1).

In addition, the skin cell death inhibitory effect and skin stimulation alleviating effect to reduce the skin irritation caused by the cosmetic substrate is excellent.

Hereinafter, the present invention will be described in more detail with reference to Examples. It should be noted, however, that these examples are only illustrative examples of the present invention, and the scope of the present invention is not limited to these examples.

< Comparative example  1> Fruit Extract Extract Manufacturer

The dried and dried 500 g of dry cellar was refluxed three times for 3 hours with a hot 95% (V / V) ethanol aqueous solution, and the mixture was cooled and then filtered through Whatman # 5 filter paper. The filtered extract was concentrated under reduced pressure and lyophilized at 50 ° C. or lower to obtain 90.5 g of ethanol extract. The fruit extract was prepared using at least one solvent selected from purified water, ethanol, butylene glycol, and propylene glycol to contain 15.0% by weight of a vacuum concentrate and a freeze-dried product.

Example 1 Preparation of Fermented Fruit Extract

Ganoderma lucidum mycelium was inclined in a test tube containing potato dextrose agar medium and stored at 4 ℃ and used subcultured every month. Aseptic homogenization of mycelial subcultured in the slope medium, inoculated so that the Ganoderma lucidum mycelium 5% (v / v) to the medium mixed with the fruit extract and yeast-wort obtained in Comparative Example 1 in the fermenter The culture was carried out for 6 days under conditions of a temperature of 25 ° C., a rotational speed of 150 rpm, aeration rate of 1.5vm, and pH 7.0. After culturing, the Ganoderma lucidum mycelium was removed from the culture medium, and then fermented fruit juice extract was obtained.

Experimental Example 1 Determination of Components and Content in Fermented Fruit Extract

In Experimental Example 1, the qualitative analysis of phenolic substances, flavonoids, flavonoid glycosides, etc., and the total phenolic content and total flavonoid content among the main components of the fruit extracts prepared in Comparative Example 1 and the fermented fruit extracts prepared in Example 1 were compared. It was.

First, qualitative analysis of phenolic substances, flavonoids, flavonoid glycosides, etc. was performed as follows.

(1) Qualitative analysis of phenolic substances and flavonoids

When 1 to 2 drops of 2.5% FeCl ₃ ethanol solution was added to the extracts of Comparative Example 1 and Example 1, the color was dark green or the formation of precipitates was observed.

(2) Qualitative analysis of flavonoid glycosides

Using concentrated sulfuric acid method, 5 ml of concentrated sulfuric acid was immersed in the extracts of Comparative Example 1 and Example 1 to observe the formation of yellow and yellow-red precipitates.

(3) Qualitative analysis of sugar

Two to three drops of 5% α-naphthol alcohol solution was added to the extracts of Comparative Example 1 and Example 1, respectively, and when the concentrated sulfuric acid was added through the walls, it was observed whether reddish purple appeared at the interface.

(4) Determination of Total Phenolic Content

10 ml of distilled water was added to 1 ml of the extract of Comparative Example 1 and Example 1, 2 ml of Folin-Ciocalteu phenol reagent was added, mixed, and reacted at room temperature for 5 minutes. 2 ml of 20% sodium carbonate was added to the reaction mixture, followed by reaction at room temperature for 1 hour, and then absorbance at 725 nm was measured. At this time, tannic acid was used as an indicator.

(5) Determination of total flavonoid content

0.5 ml of extract of Comparative Example 1 and Example 1 were mixed with 0.1 ml of 10% aluminum nitrate, 0.1 ml of 1M potassium acetate, and 4.3 ml of ethanol, respectively, and reacted at room temperature for 40 minutes, and then absorbance was measured at 415 nm. At this time, the indicator material was used quercetin (quercetin).

Name of sample Total phenol content
(Μg / ml of extract) Total Flavoid Content
(Μg / ml of extract) Comparative Example 1 585 107.7 Example 1 1021 589.4

As a result of measuring the total phenol content and the total flavonoid content (Table 1), it was confirmed that the extract of Example 1 is higher than the extract of Comparative Example 1, the total phenol content and total flavonoid content, in particular the flavonoid content increased more than 5.5 times I could confirm that.

From the above results, it was confirmed that fermentation of the Jiji extract prepared by simple extraction with Ganoderma lucidum mycelium could increase the total phenolic content and the total flavonoid content, based on which Gaji extract fermented by Ganoderma lucidum mycelium An anti-aging cosmetic composition of the present invention was prepared.

< Experimental Example  2> DPPH Radicals  Antioxidant Effect Measurement Experiment

In this experimental example, the scavenging effect on the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical was measured by slightly modifying the method used by Yoshida et al. (1989) to measure the antioxidant effect of the material obtained in Example 1. . Since DPPH is purple as a stable free radical, it is a measure of the extent to which purple is removed as the sample scavenges radicals.

First, DPPH was dissolved in methanol at a concentration of 1 × 10 ⁻⁴ M and 0.15 ml of this solution was placed in a test tube. 0.15 ml of each concentration sample was added thereto, and the absorbance was measured at 565 nm after 20 minutes of reaction at room temperature. Green tea extract was used instead of the material of the example as a positive control. The free radical scavenging effect of each sample was expressed as SC ₅₀ by calculating the concentration of 50% scavenging of radicals using Equation 1. At this time, the negative control was taken as the reaction absorbance of the liquid not treated with the sample.

Name of sample SC ₅₀ (μg / ml) Example 1 100 Green tea extract 400

As shown in Table 2 fermented fruit extract of the present invention showed a very strong activity of about four times compared to the green tea extract compared to the green tea extract, a powerful antioxidant.

< Experimental Example  3> free radicals Radicals  Antioxidant Effect Measurement Experiment

This Experimental Example shows the activity produced by the xanthine-xanthine oxidase system by slightly modifying the method used by Furuno et al. (2002) to measure the antioxidant effect of the material obtained in Example 1. The scavenging effect on the superoxide radical was measured. The free radicals produced by the xanthine and xanthine oxidase reactions react with Nitro Blue Tetrazolium (NBT) to give a blue color, so the amount of blue reduced as the sample eliminates free radicals is measured. It is.

First, 3 × 10 ^-3 M xanthine, 3 × 10 ^-4 M EDTA, 7.5 × 10 ^-4 M nitroblue tetrazolium (NBT), and 0.15 mg / mL bovine serum albumin (BSA) solution were added to each concentration sample. After mixing, the mixture was reacted at room temperature for 10 minutes. Thereafter, xanthine oxidase (0.25 U / ml) solution was added to each test tube and reacted at room temperature for 20 minutes, and then absorbance was measured at 565 nm. Green tea extract was used instead of the material of the example as a positive control. The active oxygen radical scavenging effect of each sample was expressed as SC ₅₀ by calculating the concentration of 50% scavenging radicals using Equation 2. At this time, the negative control was taken as the reaction absorbance of the liquid not treated with the sample.

Name of sample SC ₅₀ (μg / ml) Example 1 50 Green tea extract 340

As shown in Table 3, the fermented fruit extract of the present invention exhibited a very strong activity of about 6.7 times as compared to the green tea extract.

From the above results, it was possible to confirm the excellent antioxidant effect of the fermented fruiting extract of the present invention, from which the excellent antioxidant effect of the present invention could be deduced.

< Experimental Example  4> Experiment to measure collagen synthesis promoting effect

In order to observe the collagen synthesis promoting effect of the samples obtained in Example 1, this experiment was carried out by directly treating human fibroblasts obtained from humans or purchased commercially.

In a 96-well plate, add DMEM (Dulbecco's Modified Eagle's Media) containing 2.5% fetal bovine serum and human fibroblasts (5000 cells / well), and incubate until 70-80% growth. It was. Then, the cells were collected for 1 day after treatment for each concentration. The amount of collagen synthesized was measured using the collagen protein measuring instrument (Catalog N .: MK 101, Takara Shuzo Co. Ltd, Japan).

In the measurement method, first, a cell culture solution collected in a 96-hole plate incubator uniformly coated with primary collagen antibody was subjected to antigen-antibody reaction for 3 hours. After 3 hours, the chromophore-bound secondary collagen antibody was placed in a 96-hole plate incubator and allowed to react for 15 minutes. After 15 minutes, the color developing agent was added to develop color at room temperature, and 1M sulfuric acid was added to stop the color development. The color of the reaction color is yellow, depending on the intensity of the reaction. A yellowish 96-hole plate incubator was measured for absorbance at 450 nm using an absorbance spectrometer, and the degree of collagen synthesis was calculated by Equation 3 below. At this time, the reaction absorbance of the cell culture without the sample was used as a control.

Sample name (µg / mL) Collagen Synthesis Effect (%) Example 1 200 40 100 15

As shown in Table 4, the fermented fruit extract of the present invention was found to have an excellent collagen synthesis promoting effect.

< Experimental Example  5> Collagenase (MMP-1)  Activity inhibition assay

This experimental example was measured in a biochemical model of screening type in vitro to observe the inhibitory effect of collagenase (MMP-1) of the samples obtained in Example 1, which is purified collagenase and its substrate It is based on the use of collagen conjugated to fluorescein (EnzChek, Collagenase Kit, Molar Proof, USA).

First, 20 µl of 0.25 mg / ml DQ collagen lysis solution and 40 µl of the concentration sample in 100 µl of reaction buffer (0.05 M Tris-HCl, 0.15 M NaCl, 5 mM CaCl ₂ , 0.02 mM NaN ₃ , pH7.3) After the addition, the mixture was well mixed with 40 µl of 0.5 Unit of collagenase and reacted for 20 minutes at dark and room temperature. Thereafter, the fluorescence value was measured at an absorption wavelength of 495 nm and an emission wavelength of 515 nm using a fluorescence spectrophotometer. Green tea extract was used instead of the material of the example as a positive control. The inhibitory effect of collagenase activity of each sample was expressed as IC ₅₀ by calculating the concentration of 50% scavenging enzyme activity using Equation 4. At this time, the negative control was used as the fluorescence value of the liquid without the sample.

Name of sample IC ₅₀ ([mu] g / ml) Example 1 100 Green tea extract 200

As shown in Table 5 fermented fruit extract of the present invention showed a very strong inhibitory activity compared to the green tea extract, a powerful collagenase activity inhibitor.

< Experimental Example  6> Collagenase (MMP-1)  Expression inhibitory effect measurement experiment

This experimental example was carried out by slightly modifying the enzyme linked immunosorbent assay (ELISA) used by Petersen et al. (1992) to inhibit the expression of collagenase, the expression of which is increased by UVA irradiation of the sample obtained in Example 1.

First, UVA was irradiated to human dermal fibroblasts with an energy of 6.3 J / cm ² using an ultraviolet chamber, and then exchanged with a medium to which the sample was added, followed by culturing for 24 hours, and then the medium was recovered and coated in a 96-hole plate incubator. The primary antibody (MMP-1 (Ab-5) monoclonal antibody) was treated and reacted at 37 ° C. for 90 minutes, and then treated with 3% BSA for 1 hour to block unbound sites. After reacting the secondary antibody anti-mouse immunoglobulin (anti-mouse IgG, whole mouse, alkaline phosphatase conjugated) for about 90 minutes, alkaline phosphatase substrate solution (1 mg / ml p-nitrophenyl phosphate in diethanolamine buffer solution) Was added and reacted at room temperature for 30 minutes. Absorbance was measured at 405 nm using an absorbance spectrometer. The degree of inhibition of MMP-1 activity was calculated by Equation 5 below. At this time, the reaction absorbance of the cell culture without the sample was used as a control.

Sample name (µg / mL) % Inhibition of expression Example 1 100 78.3 Green tea extract 100 75

As shown in Table 6, the fermented fruit extract of the present invention showed a potent collagenase expression inhibition effect similar to that of green tea extract.

< Experimental Example  7> Evaluation of Stimulation Relaxation Effect Using Cell Culture Technology

This experimental example inhibits cell death by Sodium Lauryl Sulfate (SLS), a substance that induces skin irritation using cell culture technology, in order to evaluate the stimulating relaxation effect of the sample obtained in Example 1. To evaluate the effect of stimulation to the extent that it is.

SLS is a substance that is used as a criterion for evaluation of skin irritation based on cosmetics. It is bound to cell membranes to inhibit cell metabolism and destroys cell membranes to cause skin irritation. This experimental example evaluates the effect of reducing cell death by SLS when SLS and sample are added to human fibroblasts at the same time. Hs68 fibroblasts used in this experiment were used as human dermal dermal cells from ATCC (American Type Culture Collection, Accession No. 1635). Stimulation relaxation evaluation experiment using the cell culture technology was carried out as follows.

First, human-derived fibroblasts were dispensed at a concentration of 1 × 10 ⁵ per well in a 96-well plate incubator, and cultured for 24 hours, followed by treatment with 0.01% SLS alone, 0.01% SLS and a sample (0.01%) at the same time for 24 hours. While further cultured. After incubation, MTT (Sigma) reagent was added and incubated for 4 hours, the culture solution was removed, 1N NaOH / isopropanol solution was added, stirred for 20 minutes, and the absorbance was measured at 565 nm with an absorbance spectrometer. The effect of inhibiting apoptosis was measured.

Sample name (µg / mL) Fibroblast survival rate (%) Example 1 (100) + SLS (100) 70 SLS (100) 23

As shown in Table 7, the fermented fruiting extract was found to be a stimulating substance that reduces skin irritation that can occur with the cosmetic base when prescribed with the cosmetic base by inhibiting apoptosis caused by SLS.

< Example  2>

This Example prepared the cosmetics containing the fermented fruit extract obtained in Example 1 to evaluate the effect of improving skin wrinkles in humans by performing a comparative experiment with Comparative Example 1.

The cosmetics used in the comparative experiments are in cream form and their compositions are shown in Table 8. First, phase B) recorded in Table 8 was heated and stored at 70 ° C., phase B) was added to phase A) and pre-emulsified, then emulsified uniformly with a homomixer, and then gradually cooled to a cream (Example 2, comparison). Example 2) was prepared.

Raw material Example 2 Comparative Example 2 end Stearyl alcohol 8 8 Stearic acid 2 2 Stearic Acid Cholesterol 2 2 Squalane 4 4 2-octyldodecyl alcohol 6 6 Polyoxyethylene (25 mol addition) alcohol ester 3 3 Glyceryl monostearate Aster 2 2 I Fermented Fruit Extract One - Propylene glycol 5 5 Purified water Suitable amount Suitable amount

Note) Unit: Weight%

20 experimenters (women aged 20-35 years) were applied to the cream prepared in Example 2 on the right side of the face and the cream prepared in Comparative Example 2 on the left side of the face for two consecutive days, twice daily.

After completion of the experiment, the effect of improving skin wrinkles is obtained by using a silicone resin copy (replica) before and after using the product for 2 months, using a skin fine wrinkle device (SV600, C + K, Germany) and a skin imaging analyzer. The state of wrinkles on the eyes was compared.

Table 9 compares the average eye wrinkle depth and wrinkle reduction rate of the experimenter using the cream prepared in Example 2 with the experimenter using the cream of Comparative Example 2. As shown in Table 9, it can be seen that the skin wrinkle improvement effect is excellent in the facial periphery of the experimenter applying the cream containing the fermented fruit extract.

Evaluation items Example 2 Comparative Example 2 Before use 2 months later Before use 2 months later Average fine wrinkle depth 0.201 0.141 0.200 0.195 Wrinkle Reduction (%) 29.9% 2.5%

n = 20, p <0.01

< Experimental Example  8> SLS Skin irritation

This experimental example evaluates the stimulating alleviation effect of the cosmetic containing the fermented fruiting extract obtained in Example 1 by human patch experiment. This evaluation is a result of evaluation obtained by directly applying the result obtained in Experiment 7 to the human body. SLS (sodium lauryl sulfate) that causes irritation in general cosmetic prescriptions (cream, lotion, skin, essence) is mixed with the product prepared in Example 2 for 24 hours, 48 hours, 72 hours The effects of stimulating relaxation were evaluated.

FINN CHAMBER (FINLAND) was applied to the upper arm of 50 healthy men and women aged 20-50 years by 0.2 mg each, and the acute irritation index was evaluated after 24 hours. After the evaluation, the same amount of product was applied again to the same site again, and the delayed irritation index after 48 hours and 72 hours was evaluated.

The test resulted in no skin redness even after 24 hours, 48 hours, 72 hours of patching the product containing the fermented fruiting extract.

The results of this evaluation indicate that fermented fruiting extracts have a significant effect on reducing skin irritation by substrates (surfactants, fragrances, alcohols) that cause irritation when used in cosmetics.

< Example  3> Preparation of Toilet Water Containing Fermented Fruit Extract

To 8 g of 95% ethanol, 0.05 g of polypyrrolidone, 0.1 g of polyol alcohol, 0.2 g of polyoxyethylene monooleate, 0.2 g of fragrance, 0.1 g of paraoxybenzoic acid methyl ester, a small amount of antioxidant, and a small amount of pigment are dissolved. The mixed solution was added to 0.05 g of the fermented fruiting gland extract obtained in Example 1 and 5 g of glycerin in 85.33 g of purified water, followed by stirring to obtain a skin lotion having a skin improving effect.

< Example  4> Preparation of Latex Containing Fermented Fruit Extract

1.2 g of cetyl alcohol, 10 g of squalane, 2 g of vaseline, 0.2 g of paraoxybenzoic acid ethyl ester, 1 g of glycerin monoester acetate, 1 g of polyoxyethylene (20 mole addition) monooleate and 0.1 g of fragrance at 70 ° C The mixture was dissolved by heating, followed by heating and dissolving 0.05 g of the fermented fruit cell extract obtained in Example 1, 5 g of dipropylene glycol, 2 g of polyethylene glycol 1500, 0.2 g of triethanolamine, and 76.2 g of purified water at 75 ° C. Both were emulsified by mixing and then cooled to prepare an emulsion having a water / medium-based skin improvement effect.

< Example  5> Containing Fermented Fruit Extract Essence  Produce

5 g of 95% ethyl alcohol, 1.2 g of polyoxyethylene sorbitan monooleate, 0.3 g of kitulose, 0.2 g of sodium hyaluronate, 0.2 g of vitamin E-acetate, 0.2 g of sodium licoate, 0.1 g of paraoxybenzoic acid ester, 1 g of each fermented fruiting extract obtained in Example 1 and an appropriate amount of pigments were mixed to obtain a cosmetic liquid having a skin improving effect.

Claims (7)

Extracting an extract from Poncirin trifoliata ;
Ganoderma lucidum ( Ganoderma) in the extract lucidum ) Mycelium or Ganoderma lucidum ( Ganoderma) lucidum ) mixing the whole culture solution of the mycelium, and then main culture; And
After the main culture, Ganoderma lucidum ( Ganoderma) from the main culture lucidum ) to remove the mycelium; anti-aging functional cosmetic composition comprising the filtrate obtained from the active ingredient. The method according to claim 1,
Cosmetic composition, characterized in that using the aqueous solution containing 40 ~ 95% (v / v) ethanol as the extraction solvent during the extraction. The method according to claim 1,
The content of the filtrate is a cosmetic composition, characterized in that 0.001 to 30.0% by weight based on the entire cosmetic composition. The method according to claim 1,
The extract is a cosmetic composition, characterized in that concentrated under reduced pressure or lyophilized after extraction The method of claim 4,
The concentrated or lyophilized extract is a cosmetic composition, characterized in that dissolved 0.001 ~ 30.0% by weight in at least one solvent selected from purified water, ethanol, butylene glycol and propylene glycol. The method according to claim 1,
The culture step is to adjust the pH of the culture medium to 5.0 ~ 7.0, inoculated so that the Ganoderma lucidum mycelium 4 ~ 6% (v / v), the temperature 22 ~ 32 ℃, rotation speed 120 ~ 180rpm, Cosmetic composition, characterized in that incubated for 5-7 days at 1.2-1.8vm conditions. The method according to claim 1,
The cosmetic composition is a lotion, gel, water-soluble liquid, cream, essence, other oil-in-water (O / W) and water-in-oil (W / O) type base cosmetic formulation and oil-in-water or water-in-oil makeup base, foundation, skin cover Cosmetic composition, characterized in that selected from color cosmetic formulations, such as lipstick, lip gloss, face powder, two-way cake, eye shadow, cheek color and eyebrow pencils.