COSMETIC COMPOSITION HAVING ANTI-OXIDATION, SHITENING, AND WRINK-ameliorating EFFECTS COMPRISING CONCENTRATED FERMENTATION

Abstract

The present invention discloses a cosmetic composition and, more specifically, discloses a cosmetic composition capable of providing skin improving effects such as oxidation resistance, whitening, and wrinkle improvement by comprising a concentrated fermented product prepared by concentrating a mixed extract extracted from the root of kudzu vine and lentil seeds and fermenting the concentrated extract with a Lactobacillus strain.

Description

Technical Field

[0001] The present invention discloses a cosmetic composition, and more specifically, discloses a cosmetic composition comprising a concentrated fermentation product prepared by concentrating a mixed extract extracted from kudzu root and lentil seeds and fermenting the concentrated extract with a Lactobacillus strain, which can provide skin improvement effects such as anti-oxidation, whitening, and wrinkle improvement. Background Art

[0002] People living in modern society are experiencing accelerated skin aging due to environmental factors and lifestyle habits, so efforts have been made to develop various cosmetics that can improve skin health and prevent aging. In particular, cosmetics with skin-improving effects such as anti-oxidation, whitening, and wrinkle improvement have received great attention.

[0003] Most of the main functional ingredients of this cosmetic are plant extracts, among which Pueraria lobata root and Glycine Soja (Soybean) seed are well known for their antioxidant and skin improvement effects.

[0004] In Korea, kudzu root is also affectionately called kudzu strips and has been used as a traditional Chinese medicine for a long time. Its main ingredients include isoflavones and flavonoid compounds, which can enhance the antioxidant effect and prevent skin aging. Lentil seeds are also rich in polyphenols and proteins, which enhance skin elasticity and help whiten the skin.

[0005] However, the efficacy of existing cosmetic compositions using kudzu root and lentil seed extracts is limited, and extraction methods such as ultrasonic extraction and further concentration need to be considered in order to enhance the efficacy of functional ingredients and improve extraction efficiency.

[0006] Furthermore, there is a need to develop a new composition that utilizes a mixed extract of kudzu root and lentil seeds to perform a fermentation process to increase the bioavailability of functional ingredients and greatly enhance the skin improvement effect.

[0007] Prior art literature

[0008] Patent Literature

[0009] (Patent Document 0001) China Publication Patent Gazette No. 201410152637.2 (2014.04.16) "KuZhu Tiao Moisturizing Cream and Preparation Method thereof" Summary of the invention

[0010] In order to solve the above problems, the present invention aims to provide a cosmetic composition, which concentrates a mixed extract of kudzu root and lentil seeds and ferments it with a Lactobacillus strain to have antioxidant, whitening and wrinkle-improving effects.

[0011] In order to achieve the above object, the present invention comprises a concentrated fermentation product obtained by fermenting a concentrated solution obtained by ultrasonically extracting a mixed extract of kudzu root and lentil seeds with a Lactobacillus strain.

[0012] Furthermore, the mixed extract is prepared by subjecting a mixture of the kudzu root and the lentil seeds in a weight ratio of 3:7 to 7:3 to ultrasonic extraction in an extraction solvent at 30 to 50 Hz, 500 to 700 Watt, and 30 to 50° C. for 1 to 4 hours.

[0013] Furthermore, the extraction solvent is a solvent selected from the group consisting of water, anhydrous or hydrous lower alcohols having 1 to 4 carbon atoms, ethyl acetate, glycerol and ethylene glycol, or a mixture of two or more thereof and 1,2-hexanediol.

[0014] Furthermore, the concentrated solution is concentrated to 1 to 5 weight % relative to the weight of the mixed extract and concentrated to 40 to 60 weight % relative to the weight of the mixture to have a concentration yield of 40 to 60%.

[0015] Moreover, the Lactobacillus strain is Lactobacillus rhamnosus, and the concentrated fermentation product is prepared by mixing 1 to 10 parts by weight of the concentrated solution and 90 to 99 parts by weight of purified water, and then allowing the air in the fermentation tank to circulate at 20 to 40° C., pH 5.5 to 6.5, and 300 to 500 rpm at a volume flow rate of 3 L / min for 1 to 3 days.

[0016] The cosmetic composition comprising the concentrated fermented product of the mixed extract of kudzu root and lentil seeds according to the present invention and having antioxidant, whitening and wrinkle-improving effects provides skin-improving effects such as antioxidant, whitening and wrinkle-improving effects by concentrating the mixed extract of kudzu root and lentil seeds and further fermenting it using a specific strain. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 is a graph showing the results of a cell viability experiment.

[0018] Figure 2 It is a graph showing the results of a DPPH radical scavenging activity experiment.

[0019] Figure 3 is a graph showing the results of an ABTS free radical scavenging activity experiment.

[0020] Figure 4 is a graph showing the results of a tyrosinase activity inhibition experiment.

[0021] Figure 5 is a graph showing the results of a melanin content suppression experiment.

[0022] Figure 6 It is a graph showing the results of the MMP-1 content inhibition experiment.

[0023] Figure 7 It is a graph showing the results of a collagen biosynthesis rate experiment. DETAILED DESCRIPTION

[0024] The following detailed description of the present invention is an embodiment of the present invention that can be implemented and refers to the accompanying drawings shown as examples of the embodiment. These embodiments will be described in detail in a manner that allows those skilled in the art to fully implement them. Although the various embodiments of the present invention are different from each other, it should be understood that they are not necessarily mutually exclusive. For example, the specific shapes, structures and characteristics recorded in a certain embodiment can be implemented in other embodiments without departing from the spirit and scope of the present invention. Moreover, the position or configuration of individual elements in the embodiments recorded separately can be changed without departing from the spirit and scope of the present invention.

[0025] Therefore, the detailed description below should not be regarded as limiting. As long as it can be properly explained, the scope of the present invention can only be limited by all equivalent ranges and the appended claims that are equivalent to the content requested by the claims. Similar symbols in the figures indicate that their functions are the same or similar in all aspects.

[0026] The terms used in the present invention are selected from the currently widely used general terms as much as possible after considering their functions in the present invention, but they may change according to the intentions or conventions of technicians in the field or the emergence of new technologies. In addition, in certain cases, the terms arbitrarily selected by the applicant are also used, and their meanings will be recorded in detail in the corresponding invention content. Therefore, the terms used in the present invention cannot be defined only by the name of the term, but should be defined according to the meaning of the term and the overall content of the present invention.

[0027] When it is referred to in the present invention that a part “includes” a certain element, it means that it does not exclude other elements but can also include other elements unless otherwise specifically described.

[0028] The following is a detailed description of a cosmetic composition having antioxidant, whitening and wrinkle-improving effects comprising a concentrated fermentation product of a mixed extract of kudzu root and lentil seeds according to the present invention.

[0029] The cosmetic composition having antioxidant, whitening and wrinkle-improving effects according to the present invention, which comprises a concentrated fermentation product of a mixed extract of kudzu root and lentil seeds, comprises a concentrated fermentation product, wherein the concentrated fermentation product is fermented with a Lactobacillus strain into a concentrated solution, wherein the concentrated solution is a mixed extract of kudzu root and lentil seeds extracted by ultrasonic extraction.

[0030] Here, the Pueraria lobata root, also called Pueraria lobata, is a perennial vine belonging to the Leguminosae family and has long been widely used as a medicinal plant. The main components of Pueraria lobata root include isoflavones such as pulerin and daidzein and flavonoid compounds such as catechins and quercetin, which prevent skin aging and help improve wrinkles through a strong antioxidant effect. In addition, Pueraria lobata root promotes microcirculation of the skin to give it vitality and helps inhibit pigmentation to provide a whitening effect.

[0031] The lentil seeds (Glycine Soja (Soybean) seeds) are the seeds of lentil (Glycine Soja), and contain various bioactive substances such as polyphenols, proanthocyanidins, B vitamins, and phytosterols that have antioxidant effects and skin-improving effects. Lentil seeds can exert antioxidant effects, enhance skin elasticity, and maintain moisturizing effects through these ingredients. In particular, the polyphenol components contained in lentil seeds inhibit oxidative stress on the skin and help prevent skin aging, and inhibit melanin production and can exert whitening effects.

[0032] The mixed extract can be prepared by subjecting the mixture of the kudzu root and the lentil seeds to ultrasonic extraction in the presence of an extraction solvent.

[0033] Specifically, the mixture can be prepared by drying and crushing kudzu root and lentil seeds and mixing them in a weight ratio of 3:7 to 7:3. Preferably, kudzu root and lentil seeds are dried and crushed and mixed in equal amounts.

[0034] Next, the mixed extract is prepared by ultrasonically extracting the mixture in an extraction solvent at 30-50 Hz, 500-700 Watt, and 30-50° C. for 1-4 hours. Preferably, the mixture is extracted at 40 Hz, 700 Watt, and 40° C. for 4 hours.

[0035] The ultrasonic extraction conditions refer to the optimum conditions for extracting effective ingredients with high efficiency by rapidly destroying the cell walls of kudzu root and lentil seeds using the acoustic cavitation effect generated when ultrasonic waves are transmitted to liquid.

[0036] At this time, the extraction solvent is a solvent selected from the group consisting of water, anhydrous or hydrous lower alcohols with carbon numbers of 1 to 4, ethyl acetate, glycerol and ethylene glycol, or a mixture of two or more thereof and 1,2-hexanediol.

[0037] Specifically, the extraction solvent may be a mixture of 75 parts by weight of purified water, 15 parts by weight of glycerol, and 5 parts by weight of 1,2-hexanediol.

[0038] The concentrated solution is obtained by concentrating the mixed extract, and the concentrated solution can be concentrated to 1 to 5 weight % of the mixed extract and to 40 to 60 weight % of the mixture to have a concentration yield of 40 to 60%.

[0039] Further, preferably, the concentrate is concentrated to 1.25% by weight compared to the weight of the mixed extract and is concentrated to 50% by weight compared to the weight of the mixture so as to be concentrated with a concentration yield of 50%.

[0040] For example, if 250 g of a mixture of kudzu root and lentil seeds is added to 9750 g of an extraction solvent and extracted to prepare 10 kg of a mixed extract, it is preferably concentrated to obtain 125 g of a concentrate which is 1.25 wt % of the weight of the mixed extract. In this case, 125 g of the concentrate is obtained at a concentration yield of 50% to obtain 50 wt % of the weight of 250 g of the mixture.

[0041] Therefore, preferably, the mixed extract of the present invention is a mixture of the mixture and the extraction solvent in a weight ratio of 1:40.

[0042] The concentration process can be carried out by various known concentration methods. Preferably, a reduced pressure concentration method is used to prevent the effective ingredients from being destroyed.

[0043] The concentrated fermented product increases the biological activity of the effective ingredients and enhances the skin improvement effect by fermenting the concentrated liquid with the Lactobacillus strain.

[0044] The Lactobacillus strain used for the fermentation is a strain that is safe for the skin and has excellent efficacy, and preferably, Lactobacillus rhamnosus can be used.

[0045] Lactobacillus rhamnosus metabolizes the active ingredients of the mixed extract to enhance antioxidant activity, whitening effect, and wrinkle improvement effect, and can generate various secondary metabolites that are beneficial to the skin during the fermentation process.

[0046] Examples of such secondary products include lactic acid that maintains the weak acidity of the skin, phytochemicals such as polyphenols and isoflavones that are metabolized to be converted into phytochemical metabolites with a higher bioavailability such as equol, and gamma-aminobutyric acid (GABA) generated by the metabolism of glutamate.

[0047] At this time, the concentrated fermented product can be prepared by mixing 1 to 10 parts by weight of the concentrate and 90 to 99 parts by weight of purified water, and then allowing the air in the fermentation tank to circulate at a volume flow rate of 3 L / min at 20 to 40° C., pH 5.5 to 6.5, and 300 to 500 rpm for 1 to 3 days for fermentation.

[0048] Furthermore, preferably, the concentrated fermented product is prepared by fermenting 8 parts by weight of the concentrate and 92 parts by weight of purified water in a fermentation tank at 30° C., 6.0 pH, and 400 rpm at a volume flow rate of 3 L / min for 2 days.

[0049] Here, the air in the fermentation tank is circulated at a volume flow rate of 3L / min in order to provide a suitable growth environment for the facultative anaerobic bacteria, Lactobacillus rhamnosus, and to release the heat generated by the metabolic activity of the strain during the fermentation process to keep the temperature in the fermentation tank constant, thereby allowing the strain to maintain a certain activity and providing a basis for uniform quality of the fermentation product.

[0050] In this case, the concentrated fermented product is preferably freeze-dried and powdered to a concentration of 50% by weight based on the weight of the concentrated solution after the fermentation process is completed, so that it can be easily applied to cosmetics of various dosage forms.

[0051] The cosmetic composition of the present invention comprising the concentrated fermentation product can be formulated into toner, lotion, cream, foundation cream, essence lotion, gel, facial mask, emulsified UV protection cream, emulsified foundation cream, emulsified base makeup, oil-powder foundation cream, double-effect powder or pressed powder.

[0052] These dosage forms may contain, in addition to the cosmetic composition of the present invention, commonly used ingredients, for example, general auxiliary agents such as antioxidants, stabilizers, solubilizers, vitamins, pigments, colors and fragrances, and carriers.

[0053] For example, for an ointment, cream or gel containing the cosmetic composition according to the present invention, the carrier component can be selected from one or more of animal oil, vegetable oil, wax, paraffin, starch, tragacanth gum, cellulose derivatives, polyethylene glycol, silicone, bentonite, silicon dioxide, talc or zinc oxide.

[0054] The following examples, comparative examples and experimental examples illustrate in detail the effects of the cosmetic composition having antioxidant, whitening and wrinkle-improving effects comprising the concentrated fermented product of the mixed extract of kudzu root and lentil seeds of the present invention.

[0055] First embodiment. The cosmetic composition of the present invention

[0056] Mixed extraction process: 250 g of a mixture of dried and crushed kudzu root and lentil seeds was mixed with 9.75 kg of an extraction solvent (purified water: glycerol: 1,2-hexanediol, weight ratio 75:15:5) and ultrasonically extracted at 40 Hz, 700 Watt, 40°C for 4 hours to prepare 10 kg of a mixed extract.

[0057] Concentration process - The mixed extract was filtered to remove solids and then concentrated under reduced pressure to collect 125 g of concentrate.

[0058] Fermentation process: 80g of the concentrate and 920g of purified water were mixed and inoculated with Lactobacillus rhamnosus and fermented at 30°C, 6.0 pH, 400 rpm with air circulating at a volume flow rate of 3L / min in the fermentation tank for 2 days.

[0059] Drying process - 40 g of the cosmetic composition was prepared by freeze drying.

[0060] Comparative Example 1: Example excluding the fermentation process

[0061] The cosmetic composition of the first comparative example was prepared by eliminating the fermentation process in the preparation process of the first embodiment and changing the drying process to a process of freeze-drying the concentrate to prepare 62.5 g of the cosmetic composition.

[0062] Second comparative example: Example in which ultrasonic extraction was replaced by hot water extraction

[0063] The cosmetic composition of the second comparative example was prepared by changing the mixed extraction process in the preparation process of the first embodiment to the following process, in which 250 g of a mixture of equal amounts of dried and crushed kudzu root and lentil seeds was extracted with hot water at 40° C. for 4 hours in 9.75 kg of an extraction solvent (purified water: glycerol: 1,2-hexanediol, weight ratio 75:15:5) to prepare 10 kg of a mixed extract.

[0064] Comparative Example 3: Example of using different hot water extraction conditions and changing the extraction solvent

[0065] The cosmetic composition of the third comparative example was prepared by changing the mixed extraction process in the preparation process of the second comparative example to the following process: 250 g of a mixture of equal amounts of dried and crushed kudzu root and lentil seeds was extracted with 9.75 kg of 50% ethanol at 70°C hot water for 4 hours to prepare 10 kg of a mixed extract.

[0066] The cosmetic compositions of the first embodiment and the first to third comparative examples were mixed with purified water at various weight % to prepare samples of various concentrations.

[0067] Experimental Example 1: Confirmation of non-cytotoxicity (MTT assay)

[0068] 1) Experimental methods

[0069] The effects of the samples prepared in the first embodiment and the first to third comparative examples on cell proliferation were confirmed. In order to confirm cell proliferation and toxicity, the following method was used for the experiment.

[0070] Raw 264.7 cells (murine macrophage cell line) were cultured at 1×10 5 cells / ml were divided into 96-well microplates and cultured in a thermostatic chamber for 30 minutes. The samples of the first embodiment and the first to third comparative examples were added at various concentrations and cultured for 72 hours.

[0071] After 72 hours of culture, thiazoline blue was added and culture was continued for 4 hours. After the culture medium was completely poured out, the reaction termination solution was poured into each well of the microplate and stirred for 5 minutes, and then the absorbance was measured at 570nm. The control group was injected with 10% fetal bovine serum (FBS) culture medium according to the sample injection amount and cultured at the same time under the optimal conditions for cell growth. The cell proliferation rate of the experimental group injected with the sample was calculated by setting the cell proliferation of the control group as 100%.

[0072] The cell proliferation effect was calculated using the following mathematical formula 1, and the results are listed in the following Table 1 and Figure 1 , in Table 1 and Figure 1 The relative cell proliferation effect calculated by Mathematical Formula 1 (eg, 3%) is added to the control group reference value (100%) to show the total cell proliferation amount (eg, 103%) compared to the control group as a percentage.

[0073]

Mathematical formula 1

[0074]

[0075] 2) Experimental results

[0076]

Table 1

[0077]

[0078] As shown in Table 1, most of the experimental results showed no significant cytotoxicity. However, at high concentrations, the third comparative example showed a tendency to sharply reduce the cell proliferation effect compared to the control group, which is probably because the extraction solvent was changed to 50% ethanol.

[0079] Second Experimental Example: Confirmation of Antioxidant Efficacy (DPPH Radical Scavenging Activity)

[0080] 1) Experimental methods

[0081] The effects of the samples prepared in the first embodiment and the first to third comparative examples on the scavenging of free radicals were confirmed. In order to confirm the effects on the scavenging of free radicals, the following method was used to conduct the experiment.

[0082] The experiment was conducted using the DPPH (1,1-diphenyl-2-trinitrophenylhydrazine) method (see: Blois. MS Nature 181, 1190, 1958). DPPH (control group) and 100 μg / ml Ascorbic Acid (positive control group) were products of SIGMA. 1 ml of 0.2 mM DPPH methanol solution was added to each of the first embodiment and the first to third comparative examples and 2 ml of the positive control group with different concentrations and stirred, and then reacted at room temperature for 10 minutes. After that, the absorbance was measured at 517 nm using a microplate reader. At this time, a blank test was conducted to compensate for the background signal by replacing each sample with purified water. The blank test results confirmed that the background signal remained constant before conducting the experiment.

[0083] The free radical scavenging activity was measured using the following mathematical formula 2 and the results are shown in Figure 2 With Table 2.

[0084]

Mathematical formula 2

[0085]

[0086] 2) Experimental results

[0087]

Table 2

[0088]

[0089] As can be seen from Table 2, the DPPH free radical scavenging activity (%) shows a tendency to gradually increase with the sample concentration, and the first embodiment shows a higher scavenging activity as a whole in all concentration ranges compared with the first to third comparative examples.

[0090] Furthermore, the first example shows a tendency that the scavenging activity increases more rapidly as the concentration increases. That is, the first example shows superior antioxidant activity compared to the first to third comparative examples.

[0091] Experimental Example 3: Confirmation of Antioxidant Efficacy (ABTS Free Radical Scavenging Activity)

[0092] 1) Experimental methods

[0093] In order to further analyze the antioxidant capacity, the ABTS free radical scavenging method was used. Potassium persulfate and 7mM ABTs were mixed and used in a dark room at room temperature for about 16 hours to form ABTS+. The absorbance was measured at 415nm and diluted with ethanol to a value of 0.70±0.02. 100μg / ml Ascorbic Acid (positive control group) and 5μl of the samples of the first embodiment and the first to third comparative examples at each concentration were mixed with 95μl of the diluted ABTS solution, and the mixture was reacted for 15 minutes, and then the absorbance was measured using a microplate reader (415nm).

[0094] At this time, the radical scavenging effect was calculated by the above-mentioned Mathematical Formula 2, and the ABTs solution in a state where no sample was added was set as a control group.

[0095] The experimental results are listed in the following Table 3 and Figure 3 .

[0096]

Table 3

[0097]

[0098] As can be seen from Table 3, the first example exhibits higher ABTS radical scavenging activity in all concentration ranges compared to the first to third comparative examples. At this time, the reactivity of ABTS is higher than that of DPPH in the second experimental example, which shows a tendency to increase slowly with concentration, and it can be seen that the comparison in the low concentration range should be effective.

[0099] In particular, at a concentration of 0.5%, the free radical scavenging activity of the first embodiment is 6.1% higher than that of the first comparative example which does not include the fermentation process, confirming that the fermentation process has a positive effect on the antioxidant efficiency. Even when compared with the second comparative example and the third comparative example, they also show 4% and 2.7% higher free radical scavenging activities, respectively, confirming once again that the fermentation process and ultrasonic extraction are very important.

[0100] Experimental Example 4: Confirmation of whitening efficacy (tyrosinase activity inhibition rate)

[0101] 1) Experimental methods

[0102] The tyrosinase activity inhibition test is a test to confirm the melanin activity inhibition effect of the corresponding substance, and the measurement is performed by the Yagi method such as measuring DOPA chrome using the colorimetric method.

[0103] In this experiment, a reaction solution was prepared by adding 40 μl of 1.5 mM L-Tyrosine solution to 120 μl of 0.1 M phosphate buffer (pH 6.5) in a 96-well plate. After 10 μl of the sample solutions of the first embodiment and the first to third comparative examples were added to the reaction solution according to the concentrations, the reaction was allowed to proceed for 5 minutes. Then, 10 μl of mushroom tyrosinase (100 unit / ml) was added and the enzyme reaction was carried out at 37° C. for 10 minutes. After 10 minutes, the dopachrome generated in the reaction solution was measured at 490 nm using a microplate counter.

[0104] At this time, the control group used a sample without adding the sample solution, and the blank sample solution used a sample without adding the sample solution and mushroom tyrosinase.

[0105] The tyrosinase activity inhibition rate was calculated using the following mathematical formula 3, and the results are shown in the following Table 4 and Figure 4 .

[0106]

Mathematical formula 3

[0107]

[0108] 2) Experimental results

[0109]

Table 4

[0110]

[0111] As can be seen from Table 4, the first example exhibits superior tyrosinase activity inhibition rates in all concentration ranges compared to the first to third comparative examples.

[0112] Tyrosinase is an enzyme that plays a key role in the melanin production process. If its activity is inhibited, the production of melanin is reduced, which helps to maintain bright skin. Therefore, it can be seen that the first embodiment can provide a better whitening effect than the first to third comparative examples.

[0113] Experimental Example 5. Confirmation of whitening efficacy (melanin content inhibition rate)

[0114] 1) Experimental methods

[0115] B16F10 mouse melanoma cells were cultured at 3.5×10 4 After the cells were divided into 6-well plates and cultured for 24 hours, α-MSH (melanocyte stimulating hormone) and the samples of the first example and the first to third comparative examples prepared at various concentrations were treated in each well.

[0116] After 48 hours of culture, the culture medium was removed by washing twice with phosphate buffer saline (PBS), and the adhered cells were transferred to an EP tube and centrifuged to obtain a cultured cell pellet. The shape of the precipitated B16F10 melanoma cells was photographed, and melanin was dissolved in a 1N NaOH solution with DMSO at 60°C for 1 hour, and then 200μl of each strain was divided in a 96-well plate and the absorbance was measured at 405nm. At this time, the control group was set without adding the sample.

[0117] The melanin content inhibition rate was calculated using the following mathematical formula 4, and the experimental results are listed in the following Table 5 and Figure 5 .

[0118]

Mathematical formula 4

[0119]

[0120] 2) Experimental results

[0121]

Table 5

[0122]

[0123]

[0124] It can be seen from Table 5 that the first embodiment has a superior melanin content suppression capability compared to the first to third comparative examples.

[0125] That is, in cells induced to produce melanin by α-MSH (melanocyte-stimulating hormone), the first embodiment effectively inhibits the production of melanin and can provide a better skin whitening effect.

[0126] Experimental Example 6: Confirmation of wrinkle improvement effect (Evaluation of MMP-1 production inhibition)

[0127] HaCaT and CCD-986Sk cells were adjusted to 5×10 5 The cells were divided into 6-well plates at a concentration of 2 ml and cultured at 37°C with 5% CO2 for 24 hours.

[0128] When the cells grew to about 80% or more, 20 μl of α-MSH (10 mg / ml) was added to each well to a final concentration of 100 μg / ml. Then, 50 μl of the samples of the first example and the first to third comparative examples prepared at various concentrations were added to each well. At this time, the control group was set up with the condition of adding only α-MSH without adding the sample.

[0129] All experimental groups were cultured under the same conditions (5% CO2, 37°C) for 24 hours, and RNA was extracted using Trizol reagent. cDNA was synthesized from the extracted RNA, and GAPDH was quantitatively compared using housekeeping genes.

[0130] The expression level of MMP-1 was measured by RT-PCR analysis, and the band density of the amplified PCR product was quantified by 2% agarose gel electrophoresis. The RT-PCR conditions were denaturation at 95°C for 2 minutes, annealing at 60°C for 30 seconds, and extension at 72°C for 1 minute, and 40 cycles were performed.

[0131] At this time, the positive control group was treated with adenosine at a concentration of 100 μg / ml, and the MMP-1 production inhibition rate was calculated according to the following mathematical formula 5. The results are shown in the following Table 6 and Figure 6 .

[0132]

Mathematical formula 5

[0133]

[0134] 2) Experimental results

[0135]

Table 6

[0136]

[0137]

[0138] The MMP-1 content inhibition rates of the first embodiment and the first to third comparative examples all increased in a concentration-dependent manner. In particular, the first embodiment showed a higher inhibition rate than the first to third comparative examples in all concentration ranges, confirming that it has excellent skin wrinkle improvement efficacy.

[0139] Experimental Example 7. Confirmation of wrinkle improvement effect (Evaluation of collagen biosynthesis)

[0140] 1) Experimental methods

[0141] Human dermal fibroblasts (HDF) were cultured at 5×10 5 The cells were divided into 24-well plates at a concentration of cells / well and cultured for 24 hours under 5% CO2 and 37°C. The samples of the first embodiment and the first to third comparative examples were cultured in DMEM medium without FBS for 24 hours, and the control group was set as a sample that was not treated under the same conditions.

[0142] After the incubation, the supernatant of each well was pooled and the amount of synthesized collagen (ng / ml) was measured using Collagen type I C-peptide enzyme immunoassay (EIA) kit (Takara, Japan). The collagen synthesis increase rate (%) was calculated according to the following mathematical formula 6. Then, the collagen biosynthesis rate of the non-treated group was set as 100% for criterion, and the final biosynthesis rate after adding the increase rate of each experimental group to this criterion was listed in the following Table 7 and Figure 7 .

[0143]

Mathematical formula 6

[0144]

[0145] 2) Experimental results

[0146]

Table 7

[0147]

[0148]

[0149] As shown in Table 7, the first example exhibits excellent collagen biosynthesis rate in all concentration ranges. Therefore, it can be seen that the first example of the cosmetic composition of the present invention can provide a better wrinkle improvement effect than the first to third comparative examples.

[0150] In summary, according to the cosmetic composition of the present invention, kudzu root and lentil seeds are ultrasonically extracted and concentrated and fermented with Lactobacillus rhamnosus strains to provide excellent antioxidant, whitening and wrinkle improvement effects for the skin.

[0151] The following is a cosmetic composition comprising a concentrated fermented product of a mixed extract of kudzu root and lentil seeds according to the present invention, which has antioxidant, whitening and wrinkle-improving effects, based on the results of the above-mentioned multiple experimental examples. However, the cosmetic composition of the present invention is not limited to the following dosage form examples.

[0152] Dosage Form Example 1. Preparation of Lotion

[0153] The formulation of the lotion (body lotion) containing the cosmetic composition of the first embodiment is as follows.

[0154]

Table 8

[0155]

[0156] <Preparation method>

[0157] After adding No. 2, 3, 4, and 8 to No. 11 and stirring to dissolve them, heat No. 5 to about 60°C and dissolve it, then add No. 10 to dissolve it, and then add No. 11. Finally, add No. 6, 7, and 9, and then add No. 1, stir it thoroughly, and then mature it.

[0158] Dosage Form Example 2. Preparation of Nutritional Emulsion

[0159] The formulation of the nourishing emulsion in the cosmetic composition of the first embodiment is as follows.

[0160]

Table 9

[0161]

[0162] <Preparation method>

[0163] While mixing and stirring 10, 11, 12, 13, and 16, heat them to 80-85°C and put them into the preparation device. Then, use an emulsifier to heat 2, 3, 4, 5, 6, 7, 8, 9, and 12 to 80-85°C and dissolve them, and then emulsify them. After emulsification, while stirring with a stirrer, cool to 50°C, add No. 15, cool to 45°C, then add No. 14, add No. 1 at 35°C, cool to 25°C, and then mature.

[0164] Dosage Form Example 3. Preparation of Nutritional Cream

[0165]

Table 10

[0166]

[0167] <Preparation method>

[0168] While mixing and stirring 12, 13, 14, and 16, heat them to 80-85°C and put them into the preparation device. Then, using an emulsifier, heat 2, 3, 4, 5, 6, 7, 8, 9, 10, and 11 to 80-85°C and dissolve them, emulsify them, and then add 15 and stir them, and then add them into the preparation device to emulsify them. After emulsification, cool them to 35°C while stirring them with a stirrer, add No. 1, and then cool them to 25°C and mature them.

[0169] The foregoing text describes the present invention in conjunction with the accompanying drawings, but it is only one embodiment of the various embodiments of the subject matter of the present invention. Its purpose is to allow those with general knowledge in the technical field to which the present invention belongs to easily understand it. Obviously, the present invention is not limited to the described embodiments. Therefore, the protection scope of the present invention should be explained according to the claims. Without departing from the subject matter of the present invention, all technical spirits within the equivalent range achieved by means of change, replacement, substitution, etc. should be interpreted as rights belonging to the present invention. Moreover, some elements in the drawings are only used for more clear explanations and their actual situation is overly exaggerated or overly reduced.

Claims

1. A cosmetic composition comprising a concentrated fermentation product of a mixed extract of kudzu root and lentil seeds and having antioxidant, whitening and wrinkle-improving effects, characterized in that: The invention comprises a concentrated fermentation product obtained by fermenting the following concentrated solution with a strain of Lactobacillus, wherein the concentrated solution is a mixed extract obtained by ultrasonically extracting kudzu root and lentil seeds.

2. The cosmetic composition according to claim 1, characterized in that The mixed extract is prepared by subjecting a mixture of the kudzu root and the lentil seeds in a weight ratio of 3:7 to 7:3 to ultrasonic extraction in an extraction solvent at 30 to 50 Hz, 500 to 700 Watt, and 30 to 50° C. for 1 to 4 hours.

3. The cosmetic composition according to claim 2, characterized in that The extraction solvent is a solvent selected from the group consisting of water, anhydrous or hydrous low-carbon alcohols with carbon numbers of 1 to 4, ethyl acetate, glycerol and ethylene glycol, or a mixture of two or more substances and 1,2-hexanediol.

4. The cosmetic composition according to claim 1 or claim 2, characterized in that: The concentrate is concentrated to 1 to 5% by weight relative to the weight of the mixed extract and concentrated to 40 to 60% by weight relative to the weight of the mixture to have a concentration yield of 40 to 60%.

5. The cosmetic composition according to claim 1, characterized in that The Lactobacillus strain is Lactobacillus rhamnosus, The concentrated fermented product is prepared by mixing 1 to 10 parts by weight of the concentrated solution and 90 to 99 parts by weight of purified water, and then circulating the air in the fermentation tank at 20 to 40° C., pH 5.5 to 6.5, 300 to 500 rpm and a volume flow rate of 3 L / min for 1 to 3 days for fermentation.

Images