KR101730352B1 - Antioxidant composition for cosmetics composite, cosmetics composite containing the same, and Manufacturing method thereof - Google Patents

Abstract

The present invention relates to an antioxidant composition, a cosmetic material containing the same, and a manufacturing method thereof. More specifically, an antioxidant is produced by mixing an antioxidant composition which contains an Nrf2 activator and 4 kinds of natural ingredients in an optimum composition, and the antioxidant can be utilized as health functional food, skin ointment, cosmetics, hair products including shampoo and the like, and various products. For an example, a cosmetic material can be produced by using the antioxidant, and various products with a skin active oxygen removal function, a skin elastic improvement effect, and a skin whitening effect can be produced.

Description

[0001] The present invention relates to an antioxidant composition for cosmetics, a cosmetic composition containing the antioxidant composition and a process for producing the same, and an antioxidant composition for cosmetics,

The present invention relates to an antioxidant composition comprising an Nrf2 activator and a natural extract which have high activity of scavenging stress, which effectively restores stressed and tired skin, a cosmetic using the same, and a method for producing the same.

It is 3 ~ 10cm in diameter and 0.5 ~ 3cm in thickness. It is almost circular in shape and has a diameter of 1 ~ 3cm and a length of 5 ~ 8cm. Sometimes, There is also a vertical split. The cross section is grayish brown to yellowish brown, and the brown layer is clear in the vicinity of the formation layer. And the side is dark brown, with horizontal wrinkles, peeled tan, the neck diameter is about 3-4 times the skin, and the outer surface is very hard but not fibrous. And it is a medicinal herb that has a characteristic odor and has a characteristic that it is chewy and chewy when it is chewed, and it is colored yellow. The composition for preventing and treating diabetes using such an extract of Longevity (Korean National Publication No. 2005-0003669), a plant disease control activator (Republic of Korea Publication No. 2010-0048351), constipation treatment (Korean Registration No. 1286467) Technology has been known, but there is no known technology for skin remedy using a lyophilized extract.

It is 3 ~ 10cm in diameter and 0.5 ~ 3cm in thickness. It is almost circular, and it is 1 ~ 3cm in diameter and 5 ~ 8cm in length. Sometimes it is vertical . The cross section is grayish brown to yellowish brown, and the brown layer is clear in the vicinity of the formation layer. And the side is dark brown, with horizontal wrinkles, peeled tan, the neck diameter is about 3-4 times the skin, and the outer surface is very hard but not fibrous. And it is a medicinal herb that has a characteristic odor and has a characteristic that it is chewy and chewy when it is chewed, and it is colored yellow. This composition was used as a composition for preventing and treating diabetes using an extract of this species (Korean National Publication No. 2005-0003669), a plant disease control activator (Republic of Korea Publication No. 2010-0048351), constipation treatment (Korean Registration No. 1286467) Technology has been known, but there is no known technology for skin remedy using a sardine extract.

Active oxygen is a substance that is constantly produced in the normal metabolic process of the human body. In general, about 2 to 5% of the oxygen we breathe is converted to active oxygen. This active oxygen is dangerous because of the strong oxidizing action. Excessive active oxygen in the body damages cells, proteins, and DNA, causing abnormalities in cell structure, function, and signal transduction, ultimately leading to inflammation or aging in the body. The ultraviolet rays of the sun, which is one of the causes of active oxygen, cause reactive oxygen species (ROS) and cause diseases such as erythema, water, ultraviolet rays, photoaging and skin cancer. ROS, including superoxide anion (O ₂ ^- ), H ₂ O ₂ (hydrogen peroxide) and OH (hydroxy radical), is continuously produced through respiration and metabolism. In the body, SOD (superoxide dismutase), Gpxes (enzymes such as glutathione peroxidase and catalase) are used to prevent ROS damage. However, excessive generation of ROS by the sunlight exceeds the antioxidative defense ability in vivo, and oxidative stress is generated. That is, it reacts with DNA, cell membrane and protein to cause damage to cells and tissues, which is known to promote various inflammatory diseases, Parkinson's disease, photoaging, and cancer development.

Thus, Nrf2, which is distributed in the epidermal layer of the skin, is a transcription factor that acts on the removal of active oxygen generated from the inside and the outside of the human body. Activation of Nrf2 can prevent skin aging, skin cells and skin tissue damage.

The inventors of the present invention have continuously studied and found that the extract of L. sativa and the species of R. radivifolia promote the activity of Nrf2 to remove active oxygen causing aging of the skin, thereby protecting and improving the skin, When they are mixed in an appropriate amount, the synergistic effect of them increases the active oxygen scavenging ability, thereby completing the present invention. In addition, if the Nrf2 activator alone is used, skin irritation may be induced due to a strong antioxidative effect. However, when the Nrf2 activator is mixed with a specific natural extract, the skin trouble caused by skin irritation can be minimized while maximizing the antioxidant effect The present invention has been completed. That is, the present invention provides an antioxidant composition comprising the Nrf2 activator and a natural mixed extract, a cosmetic comprising the same, and a method for producing the same.

In order to solve the above problems, the present invention provides a pharmaceutical composition comprising: an activator of Nrf2, And a mixed natural extract comprising a mushmundong extract, a mushroom extract, a lotus extract and a chaga mushroom extract.

The present invention also relates to the above antioxidant composition; Aqueous solution; Emulsion; Spices; And purified water.

The present invention also provides a method for producing the cosmetic composition.

The present invention also provides a cosmetic comprising the cosmetic.

The antioxidant composition of the present invention assists the activity of Nrf2 distributed in the skin epidermal layer to greatly enhance the active oxygen elimination performance in the skin, thereby improving skin elasticity, excellent antioxidative effect and whitening effect, and restoring vigorous and dull skin The antioxidant composition of the present invention can be applied to various products such as a health functional food, a skin ointment, a cosmetic product, a hair product such as a shampoo, etc. Specifically, Cosmetics incorporating cosmetics can be applied to the skin at any time without regard to the time of use, and there is no side effect of skin troubles or it can be minimized.

FIGS. 1A and 1B are graphs of RFU changes according to the Trolox concentration of the positive control group performed in Experimental Examples 1-1 and 1-2, respectively.
2A and 2B are graphs of netAUC standard curves according to the positive control Trolox concentration in Experimental Examples 1-1 and 1-2, respectively.
FIG. 3A is a graph of RFU change according to concentration of the crude extract obtained in Experimental Example 1-1. FIG.
FIG. 3B is a graph of RFU change according to the concentration of the species rhubarb extract obtained in Experimental Example 1-2. FIG.
FIG. 4A shows the results of measurement of the oxygen radical scavenging activity (netAUC) by the concentration of the lyophilized extract obtained in Experimental Example 1-1.
FIG. 4B shows the results of measurement of the oxygen radical scavenging activity (netAUC) by the concentration of the species interaction extract in Experimental Example 1-2.
FIG. 5 shows the results of the measurement of the activity of luciferase in Experimental Example 2, wherein FIG. 5A shows the results of the measurement of the lyophilized extract and FIG. 5B shows the results of the measurements of the lycopene extract.
FIG. 6 is a cytotoxicity measurement result of Experimental Example 3, FIG. 6A is a cytotoxicity measurement result of the lyophilized extract, and FIG. 6B is a cytotoxicity measurement result of a species lyophilized extract.
FIG. 7 is a result of quantifying a protein through Western blot performed in Experimental Example 4, wherein FIG. 7A is a quantification result of a lyophilized extract, and FIG. 7B is a quantitative result of a rhododendron extract.
8A to 8C are the results of measurement of mRNA (messenger RNA) levels of Nrf2, HO-1 and NQ01 of the prey extract by PCR (Real Time PCR) performed in Experimental Example 5, This is the result of measurement of mRNA (messenger RNA) level of Nrf2, HO-1 and NQ01 of the rhubarb extract.
FIG. 9 shows the results of the oxidative stress defense test of the extracts of L. sativa L. and P. lanceolata, and FIG. 9C (extract of L. sativa) and FIG. 9C 8-hydroxy deoxyguanosine levels were measured. Fig. 9b (stool extract) and Fig. 9d (bell pepper extract) were photomicrographs of intracellular 4-HNE (4-hydroxynonenal) wool.
Fig. 10 is an optical microscope photograph showing the removal of active oxygen from the species rhubarb extract obtained in Experimental Example 7. Fig.
FIG. 11 shows the DPPH radical scavenging activity of the crude extract and the sardine extract obtained in Experimental Example 8. FIG. 11A shows the results of the measurement according to the concentration of the crude extract and FIG. 11B shows the results according to the concentrations of the sardine extract.

Hereinafter, the present invention will be described in detail.

The antioxidant composition of the present invention comprises an activator of Nrf2; And the mixed natural extract at a weight ratio of 1: 1.5 to 4, preferably 1: 1.8 to 3.5, more preferably 1: 2.0 to 3.2.

First, the activator of Nrf2 among the components of the present invention will be described. The activator of Nrf2 includes a serpentinized extract and a sardine extract.

Preparing a raw material by mixing 4 to 10 parts by weight of shelf-wise with 100 parts by weight of purified water; Extracting the raw material by hot water extraction at 90 to 100 ° C for 4 to 8 hours to obtain an extract; Preparing a primary filtrate obtained by first filtering the extract; Adding 1,2-hexanediol and ethylhexyl glycerin to the primary filtrate and stirring the mixture, and then subjecting the filtrate to secondary filtration to prepare a lyophilized extract.

In the preparation of the lyophilized extract, the primary filtering may be performed with a filter membrane having an average pore size of 20 to 30 탆, and preferably filtering with a filter membrane having an average pore size of 23 to 27 탆.

The secondary filtering may be performed with an average pore size of 8 to 15 μm filter layer, preferably 8 to 12 μm filter layer.

The activator of Nrf2, which was solely applied with the above-described lyophilized extract, was converted into Trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid) equivalents as a control group, (ORAC) value of 90-110 mM TE / g, preferably 92-105 mM TE / g, more preferably 95-102 mM TE / g, .

Preparing a raw material by mixing 4 to 10 parts by weight of a rhizome with respect to 100 parts by weight of purified water; Extracting the raw material by hot water extraction at 90 to 100 ° C for 4 to 8 hours to obtain an extract; Preparing a primary filtrate obtained by first filtering the extract; A step of adding 1,2-hexanediol and ethylhexyl glycerin to the primary filtrate and stirring the mixture, and then subjecting the filtrate to a secondary filtration to prepare a sardine extract.

The primary filtering may be performed with a filter membrane having an average pore of 20 to 30 탆, and preferably a filtering membrane having an average pore of 23 to 27 탆.

The secondary filtering may be performed with an average pore size of 8 to 15 μm filter layer, preferably 8 to 12 μm filter layer.

When the activator of Nrf2, which is solely applied to the spermatogonia extract alone, was converted into the equivalent amount of Trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid) (ORAC) value of the extract of Bellflower seeds at a concentration of 0.00625% by volume is 580 to 650 mM TE / g, preferably 600 to 640 mM TE / g, more preferably 610 to 635 mM TE / g Lt; / RTI >

The Nrf2 activator has a lipase activity value of 2,2-diphenyl-1-picrylhydrazyl (DPPH) at a concentration of 0.027 vol% (solvent purified water) It may have a _50% ((IC 50). Further, as the Nrf2 activator component case containing the longitudinal rhubarb extract alone, or species rhubarb extract concentration is 0.174% by volume (solvent purified) one time, DPPH radical scavenging activity value Can have 50% (IC ₅₀ ).

However, the use of a combination of the supernatant extract and the sardine sulphate extract as a component of the Nrf2 activator significantly increases the DPPH radical scavenging activity. As a specific example, when the Nrf2 activator contains a lyophilized extract of 0.027 vol% (solvent purified water) at a concentration of 0.174 vol% (solvent purified water) at a 1: 1 weight ratio, the DPPH radical scavenging activity value 60 to 65%, and can have a higher DPPH radical scavenging activity value when the total amount of the extracts is the same as that of the case of using the extracts of L. ssp.

The Nrf2 activator may contain the lyophilized extract and the species rhubarb extract in a weight ratio of 1: 0.5 to 2, preferably 1: 0.8 to 1.8, more preferably 1: 1 to 1.5, When the extract is used in an amount of less than 0.5 parts by weight, the DPPH radical scavenging activity can not be improved more than when the extract is used, and even if the scurvy radical extract is used in an amount exceeding 2 parts by weight, the DPPH radical scavenging activity Therefore, it is preferable to mix them at a weight ratio within the above range.

The concentration of the lyophilized extract is 0.005-0.30% (volume% based on water as a solvent) and the concentration of the extract is 0.01-0.4% (volume% relative to water as the solvent) Activity and DPPH radical scavenging activity.

Next, the mixed natural extract as the antioxidant composition of the present invention will be described.

Liriope platyphylla is a perennial plant belonging to the lily family and grows in a shady place, mainly in Korea, Japan, China, and Taiwan. Leaves gather from short, coarse rootstocks to form an abandon, and often the root ends grow like peanuts. Flowers bloom in May ~ June, purplish and 3 ~ 5 on the nodes of water inflorescence. It is top of the ovary. The fruit is round and round, and the peel is peeled off early, so the seed is exposed and black. Root of root is used as anti-inflammatory, tonic, broad-spectrum expectorant, and cardiotonic agent. A preferred site for use in accordance with the present invention is the germinal seed region. In addition, the present invention introduces the extract of McMundong as the above-mentioned mixed natural extract component.

Melanin is converted from tyrosine to dopa and dopaquinone by tyrosinase action in melanocytes, and is produced through dopachrome. This melanin exists in the skin and protects the body from ultraviolet light and has an important function to regulate hormone secretion in the body. However, the over-produced melanin is known to form spots and freckles, promotes skin aging, and plays an important role in skin cancer induction. The McMundong seed extract inhibits such tyrosinase activity, have.

In addition, the above-mentioned McMundong seed extract can be prepared according to a conventional solvent extraction method. In a preferred embodiment, a 70% ethanol aqueous solution is added to McMundong seed, which is dried by shade drying, and then cooled at room temperature for 48 hours. The crude extract can be obtained by concentration. The crude extract thus obtained is dissolved in a sufficient amount of 80% methanol aqueous solution and placed in a separating funnel. The same amount of hexane is added thereto, and the mixture is left to shake. The hexane layer and the lower portion are separated into 80% methanol layer in a few minutes. Leave for more than an hour. Separate the separated hexane layer and repeat the dispensing with hexane added to the 80% methanol layer again. For the second time, the separated hexane layer is taken, and hexane is added again to the 80% methanol layer. The hexane layers thus separated three times may be combined and concentrated under reduced pressure to give a hexane fraction. The 80% methanol layer is concentrated to a 30% methanol solution, this time being dispensed with chloroform. Chloroform is heavy and forms a lower layer, and the 30% methanol layer is the upper part. The chloroform layer was separated and the same chloroform layer was separated by repeating the same procedure twice. The filtrate was concentrated under reduced pressure to give a chloroform fraction.

Then, the 30% methanol layer is concentrated, the methanol is completely removed, and the mixture is distributed three times with the same amount of butanol and water. The isolated butanol layers are combined and concentrated under reduced pressure to give a butanol fraction. This extraction method can be used to produce McMundong seed extract.

Next, the moxa extract, which is one of the components of the mixed natural extract, is able to inhibit skin troubles by imparting antioxidant and anti-inflammatory effects. Ecklonia Stolonifera OKAMURA and Ecklonia Cave are perennial brown algae which are distributed in Korea and Japan. (Arch, Pharm. Res. 19, p223-227, 1996), antimutagenic activity (Lee, JH et al. JH et al., J. Food Sci. Nutr. 1, pp. 64-68, 1996; Lee, JH et al., Nat. Prod. Sci 4, p105-114, 1998; Han ES et al., Environ. (Park, DC, et al., Fish. Schi.), Inhibitory effects on food intake (Taniguchi K. et al., Nippon Suisan Gakkaishi, 57, p2065-2071, 1991) and tyrosinase inhibitory activity Tech. 3, p195-199, 2000).

The extract of Momogipy is a crude extract or a nonpolar solvent soluble extract, and the crude extract includes extracts soluble in water, ethanol, methanol, lower alcohol such as butanol or a mixed solvent thereof, preferably ethanol, and nonpolar solvent- hexane, ethyl acetate, n-hexane, dichloromethane, chloroform or ethyl acetate, preferably n-hexane or ethyl acetate. In one preferred embodiment of preparing the mugwort extract, the mugwort is natural dried in a shade, ground and pulverized. Next, a volume of water of about 3 to 20 times, preferably 3 to 5 times the weight of the powdered dried mushroom phloem, a lower alcohol such as methanol, ethanol and butanol, or a mixing ratio of 1: 0.1 to 10 The mixture is subjected to hot water extraction, cold extraction, reflux cooling extraction or ultrasonic extraction at an extraction temperature of 20 to 100 ° C, preferably 20 to 70 ° C for 0.5 hours to 2 days, preferably 1 hour to 1 day After extraction by filtration, the filtrate is concentrated under reduced pressure at 20 to 100 DEG C, preferably 40 to 70 DEG C by using a vacuum rotary condenser, and water, a lower alcohol Or a mixture of these solvents can be obtained.

The mushroom extract is suspended in water and then extracted with a solvent in the order of n-hexane, dichloromethane, ethyl acetate and n-butanol to obtain a herbicidal non-polar solvent-soluble extract of the present invention, preferably n-hexane or ethyl acetate Hexane, water and ethanol are mixed at a predetermined ratio, preferably about 10: 9: 1, to a n-hexane soluble fraction and water Soluble fractions and extracting the water-soluble fraction with dichloromethane to obtain a water-soluble fraction and a dichloromethane-soluble fraction, wherein ethyl acetate is added to the ethyl acetate-soluble fraction and the water-soluble fraction Finally, the water-soluble fraction was extracted with butanol to obtain a water-soluble fraction It can be obtained hoekmul and water-soluble fractions.

The non-polar solvent-soluble fraction may be subjected to silica gel column chromatography (SiO ₂ column chromatography), and may be subjected to column chromatography using a mixed solvent of ethyl acetate and methanol, preferably a mixed solvent of ethyl acetate and methanol, , Preferably at a rate of about 1000 mL per hour, to yield 20 fractions from F1 to F20. The F1 fraction was further purified by silica gel column chromatography using a mixed solvent of hexane: ethyl acetate at a ratio of about 1: 1, followed by separation using a 20 to 100% methanol solution by concentration gradient elution in column chromatography, To isolate and identify the compounds.

The content of the mushroom extract, which is one of the components of the mixed natural extract, is preferably 0.2 to 0.6, preferably 0.3 to 0.5, by weight based on the weight of the mushmundong extract, wherein the mushroom extract is less than 0.2 weight ratio There is a problem that the anti-inflammatory effect and the skin trouble-preventing effect become small, and when it is used in excess of 0.6 weight ratio, there may be a problem that compatibility with other extract components is inferior.

Next, the lotus extract, which is one of the components of the mixed natural extract, not only reduces oxidative stress caused by the sunlight (skin irritation mitigation effect), but also enhances the whitening effect by increasing the antioxidant effect. The lotus is a perennial herb with a buttercup (Nerumbo nucifera). The lotus is mainly used for ornamental purposes, but its application has not been studied much, especially the leaves contain alkaloids and tannins, so they are used for astringent and hemostatic agents or for treatment of nocturnal encephalopathy in the private sector. The underground stem is called Lotus root, it contains asparagine, it is edible, and it is also used as a nourishing medicine. The fruit is called the federation, the seed and the lady is known to be good for women. Also, Yeonhyupju, which is a soaked liquor using fried lotus root leaves or lotus leaf, is a part of Korean traditional food.

Preparing a raw material by mixing 10 to 15 parts by weight of lotus powder with 100 parts by weight of purified water; Extracting the raw material by hot water extraction at 90 to 100 ° C for 4 to 8 hours to obtain an extract; Preparing a primary filtrate obtained by first filtering the extract; A step of adding 1,2-hexanediol and ethylhexyl glycerin to the primary filtrate and stirring the mixture, and then subjecting the filtrate to a secondary filtration to prepare a sardine extract.

In the preparation of the lotus leaf extract, the primary filtering may be performed with a filter membrane having an average pore size of 20 to 30 μm, and preferably filtering with a filter membrane having an average pore size of 23 to 27 μm.

The secondary filtering may be performed with an average pore size of 8 to 15 μm filter layer, preferably 8 to 12 μm filter layer.

The content of the lotus extract, which is one of the components of the mixed natural extract, is preferably 2 to 4 parts by weight, preferably 2.3 to 3.5 parts by weight, based on 1 part by weight of the extract of Mukmunjong Seed Extract. When the amount of the lotus extract is less than 2 parts by weight The skin irritation-mitigating effect can be hardly observed and the whitening-enhancing effect can be lowered. If it is used in an amount exceeding 4 parts by weight, the compatibility with other extract components may be deteriorated and there is no further skin irritation mitigation effect It is preferable to use it within the above range.

The origin of the mushroom is black birch. If you look at the tea mushrooms native to the Kamchatka peninsula, they are attached to a living birch like a black hump, and this black hump grows gradually, and the birch is gradually dying. Fruiting bodies are formed in the bark of birch, and the epidermis is black and sclerotia is formed with yellow tissue. In particular, it is presumed that chaga mushrooms produce yellow pigments and these pigments are related to various physiological activities of chaga mushrooms. In addition, the yellow colorant may contain antioxidant compounds such as inonoblin A, inonoblin B, phelligridin D, phelligridin E, and phelligridin G, G), which is a herbal medicine component having an excellent antioxidative effect.

The present invention relates to a method of preparing a raw material by mixing 0.5 to 5 parts by weight of a mushroom powder with respect to 100 parts by weight of purified water, wherein the extract of the mushroom is introduced as one of the components of the mixed natural extract. Extracting the raw material by refluxing cooling extraction at 30 to 50 ° C for 2 to 3 hours to obtain an extract; Preparing a primary filtrate obtained by first filtering the extract; A step of adding 1,2-hexanediol and ethylhexyl glycerin to the primary filtrate and stirring the mixture, and then subjecting the filtrate to a secondary filtration to prepare a tea mushroom extract.

The primary filtering may be performed with a filter membrane having an average pore of 20 to 30 탆, and preferably a filtering membrane having an average pore of 23 to 27 탆.

The secondary filtering may be performed with an average pore size of 8 to 15 μm filter layer, preferably 8 to 12 μm filter layer.

The content of chaga mushroom extract, which is one of the mixed natural extract components, is preferably 0.1 to 0.5 parts by weight, preferably 0.2 to 0.4 parts by weight based on 1 part by weight of the extract of mackundo seed extract, wherein the content of the lotus extract is less than 0.1 part by weight , The antioxidant-promoting effect is weak, and if it is used in excess of 0.5 weight ratio, the price competitiveness of the finished product may be deteriorated due to the high price.

The cosmetic of the present invention uses an antioxidant including the antioxidant composition described above, and the antioxidant; Aqueous solution; Emulsion; Spices; And purified water.

The cosmetic of the present invention comprises 15 to 20% by weight of the aqueous liquid, 18 to 25% by weight of the emulsion, 5 to 18% by weight of the antioxidant, 0.05 to 1% by weight of the perfume, , 16 to 19.5% by weight of the aqueous phase, 18.5 to 23% by weight of the emulsion, 6 to 15% by weight of the antioxidant, 0.1 to 0.5% by weight of the perfume and remaining water.

If the emulsion is less than 18% by weight, it may be difficult to keep the skin moisturized due to oil content. If it exceeds 25% by weight, excessive oil component may be contained to cause problems such as skin respiration disturbance and oily skin porridge. have. If the activator of Nrf2 is less than 1% by weight, the content of the activator of Nrf2 may be too small to exhibit the effect of removing active oxygen. If the activator of Nrf2 is more than 7% by weight, the compatibility with other components may deteriorate. And it is not economical since the increase of the Nrf2 activation effect due to overuse is almost negligible.

Perfume 85147, Perfume 25399 and the like can be used as a specific example of the cosmetic ingredient.

[Water solution]

Among the cosmetic ingredients of the present invention, the aqueous liquid includes a skin conditioner, a sequestering agent, a thickener, a moisturizer, a viscosity reducing agent, a sterilizing preservative and a solvent.

The skin conditioning agent may be selected from the group consisting of sodium hyaluronate, hyaluronic acid, licorice powder, and hydrolite. The hyaluronic acid, Licorice powder and hydrolite, and more preferably hyaluronic acid, licorice powder and hydrolite may be mixed and used. When mixed with hyaluronic acid, licorice powder and hydrolite, the amount of hyaluronic acid, licorice powder and hydrolite is adjusted to 1: 0.01 to 0.2: 1 to 3 by weight, preferably 1: 0.02 to 0.15: It is preferable to mix them at a weight ratio of 1.2 to 2.7.

The metal ion sequestering agent in the aqueous liquid component serves to prevent deterioration by metal ions contained in the preparation and is used in an amount of 0.1 to 2 parts by weight based on 100 parts by weight of the skin conditioner, May be used in an amount of 0.2 to 1.5 parts by weight. If the amount of the sequestering agent is less than 0.1 part by weight, the metal ion blocking effect may be deteriorated. If the amount is more than 2 parts by weight, It is recommended to use it within the above range. The metal ion sequestrant may be any of those commonly used in the art, and preferably disodium EDTA, dipotassium EDTA, trisodium dithiate, tetrasodium dithiothreitol, And rice phytate may be used singly or in combination of two or more.

Next, the thickening agent of the aqueous liquid component serves to increase the skin adhesion property by imparting an appropriate viscosity and tackiness to the preparation. The amount of the thickening agent to be used is 0.5 to 2 parts by weight, preferably 0.5 to 2 parts by weight per 100 parts by weight of the skin conditioner If the amount of xanthan gum used is less than 0.5 parts by weight, the amount of the xanthan gum used is too small to be used. If the amount of xanthan gum used is more than 2 parts by weight, the viscosity of the preparation becomes too high, There may be a problem in that the stickiness is increased too much and the merchantability is lowered. Therefore, it is preferable to use within the above range. As the thickening agent, those generally used in the art can be used, and preferably, one or two or more selected from the group consisting of antimicrobial agent, guarhydroxytrimonium chloride and hydroxyethylcellulose can be used in combination.

Next, the above-mentioned moisturizing agent of the aqueous liquid component serves to enhance skin moisturization together with the water-evaporating and blocking agent of the emulsion. The amount of the moisturizing agent to be used is 40 to 80 parts by weight, preferably 42 to 80 parts by weight per 100 parts by weight of the skin conditioner If the amount of the moisturizing agent is less than 40 parts by weight, the skin moisturizing effect may be deteriorated. If the amount of the moisturizing agent is more than 80 parts by weight, there may be an uneconomical problem because there is no moisturizing effect. It is good to use. As the moisturizing agent, methylpropanediol and glycerin may be used alone or in admixture of two or more.

Next, the viscosity reducing agent is used to increase the adhesive force with the skin and the skin absorbency while maintaining an appropriate viscosity. The amount of the viscosity reducing agent is 45 to 90 parts by weight, If the amount of the viscosity reducing agent is less than 45 parts by weight or more than 90 parts by weight, the viscosity of the preparation may be too high or too low, There may be a problem that the adhesive strength is too low or the tackiness becomes too large and the feeling of use is lowered. The viscosity reducer may be a conventional viscosity reducer used in the art, preferably a di (C2-C5 alkylene) glycol, more preferably a dipropylene glycol .

Next, among the aqueous liquid components, the sterilizing preservative is used to prevent the cosmetic from being contaminated by microorganisms and the like. Any sterilizing preservative that is not prohibited to be used as a cosmetic material can be used without any particular limitation. Etc. may be used. The use amount of the skin conditioning agent may be 2 to 10 parts by weight, preferably 4 to 8 parts by weight, based on 100 parts by weight of the skin conditioner. If the amount is less than 2 parts by weight, the effect of preventing microbial contamination may be insufficient And use of more than 10 parts by weight may cause skin troubles due to excessive use, so that it is preferable to use within the above range.

Next, as the above-mentioned solvent for the aqueous liquid component, one or a mixture of two or more selected from hexanediol, propylene glycol, butylene glycol and pentylene glycol may be used in combination, and hexanediol and Butylene glycol, and more preferably, 1,2-hexanediol and 1,3-butylene glycol can be mixed and used. The solvent may be used in an amount of 30 to 60 parts by weight, preferably 35 to 50 parts by weight, based on 100 parts by weight of the skin conditioner. If the amount of the solvent is less than 30 parts by weight, And it is uneconomical to use more than 60 parts by weight.

[Emulsion]

Among the cosmetic ingredients of the present invention, the emulsion contains a moisture-blocking agent, tocopheryl acetate, a skin conditioner, an emulsifier, a skin-softening agent, a nonionic surfactant and a film former.

The moisture-blocking agent in the oily component serves to maintain an appropriate moisture content of the cosmetic and to maintain the proper moisture content together with the moisturizing agent when applying the prepared preparation to the skin, and it is possible to use those generally used in the art, Caprylic / Capric Triglyceride oil, MCT oil, and squalane. More preferably, the MCT oil and squalane are mixed in an amount of 5 to 10: 1 It is preferable to mix them in a weight ratio.

Next, the tocopheryl acetate in the oily component serves to prevent the preparation from being oxidized and deteriorated over time. The tocopheryl acetate is used in an amount of 0.1-5 parts by weight, preferably 0.1-5 parts by weight, per 100 parts by weight of the moisture- 0.4 to 2 parts by weight may be used. If the amount of tocopheryl acetate used is less than 0.1 part by weight, the amount of tocopheryl acetate used may be too small to prevent oxidation. If the amount of tocopheryl acetate exceeds 5 parts by weight, there may be a problem of irritation to the skin. It is good to use.

Next, the skin conditioning agent serves to supply nutrients to the skin. The skin conditioning agent may be a mixture of two or more species selected from shea butter, coconut oil, macadamia nut oil and jojoba oil. , Preferably a mixture of shear fat and coconut oil at a weight ratio of 1: 0.4 to 0.8. The skin conditioning agent may be used in an amount of 10 to 35 parts by weight, preferably 13 to 30 parts by weight, based on 100 parts by weight of the water vapor barrier. If the amount of the skin conditioning agent is less than 10 parts by weight, If the content exceeds 35 parts by weight, there may be a problem that the use amount thereof is excessively large, so that it may remain on the surface of the skin after the skin is partially absorbed, so it is recommended to use within the above range.

Next, the emulsifier among the emulsion components is used for improving the compatibility between the emulsion compositions and enhancing the emulsifying power, and it is possible to use those generally used in the art, preferably cetearyl glucoside and cetearyl alcohol), cetearyl alcohol, glyceryl stearate, phage-6 sorbitan stearate, phage-20 stearate, phage-40 stearate and phage- Two or more of them may be used in combination. More preferably, they may be used alone or in combination of two or more selected from the group consisting of cetearyl glucoside-cetearyl alcohol and cetearyl alcohol. The emulsifier may be used in an amount of 30 to 60 parts by weight, preferably 40 to 55 parts by weight, based on 100 parts by weight of the skin conditioner. If the amount of the emulsifier is less than 30 parts by weight, And it is uneconomical to use more than 60 parts by weight.

Next, the skin-softening agent of the oily component serves to soften rough skin. The amount of the skin-softening agent to be used is 2 to 10 parts by weight, preferably 3 to 7.5 parts by weight based on 100 parts by weight of the skin conditioner If the amount of the skin-softening agent is less than 2 parts by weight, the effect may be insignificant. If the content of the skin-softening agent is more than 10 parts by weight, the amount of the skin-softening agent may be excessive, Therefore, it is recommended to use within the above range. The skin-softening agent may be used alone or in admixture of one or more kinds selected from an oily agent containing at least one intermediate chain monoglyceride, glyceryl caprate, polyglyceryl-2-laurate and polyglyceryl-10- And may be used in admixture. Preferably, one or more of an oil-in-water agent containing at least one intermediate chain monoglyceride and polyglyceryl-10-laurate may be used alone or in combination. Specific examples of the oily agent containing at least one intermediate chain monoglyceride include Alcoline MCM (AKOLINE MCM) and the like.

Next, the nonionic surfactant among the emulsion components serves as a solubilizing agent for the compositions together with the emulsifier. The nonionic surfactant used in the art can be used. Specific examples thereof include Tween 20 Tween 40 and Tween 60 may be used singly or in combination of two or more. The nonionic surfactant may be used in an amount of 7 to 20 parts by weight, preferably 10 to 15 parts by weight, based on 100 parts by weight of the skin conditioner.

Next, the film-forming agent in the oily component serves to protect the skin from external unnecessary pollutants after the preparation is applied to the skin. The film-forming agent can be a conventional one used in the art. Plus 400 (sepiplus 400). The amount of the film-forming agent to be used may be 1.5 to 8 parts by weight, preferably 2.5 to 7 parts by weight, of the film-forming agent per 100 parts by weight of the skin conditioner. If the amount is less than 1.5 parts by weight, The effect of using it can not be seen. If it is used in an amount exceeding 8 parts by weight, solid contents in the emulsion liquid component may be generated.

Further, the emulsion may further contain a thickening stabilizer. The amount used may be 1 to 10 parts by weight, preferably 2 to 8 parts by weight, based on 100 parts by weight of the skin conditioner. If the amount is more than 10 parts by weight, It is recommended to use it within the above range. As the above-mentioned thickening stabilizer, one or two kinds selected from sodium polyacrylate, polyacrylate, polyisobutene, carbomer, polysorbate 20, polysorbate 50, polysorbate 60 and polysorbate 80 Polyisobutene and polysorbate 80 may be used singly or in combination of two or more.

Hereinafter, a method for producing the cosmetic of the present invention will be described.

The cosmetic preparation of the present invention is prepared by preparing each of an emulsion and an aqueous liquid and then emulsifying them under specific conditions of temperature. Adding the aqueous phase to the emulsion and stirring to prepare a mixture; Mixing the mixture, the antioxidant mixed with the antioxidant composition, the perfume and the purified water at 40 to 50 ° C, and stirring the mixture at a stirring speed of 2,000 to 3,000 rpm.

The emulsion of step 1 is prepared by mixing a skin conditioner, tocopheryl acetate, an emulsifier, a skin softening agent, a water-evaporation blocking agent and a nonionic surfactant at a temperature of 60 to 75 ° C, preferably 65 to 72 ° C To prepare a solution, and then dispersing and stirring the film-forming agent in the solution.

The aqueous phase solution of Step 1 may be prepared by mixing a skin conditioner, a sequestering agent, a thickening agent, a moisturizer, a viscosity reducing agent, a sterilizing preservative, and a solvent, followed by heating and stirring at 50 to 60 ° C.

The components used in the emulsions and aqueous solutions and their amounts used are the same as described above.

In the second step, the aqueous phase may be added to the emulsion, followed by emulsification at 70 to 80 ° C at a stirring speed of 2,000 to 3,000 rpm for 1 to 4 minutes to prepare a mixture.

In addition, the three-stage lyophilized extract and the species-specific rhubarb extract may be prepared by the method described above.

The cosmetic of the present invention thus prepared may have a pH of 5.4 to 6.2, preferably a pH of 5.8 to 6.0, and more preferably a pH of 5.85 to 5.95.

Various types of cosmetics can be manufactured using the above-described cosmetics of the present invention. For example, cosmetics such as cream type and serum type can be manufactured.

Hereinafter, the present invention will be described in more detail based on examples. However, the scope of the present invention should not be construed as being limited by the following examples.

[Example]

Example 1: Preparation of a shelf-life extract

5 parts by weight of shelf-life was mixed with 100 parts by weight of purified water to prepare a raw material.

Next, the raw material was subjected to hot water extraction at 90 to 100 DEG C for 4 to 8 hours to obtain an extract.

Next, the above extract was filtered with a filter membrane having an average pore size of 25 mu m to obtain a primary filtrate.

Then, 5 ml of 1,2-hexanediol and 0.1 ml of ethylhexyl glycerin were added to 94.9 ml of the filtrate, and the mixture was sufficiently stirred. Then, the filtrate was subjected to secondary filtration with a filter membrane having an average pore size of 10 탆 To obtain a starch extract.

Example 2: Preparation of bell pepper extract

And 5 parts by weight of sardine sulphate was mixed with 100 parts by weight of purified water to prepare a raw material.

Next, the raw material was subjected to hot water extraction at 90 to 100 DEG C for 4 to 8 hours to obtain an extract.

Next, the above extract was filtered with a filter membrane having an average pore size of 25 mu m to obtain a primary filtrate.

Then, 5 ml of 1,2-hexanediol and 0.1 ml of ethylhexyl glycerin were added to 94.9 ml of the filtrate, and the mixture was sufficiently stirred. Then, the filtrate was subjected to secondary filtration with a filter membrane having an average pore size of 10 탆 To thereby prepare a seed rhubarb extract.

Experimental Example 1-1: Determination of antioxidative and oxygen radical scavenging activity of the lyophilized extract

The antioxidant and oxygen radical scavenging activity of the lyophilized extract prepared in Example 1 was measured by a Korean dermatological research institute (test period from 2015.11.27 to 2016.1.14).

Antioxidant activity (ORAC-FL assay, Oxygen Radical Antioxidant Capacity) The fluorescence intensity was measured by reacting Fluorescein, a test substance with AAPH, a peroxyl radical (Excitation: 485 nm, Emmision: 525 nm). Specifically, the sample was subjected to dilution to a final concentration of 0.0007813-0.125% by volume, and Trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid) was used as a control.

25 μl of each test substance was dispensed into a black 96-well plate, and 150 μl of fluorescein was added thereto, followed by reaction at 37 ° C for 30 minutes. Next, 25 μl of AAPH (2-amidinopropane) dihydrochloride was dispensed into each well, and the fluorescence was measured at excitation 485 and emission 528 nm (60 minutes , Interval 3 minutes).

The resulting calculation was expressed as net AUC corresponding to the area between the blank and the sample graph, minus the blank fluorescence at the sum of the fluorescence intensity over time. Trolox equivalents (in mM TE / g) were calculated from Trolox equivalents (net AUC values) of corresponding concentrations using Trolox standard curves. In addition, the ORAC value (value) was converted to the Trolox equivalent value per 1 g of the sample.

The net AUC (area under the curve) was measured in a positive control group, Trolox, at a concentration range of 5 to 50 uM (see FIG. 1A), and a net AUC standard curve by concentration was prepared (see FIG.

FIG. 3A shows a net AUC measurement graph of the lyophilized extract, and the results of measurement of the oxygen radical scavenging activity of the lyophilized extract are shown in Table 1 and FIG. 4A.

Leaf extract
Treatment concentration (vol%) Trolox equivalent (mM TE / g) Primary Secondary Third Average Standard Deviation 0.0125 113.87 111.09 107.34 110.77 3.28 0.0250 99.69 96.41 97.14 97.75 1.72 0.0500 87.21 86.21 80.03 84.49 3.89 0.1000 66.06 66.15 64.48 65.57 0.94

As can be seen from Table 1 and FIG. 4-1, the L. sativa extract showed an ORAC value of 97.75 mM TE / g when treated at a concentration of 0.025% by volume and had excellent antioxidative activity (peroxyl radical scavenging activity) .

Experimental Example 1-2: Determination of antioxidative and oxygen radical scavenging activity of the seedlings

The antioxidant and oxygen radical scavenging activity of the seedlings obtained in Example 2 was measured by the Korean Dermatological Research Institute (test period 2016.2.15 to 2016.2.26).

Antioxidant activity (ORAC-FL assay, Oxygen Radical Antioxidant Capacity) The fluorescence intensity was measured by reacting Fluorescein, a test substance with AAPH, a peroxyl radical (Excitation: 485 nm, Emmision: 525 nm). Specifically, the sample was subjected to dilution with a final concentration of 0.0007813-0.125% by volume, and Trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid) was used as a control.

25 μl of each test substance was dispensed into a black 96-well plate, and 150 μl of fluorescein was added thereto, followed by reaction at 37 ° C for 30 minutes. Next, 25 μl of AAPH (2-amidinopropane) dihydrochloride was dispensed into each well and fluorescence was measured at excitation 485 nm and emission 528 nm (60 minutes, interval 3 minute).

The resulting calculation was expressed as net AUC corresponding to the area between the blank and the sample graph, minus the blank fluorescence at the sum of the fluorescence intensity over time. Trolox equivalents (mM TE / g) were calculated from Trolox equivalents (net AUC values) using the Trolox standard curve. In addition, the ORAC value (value) was converted to the Trolox equivalent value per 1 g of the sample.

The net AUC (area under the curve) was measured in a positive control group, Trolox, at a concentration range of 2.5 to 50 uM (see FIG. 1b), and a net AUC standard curve for each concentration was prepared (see FIG.

FIG. 3B is a graph showing the net AUC measurement of the sardine extract, and the results of measurement of the oxygen radical scavenging activity of the sardine extract are shown in Tables 2 and 4B.

Sardine extract
Treatment concentration (vol%) Trolox equivalent (mM TE / g) Primary Secondary Third Average Standard Deviation 0.00078 567.08 877.76 688.24 711.03 156.59 0.00156 600.76 599.16 452.49 550.80 85.15 0.00313 678.48 643.87 621.37 647.90 28.77 0.00625 636.92 621.71 608.41 622.35 14.27 0.01250 518.80 491.23 471.35 493.79 23.83

As shown in the results of Table 2 and FIG. 4B, the sardine extract showed an ORAC value of 622.35 mM TE / g when treated at a concentration of 0.00625% by volume, showing excellent antioxidative activity (peroxyl radical scavenging activity) there was.

Experimental Example 2: Determination of luciferase activity of the lyophilized extract

(1) HaCaT reporter cell (HaCaT-ARE-GFP-luciferase cell) was prepared by inserting luciferase as an antioxidant response element into human skin cell line HaCaT cell, The activity of azeotropes was measured. This cell line is a measurement that can detect the activity of Nrf2 quickly because GFP or luciferase is expressed when ARE is activated by Nrf2.

The luciferase activity was determined by incubating HaCaT-ARE-GFP-luciferase cells with RPMI 1640 media 0 ug / ml (control, control), the lyophilized extract of Example 1 (20 ug / ml, 50 (10 μg / ml, 100 μg / ml, 200 μg / ml) and brotholylazoid (sulforaphane, Positive Control, positive group) were incubated for 24 hours, Protein quantification of lysate and luciferase activity were measured and data were processed.

As a result of the measurement, it was confirmed that a natural product having an activity inducing effect of 70% or more in comparison with the same amount of the L. monocytogenes extract (see Fig. 5A).

(2) In the same manner, the activity of luciferase activity was measured using the endosulfur extracts of Example 2 (20 ug / ml, 50 ug / ml, 100 ug / ml and 200 ug / ml) A natural product having an activity inducing effect of 70% or more upon comparing the amounts of the extracts was confirmed (see FIG. 5B).

Experimental Example 3: Measurement of cytotoxicity of the extract of L. sativa and the species of R.

To determine the cytotoxicity of the extracts of Example 1 and Example 2 when the concentrations of the extracts were increased, cytotoxicity was measured using an MTT assays, and HaCaT cells The absorbance (570 nm) of the extract was measured after 24 hours and 48 hours of incubation for 4 hours in tetrazolium. The results of the lyophilized extract were shown in FIG. 6A, The results of the rhubarb extract are shown in FIG. 6B. 6A and 6B, it was confirmed that both the L. monocotyledonous extract and the R. sardinum extract had no cytotoxicity.

Experimental Example 4: Western blot measurement

HO-1 and NQO1, which are known to be electron-modulated by Nrf2, were subjected to Western blotting using the polyclonal antibodies of the lyophilized extract of Example 1 and the lysozyme extract of Example 2, respectively, , And the results are shown in Fig. 7a (Sambrook extract) and Fig. 7b (Safflower extract). At this time, the concentration of each of the L. monocytogenes extract and the species lysine extract was 200 ug / ml.

In this study, we investigated the effect of the NF-E2-related factor-2 (NF-2) transcription factor promoter) that has an ARE sequence. Thus, Nrf2 increases the expression of Phase II protein, which removes reactive oxygen through ARE binding, at transcriptional level and protects it from active oxygen. As shown in FIG. 7-2, Western blotting results show that transcription factor Are derived.

Experimental Example 5: Real-time RT Real time RT PCR measurement

It is known that enzymes involved in the Phase II metabolic system react with this reactive oxygen to increase its level when active oxygen is generated. At this time, the increase of the protein involved in the modulation of the Phase II metabolism is regulated in the transcription stage, and transcription factors regulating it are Nrf2, HO-1 and NQO-1.

Real-time RT-PCR was used to confirm the increase in mRNA level. RNA was extracted from HaCaT cells (20 ug / ml) or bovine rhubarb extract (20 ug / ml) by incubation (time lapse incubation), and the extracted RNA was synthesized again with cDNA. MRNA levels were measured with RT PCT.

The results for the roots of the sulfur extracts are shown in FIG. 8a (Nrf2), FIG. 8b (HO-1) and FIG. 8c (NQ01), respectively. 8A to 8C, it was confirmed that the phase II mRNA levels of Nrf2, HO-1 and NQO-1 were increased with time.

8D (Nrf2), FIG. 8E (HO-1) and FIG. 8F (NQ01), respectively. 8d to 8f, it was confirmed that the phase II mRNA levels of Nrf2, HO-1 and NQO-1 were increased with time.

Experimental Example 6: Oxidative Stress Deficiency Test Measurement

HaCaT cells (control, control), HaCaT cells treated with TPA (Phorbol 12-myristate 13-acetate) only and HaCaT cells treated with TPA and the lyophilized extract of Example 1 (SBH, 20 ug / ml) (JDH, 20 ug / ml) of Haemophilus influenzae (Haemophilus influenzae).

9A and 9B show the measurement results of the lyophilized extract, and Figs. 9C and 9D show the results of the measurement of the lybafil extract.

9A and 9C show DNA damage to HaCaT cells in order to examine whether DNA damage is caused by oxidative stress. The levels of 8-OH-dG (8-hydroxy-deoxyguanosine) in cells were measured by DNA staining. And FIG. 9D shows DNA 4-hydroxynonenal levels measured in HaCaT cells by DNA staining to determine whether lipid peroxidation occurs.

9A and 9B, it can be confirmed that the treatment with TPA alone and the treatment with TPA together with the starch extract (SBH) have an oxidative stress defense.

9 (c) and 9 (d), it can be confirmed that the treatment with TPA alone and the treatment with TPA and oxalic acid extract (JDH) have an oxidative stress defense.

Experimental Example 7: Antioxidant measurement of sardine seed extract

HaCaT cells were treated with TPOR (Phorbol 12-myristate 13-acetate) to induce reactive oxygen species (ROS) and then treated with the seeds of Raphanus sativus extract (JDH, 20 ug / ml) To confirm whether or not there is any. At this time, the active oxygen level was measured by DCFDA (Cellular Reactive Oxygen Species Detection Assay). DCFDA itself can pass through the cell membrane. When DCFDA meets the active oxygen species, DA is cut off, (trafficking). Therefore, it can be confirmed that the sardine extract is inhibiting active oxygen. As shown in FIG. 10, when TPA alone was used, it was confirmed that the fluorescent substance was not sharply inhibited by the active oxygen. On the contrary, in the case of TPA + JDH, the fluorescent substance did not emit light, and thus it was confirmed that the extract of the rhizome of the species inhibits the active oxygen.

Experimental Example 8: Determination of DPPH radical scavenging activity of the extract of L. sativa and the species R.

(2,2-Diphenyl-1-picrylhydrazyl) radical scavenging activity of the crude extract prepared in Example 1 and the crude scutellariae extract prepared in Example 2, The scavenging activity was measured (test period 2016.1.25 ~ 2016.2.5).

Samples were diluted to a suitable concentration range of about 50% free radical scavenging activity through preliminary concentration experiments and ascorbic acid was used as a control.

250 μl of 0.1 mM DPPH and 200 μl of EtOH were added to each test substance, followed by reaction at 4 ° C. for 30 minutes. Then, the absorbance at 520 nm was measured to determine the concentration of DPPH radicals. The DPPH radical scavenging activity of Respectively.

[Equation 1]

DPPH scavenging activity (%) = 100 (%) - (absorbance of the test substance added group / absorbance without addition) 100 (%)

IC ₅₀ : concentration of sample required to clear 50% of DPPH radicals

(1) Measurement of DPPH radical scavenging activity of positive control group ascorbic acid

The results of the DPPH radical scavenging activity test using ascorbic acid known to have antioxidant activity as a positive control are shown in Table 3. As a result, the concentration-dependent DPPH radical scavenging activity (IC ₅₀ : 3.72 ug / ml) was observed in the test concentration range, and it was confirmed that there was no problem in the present test procedure.

Free radical scavenging (% of positive control) Concentration (ng / ml) Primary average Secondary mean Tertiary average Average Standard Deviation 0 0 0 0 0 0 0.078125 3.78 0.67 1.41 1.95 1.62 0.15625 4.09 -1.51 4.08 2.22 3.23 0.3125 8.35 3.02 6.12 5.83 2.68 0.625 14.80 8.72 13.19 12.24 3.15 1.25 28.66 24.16 27.63 26.82 2.36 2.5 42.99 39.43 45.53 42.65 3.06 5 56.69 52.18 59.18 56.02 3.55

(2) Measurement of DPPH radical scavenging activity of the lyophilized extract

The antioxidant activity of the fleet flower extract, it was confirmed through the DPPH radical scavenging activity test (Table 4) As a result, the fleet flower extract is 0.00156% by volume to 0.05 in vol.% Concentration dependent and (see Fig. 11a), IC ₅₀ 0.027% And showed excellent DPPH radical scavenging activity.

Free radical scavenging (% of positive control) Concentration (ng / ml) Primary average Secondary mean Tertiary average Average Standard Deviation 0 0 0 0 0 0 0.00156 9.38 6.53 8.27 8.06 1.44 0.00313 15.94 10.44 15.62 14.00 3.09 0.00625 28.28 23.49 28.79 26.85 2.92 0.0125 41.25 36.87 42.11 40.08 2.81 0.025 52.66 48.61 54.52 51.93 3.02 0.05 56.41 54.81 58.50 56.57 1.85 0.1 55.47 53.18 54.98 54.54 1.20

(3) Measurement of DPPH radical scavenging activity of sardine extract

The results are shown in Table 5. The antioxidant activity of the extract was found to be dependent on the concentrations in the range of 0.00313 to 0.25% by volume (see FIG. 11b) and the IC _{50 of} 0.174% DPPH radical scavenging activity, indicating that it has excellent antioxidative activity.

Free radical scavenging (% of positive control) Concentration (ng / ml) Primary average Secondary mean Tertiary average Average Standard Deviation 0 0 0 0 0 0 0.00156 2.83 -0.49 1.69 1.34 1.69 0.00313 3.62 -1.97 3.38 1.68 3.16 0.00625 6.60 0.82 6.62 4.68 3.34 0.0125 13.84 5.91 11.38 10.38 4.06 0.025 20.75 17.08 20.62 19.48 2.08 0.05 32.70 28.24 33.54 31.50 2.85 0.1 44.81 38.26 45.54 42.87 4.01 0.125 48.00 50.23 49.25 49.16 1.12 0.25 54.46 56.43 57.01 55.97 1.34

(4) Determination of DPPH radical scavenging activity of the extract of L. sativa and the species R.

The IC ₅₀ and IC _{50 values} of IC ₅₀ and IC _{50 were} 0.174% and 0.174%, respectively, based on the DPPH radical scavenging activity of each of the above extracts.

Thus, the crude extracts of 0.027% by volume and the species of radishes of 0.174% by volume were mixed at a weight ratio of 1: 0.1 to 2.5, and their DPPH radical scavenging activity was measured. The results are shown in Table 6 below.

Luteinization extract: Weight ratio of bell pepper extract Free radical scavenging (% of positive control) 1: 0.2 50.53 1: 0.35 51.83 1: 0.5 58.04 1: 0.8 62.84 1: 1 66.53 1: 1.5 67.25 1: 2.0 68.02 1: 2.5 66.35

The DPPH radical scavenging activity was significantly increased when the lyophilized extract and the species rhubarb extract were used at a weight ratio of 1: 0.5 or more, and even when the species was used in excess of 2 weight%, DPPH radical scavenging activity We can confirm that there is no further increase. Through this, it was confirmed that the use of a mixture of the L. monocotyledonous extract and the bovine rhododendron extract at a weight ratio of 1: 0.5 ~ 2.0 is advantageous in terms of DPPH scavenging activity.

Example 3: Preparation of mixed natural extracts

(1) production of McMundong seed extract

The dried powder of McMundong seed was added with a 70% by volume aqueous ethanol solution, and the mixture was cooled at 26 to 27 ° C for 48 hours, filtered and concentrated under reduced pressure to obtain crude extract. Next, the crude extract was dissolved in an aqueous methanol solution of 80 vol% concentration, placed in a separating funnel, equilibrated with hexane, and left to vigorously shake. Next, the hexane layer was separated into a hexane layer on the top and the methanol layer on the bottom in an amount of 80% by volume within a few minutes, and further left for 1 hour or more for complete separation. Separately, the separated hexane layer is taken separately, and hexane is added to the methanol layer of 80 vol% concentration, and the distribution is repeated. The separated hexane layer is also taken for the second time, and hexane is added to the methanol layer of 80 vol.% Concentration and distributed. The hexane layers thus separated three times were combined and concentrated under reduced pressure to carry out hexane fractionation. Next, the methanol layer of 80 vol% concentration was concentrated to a methanol solution of about 30 vol% concentration, and this time, it was distributed with chloroform. Chloroform had a large specific gravity and formed a lower layer. The methanol layer having a concentration of 30% by volume became the upper portion. The chloroform layer was separated, and the chloroform layers were separated by repeating the same two times. The separated chloroform layers were combined and concentrated under reduced pressure to give a chloroform fraction. Next, the methanol layer having a concentration of 30% by volume is concentrated to completely remove the methanol, and then is distributed three times with the same amount of butanol and water. The isolated butanol layers were combined and concentrated under reduced pressure to give a butanol fraction, which was then extracted.

(2) Production of mugi extract

Momopsis is dried naturally in the shade, ground and pulverized. Subsequently, the mixture was subjected to reflux cooling extraction and extraction three times consecutively for 12 hours at an extraction temperature of 50 캜 with a mixed solvent having water and ethanol in a volume ratio of 1: 0.3, which was 4.2 times as much as the weight of dried powdery moth, , The mixture was filtered under reduced pressure, and the filtrate was concentrated under reduced pressure at 45 ° C in a vacuum rotary condenser to obtain a moxiflox extract soluble in a mixed solvent.

Next, after suspending the above-mentioned mythic mushroom extract in water, n-hexane, water and ethanol are mixed at a ratio of about 10: 9: 1 to the mushroom extract to obtain a n-hexane soluble fraction and a water soluble fraction, The water-soluble fraction was again extracted with dichloromethane to obtain a water-soluble fraction and a dichloromethane-soluble fraction.

Among them, the water soluble fraction was used as a mugwort extract.

(3) Production of lotus extract

13 parts by weight of dried lotus flower powder was mixed with 100 parts by weight of purified water to prepare a raw material.

Next, the raw material was subjected to hot water extraction at 85 캜 for 4 hours to obtain an extract.

Next, the above extract was filtered with a filter membrane having an average pore size of 25 mu m to obtain a primary filtrate.

Then, 5 ml of 1,2-hexanediol and 0.1 ml of ethylhexyl glycerin were added to 94.9 ml of the filtrate, and the mixture was sufficiently stirred. Then, the filtrate was subjected to secondary filtration with a filter membrane having an average pore size of 10 μm To produce a lotus extract.

(4) Production of chaga mushroom extract

To 100 parts by weight of purified water, 2.2 parts by weight of dried mushroom powder was mixed to prepare a raw material.

Next, the raw material was subjected to reflux cooling extraction at 35 DEG C for 6 hours to obtain an extract.

Next, the above extract was filtered with a filter membrane having an average pore size of 25 mu m to obtain a primary filtrate.

Then, 5 ml of 1,2-hexanediol and 0.1 ml of ethylhexyl glycerin were added to 94.9 ml of the filtrate, and the mixture was sufficiently stirred. Then, the filtrate was subjected to secondary filtration with a filter membrane having an average pore size of 10 μm To produce chaga mushroom extract.

(5) Preparation of mixed natural extract

Mixed natural extracts were prepared by mixing 1: 0.4: 3.2: 0.2 weight ratio of the extracts of mukmun-dong seed extract, mugwort extract, lotus extract and chaga mushroom.

Example 4-1: Preparation of antioxidants

The lyophilized extract of Example 1 and the species lyophilized extract of Example 2 were mixed at a weight ratio of 1: 1.3 and sufficiently stirred to prepare an Nrf2 activator.

Next, the Nrf2 activator and the mixed natural extract in Example 3 were mixed and stirred at a weight ratio of 1: 2.2 to prepare an antioxidant.

Examples 4-2 to 4-3 and Comparative Examples 4-1 to 4-4: Preparation of antioxidants

The antioxidant was prepared in the same manner as in Example 4-1, except that the antioxidant was prepared so as to have the composition shown in Table 7 below, and the antioxidant was prepared in the same manner as in Example 4-2 to Example 4-3 and Comparative Example 4-1 to Comparative Example 4-4.

division Nrf2 activator ⁽¹⁾ and mixed natural extract weight ratio Mixture of natural extracts of mokmun-dong, mugwipi, lotus and chaga mushroom extract Mr. McMundong Kempie Lotus Mushroom Example 4-1 1: 2.2 One 0.4 3.2 0.2 Example 4-2 1: 3.2 One 0.4 3.2 0.2 Example 4-3 1: 2.2 One 0.5 2.6 0.4 Comparative Example 4-1 1: 2.2 One - 3.2 0.2 Comparative Example 4-2 1: 2.2 One 0.4 One 0.2 Comparative Example 4-3 1: 2.2 One 0.4 3.2 0.05 Comparative Example 4-4 1: 4.2 One 0.4 3.2 0.2 (1): The Nrf2 activator is a mixture of Example 1 and Example 2 at a weight ratio of 1: 1.3.

Production Example 1: Preparation of cosmetic material

(1) Production of aqueous liquid

After the components shown in Table 8 were completely dispersed in a solvent, purified water was added, and the mixture was heated and stirred at 56 to 57 ° C to be completely dissolved to prepare an aqueous liquid.

Water solution division Kinds Weight portion Skin Conditioning Hyaluronic acid (1 volume% concentration) 39.2 Licorice powder 2 Hydrolite-5 58.8 Metal ion sequestrant EDTA-2NA 0.39 Thickener Kantrol F 0.98 Moisturizer glycerin 49.06 Viscosity reducer Dipropylene glycol 58.83 solvent 1,2-hexanediol 19.61 1,3-butylene glycol 19.61 Sterilization preservative Clofenine 5.88

 (2) Production of Emulsion

The ingredients of Table 9 below were mixed and mixed and dissolved while stirring with an agitator at 68 ° C. Then, a film-forming agent was added thereto and dispersed and mixed evenly to prepare an emulsion.

Emulsion division Kinds Amount used (parts by weight) Skin Evaporation Blocker MCT oil 87.5 Squalane 12.5 Tocopheryl acetate Vit-E Acetate 0.625 Skin Conditioning Sheer butter 12.50 Coconut oil 6.25 Emulsifier Cetearyl glucoside and cetearyl alcohol
(Emulgade PL 68/50) 18.75 Cetearyl alcohol (KALCOL 6850) 25 Skin softening agent One or more intermediate chain monoglycerides
Includes AKOLINE MCM 5 Nonionic surfactant Tween 60 12.5 Film forming agent SEPIPLUS 400 3.75

(3) Production of cosmetics

The aqueous phase was slowly added to the emulsion prepared above, and the mixture was stirred at a stirring speed of 2,500 rpm for 2 minutes at 74 to 75 ° C using a homomixer. Antioxidant And perfume (Perfume 85147) were added, and the mixture was slowly stirred to prepare a cosmetic. The cosmetic preparation was a serum type and had a pH of 5.98.

The resulting preparation contained 17.90% by weight of the aqueous solution, 20.70% by weight of the emulsion, 10.68% by weight of the antioxidant of Example 4, 0.28% by weight of fragrance and a balance of water.

Production Example 2 to Production Example 3 and Comparative Production Example 1 to Comparative Production Example 5: Production of cosmetic material

Cosmetic preparations were prepared in the same manner as in Preparation Example 1, except that the antioxidants shown in Table 10 were used instead of the antioxidants used in Example 4-1, respectively.

division Antioxidant Production Example 1 Example 4-1 Production Example 2 Example 4-2 Production Example 3 Example 4-3 Comparative Preparation Example 1 Comparative Example 4-1 Comparative Production Example 2 Comparative Example 4-2 Comparative Production Example 3 Comparative Example 4-3 Comparative Production Example 4 Comparative Example 4-4 Comparative Preparation Example 5 -

Experimental Example 9: Measurement of skin irritation property of cosmetic material

A human patch test was performed to evaluate the degree of irritation to the skin. The skin patch test was carried out on 20 healthy persons aged 20 years or older. The cosmetic preparation of Preparation Example 1 was attached to the inner part of the upper abdomen using a Finn chamber for 48 hours, After the passage of time, secondary readings were performed to determine skin reaction results. The evaluation criteria and the calculation of the irritation index are shown in Table 11, and the results are shown in Table 10.

division weight Evaluation standard - 0 No reaction ± 0.5 Suspicious Positive Response + One Weakly positive reaction ++ 2 Strong positive reaction +++ 3 Polar positive reaction [Equation 1]
Skin irritation index =? (Weight x reaction factor) / (maximum weight x total subjects) x 100

division Skin irritation index Degree of stimulation Production Example 1 0.2 No stimulation Production Example 2 0.3 No stimulation Production Example 3 0.3 No stimulation Comparative Preparation Example 1 0.9 No stimulation Comparative Production Example 2 0.1 No stimulation Comparative Production Example 3 0.2 No stimulation Comparative Production Example 4 0.7 No stimulation Comparative Preparation Example 5 1.4 Very slight irritation

As a result of the measurement of the skin irritation shown in Table 12, no skin irritation was found in all of the preparation examples and the comparative preparation examples. However, when the preparation example 1 was compared with the comparative preparation example 1 in which the mugwort extract was not used, it was confirmed that the skin trouble-preventing effect tended to be lowered. In addition, the skin irritation index was somewhat increased in Comparative Example 4 in which the mixed natural extract was used in an amount of 4% by weight, and in Comparative Example 2 in which the mixed natural extract was used in a weight ratio of 3.2.

In addition, when Production Example 1 and Production Example 3 were compared, Production Example 1 having a high content of lotus extract had a slightly lower skin irritation index than Production Example 3.

In Comparative Preparation Example 5 in which the mixed natural extract was not used, the skin irritation index tended to be greatly increased as compared with Production Examples 1 to 3.

Experimental Example 10: Skin whitening clinical test of cosmetics

Clinical trials were conducted to confirm the skin whitening effect of the cosmetic preparations prepared in Production Example.

Twenty women in their 20s and 40s were divided into 4 groups of 5 persons each for Comparative Example 1 and Examples 1 and 3 and used for 4 weeks after morning washing and evening washing twice a day.

Before and after use, the degree of brightness of the skin was measured using a chromameter (Minolta Co., Ltd., Japan), and the value of ΔL * (whiteness) 2). The measured? L * values are shown in Table 11.

&Quot; (2) "

ΔL * = (skin L * value before use of skin L * value after 4 weeks)

division DELTA L * Production Example 1 +8.6 Production Example 2 +8.9 Production Example 3 +7.5 Comparative Preparation Example 1 +5.9 Comparative Production Example 2 +4.1 Comparative Production Example 3 +4.3 Comparative Production Example 4 +8.7 Comparative Preparation Example 5 +3.2

In the skin whitening measurement results of Table 13, comparative preparation example 5 in which the mixed natural extract was not used exhibited a whitening effect by the Nrf2 activator. However, in the production examples 1 to 3 and the comparative preparation example When compared with 1 to 4, it was confirmed that the whitening effect was relatively low.

In Comparative Preparation Example 1 in which no mugwort extract was used, Comparative Preparation Example 2 in which less than 2 parts by weight of lotus extract was used, and Comparative Production Example 3 in which less than 0.1 part by weight of tea mushroom were used, , But the whitening effect was relatively lower than that of Preparation Examples 1 to 3, and this is because the effect of the Nrf2 activator to increase the antioxidation due to the use of the mixed natural extract was insufficient.

Further, in Comparative Preparation Example 4 in which the mixed natural extract was used in an amount of 4 parts by weight, there was no further whitening effect when compared with Preparation Example 2 in which 3.2 parts by weight of the mixed natural extract was used.

Through the above Examples and Experimental Examples, it was confirmed that the antioxidants including the Nrf2 activator and the mixed natural extract and the cosmetic preparation prepared therefrom exhibit excellent antioxidative and whitening effects. Through this, it is considered that the antioxidant of the present invention can be applied not only to cosmetics but also various products such as health functional foods, skin ointment products, hair products such as shampoo, and the like.

Claims (11)

An Nrf2 activator and a mixed natural extract,
Wherein the Nrf2 activator comprises a lyophilized extract and a seed rhubarb extract,
The antioxidant composition for cosmetics according to any one of claims 1 to 3, wherein the mixed natural extract comprises matsumundong seed extract, mugwort extract, lotus extract and chaga mushroom extract. [Claim 2] The method according to claim 1, wherein the Nrf2 activator comprises a lyophilized extract and a seed rhubarb extract at a weight ratio of 1: 0.5 to 2,
Wherein the mixed natural extract comprises 1: 0.2 ~ 0.6: 2 ~ 4: 0.1 ~ 0.4 weight ratio of makkun-dong extract, mugwort extract, lotus extract and chaga mushroom extract. The antioxidant composition for cosmetics according to claim 1, wherein the concentration of the lyophilized extract is 0.005 to 0.30% by volume and the concentration of the extract is 0.01 to 0.4% by volume. The antioxidant composition for cosmetics according to claim 1, wherein the antioxidant composition for cosmetics comprises the Nrf2 activator and the mixed natural extract at a weight ratio of 1: 1.5-4. An antioxidant comprising a composition of any one of claims 1 to 4; Aqueous solution; Emulsion; Spices; And purified water. 6. The method of claim 5,
Wherein the cosmetic comprises 5 to 18% by weight of an antioxidant, 15 to 20% by weight of an aqueous solution, 18 to 25% by weight of an emulsion, 0.05 to 1% by weight of a perfume, and purified water of a remaining amount. The aqueous liquid preparation according to claim 5, wherein the aqueous liquid comprises a skin conditioning agent, a sequestering agent, a thickener, a moisturizer, a viscosity reducing agent, a sterilizing preservative and a solvent,
Wherein the emulsion contains a moisture-blocking agent, tocopheryl acetate, a skin conditioner, an emulsifier, a skin-softening agent, a non-ionic surfactant and a film-forming agent. [Claim 10] The composition according to claim 7, wherein the aqueous liquid comprises 0.1 to 2 parts by weight of a sequestering agent, 0.5 to 2 parts by weight of a thickener, 40 to 80 parts by weight of a moisturizer, 45 to 90 parts by weight of a viscosity reducing agent 2 to 10 parts by weight of a sterilizing preservative, and 30 to 60 parts by weight of a solvent. 8. The method of claim 7,
The emulsion contains 0.1 to 5 parts by weight of tocopheryl acetate, 10 to 35 parts by weight of a skin conditioning agent, 30 to 60 parts by weight of an emulsifier, 2 to 10 parts by weight of a skin-softening agent, 7 to 20 parts by weight of an active agent and 1.5 to 8 parts by weight of a film former. The cosmetic according to claim 5, wherein the cosmetic has a pH of 5.85 to 6.10. A first step of preparing an aqueous liquid and an emulsion respectively;
Adding the aqueous phase to the emulsion and stirring to prepare a mixture;
Mixing the above mixture with an antioxidant, a perfume and a purified water comprising the composition of any one of claims 1 to 4 at 40 to 50 ° C and stirring at a stirring speed of 2,000 to 3,000 rpm; Lt; / RTI >
Wherein the Nrf2 activator comprises a lyophilized extract and a seed rhubarb extract,
Wherein the mixed natural extract comprises matsumundong seed extract, mugwort extract, lotus extract, and chaga mushroom extract.

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