KR20180077927A - Composition for whitening, antioxidizing or antibacterial comprising Paeonia lactiflora extract or fractions thereof - Google Patents

Abstract

The present invention relates to a composition for whitening, antioxidant or antimicrobial activity comprising an extract of Peony root or a fraction thereof as an active ingredient. The peony extract of the present invention or its fraction has an excellent antitussive effect against tyrosinase activity, excellent DPPH and ABTS free radical scavenging activity, and an antimicrobial activity against bacteria including E. coli and S. aureus . Based on the whitening, antioxidant, and antibacterial activity as described above, the composition of the present invention can be utilized as a cosmetic composition for whitening or antiaging, and furthermore can be used as a cosmetic composition for skin whitening, antioxidant or antibacterial activity, Additives and the like.

Description

TECHNICAL FIELD [0001] The present invention relates to a composition for whitening, antioxidant or antimicrobial use, which comprises an extract of Peony root or a fraction thereof as an active ingredient.

The present invention relates to a composition for whitening, antioxidant or antimicrobial activity comprising an extract of Peony root or a fraction thereof as an active ingredient.

As the aging of human skin progresses, the secretion of various hormones that regulate metabolism is reduced, and the function and activity of immune cells and skin cells are lowered. As a result, the biosynthesis of immune proteins and biocompatible proteins necessary for the living body is reduced, Intrinsic aging occurs, and as a result, the content of ultraviolet rays reaching the surface of the sunlight increases due to destruction of the ozone layer, and as the environmental pollution is further intensified, free radicals and active noxious oxygen are increased, Not only the thickness is reduced, the wrinkles are increased, the elasticity is reduced, the skin color is also dullly improved, skin troubles are frequent, and spots, freckles and black spots are also increased.

The field of cosmetics is a typical field where new technology can be converged. It is no exaggeration to say that the topic of cosmetics in the 21st century is functional cosmetics. Interest and consumption of functional cosmetics are increasing with time. According to the report of the Korea Health Industry Promotion Agency (KHIDI) in 2013, the cosmetics industry is seen as a promising industry with high growth potential. The trend is to pursue beauty, increase the economically active population of women, Despite the global economic crisis, the global cosmetics market is forecast to grow by 3.9% YoY to US $ 249.5bn in 2013 and continue to grow steadily to US $ 308.9bn in 2016. Among them, there is a high interest in functional cosmetics having various functions such as whitening, anti-aging, wrinkle improvement, and ultraviolet shielding.

In particular, recently, problems arise due to genetic modification of cosmetic ingredients and environmental hormones, so that functional cosmetics based on extracts of natural substances that can be safely used without being harmful to the human body in the long term are increasingly expected. Peony (藥药, Jakyak, Paeoniae radix) is one of the most important medicinal herbs as a root of the medicinal herb and its plants. Leaves are alternate phyllotaxis, petiole is long, 2 times exudate leaf, small leaf is elliptical, and there is no sawtooth. Flowers bloom one at the end of the stem and bloom in May-June. Herbal medicine is dried root, and the plant grows in the shade of a mountain tree. It is distributed all over Korea. In one branch, it is collected in March and August, and it is dried in the sun to produce a mixture of algae, blood 补血 補, blood 补血, Pain). In the oriental medicine, it has been used as convergence relaxation, jinbyeong, and analgesic (pain), and the stamens have been used as bacillus. Fruits are 1 ~ 3 corolla, and horny, red mature seeds and black mature seeds are exposed. The breeding is done in the form of a minus or seed. The medicinal peony is a medicinal plant belonging to the Ranuculaceae Paeonia, which is divided into herbaceous peony and tree-like peony. Paeonia lactiflora Pall, Paeonia obovata Max., And Paeonia japonica Miyabe et Takeda are classified as phytophagous cultivated in Korea.

The present inventors have sought to develop an effective substance having excellent antibacterial, antioxidant and whitening effect, which can be safely used without adverse effects on skin, and which has excellent product stability, as a natural material. As a result, it was confirmed that the composition containing the peony extract or fraction thereof as an active ingredient had a remarkable antioxidant or whitening effect, and thus the present invention was completed.

KR Patent No. 10-1671852

An object of the present invention is to provide a composition for antioxidant, antibacterial or whitening comprising an extract of Peony root or a fraction thereof as an active ingredient.

In order to achieve the above object, the present invention provides a cosmetic composition for skin whitening or anti-aging comprising peony extract or fractions thereof as an active ingredient.

The present invention also provides a health functional food composition for improving skin condition comprising an extract of peony root extract or a fraction thereof as an active ingredient.

In addition, the present invention provides an antimicrobial composition comprising a peony root extract or a fraction thereof as an active ingredient.

The present invention also provides a composition for external application for skin for antimicrobial use comprising an extract of Peony root or a fraction thereof as an active ingredient.

The present invention also provides an antimicrobial food additive comprising an extract of Peony root or a fraction thereof as an active ingredient.

The peony extract of the present invention or its fraction has an excellent antitussive effect against tyrosinase activity, excellent DPPH and ABTS free radical scavenging ability, and has an antimicrobial activity effect against bacteria including E. coli and S. aureus . Based on the whitening, antioxidant, and antibacterial activity as described above, the composition of the present invention can be utilized as a cosmetic composition for whitening or antiaging, and furthermore can be used as a cosmetic composition for skin whitening, antioxidant or antibacterial activity, Additives and the like.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a schematic diagram illustrating the extraction and fractionation process of the peony root extract and fractions thereof according to the present invention. FIG.
FIG. 2 is a graph showing the result of confirming the inhibitory effect of tyrosinase activity to confirm the whitening effect of arbutin, which is a control group.
FIG. 3 is a graph showing the result of confirming the inhibitory effect of tyrosinase activity in order to confirm the whitening effect of the fragrant chloroform fraction.
4 is a graph showing the results of confirming the inhibitory effect of tyrosinase activity in order to confirm the whitening effect of the ethyl acetate fraction of Peony root.
FIG. 5 is a graph showing the results of confirming the effect of inhibiting tyrosinase activity in order to confirm the whitening effect of the butanol fraction of Peony root.
FIG. 6 is a graph showing the result of confirming the inhibitory effect of tyrosinase activity to confirm the whitening effect of the peony root fraction. FIG.
FIG. 7 is a graph showing the results of confirming the cytotoxicity of the fractions of ethyl acetic acid peanut by MTT analysis. FIG.

The present invention provides an antioxidant, antibacterial or whitening composition comprising an extract of Peony root or a fraction thereof as an active ingredient.

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

The present invention provides a cosmetic composition for skin whitening or antiaging treatment comprising an extract of peony root extract or a fraction thereof as an active ingredient.

In the present invention, the peony root extract is a stem, leaf or root extract, preferably a stem extract, but is not limited thereto.

In the present invention, the peony root extract may be obtained by extracting, isolating and fractionating the peony root using the extraction, separation and fractionation methods known in the art. The "extract" as defined in the present invention is obtained by extraction treatment from a peony root extract using an appropriate solvent and includes, for example, a crude extract of Peony root, a polar solvent extract or a nonpolar solvent soluble extract, Soluble extract is preferable.

In the present invention, the peony root extract may be extracted according to various extraction solvents and extraction methods. Examples of suitable solvents for extracting peony root extract include water or an organic solvent such as water, methanol C ₁ -C ₄ alcohols such as methanol, ethanol, propanol, isopropanol, butanol and the like or a mixed solvent thereof may be used. Preferably, ethanol can be used as an extraction solvent for the extract of the present invention, more preferably 70 to 90% ethanol, and most preferably 80% ethanol can be used, but the present invention is not limited thereto. When preparing a peony root extract using 80% ethanol, an extract comprising a combination of active ingredients which can sufficiently dissolve all of the fat soluble and water soluble active ingredients in the peony root extract and exhibit optimal activity for antibacterial, antioxidative and whitening activity can do.

The extraction temperature is preferably room temperature, more preferably 20 to 100 ° C, but is not limited thereto. Examples of the extraction method include hot water extraction, cold extraction, reflux cooling, solvent extraction, steam distillation, ultrasonic extraction, elution, and compression.

After the extract is obtained using water or an organic solvent as described above, a liquid material can be obtained by freezing, heating and filtering at room temperature by a conventional method known in the art, or a liquid material can be obtained by further evaporating, spray drying or freeze- You may.

In the present invention, the peony extract can be obtained by further distillation and fractionation. Specifically, it can be obtained by silica gel column chromatography, thin layer chromatography, high performance liquid chromatography performance liquid chromatography, and the like. The fraction may be further fractionated using thin layer chromatography, but the present invention is not limited thereto, and the fraction may be fractionated using an appropriate solvent Any method of obtaining additional fractions can be used.

In the present invention, the solvent used for obtaining the fractions may be any one or more selected from the group consisting of chloroform, ethyl acetic acid, butanol, and water. Preferably, chloroform, ethyl acetic acid, and butanol may be used. Ethyl acetic acid may be used, but is not limited thereto.

In the present invention, the peony extract or the fraction thereof is contained in an amount of 0.001 to 60% by weight with respect to the whole cosmetic composition. When the peony extract has a weight percentage of less than 0.001, it may be difficult to achieve the desired effect. If it has an excess weight percentage, it may be cytotoxic and difficult to formulate the formulations in the manufacture of the product.

In the present invention, the cosmetic composition has anti-aging or whitening effect as a fundamental cause of damaging the skin, and the 'whitening' means whitening the skin.

Melanin is produced by a complex process from tyrosine by the action of tyrosinase present in the pigment cells. The resulting melanin is transferred to the skin cells, and melanin is lost with epidermal detachment, and the circulating action appears to disappear. That is, when the skin is exposed to ultraviolet rays, tyrosinase is activated. The tyrosinase acts on tyrosine present in the skin tissue to induce an oxidation process of generating dopa (DOPA) and dopaquinone. As a result, Melanin is synthesized in melanoma, and synthesized melanin is transferred to keratinocyte which is keratinocyte of skin and reaches skin surface by keratinization process to protect skin from ultraviolet rays. However, when the melanin is locally over-synthesized or the skin physiological functions are deteriorated due to skin lesions and aging, melanin is deposited on the skin surface and causes various pigment deposits such as spots and freckles. Is characterized by inhibiting the activity of tyrosinase and inhibiting the mechanism by which melanogenesis is suppressed from melanosomes in melanocytes, which are skin pigment cells, to have a whitening effect, and thus the present invention is not limited thereto.

In the present invention, the anti-aging properties of the cosmetic composition of the present invention can be applied to various diseases, diseases or abnormal conditions which can be prevented or treated by inhibiting or eliminating oxidation states as well as anti-aging effects obtained from antioxidative effects. Antioxidant related diseases that can be prevented or treated by the composition of the present invention include, but are not limited to, atherosclerosis, coronary artery disease, coronary artery restenosis, reperfusion injury, neurodegenerative diseases such as Parkinson's disease or Alzheimer's disease, stroke, But are not limited to, cardiovascular disease, osteoporosis, central nervous system disorders, peripheral vascular disease, and dyspnea.

The association of many disease states with free radical damage has been well established and many cellular constituents including enzymes, ion channels, structural proteins and membrane lipids are potential targets for reactive free radical species (Rice-Evans C , Mol Aspects of Med 13 (1): 1-111 (1992)). The reaction of these potential targets with free radicals impairs a certain range of cellular function, causing pathological changes and ultimately killing the cells. In addition, various diseases caused by physiological oxidation are described in U.S. Patent No. 5750351; 5773209; 5773231 and 5846959.

In the present invention, the cosmetic composition may further contain at least one known active ingredient having skin whitening or anti-aging effect together with the above dried apricot extract or its fraction.

In the present invention, the components contained in the cosmetic composition include components commonly used in cosmetic compositions in addition to the peony extracts or fractions thereof as an active ingredient, and include conventional components such as antioxidants, stabilizers, solubilizers, vitamins, Phosphorous additive, and carrier. In addition, the cosmetic composition may further include a skin absorption promoting substance to improve its effect.

In the present invention, the cosmetic composition may be prepared in any form conventionally produced in the art, for example, a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, a powder, a soap, But are not limited to, cleansing, oils, powder foundations, emulsion foundations, wax foundations and sprays. More specifically, it can be manufactured in the form of a soft lotion, a nutritional lotion, a nutritional cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray or a powder.

When the formulation of the cosmetic composition in the present invention is a paste, a cream or a gel, an animal oil, a vegetable oil, a wax, a paraffin, a starch, a tracer, a cellulose derivative, a polyethylene glycol, a silicone, a bentonite, Etc. may be used.

In the present invention, when the cosmetic formulation is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In particular, in the case of a spray, , Propane / butane or dimethyl ether.

In the present invention, when the cosmetic preparation is a solution or an emulsion, a solvent, a dissolving agent or an emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, , 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid esters of sorbitan.

In the case where the formulation of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.

In the present invention, when the cosmetic formulation is an interface-active agent-containing cleansing, the carrier component is selected from the group consisting of aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, Fatty acid amide ether sulfate, alkylamidobetaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester.

The present invention also provides a health functional food composition for improving skin condition comprising an extract of peony root extract or a fraction thereof as an active ingredient.

The improvement of the skin condition makes the condition of the skin health and beauty including the skin tone, wrinkles of the skin, elasticity of the skin and skin texture better, and specifically, it is for whitening or antioxidation for improving the skin condition But is not limited thereto.

In the present invention, the health functional food refers to a food group imparted with added value to function or express the function of the food by physical, biochemical or biotechnological techniques, or to control the bio-defense rhythm of the food composition, And restoration of the body to control the function of the body is designed to fully express the body, long-term consumption should be harmless to human body.

In the present invention, the health functional food may include food-acceptable food supplementary additives, and may further include appropriate carriers, excipients and diluents conventionally used in the production of health functional foods.

When the health functional food composition of the present invention is used as a food additive, the composition may be added as it is, or may be used together with other food or food ingredients, and may be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (prevention, health or therapeutic treatment). In general, the composition of the present invention is added in an amount of not more than 15% by weight, preferably not more than 10% by weight based on the raw material, in the production of food or beverage. However, in the case of long-term intake for the purpose of health and hygiene or for the purpose of controlling health, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range.

There is no particular limitation on the kind of the health functional food. Examples of health functional foods to which the above substances can be added include dairy products including ice cream, various soups, drinks, tea, drinks, alcoholic beverages and vitamin complexes. And foodstuffs designed to fully express in vivo the function of controlling the body in terms of regulation of bio-defense rhythm of food composition, prevention and recovery of disease, and the like.

In addition to the above, the health functional food composition of the present invention may contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, , Alcohols, carbonating agents used in carbonated drinks, and the like. In addition, the food composition of the present invention may comprise flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components may be used independently or in combination. Although the ratio of such additives is not critical, it is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

In addition, the present invention provides an antimicrobial composition comprising a peony root extract or a fraction thereof as an active ingredient.

In the present invention, 'antimicrobial' or 'antimicrobial activity' refers to a property of resisting microorganisms such as bacteria or fungi. More specifically, antimicrobial activity means a property of inhibiting growth or proliferation of bacteria.

In the present invention, the 'antimicrobial composition' is a composition having an activity of inhibiting the growth of microorganisms such as bacteria or fungi, and may include all forms used in various fields requiring an antimicrobial effect. Examples thereof include medicines, A skin external preparation, a food additive or a feed additive. Specifically, it can be used for medicines such as antibiotics and antifouling agents, in foodstuffs for preservation or antibacterial purposes, in cosmetics and household goods, for anti-acne products, and in products directly related to microorganisms. It is not.

The antimicrobial effect of the composition of the invention is the effect that appears in common in all kinds of bacteria, without the cover-positive gram-negative and gram, preferably a representative of Gram-negative bacteria Escherichia coli (E. coli) and Pseudomonas aeruginosa (P.aeruginosa), Gram-positive bacteria representative But not limited to, S. aureus and S. epidermidis .

As described above, when the present invention is used as a medicament, the composition of the present invention may further comprise pharmaceutically acceptable additives. Examples of the pharmaceutically acceptable additives include starch, gelatinized starch, microcrystalline cellulose, Lactose, colloidal silicon dioxide, calcium hydrogen phosphate, lactose, mannitol, sugar, gum arabic, pregelatinized starch, corn starch, powdered cellulose, hydroxypropylcellulose, opaques, starch glycolate sodium, carnauba wax, synthetic Aluminum silicate, stearic acid, magnesium stearate, aluminum stearate, calcium stearate, and white sugar may be used. The pharmaceutically acceptable additives according to the present invention are preferably included in the composition in an amount of 0.1 to 90 parts by weight, but not limited thereto.

In the present invention, the composition may be administered orally or parenterally in various clinical formulations. In the case of formulation, a diluent such as a filler, an extender, a binder, a wetting agent, a disintegrant, (Remington ' s Pharmaceutical Science, recently published by Mack Publishing Company, Easton PA) is preferably used. Examples of carriers, excipients and diluents that can be included in the composition include lactose, dextrose, sucrose, oligosaccharide, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil and the like.

The solid preparations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose, Lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Examples of the liquid preparation for oral administration include suspensions, solutions, emulsions and syrups. In addition to water and liquid paraffin which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like May be included.

The preparation for parenteral administration includes sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used as the non-aqueous solvent and suspension agent. As a base for suppositories, witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like can be used. The parenteral administration can be carried out using an external or intraperitoneal injection, intramuscular injection, subcutaneous injection, intravenous injection, intramuscular injection or intra-thoracic injection.

The composition of the present invention may be administered in a pharmaceutically effective amount.

The term "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment and the effective dosage level will vary depending on the species and severity, age, sex, Activity, sensitivity to the drug, time of administration, route of administration and rate of release, duration of treatment, factors including co-administered drugs, and other factors well known in the medical arts. However, for the desired effect, the composition of the present invention is preferably administered at 0.0001 to 100 mg / kg per day, preferably 0.001 to 100 mg / kg per day. The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. And can be administered singly or multiply. It is important to take into account all of the above factors and to administer the amount in which the maximum effect can be obtained in a minimal amount without adverse effect, and can be easily determined by those skilled in the art.

The term "administering" as used in the present invention means providing a certain composition of the present invention to a subject in any suitable manner.

In the present invention, the antimicrobial composition can be administered to a subject in various routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine dural or intracerebral injection.

The antimicrobial composition of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy, and biological response modifiers for antibacterial activity.

The above description of the composition of the present invention is omitted in order to avoid excessive complexity due to the overlapping description, and terms not otherwise defined in the present specification mean the terms commonly used in the technical field of the present invention .

Hereinafter, preferred embodiments and examples of the present invention will be described in order to facilitate understanding of the present invention. However, the following examples and preparation examples are provided only for the purpose of easier understanding of the present invention, and thus the content of the present invention is not limited thereto.

Example  1. Peony extract and its Fraction  Produce

1-1. Manufacture of peony extracts

The peony used in this experiment ( Paeonia lactiflora Pall. ) Was purchased from the Jirisan Herb Goal, which specializes in domestic wild and native herbs in Sancheong - gun, Kyongsang Province. To 5 kg of the shredded peony, 80% (v / v) ethanol was added and extracted three times at room temperature for three times to obtain an extract, which was then filtered using 2 to 3 μm filter paper. The filtered peach ethanol extract was concentrated using a vacuum concentrator.

1-2. Peony Fraction  Produce

The peony extracts prepared in Example 1-1 were further fractionated in the order of chloroform, ethyl acetate, butanol and water to obtain fractions. More specifically, sequential solvent fractions were obtained in the order shown in FIG.

Example  2. Peony extract and its Fraction  Confirm whitening effect

The effect of inhibiting tyrosinase activity was confirmed in order to confirm the whitening effect of the peony extract and peony root fraction prepared in Example 1. Specifically, the tyrosinase activity was determined by calculating the amount of DOPA chromium, an oxide oxidized by tyrosine, with L-tyrosine at 490 nm by absorbance measurement. To this end, a solution was prepared so as to have a final volume of 500 μl containing 50 mM sodium phosphate buffer (pH 7.2), 8 mM L-DOPA (0.36 mM), mushroom tyrosinase 30 U / ml, After that, the activity of tyrosinase was measured by measuring the absorbance with UV-VIS Spectrophotometer (Perkin-Elmer, USA) at room temperature and 475 nm wavelength. The results are shown in the graphs showing the correlation between concentrations of peony extract and peony fractions and tyrosinase activity, which are shown in FIGS. 2 to 6, respectively, and IC ₅₀ values are derived from these values. Arbutin was used as a control group.

As shown in Figs. 2 to 6 and Table 1, the IC ₅₀ values of arbutin (control), fructose chloroform fraction, fructose ethylacetic acid fraction, fructose butanol fraction and fructose fructose fraction were 2.77%, 0.077%, 0.0154%, 0.265% And 1.28%. In other words, the tyrosinase activity inhibition effect was found to be about 180 times higher on the arbutin than on the arbutin, about 36 times higher for the chrysanthemum fraction, about 10 times higher for the peanut butter fraction, , Indicating that the peony fractions showed excellent whitening activity inhibitory effect. In particular, it was confirmed that the ethyl acetic acid fraction had the most excellent tyrosinase activity inhibitory effect.

Example  3. Peony extract and its Fraction  Identify antioxidant and anti-aging effects

3-1. DPPH (1,1- 피덴 -2- picrylhydrazyl Free radical Scatters  Measure

DPPH free radical scavenging activity was measured in order to confirm the antioxidative effect of the extracts of the peony root extract prepared in Example 1 and the fractions thereof. First, peony extract or peony fractions and 0.1 mM DPPH solution were mixed and allowed to react in a dark room for 20 to 30 minutes. The absorbance was measured at 517 nm using an ELISA microplate reader (Bio-Rad, USA), and the IC ₅₀ value was calculated based on the absorbance. Arbutin and L-ascorbic acid were used as controls. The results are shown in Table 2.

As shown in Table 2, it was confirmed that the peony extract, peach chloroform fraction, peanut ethyl acetic acid fraction and peony butanol fraction had high DPPH radical scavenging activity of 0.0094%, 0.0069%, 0.0005% and 0.0062%, respectively. As compared to the control group L-ascorbic acid, the DPPH radical scavenging activity was about 2.5 times higher than that of the control group, the DPPH radical was higher than the DPPH radicals by about 3.4 times, the peanut ethyl acetic acid fraction was about 45.7 times and the peanut fraction was 3.8 times higher It was confirmed that it had an elimination activity. It was also confirmed that DPPH radical scavenging activity was higher than that of arbutin, which is another control group, by about 5.7 times in the peony extract, 7.9 times in the peony chloroform fraction, 105.7 times in the peony root ethyl acetate fraction and 8.8 times in the peony root butanol fraction. This indicates that the peony extract and peony fractions have excellent antioxidative activity, and the peanut ethyl acetic acid fraction has the highest antioxidant activity.

3- 2. ABTS (2,2-azino-bis (3- ethylbenzthiazoline -6- ulfonic  acid) diam - monium  salt) Free radical scavenging ability measurement

The free radical scavenging activity of ABTS was measured in order to confirm the antioxidative effect of the extracts and fractions of peony root extract prepared in Example 1. First, 7.4 mM ABTS and 2.6 mM calcium sulfate were mixed and left in a dark room at room temperature for 24 hours. The resulting ABTS free radical was diluted with ethanol to an absorbance of 0.7 at a wavelength of 734 nm. The diluted ABTS free radicals were treated with peony extract or peony fractions and the absorbance was measured at 734 nm using an ELISA microplate reader (Bio-Rad, USA), and IC ₅₀ values were derived therefrom. Arbutin and L-ascorbic acid were used as controls. The results are shown in Table 3.

As shown in Table 3, the ABTS free radical scavenging activity, which is concentration-dependent (concentration change of 0.25% to 0.0025%), of the peony extract and peony fractions was confirmed. The IC ₅₀ values of the peony extract, peanut fraction, ethyl acetate fraction and peony butanol fraction were 0.038%, 0.012%, 0.002% and 0.023%, respectively. Compared with the ascorbic acid used as the control group, , And the ABTS radical scavenging ability was about 5 times higher in the peach chloroform fraction, about 23.1 times in the crude ethyl acetate fraction and about 2.5 times higher in the peach butanol fraction, and the highest ABTS radical scavenging activity was observed in the ethyl acetate fraction . This indicates that the peony extract and peony fractions have a high antioxidative activity, and the peanut ethyl acetic acid fraction has the highest antioxidant activity.

Example  4. Peony extract and its Fraction  Confirmation of antibacterial effect

A well diffusion assay was performed to confirm the antimicrobial effect of the peony extract and the peony extract prepared in Example 1. First, the strains shown in Table 4 below were suspended in distilled water, plated on a solid medium, and cultured in an incubator at 37 ° C for 24 hours. For the well diffusion analysis, first, a single colony of the microorganisms cultured in the solid medium was inoculated into the liquid medium, and then cultured in an incubator stirred at 200 rpm at 37 캜 for 24 hours. Each of the above extracts and fractions was divided into 2 쨉 l / well and incubated at 37 째 C in an incubator (Sanyo Incubator MIR-253) for 24 hours And a clear growth zone was observed as a result. The results are shown in Table 5.

As shown in Table 5, the fructose ethylacetic acid fractions, peach butanol fractions, and peony extracts showed strong inhibitory activity against S. aureus and S. spidermidis , and in the peach chloroform fraction, , Respectively. In addition, it was confirmed that peach acetic acid fraction, peanut butanol fraction and peony extract had strong inhibitory activity against P. aeruginosa , and it showed moderate inhibitory activity in peanut fraction of chloroform. In E. coli , , The peanut ethyl acetic acid fraction and peony root extract showed the strongest inhibitory activity. Particularly, the highest antimicrobial activity was obtained in the ethyl acetate fraction of Peony root in all the above strains. The results show that the peony extract and peony fractions have a high antimicrobial effect, indicating that these substances are suitable for use as cosmetics or foods for use on the skin.

Example  5. Peony Fraction  Cytotoxicity check

(MTT [3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl-tetrazolium bromide] analysis was performed to confirm the cytotoxicity of the fractions of ethyl acetic acid prepared in Example 1-2. The cells used in the experiment were suspended in DMEM containing 10% fetal bovine serum and 1% penicillin streptomycin as mouse melanoma cells (B16F10) purchased from Korean Cell Line Bank, After centrifugation at 1000 rpm for 3 minutes at 25 ° C, it was suspended in the medium and incubated in a 5% carbon dioxide incubator for 24 hours before being used in the experiment. Next, mouse melanoma cells (B16F10) were cultured in 9 well plates at 4 cells / ml to confirm cytotoxicity in the cells. After 24 hours, the medium was removed and the fractions of ethyl acetate were treated with concentrations (0.1 μl / ml, 0.05 μl / ml, 0.025 μl / ml, 0.013 μl / ml, 0.006 μl / ml and 0.003 μl / ml) And cultured in a 5% carbon dioxide incubator at 37 캜 for 48 hours. After this, 5 mg / ml MTT reagent was added and incubated for an additional 3 hours in a 5% CO 2 incubator. Thereafter, it was centrifuged at 1000 rpm at 25 ° C for 3 minutes. The supernatant was removed so that the formazan formed on the bottom of the well was not dispersed, and 1: 1 DMSO: ethanol was added to the supernatant- Absorbance was measured at 540 nm wavelength using an ELISA microplate reader (Bio-Rad, USA). The results are shown in Fig.

As shown in Fig. 7, the fragrant ethyl acetic acid fractions showed high cell viability. This indicates that the peanut ethyl acetic acid fraction is a safe substance free from cytotoxicity, and at the same time, it is a biocompatible material suitable for use as a cosmetic and food product.

Taken together, the peony extracts or fractions thereof of the present invention are effective for inhibiting tyrosinase activity, exhibiting an antimicrobial activity effect in bacterium including DPPH, ABTS free radical scavenging ability, E. coli , S. aureus , Or an antibacterial effect. Accordingly, the present invention can be utilized as a cosmetic composition for preventing whitening or aging, and further useful as a health functional food related to whitening, antioxidant or antibacterial, composition for antibacterial, external preparation for skin, food additive and the like.

Examples of formulations for the composition of the present invention are illustrated below.

Formulation Example 1. Preparation of cosmetic preparation

1. Flexible longevity manufacturing

Peony extract or its fraction 1%

5.2% by weight of 1,3-butylene glycol, 1.5% by weight of oleyl alcohol,

3.2% by weight of ethanol

Polysorbate 20 3.2 wt%

Benzophenone-9 2.0 wt%

1.0% by weight of a carboxyl vinyl polymer, 3.5% by weight of glycerin,

A minor amount of fragrance, a small amount of preservative, and a remaining amount of purified water were mixed to prepare a softening water by a conventional method.

2. Milk lotion manufacturing

Peach extract or fractions thereof 0.1%

Glycerin 5.1 wt%

Propylene glycol 4.2 wt%

3.0% by weight of tocopheryl acetate

Liquid paraffin 4.6 wt%

1.0% by weight triethanolamine

Squalane 3.1 wt%

Macadamia nut oil 2.5 wt%

Polysorbate 60 1.6 wt%

1.6% by weight of sorbitan sesquiterate

Propyl paraben 0.6 wt%

Carboxyl vinyl polymer 1.5 wt%

A minor amount of fragrance, a small amount of preservative, and a remaining amount of purified water were mixed to prepare an milk lotion by a conventional method.

3. Nutritional cream manufacturing

0.5% < tb > < tb >

Glycerin 4.0 wt%

Vaseline 3.5 wt%

2.1% by weight triethanolamine

Liquid paraffin 5.3 wt%

Squalane 3.0 wt%

Wax 2.6 wt%

Tocopheryl acetate 5.4 wt%

Polysorbate 60 3.2 wt%

Carboxyl vinyl polymer 1.0 wt%

3.1 weight percent sorbitan sesquioleate

A minor amount of fragrance, a small amount of preservative, and a remaining amount of purified water were mixed to prepare a nutritional cream by a conventional method.

4. Massage cream manufacturing

0.5% < tb > < tb >

Glycerin 4.0 wt%

3.5% by weight of petrolatum, 0.5% by weight of triethanolamine,

Liquid paraffin 24.0 wt%

Squalane 3.0 wt%, wax 2.1 wt%

0.1% by weight of tocopheryl acetate

Polysorbate 60 2.4 wt%

Carboxyl vinyl polymer 1.0 wt%

2.3 weight% sorbitan sesquioleate

A minor amount of fragrance, a small amount of preservative and a remaining amount of purified water were mixed to prepare a massage cream by a conventional method.

Formulation Example 2 Preparation of food preparation

1. Manufacture of health food

Peony extract or its fractions 100 mg

Vitamin mixture quantity

70 g of vitamin A acetate

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

0.15 mg of vitamin B2

Vitamin B6 0.5 mg

0.2 g of vitamin B12

Vitamin C 10 mg

Biotin 10 g

Nicotinic acid amide 1.7 mg

Folate 50 g

Calcium pantothenate 0.5 mg

Mineral mixture quantity

1.75 mg of ferrous sulfate

0.82 mg of zinc oxide

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Secondary calcium phosphate 55 mg

Potassium citrate 90 mg

Calcium carbonate 100 mg

Magnesium chloride 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixture is comparatively mixed with a composition suitable for health food as a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional method for producing healthy foods , Granules can be prepared and used in the manufacture of health food compositions according to conventional methods.

2. Manufacture of health drinks

Peony extract or its fractions 100 mg

Vitamin C 15 g

Vitamin E (powder) 100 g

19.75 g of ferrous lactate

3.5 g of zinc oxide

Nicotinic acid amide 3.5 g

Vitamin A 0.2 g

Vitamin B1 0.25 g

Vitamin B2 0.3 g

Water quantification

The above components were mixed according to a conventional health drink manufacturing method, and the mixture was stirred and heated at 85 for about 1 hour. The resulting solution was filtered, sterilized in a sterilized 2 L container, sealed, ≪ / RTI >

Although the compositional ratio is relatively mixed with a component suitable for a favorite drink, it is also possible to arbitrarily modify the compounding ratio according to the regional or national preference such as the demand class, the demanding country, and the use purpose.

Formulation Example 3. Preparation of pharmaceutical preparations

1. Manufacturing of powder

Peony extract or its fractions 20 mg

Lactose 100 mg

Talc 10 mg

The above components can be mixed and filled in an airtight container to prepare powders.

2. Preparation of tablets

Peony extract or its fractions 10 mg

Corn starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above components, tablets may be prepared by tableting according to a conventional method for producing tablets.

3. Preparation of capsules

Peony extract or its fractions 10 mg

Crystalline cellulose 3 mg

Lactose 14.8 mg

Magnesium stearate 0.2 mg

The above components may be mixed and filled in a gelatin capsule according to a conventional method for preparing a capsule, thereby preparing a capsule.

4. Preparation of injections

Peony extract or its fractions 10 mg

180 mg mannitol

Sterile sterilized water for injection 2974 mg

Na ₂ HPO ₄ 2H ₂ O 26 mg

(2 ml) per 1 ampoule according to the usual injection preparation method.

5. Manufacture of liquids

Peony extract or its fractions 20 mg

10 g per isomer

5 g mannitol

Purified water quantity

Each component is added to and dissolved in purified water according to a conventional liquid preparation method, and the lemon flavor is added in an appropriate amount, then the above components are mixed, and then purified water is added. The total amount of the liquid agent added to the purified water is adjusted to 100 ml, and the liquid agent can be finally prepared by filling it in a brown bottle and sterilizing it.

Claims (10)

A cosmetic composition for skin whitening or antiaging treatment comprising peony extract or fraction thereof as an active ingredient.
The method according to claim 1,
Wherein the peony extract is a peony root extract.
The method according to claim 1,
The cosmetic composition according to claim ₁ , wherein the crude extract is extracted with water, a C ₁ -C ₄ alcohol or a mixed solvent thereof.
The method according to claim 1,
Wherein the fraction is a fraction obtained by fractionating with at least one solvent selected from the group consisting of chloroform, ethyl acetic acid, butanol and water.
The method according to claim 1,
The cosmetic composition according to claim 1, wherein the peony extract or the fraction thereof is contained in an amount of 0.001 to 60% by weight based on the whole cosmetic composition.
A health functional food composition for improving skin condition comprising peanut extract or fraction thereof as an active ingredient.
The method according to claim 6,
Wherein the skin condition improving agent is for whitening or antioxidant.
An antimicrobial composition comprising a peony root extract or a fraction thereof as an active ingredient.
A composition for external application for skin for antimicrobial use comprising a peony extract or a fraction thereof as an active ingredient.
An antimicrobial food additive containing a peony root extract or a fraction thereof as an active ingredient.

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