KR101425560B1 - Composition for antioxidant comprising extracts or its fractions of Elaeocarpus petiolatus as an active ingredient - Google Patents

Abstract

The present invention relates to an antioxidant composition comprising an Elaeocarpus petiolatus extract or a fraction thereof as an active ingredient, and more particularly, to a composition for preventing aging comprising Eliocaccus petiatus extract or a fraction thereof of the present invention in a concentration- DPPH free radical scavenging effect and ABTS free radical scavenging effect and exhibits a remarkable superoxide radical scavenging effect and excellent antioxidative effect, it can be effectively used as an active ingredient of a cosmetic composition for preventing aging, a pharmaceutical composition and a health food.

Description

[0001] The present invention relates to an antioxidant composition containing an Eliocaccus pentylatorus extract or a fraction thereof as an active ingredient,

The present invention relates to an antioxidant composition, and more particularly to a composition for preventing aging which comprises Elaeocarpus petiolatus extract or fractions thereof as an active ingredient, and to a pharmaceutical composition and a health food.

Free radicals that cause aging and disease in humans are known to form in biological reactions during normal metabolic processes in the body, causing harmful toxins to cells and tissues and causing various diseases (Folin O & Denis W, 1912, J Biol Chem 12: 239-249). Physiological actions to inhibit such harmful free radicals include electron donating ability to inhibit oxidation by donating electrons to oxidative free radicals and superoxide anion (SOD), which has a similar reactivity with superoxide dismutase (SOD) It has the role of converting radical (superoxide anion radical) into normal oxygen. Several physiologically active ingredients with strong antioxidant effects contribute to prevention of ulcers and seizures caused by oxidative stress, regulation of gastric secretion and prevention of diarrhea, prevention of diabetes, prevention and reduction of cancer, heart disease and degenerative diseases (Azuma K et < RTI ID = 0.0 > al ., < / RTI > 1999, J Agric Food Chem 47: 3963-3966, Ham SS et al ., 1997, J Food Sci Nutr 2: 155-161). Recently, as interest in health has increased, a resource containing a large amount of a functional substance having various effects such as anticancer, antiallergic, anti-obesity, antioxidant and antibacterial activity has been selected from various plant resources, (Shin DB, et < RTI ID = 0.0 > al ., 2006, Food Sci Biotechnol., 15: 357-362, Park et al al ., 2004, Korean J Nutr, 37: 364-372, Kim et al ., 2006, Kor J Food Sci. Technol. 38: 799-804, Ali KA et al ., 2005, Food Chem., 89: 27-36).

Superoxide dismutase, an antioxidant, breaks down the active oxygen generated by the respiration process of cells, ie, superoxide radicals into hydrogen peroxide and oxygen. Hydrogen peroxide reacts with metal ions such as iron or copper ions to generate stronger active oxygen called hydroxy radicals. Catalase and glutathione peroxidase, antioxidants that remove hydrogen peroxide before it reacts with metal ions, decompose hydrogen peroxide into water and carbon dioxide. The antioxidant that removes hydrogen peroxide can not completely remove 5% of hydrogen peroxide, which reacts with metal ions in the body to turn into hydroxy radicals, aging our bodies and causing them to die. Frequent exposure to radiation or ultraviolet light changes the hydrogen peroxide into hydroxy radicals to promote senescence. An antioxidant is a method of delaying this. Superoxide radicals (O ^{2 -} ) themselves are relatively weak in reactivity, but they are easily converted to H ₂ O ₂ , resulting in hydroxyl radicals that are highly reactive or react with nitric oxide (NO) (Peroxynitrite) (ONOO), which causes oxidation of SH-group, nitration of protein tyrosine, lipid peroxidation, and DNA damage.

In addition, when aging progresses, melanin, which is a pigment that blocks ultraviolet rays coming from sunlight, is deposited on the skin surface when the physiological function of the skin is lowered, causing spots, freckles and various pigment deposits (Hill, HX, et al, 1997, Pigment Cell Res. 10, 158).

Antioxidants are widely distributed in copper and plants. Many antioxidants such as phenolic compounds, flavonoids, tocopherols, vitamin C and selenium in fruits and vegetables can delay or prevent the oxidation of fats and cause cancer and cardiovascular diseases Prevention, and delaying the disease. Tocopherol and vitamin C, which are natural antioxidants in the vegetable field, are widely used in foods, medicines and cosmetics. The rancidity of foods containing oil or fat is mainly caused by binding with oxygen in the air. To prevent this, many synthetic or natural antioxidants have been developed. However, synthetic antioxidants are used in practice because of their effectiveness and economy. Examples of synthetic antioxidants include Butylated Hydroxy Toluene (BHT), Butylated Hydroxy Anisole (BHA), and Propyl Gallate. Synthetic antioxidants are widely used in commercial foods because of their excellent antioxidant ability and low cost, but their use is regulated due to concerns about safety, such as human side effects. Natural antioxidants such as tocopherol are safe for the human body, but have the disadvantage that they have the ability to inhibit oxidative chain reaction and are expensive. Therefore, in recent years, researches for the development of natural antioxidants which are more safe and have excellent antioxidative effects have been actively conducted.

On the other hand, Elaeocarpus petiolatus is distributed in tropical and subtropical evergreen trees and shrubs in Elaeocarpus, India, Malaysia, southern China, Japan, Australia, New Zealand, Fiji and Hawaii. . It has been reported that Eliocaccus persiphatus is effective in preventing skin dryness in Japan and improving wrinkles. Other species of Elieocarpus have been reported to promote hair growth in Japan Have been reported. In Europe and the United States, it has been reported that the tannin components extracted from the above-described plants inhibit the reverse transcriptase of human retroviruses and exhibit an antiviral effect. However, Elaeocarpus petiolatus of the present invention is not described, and its antioxidative effect is not yet known.

Thus, the inventors of the present invention found that the extract of Eliocaffus pentylatus or its fractions exhibited excellent DPPH (1,1-diphenyl-2-picryl-hydrazyl) free radical scavenging effect, ABTS , 2'-azinobis- (3-ethylbenzothiazoline-6-sulphonic acid)) free radical scavenging effect and superoxide radical scavenging effect, and thus it can be effectively used as an active ingredient of an anti- The present invention has been completed.

An object of the present invention is to provide an anti-aging cosmetic composition, a pharmaceutical composition and a health food containing Elaeocarpus petiolatus extract or a fraction thereof as an active ingredient.

In order to accomplish the above object, the present invention relates to a method for producing an Elaeocarpus petiolatus extract or a fraction thereof as an active ingredient.

In addition, the present invention provides a pharmaceutical composition for preventing aging comprising Eliocaffus pentylatorus extract or a fraction thereof as an active ingredient.

In addition, the present invention provides an anti-aging health food containing Eliocaccus pentioatus extract or a fraction thereof as an active ingredient.

The Elaeocarpus < RTI ID = 0.0 > petiolatus extract or a fraction thereof has an excellent antioxidative effect by showing DPPH free radical generation inhibitory effect, ABTS free radical generation inhibitory effect and superoxide radical inhibitory effect in a concentration dependent manner, and thus it is possible to provide a cosmetic composition, It can be usefully used as an active ingredient of health food.

Figure 1 is a graphical representation of Elaeocarpus < RTI ID = 0.0 > petiolatus methanol extract of the present invention showed DPPH free radical scavenging effect.
FIG. 2 is a graph showing the ABTS elimination effect of the Elyokapherpeticia methanol extract. FIG.
FIG. 3 is a graph showing the effect of SOD (superoxide dismutase) activity of the Eliocaccus petioatum methanol extract. FIG.
FIG. 4 is a graph showing a method for producing a solvent-separated fraction from an Eliocaccus petioatum methanol extract. FIG.
FIG. 5A is a graph showing the DPPH free radical scavenging effect of the Eliocaccus pentioatus fraction. FIG.
FIG. 5B is a graph showing the ABTS free radical scavenging effect of the Eliocaffus petioleus fraction. FIG.
FIG. 6 is a graph showing the effect of the Eliocaccus pentioatus fraction and the coexisting heterologous extract on the amount of melanin production. FIG.

Hereinafter, the present invention will be described in detail.

The present invention provides an anti-aging cosmetic composition containing Elaeocarpus petiolatus extract or a fraction thereof as an active ingredient.

The aging is preferably, but not exclusively, skin aging.

Preferably, the Eliocaccus pentylatorus extract or fraction thereof is produced by a manufacturing method comprising the following steps, but not limited thereto:

1) extracting Elaeocarpus petiolatus with an extraction solvent;

2) filtering the extract of step 1);

3) concentrating the filtered extract of step 2) under reduced pressure and then drying to prepare an extract of Eliocaffus pentiaratus; And

4) Extracting the Eliocaffus pentryatus extract of step 3) with an organic solvent to prepare an Eliocaccus pentiolatus fraction.

In this method, Elaeocarpus petiolatus in step 1) can be used without limitation, such as cultivated or commercially available. It is most preferable that the Eliocaccus petioleus is a stem, but not limited to, leaves, stems or roots.

In the above method, it is preferable to use water, an alcohol or a mixture thereof in the extraction solvent of the step 1). As the alcohol, C ₁ to C ₂ lower alcohol is preferably used, and as the lower alcohol, ethanol or methanol is preferably used. As the extraction method, it is preferable to use shaking extraction, Soxhlet extraction or reflux extraction, but it is not limited thereto. The extraction solvent is preferably added by 1 to 10 times, more preferably 2 to 3 times, to the dried amount of Elioca cupperiotuaratus. The extraction temperature is preferably 20 占 폚 to 100 占 폚, more preferably 20 占 폚 to 40 占 폚, and most preferably room temperature, but is not limited thereto. The extraction time is preferably 10 to 48 hours, more preferably 15 to 30 hours, most preferably 24 hours, but is not limited thereto. In addition, the extraction number is preferably 1 to 5 times, more preferably 3 to 4 times, and most preferably, 3 times, but not limited thereto.

In the above method, it is preferable to use a vacuum decompression concentrator or a vacuum rotary evaporator for the decompression concentration in step 3), but it is not limited thereto. The drying is preferably performed under reduced pressure, vacuum drying, boiling, spray drying or freeze drying, but not always limited thereto.

In the above method, the organic solvent in step 4) is preferably, but not limited to, n-hexane, chloroform, ethyl acetate or butanol. The fractions were prepared by suspending the Eliocaccus extract in water and then fractionating the fractions sequentially with normal-hexane, chloroform, ethyl acetate, n-butanol and water to obtain fraction of normal-hexane, fraction of chloroform, fraction of ethyl acetate, fraction of n-butanol Or water fraction, and it is more preferable that it is an ethyl acetate fraction, but it is not limited thereto. The fraction may be obtained by repeating the fractionation process 1 to 5 times, preferably 3 times, from the Eliocaccus petiota rose extract, and the fraction is preferably concentrated under reduced pressure, but not limited thereto.

In a specific example of the present invention, in order to confirm the antioxidative effect of the Eliocaffus pentioatus extract or its fraction, the DPPH free radical scavenging effect was evaluated by measuring 1,1-diphenyl-2-pyryl-hydrazyl (1,1 -diphenyl-2-picryl-hydrazyl, DPPH, Sigma). As a result, it was confirmed that the Elyokapherpeticia methanol extract or its fraction exhibited the DPPH free radical scavenging effect in a concentration-dependent manner 5a).

Further, in another specific embodiment of the present invention, in order to confirm the antioxidative effect of the Eliocaffus pentylatus extract or its fraction, the ABTS free radical scavenging activity according to various concentrations of the Eliocaffus pentiolatum methanol extract or its fractions As a result of the measurement of the effect, it was confirmed that the Elyokapherpeticia methanol extract or the fraction thereof showed an ABTS free radical scavenging effect in a concentration-dependent manner (see FIGS. 2 and 5B).

Further, in another specific example of the present invention, in order to confirm the SOD (superoxide dismutase) activity of the Eliocaccus pentioatus extract, the SOD kit was used to measure the superoxide radical scavenging activity of the Elioca- As a result of the measurement of the effect, it was confirmed that the Eliocaccus pentylatorus extract showed an effect of eliminating superoxide radicals in a concentration-dependent manner (see FIG. 3).

In addition, the free radical scavenging of the Elyokapus pentiolatus ethyl acetate fraction and the Elyokapus pentylatus heterologous species showed that the Elyokapus petiota et Ethylacetate fraction was superior to the extract of the same heterologous species (See Table 1 and Table 2), and it was confirmed that the Eliocaccus persethurus hexane fraction had an excellent inhibitory effect on melanin formation as compared with the Eliocaccus petiotaus heterologous extract 6).

Therefore, the Eliocaccus pentylatorus extract of the present invention or its fraction exhibits a DPPH free radical scavenging effect and an ABTS free radical scavenging effect in a concentration-dependent manner, exhibits a remarkable superoxide radical scavenging effect and exhibits an excellent antioxidative effect and a melanin formation inhibitory effect It can be used effectively as an active ingredient of a cosmetic composition for preventing aging.

The cosmetic composition may be in the form of a solution, a gel, a solid or a paste anhydrous product, an emulsion obtained by dispersing an oil phase in water, a suspension, a microemulsion, a microcapsule, a microgranule or an ionic (liposome) A cream, a skin, a lotion, a powder, an ointment, a spray, or a cone stick. It can also be prepared in the form of a foam or an aerosol composition further containing a compressed propellant.

In addition, the cosmetic composition may further contain, in addition to the Eliocaffus pentylatorus extract of the present invention or its fractions, a fatty substance, an organic solvent, a solubilizing agent, a thickening agent and a gelling agent, a softening agent, an antioxidant, a suspending agent, a stabilizer, a surfactant, a water, an ionic or nonionic emulsifier, a filler, a sequestering and chelating agent, a preservative, a vitamin, a barrier, a wetting agent, an essential oil, a dye, a pigment, a hydrophilic or lipophilic active agent, Lipid vesicles or any other ingredient conventionally used in cosmetics, as well as adjuvants commonly used in the cosmetics field.

In the cosmetic composition containing the Eliocaffus pentioatus extract or the fraction thereof of the present invention, the methanol extract of the present invention is added to the cosmetic composition usually contained in an amount of 0.1 to 50% by weight, preferably 1 to 10% by weight .

When the Eliocaccus pentylatorus extract of the present invention or a fraction thereof is used as an external preparation for skin, it may further contain a fatty substance, an organic solvent, a solubilizer, a thickening agent and a gelling agent, a softener, an antioxidant, a suspending agent, surfactants, water, ionic or nonionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents , Lipid vesicles, or any other ingredient conventionally used in external preparations for skin. In addition, the components can be introduced in amounts commonly used in the dermatology field.

In addition, the present invention provides a pharmaceutical composition for preventing aging comprising Eliocaffus pentylatorus extract or a fraction thereof as an active ingredient.

The aging is preferably, but not exclusively, skin aging.

Preferably, the Eliocaccus pentylatorus extract is a methanol extract, and the fraction is n-hexane fraction, chloroform fraction, ethyl acetate fraction, n-butanol fraction or water fraction, but is not limited thereto.

In a specific embodiment of the present invention, the Eliocaccus pentylatorus extract or its fraction exhibits a DPPH free radical scavenging effect and an ABTS free radical scavenging effect in concentration-dependent manner, exhibits a superior superoxide radical scavenging effect and has an excellent antioxidative effect It can be used as an effective ingredient of a pharmaceutical composition for preventing aging.

The composition containing the Eliocaffus pentylatorus extract of the present invention or a fraction thereof may further contain one or more active ingredients showing the same or similar functions in addition to the above components.

The composition of the present invention may further comprise a pharmaceutically acceptable additive, wherein pharmaceutically acceptable additives include starch, gelatinized starch, microcrystalline cellulose, lactose, povidone, colloidal silicon dioxide, calcium hydrogen phosphate, lactose Starch glycolate, sodium starch glycolate, carnauba wax, synthetic aluminum silicate, stearic acid, magnesium stearate, aluminum stearate, calcium stearate, calcium stearate, , White sugar, dextrose, sorbitol and talc. 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 are not limited thereto.

That is, the composition of the present invention can be administered in various formulations of oral and parenteral administration at the time of actual clinical administration. In the case of formulation, a diluent such as a filler, an extender, a binder, a wetting agent, a disintegrant, . ≪ / RTI > Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate Calcium carbonate, Sucrose, Lactose, Gelatin and the like.

In addition to simple excipients, lubricants such as magnesium stearate talc may also be used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions and syrups, and various excipients such as wetting agents, sweetening agents, fragrances, preservatives and the like may be included in addition to water and liquid paraffin, which are simple diluents commonly used . Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solutions, 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. Examples of suppository bases include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin, and the like.

The composition of the present invention may be administered orally or parenterally in accordance with the intended method, and may be administered orally, parenterally or intraperitoneally, rectally, subcutaneously, intravenously, intramuscularly, . The dosage varies depending on the patient's body weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and disease severity.

The dosage of the composition of the present invention varies depending on the patient's body weight, age, sex, health condition, diet, administration time, administration method, excretion rate and severity of the disease, and the daily dose is Elioca cupperia Preferably from 0.001 to 10 mg / kg, based on the amount of the tofu extract or fraction thereof, and can be administered 1 to 6 times a day.

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

The present invention also provides an anti-aging health food containing Eliocaccus pentioatus extract or a fraction thereof as an active ingredient.

The aging is preferably skin aging, but not always limited thereto.

Preferably, the Eliocaccus pentylatorus extract is a methanol extract, and the fraction is n-hexane fraction, chloroform fraction, ethyl acetate fraction, n-butanol fraction or water fraction, but is not limited thereto.

In a specific embodiment of the present invention, the Eliocaccus petiatura extract or its fraction exhibits a DPPH free radical scavenging effect and an ABTS free radical scavenging effect in concentration-dependent manner, exhibits a remarkable superoxide radical scavenging effect and has an excellent antioxidative effect It can be understood that it can be usefully used as an active ingredient of an anti-aging health food.

The health food of the present invention can be used as it is, or can be used in combination with other food or food ingredients, and can be suitably used according to conventional methods.

There is no particular limitation on the kind of the health food. Examples of the food to which the Eliocaccus pentylatorus extract or a fraction thereof can be added include dairy products including meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramie noodle, other noodles, gums, ice cream, Various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes, and includes all the health foods in the ordinary sense.

The health beverage composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. Such natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. Examples of sweeteners include natural sweeteners such as tau martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like. The ratio of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 of the composition of the present invention.

In addition to the above, the health food of the present invention may contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acids and salts thereof, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, , A carbonating agent used in carbonated drinks, and the like. It may also contain flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components may be used independently or in combination. The proportion of such additives is not critical, but 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.

The present invention also provides a method for inhibiting senescence comprising the step of administering to a subject a pharmaceutical composition comprising a pharmaceutically effective amount of Eliocaffus pentylatus extract or a fraction thereof as an active ingredient.

The aging is preferably, but not exclusively, skin aging.

Preferably, the Eliocaccus pentylatorus extract is a methanol extract, and the fraction is n-hexane fraction, chloroform fraction, ethyl acetate fraction, n-butanol fraction or water fraction, but is not limited thereto.

The pharmaceutically effective amount is 0.00001 to 10 mg / kg, preferably 0.0001 to 1 mg / kg, but is not limited thereto. The dose may vary depending on the weight, age, sex, health condition, diet, administration period, method of administration, rate of elimination, severity of disease, and the like of a particular patient.

The subject is a vertebrate animal, preferably a mammal, more preferably an experimental animal such as a mouse, a rabbit, a guinea pig, a hamster, a dog or a cat, and most preferably an ape-like animal such as a chimpanzee or a gorilla.

The above administration method is preferably oral or parenteral, and skin application is most preferred.

Hereinafter, the present invention will be described in detail with reference to Examples, Experimental Examples and Production Examples.

However, the following Examples, Experimental Examples and Preparation Examples are merely illustrative of the present invention, and the contents of the present invention are not limited by Examples, Experimental Examples and Production Examples.

< Example  1> Eli Ocarpus Petit Oratory  The extract and Heterogeneous species  Preparation of extract

The inventors have found that Elaeocarpus &lt; RTI ID = 0.0 &gt; 20 mg of petiolatus stem extract was purchased from the Center for Biomaterials Hub, Korea Research Institute of Bioscience and Biotechnology. In addition, it is also possible to use Elioccus persethiatus as a heterologous eelocyte E. dubius , E. stipularis , E. glaber , Russ (E. glabripetalus), Eli Oka Perth Oh Chrono Dia (E. acronodia) extracts also were purchased by each 20 mg abroad biomaterial hub center.

< Example  2> Eli Ocarpus Petit Oratory  menstruum Fraction  Produce

In order to prepare the Eliocaccus pentylatorus solvent fraction, an additional 5.22g of Eliocaccus petiota oratus methanol extract was purchased from the Center for Biomaterials Hub Center, Korea Research Institute of Bioscience and Biotechnology. The methanol extract was suspended in water, The same amount of 100% hexane was added and the mixture was left to shake for 3 times to obtain 33 mg of a hexane fraction. Thereafter, the hexane fraction was removed and chloroform was equally added to the remaining water layer. The chloroform fraction was recovered by shaking three times, and chloroform fraction was obtained in an amount of 134 mg. Ethyl acetate was added to the remaining water layer in the same amount to obtain 596 mg of ethyl acetate fraction , Butanol was equally added to the remaining water layer to obtain 859 mg of a butanol fraction in the same manner. The remaining water layer was concentrated to obtain 2.7 g of a water fraction (FIG. 4).

< Experimental Example  1> Eli Ocarpus Petit Oratory  Methanol extract DPPH Free radical  Verify erase effect

The free radical scavenging effect of the Eliocaccus pentiolatum methanol extract of Example 1 was measured by using 1,1-diphenyl-2-picryl-hydrazyl, DPPH , Sigma) method.

Specifically, to measure the effect of free radical scavenging, a 0.15 mM DPPH solution 90 well dissolved in ethanol was placed in a 96-well plate, and the methanol extracts were dispensed at concentrations of 2.5, 5, 10, 20, 40 and 80 ug / ml , Left at room temperature for 20 minutes, and then measured for absorbance at 517 nm with an ELISA reader. The free radical scavenging effect (%) was calculated by the following equation (1). At this time, the group treated with DMSO was used as a negative control, and the group treated with BHA as a synthetic antioxidant was used as a positive control.

A: Absorbance of the negative control not treated with the Eliocaccus pentogenatum methanol extract of the present invention;

B: Absorbance of the experimental group treated with the Eliocaccus pentioatum methanol extract of the present invention.

As a result, as shown in FIG. 1, the DPPH free radical scavenging effect on the EliocaPerf.Petioratum methanol extract was measured. As a result, it was found that the DPPH free radical scavenging effect depending on the treatment concentration of the EliocaPerf.Petioratum methanol extract And showed similar activity to that of the positive control group BHA (Fig. 1).

< Experimental Example  2> Eli Ocarpus Petit Oratory  Methanol extract ABTS Free radical  Verify erase effect

The ABTS free radical scavenging effect was confirmed by using the Elyokapus petioatum methanol extract of Example 1 above.

Specifically, the ABTS free radical scavenging effect was obtained by mixing ABTS and potassium persulfate to a final concentration of 7 mM and 2.45 mM, respectively, and then left in a dark place for 12 hours to form an ABTS radical. This was diluted with distilled water to an absorbance of 0.7 at 734 nm and dispensed into a 96-well plate. The sample was dispensed at a concentration of 2.5, 5, 10, 20, 40 or 80 ug / ml, And reacted for 7 minutes. Absorbance was measured using a microplate meter at 734 nm. At this time, the group treated with DMSO was used as a negative control, and the group treated with BHA as a synthetic antioxidant was used as a positive control.

As a result, as shown in FIG. 2, the ABTS free radical scavenging effect on the Elyokapherpeticia methanol extract was measured. As a result, it was found that the ABTS free radical scavenging effect depending on the treatment concentration of the Elyokapus petiatis methanol extract (Fig. 2).

< Experimental Example  3> Eli Ocarpus Petit Oratory Methanol extract SOD  activation

In order to confirm the antioxidative effect of the methanol extract of Example 1, the effect of eliminating the superoxide radical acting as a precursor of other harmful active oxygen species was measured. The superoxide radical scavenging effect can be detected by measuring the activity of superoxide dismutase (SOD), which is an enzyme that cleaves superoxide by dismutting superoxide. In the present invention, SOD kit, (Dojindo Co.) was used.

Specifically, samples of each concentration were dispensed into a 96-well plate, and water-soluble WST-1 (2- (4-indophenyl) -3- (4-nitrophenyl) -5- -Dispephenyl) -2H-tetrazolium) was added to the reaction mixture, and the SOD enzyme solution was added thereto. The mixture was reacted at 37 ° C for 20 minutes, and the absorbance of the reaction solution was measured at 450 nm using a microplate meter. The superoxide radical scavenging effect of each sample was also calculated by the reduction reduction of WST for the control group, and the concentration of the sample to which the superoxide radical was 50% erased was calculated as IC ₅₀ . At this time, the group treated with DMSO was used as a negative control, and the group treated with BHA as a synthetic antioxidant was used as a positive control.

As a result, as shown in FIG. 3, the effect of superoxide radical scavenging on the Elyokapherpeticia methanol extract was measured. As a result, it was found that the superoxide radical scavenging effect was dependent on the treatment concentration of Elyokapus petiatorum methanol extract Respectively. In addition, the IC ₅₀ value of the positive control group BHA was 60.519 ± 0.066, while the Elioccus persepturatum methanol extract was found to be 10.385 ± 0.055, and the Eliocaccus pentioatum methanol extract of the present invention was found to be more effective than the positive control group (Fig. 3).

< Experimental Example  4> Eli Ocarpus Petit Oratory Fraction DPPH  And ABTS Free radical Scatters

DPPH and ABTS free radical scavenging activity was measured for hexane, chloroform, ethyl acetate, butanol and water fractions by the solvent in Example 2, in the same manner as Experimental Example 1 and Experiment 2. Each fraction was fractionated at concentrations of 2.5, 10, 40, and 160 ug / ml, reacted at room temperature, and then absorbance was measured with a microplate absorber. At this time, the group treated with DMSO was used as a negative control, and the group treated with Vitamin C, a natural antioxidant, was used as a positive control.

As a result, as shown in FIG. 5, it was confirmed that the DPPH and ABTS free radical scavenging effects of the Elyokapus petiotaus fraction were increased depending on the treatment concentration of the fraction. Especially, the ethyl acetate fraction showed DPPH and ABTS free radicals It is confirmed that the erasing effect is excellent. In addition, when compared with Vitamin C, which is a positive control group derived from natural products, it showed excellent activity (FIGS. 5A and 5B).

< Experimental Example  5> Eli Ocarpus Petit Oratory  Ethyl acetate Fraction Eli Ocarpus copper Of heterologous extract DPPH  And ABTS Free radical Scatters  compare

As a result of the above Experimental Example 4, it was confirmed that the EliocaPerfentiotic ethyl acetate fraction showed the most excellent effect, and the fraction of Eliocaffus petioatus ethyl acetate and the fraction of Eliocaferus ethyl acetate prepared in Example 1 The free radical scavenging ability of the extracts of different species was compared.

Eli Oka Perth Petit aura tooth and Lee, Jong dongsok Eli Oka Perth Pardubice Earth (E. dubius), Eli Oka Perth styryl furanyl less (E. stipularis), Eli Oka Perth article rabeo (glaber E.), Eli Oka Perth article Russ L 'Abri Petal (E. glabripetalus), Eli Oka Perth Oh chronograph Dia (E. acronodia) the extract, respectively 10, and each divided by 40 ug / ml concentration of <experimental example 1 and experimental example 2> and DPPH and ABTS free-radical in the same manner The scavenging ability was measured. At this time, the group treated with DMSO was used as a negative control, and the group treated with Vitamin C, a natural antioxidant, was used as a positive control.

As a result, it was confirmed that the DPPH and ABTS free radical scavenging effects of the Elyokapiperentiator ethyl acetate fraction were superior to those of the heterologous kinase, as shown in the following Tables 1 and 2. In addition, it showed excellent activity when compared with Vitamin C, a positive control group derived from natural products.

DPPH scevenging (% of control) Sample concentration 10 mg / ml 40mg / ml E. petiolatus EA 52.13 ± 1.93 93.24 ± 0.01 E.dubius 35.35 ± 0.10 89.43 ± 0.88 E. stipularis 38.36 ± 0.69 90.91 ± 0.55 E.glaber 19.27 ± 0.89 55.07 ± 1.01 E.glabripetalus 15.15 ± 0.39 36.07 ± 0.46 E. acronodia 34.54 ± 0.65 88.66 ± 0.72 Vitamin C 4.59 ± 1.45 20.39 ± 1.15

ABTS scevenging  (% of control ) Sample concentration 10 mg / ml 40mg / ml E. petiolatus EA 99.84 ± 0.14 99.85 ± 0.05 E. dubius 86.19 ± 0.32 99.63 ± 0.10 E. stipularis 90.31 ± 0.31 99.23 ± 1.30 E. glaber 57.81 ± 1.20 99.03 ± 0.27 E. glabripetalus 45.76 ± 0.08 95.78 ± 0.41 E. acronodia 92.05 ± 1.07 98.66 ± 2.23 Vitamin C 16.78 ± 0.04 57.90 ± 1.09

<Experimental Example 6> Melanin production inhibitory effect of Eliocaccus sp.

In order to compare the whitening effect of the Elioca flus petioatus hexane fraction with the extract of the Eliocaffus homologous species prepared in Example 1 above, an inhibition of melanin formation was performed in animal cells.

Specifically, B16F10 cells, known as murine melanoma cell lines, were purchased from ATCC and grown in Dubecco's modified Eagle's medium (DMEM) medium containing 10% fetal bovine serum (GibcoBRL, USA) and 1% penicillin-streptomycin At 37 ° C and 5% CO ₂ . In order to quantitate melanin, 1 × 10 ⁵ cells were added to a 24-well plate, and the cells were adhered for one day. The cells were then exchanged for 1 day with FBS-containing medium. The methanol extract of E. Pellitus , , ethyl acetate fraction, or Eli Oka Perth Pardubice Earth (E. dubius), Eli Oka Perth styryl furanyl less (E. stipularis), Eli Oka Perth article rabeo (E. glaber), Eli Oka Perth article Russ L 'Abri Petal (E. glabripetalus), Eli Oka Perth Oh chronograph Guardia (and cultured for two days to dispense with E. acronodia) concentration of 40 ug / ml each of the extract. Then, the cells were washed with PBS buffer, collected, and 100 μl of 1N NaOH containing 10% DMSO was added thereto. The cells were incubated at 80 ° C. for one hour, transferred to a 96-well plate, and absorbance was measured at 405 nm. The results obtained were calculated assuming that the amount of melanin produced in the negative control treated with 0.2% DMSO was 100%.

As a result, as shown in FIG. 6, it was confirmed that the melanin formation inhibitory effect of the Elioca-fructifetriatuhexane fraction was superior to that of the heterologous kinase (FIG. 6).

< Manufacturing example  1> Eli Ocarpus Petit Oratory  Extract or its The fraction  Antioxidant as an active ingredient Cosmetic  Produce

<1-1> Manufacture of flexible lotion

The softened lotion containing the Eliocaccus pentylatorus extract or a fraction thereof as an active ingredient was prepared as shown in Table 3 below.

Raw material Content (parts by weight) The Eliocaccus pentylatorus extract of the present invention or its fraction 10.00 1,3-butylene glycol 1.00 Disodium iodide 0.05 Allantoin 0.10 Dipotassium glycyrrhizate 0.05 Citric acid 0.01 Sodium citrate 0.02 Glycereth-26 1.00 Arbutin 2.00 Hydrogenated castor oil 1.00 ethanol 30.00 Preservative a very small amount coloring agent a very small amount Flavoring agent a very small amount Purified water Balance

<1-2> Preparation of nutritional cream

The nutritional cream containing the above Eliocaffus pentylatorus extract or its fraction as an active ingredient was prepared as shown in the following Table 4. [

Raw material Content (parts by weight) The Eliocaccus pentylatorus extract of the present invention or its fraction 10.0 1,3-butylene glycol 7.0 glycerin 1.0 D-Panthenol 0.1 Plant extract 3.2 Magnesium aluminum silicate 0.3 PEG-40 stearate 1.2 Stearic acid 2.0 Polysorbate 60 1.5 Chin type glyceryl stearate 2.0 Sorbitan sesquioleate 1.5 Cetearyl alcohol 3.0 Mineral oil 4.0 Squalane 3.8 Carlyric / capric triglyceride 2.8 vegetable oil 1.8 Dimethicone 0.4 Dipotassium glycyrrhizate a very small amount Allantoin a very small amount Sodium hyaruronate a very small amount Tocopheryl acetate Suitable amount Triethanolamine Suitable amount Preservative Suitable amount Flavoring agent Suitable amount Purified water Balance

< Manufacturing example  2> Eli Ocarpus Petit Oratory  Extract or its The fraction  Active ingredient Preparation of pharmaceutical preparations

<2-1> Sanje  Produce

2 g &lt; RTI ID = 0.0 &gt; of the &lt; / RTI &gt; Eliocaccus pentioatus extract or fraction thereof

Lactose 1 g

The above components were mixed and packed in airtight bags to prepare powders.

<2-2> Preparation of tablets

100 mg &lt; / RTI &gt; of the Eliocaccus pentioatus extract or fraction thereof of the present invention

Corn starch 100 mg

100 mg of milk

Magnesium stearate 2 mg

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

&Lt; 1-3 > Preparation of capsules

100 mg &lt; / RTI &gt; of the Eliocaccus pentioatus extract or fraction thereof of the present invention

Corn starch 100 mg

100 mg of milk

2 mg of magnesium stearate

After mixing the above components, the capsules were filled in gelatin capsules according to the conventional preparation method of capsules.

&Lt; 1-4 >

1 g of the Eliocaccus pentylatorus extract of the present invention or a fraction thereof

Lactose 1.5 g

Glycerin 1 g

0.5 g of xylitol

After mixing the above components, they were prepared so as to be 4 g per one ring according to a conventional method.

<1-5> Preparation of granules

150 mg of the Eliocaccus pentylatorus extract of the present invention or its fraction

Soybean extract 50 mg

200 mg of glucose

600 mg of starch

After mixing the above components, 100 mg of 30% ethanol was added and the mixture was dried at 60 캜 to form granules, which were then filled in a capsule.

< Manufacturing example  3> Eli Ocarpus Petit Oratory  Extract or its The fraction  Active ingredient Manufacture of anti-aging health food

<3-1> Production of flour food

0.5 to 5.0 parts by weight of the Eliocaccus pentylatorus extract of the present invention or its fractions were added to wheat flour and the mixture was used to prepare breads, cakes, cookies, crackers and noodles.

<3-2> soup  And juicy ( gravies )

0.1 to 5.0 parts by weight of the Eliocaccus pentioatus extract of the present invention or its fractions were added to the soup and the juice to prepare a health improvement meat product, noodle soup and juice.

<3-3> Ground Beef  Produce

10 parts by weight of the Eliocaffus pentioatus extract of the present invention or its fraction was added to ground beef to prepare a ground beef for health promotion.

<3-4> Dairy products ( dairy products )

5 to 10 parts by weight of the Eliocaccus pentylatorus extract of the present invention or its fractions were added to milk and various dairy products such as butter and ice cream were prepared using the milk.

<3-5> Solar  Produce

Brown rice, barley, glutinous rice, and yulmu were dried by a known method and dried, and the mixture was granulated to a powder having a particle size of 60 mesh.

Black soybeans, black sesame seeds, and perilla seeds were steamed and dried by a conventional method, and then they were prepared into powder having a particle size of 60 mesh by a pulverizer.

The Eliocaccus pentioatus extract or its fraction of the present invention was concentrated under reduced pressure in a vacuum concentrator, dried by spraying, and dried with a hot-air drier, and the resulting dried product was pulverized to a size of 60 mesh with a pulverizer to obtain a dry powder.

The above-prepared cereals, seeds and the Eliocaccus pentioatus extract of the present invention or fractions thereof were prepared in the following proportions:

(30 parts by weight of brown rice, 15 parts by weight of yulmu, 20 parts by weight of barley)

Seeds (7 parts by weight of perilla, 8 parts by weight of black beans, 7 parts by weight of black sesame seeds)

(3 parts by weight) of the Eliocaccus pentylatorus extract or fraction thereof of the present invention,

(0.5 part by weight),

Rhubarb (0.5 parts by weight).

< Manufacturing example  4> Eli Ocarpus Petit Oratory  Extract or its The fraction  Production of anti-aging beverage containing active ingredient

<4-1> Health drink  Produce

(5%) of the Eliocaccus pentylatorus extract of the present invention or its fractions, such as liquid fructose (0.5%), oligosaccharide (2%), sugar (2%), Homogenously blended and sterilized instantly, and packaged in small containers such as glass bottles and plastic bottles.

<4-2> Production of vegetable juice

Vegetable juice was prepared by adding 5 g of the Eliocaccus pentylatorus extract of the present invention or a fraction thereof to 1,000 ml of tomato or carrot juice.

<4-3> Preparation of fruit juice

A fruit juice was prepared by adding 1 g of the Eliocaccus pentylatorus extract of the present invention or a fraction thereof to 1,000 ml of apple or grape juice.

As can be seen from the above, the Elaeocarpus &lt; RTI ID = 0.0 &gt; petiolatus extract or its fractions exhibit an excellent antioxidative effect and thus can be effectively used as an active ingredient of a cosmetic composition for preventing aging, a pharmaceutical composition and a health food.

Claims (12)

An antioxidant cosmetic composition comprising an ethyl acetate fraction of Elaeocarpus petiolatus extract as an active ingredient.
The antioxidative cosmetic composition according to claim 1, wherein the Eliocaccus pentylatorus extract is extracted using water, a C ₁ - C ₂ lower alcohol or a mixture thereof as a solvent.
The antioxidative cosmetic composition according to claim 2, wherein the lower alcohol is ethanol or methanol.
delete An antioxidant health food containing the ethyl acetate fraction of Elioccus persethotaurus extract as an active ingredient.
The method of claim 5, wherein the elementary Oka Perth Petit aura tooth extract antioxidant for health food characterized in that the extraction by using the lower alcohol or a mixture of water, C ₁ ~ C ₂ as the solvent.
7. The antioxidant health food according to claim 6, wherein the lower alcohol is ethanol or methanol.
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