KR101425047B1 - Composition for antioxidant comprising extract or fractions of Rhododendron album Blume as an active ingredient - Google Patents

Abstract

The present invention relates to a process for the preparation of < RTI ID = 0.0 > Rhododendron & album The present invention relates to an antioxidative composition containing an extract or a fraction thereof as an active ingredient. More specifically, the Rhodendron albumin extract of the present invention exhibits DPPH free radical scavenging effect and ABTS free radical scavenging effect in a concentration-dependent manner, , Rhododendron alum ethyl acetate and butanol fraction showed excellent antioxidative effect. Accordingly, Rhododendron alumbaum extract or its fractions can be usefully used as an active ingredient of a pharmaceutical composition for antioxidation, a health food, and a cosmetic composition.

Description

TECHNICAL FIELD The present invention relates to an antioxidant composition containing Rhododendron alumbaum extract or a fraction thereof as an active ingredient,

The present invention relates to a process for the preparation of < RTI ID = 0.0 > Rhododendron & album Blume extract or fractions thereof as an active ingredient, and a cosmetic composition for 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 suppress these harmful free radicals include electron donating ability to inhibit oxidation by donating electrons to oxidative free radicals and superoxide radical cation radicals having a very similar reactivity and destructive role to superoxide dismutase 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 al., 1999, J Agric Food Chem 47: 3963-3966, Ham SS et al., 1997, J Food Sci Nutr 2: 155-161). In addition, as interest in health has increased recently, resources that contain a large amount of functional substances having various effects such as anticancer, antiallergic, anti-obesity, antioxidant and antibacterial activity in various plant resources are selected for development as raw materials for pharmaceuticals, food additives and cosmetics Kim et al., 2006, Food Sci Biotechnol., 15: 357-362, Park et al., 2004, Korean J Nutr, 37: 364-372. , 2006, Kor J Food Sci. Technol. 38: 799-804, Ali KA et al., 2005, Food Chem., 89: 27-36).

Superoxide dismutase (SOD), an antioxidant, breaks down the active oxygen (Superoxide radical, O ^2- ) produced by the respiration process of cells into hydrogen peroxide and oxygen. The superoxide radicals themselves are relatively weak in reactivity, but they are easily converted to H ₂ O ₂ to generate hydroxyl radicals that are highly reactive or react with nitric oxide (NO) to form reactive peroxy nitrite nitrite, ONOO ^- ) to cause oxidization of SH-group, nitration of protein tyrosine, lipid peroxidation, DNA damage, and the like. Hydrogen peroxide, a decomposition product of superoxide dismutase, 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. 5% of the total hydrogen peroxide that can not be removed by antioxidants reacts with metal ions in the body to turn into hydroxy radicals, destroying cells and causing diseases. Frequent exposure to radiation or ultraviolet light stimulates senescence by converting hydrogen peroxide into hydroxy radicals, an antioxidant that can be used to delay it.

As a method of measuring free radical scavenging ability, DPPH can measure antioxidant activity from electron donating ability because it forms an irreversibly stable molecule by receiving electron or hydrogen from antioxidant substance. When it meets with antioxidant activity substance, free radical (free radical) (Jeon et al., J. Kor. Soc., Food Sic., Nutr. 38 (1)), and the electron donating ability is measured by colorimetry of this color difference. : 1-8, 2009). In addition, the ABTS method was designed to measure the physiological activity of a sample in 1993, but has since been widely applied to test food and natural water-soluble phenolic materials. The cyan radical cation (ABTS ^{· +} ) produced by the mixture of ABTS [2,2'-azinobis (3-ethyl-benzo-thiaziline-6-sulfonate)] and potassium persulfate (Miller et al., Clin. Sci . 26: 265-277, 1993). In the present invention, the electron donating ability is measured by measuring the electron donating ability.

 Antioxidants are widely distributed in plants 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 can lead to cancer, cardiovascular diseases It also plays an important role in prevention of aging by preventing and delaying. 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 they are excellent in antioxidant power and low in cost. However, their usage is regulated due to safety concerns such as human side effects. Although natural antioxidants such as tocopherol are safe for human body, they have a disadvantage that they have low 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.

Rhododendron album is a native plant of Indonesia belonging to the Azalea family. This plant is white, and in the case of variants it may be light purple. Surgery is orange or pink (Bay Science Foundation, 2008). The habitat of this plant is threatened by species reduction due to logging, crop cultivation, and expansion of infrastructure (Whitten et al. 1996). In addition, this plant has a high aesthetic value worldwide, especially in the US variant, Rhododendron I have cawtawbiencse . album The same plant has been developed and used in the landscape industry (Paghat, 2008). However, the antioxidative effect of Rhododendron album Blume extract has not been reported at present.

Therefore, the inventors of the present invention found that Rhododron aroma extract or its fractions exhibit excellent DPPH (1,1-diphenyl-2-picryl-hydrazyl) free radical scavenging effect and ABTS (2,2 ' -azinobis- (3-ethylbenzothiazoline-6-sulfonate)) free radical scavenging effect and thus has an excellent antioxidative effect. Thus, the present invention has been completed on the basis that it can be effectively used as an active ingredient of antioxidative composition.

An object of the present invention is to provide a pharmaceutical composition for antioxidation, a composition for health food, and a cosmetic composition containing Rhododendron album Blume extract or a fraction thereof as an active ingredient.

In order to achieve the above object, the present invention provides a pharmaceutical composition for antioxidant containing Rhododendron album Blume extract or a fraction thereof as an active ingredient.

In addition, the present invention provides a composition for health food for antioxidation containing Rhodendron aroma extract or a fraction thereof as an active ingredient.

In addition, the present invention provides an antioxidative cosmetic composition comprising Rhodendron aroma extract or a fraction thereof as an active ingredient.

The present invention relates to a process for the preparation of < RTI ID = 0.0 > Rhododendron & album The present invention relates to an antioxidative composition containing an extract or a fraction thereof as an active ingredient. More particularly, the present invention relates to a composition for preventing or treating diseases caused by the radical scavenging activity of DPDH free radical scavenging and ABTS free radical scavenging By having an antioxidant effect, it can be effectively used as an active ingredient of a pharmaceutical composition for antioxidation, a composition for health food, and a cosmetic composition.

Figure 1 album Rhododendron (Rhododendron album Blume extract and fractions thereof.
Figure 2 shows the TLC (Thin Layer Chromatography) of Rhododendron alboom extract and fractions:
T: Rhododendron almum extract;
H: Rhododendron alum-n-hexane fraction;
C: Rhododendron alum chloroform fraction;
E: Rhodendron A boom ethyl acetate fraction;
B: Rhododendron alphobutanol fraction; And
W: Rhododendron alum water fraction.
3 shows a CAD (Charge Aerosol Detector) of a Rhododendron alumbum extract or a fraction thereof.
4 is a graph showing DPPH free radical scavenging effect of Rhododendron alumbum extract.
FIG. 5 is a graph showing the ABTS free radical scavenging effect of Rhododendron alumbum extract. FIG.

Hereinafter, the present invention will be described in detail.

The present invention relates to a process for the preparation of < RTI ID = 0.0 > Rhododendron & album Blume) extract or a fraction thereof as an active ingredient.

The present invention also relates to the use of Rhododendron album Blume) or a fraction thereof as an active ingredient.

The rhododendron alumbaum extract ( Rhododendron album Blume) is preferably, but not limited to, prepared by a process comprising the steps of:

1) extracting Rhodendron alum with an extraction solvent;

2) filtering the extract of step 1); And

3) Concentrating the filtrate obtained in step 2) under reduced pressure and drying to prepare an extract of Rhododendron albaum.

In the above method, the rhododendron alum in step 1) may be used without limitation such as grown or commercially available. The rhododendron alum can be used as a leaf, a stem or a root.

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, a C ₁ to C ₂ lower alcohol is preferably used. As the lower alcohol, ethanol or methanol is preferably used, and methanol is most preferably used, but not limited thereto. 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 rhododendron boomer fraction, but it is not limited thereto. The extraction temperature is preferably 20 to 100 ° C, and preferably 20 to 40 ° C, but is not limited thereto. In addition, the extraction time is preferably 10 to 48 hours, more preferably 15 to 30 hours, but is not limited thereto. The number of times of extraction is preferably 1 to 5 times, more preferably 3 to 4 times, but is 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 addition, the fraction of Rhodendron boarbum extract is preferably prepared as follows, but is not limited thereto.

1) suspending Rhodendron alumbum extract in water, separating n-hexane layer by adding n-hexane;

2) adding chloroform to the remaining water layer of step 1) to separate the chloroform layer;

3) adding ethyl acetate to the remaining water layer of step 2) to separate the ethyl acetate layer;

4) separating the butanol layer by adding butanol to the remaining water layer of step 3); And

5) separating the water fraction from the remaining water layer of step 4).

In the above method, the organic solvent is preferably but not limited to n-hexane, chloroform, ethyl acetate or n-butanol. The fraction may be obtained from the Rhododendron albaum extract by repeating the fractionation process one to five times, preferably three times, followed by fractionation and concentration under reduced pressure. However, the fraction is not limited thereto.

More preferably, the organic solvent is ethyl acetate or butanol.

In a specific example of the present invention, the present inventors purchased the Rhododendron alumb extract from the overseas plant extract bank of the International Biomaterials Hub Center, Korea Research Institute of Bioscience and Biotechnology, and used the Rhododendron albumin fraction (See Fig. 1).

Further, in another specific embodiment of the present invention, TLC and UPLC irradiation of Rhododendron alumbaum extract or fractions thereof were performed (see FIGS. 2 and 3).

Further, in another specific example of the present invention, in order to confirm the antioxidative effect of Rhododendron alumbaum extract, the DPPH free radical scavenging effect was evaluated by measuring 1,1-diphenyl-2-picryl-hydrazyl (1,1- diphenyl-2-picryl-hydrazyl, DPPH, Sigma). As a result, it was confirmed that the Rhododendron alumbaum extract showed a DPPH free radical scavenging effect in a concentration-dependent manner (see FIG. 4).

Further, in another specific example of the present invention, in order to confirm the antioxidative effect of Rhododendron alumbaum extract, the ABTS free radical scavenging effect according to various concentrations of Rhododendron alumbaum extract was measured, It was confirmed that ABTS free radical scavenging effect was shown in a concentration-dependent manner (see FIG. 5).

In addition, in another specific example of the present invention, the DPDH and ABTS free radical scavenging activities of Rhodendron albumin fractions were confirmed, and Rhodendron albumin solvent fractions showed excellent DPPH and ABTS free radical scavenging effects, Dendron ABM ethyl acetate and butanol fractions were found to have significantly higher DPPH and ABTS free radical scavenging effects and also showed superior activities when compared to the groups treated with a-tocopherol and ascorbic acid, (See Table 1).

Therefore, the rhododendron alumb extract of the present invention or its fractions exhibits DPPH free radical scavenging effect and ABTS free radical scavenging effect in concentration-dependent manner and has an excellent antioxidative effect. Therefore, the pharmaceutical composition for antioxidative or anti-aging pharmaceutical composition is effective It can be seen that it can be usefully used as a component.

The composition containing the Rhodendron aroma extract or the fraction thereof of the present invention may further contain one or more active ingredients which exhibit 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 ), 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 disease, and the daily dose is Rhodendron alum Or from 0.0001 to 100 mg / kg, preferably 0.001 to 10 mg / kg, of the fraction thereof, and can be administered 1 to 6 times per 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 a method of inhibiting senescence comprising the step of administering a pharmaceutically effective amount of Rhodendron alumb extract or a fraction thereof to a subject.

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, 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.

In addition, the present invention provides a composition for health food for antioxidation containing Rhodendron aroma extract or a fraction thereof as an active ingredient.

The present invention also provides a composition for anti-aging health food comprising Rhododendron alumb extract or a fraction thereof as an active ingredient.

The composition for a health food of the present invention can be used as it is, or can be used together with other food or food ingredients, and can be suitably used according to a conventional method.

There is no particular limitation on the kind of the composition for health food. Examples of the food to which the Rhododendron albaum extract or its fractions can be added include meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, dairy products including ice- , Drinks, tea, a drink, an alcoholic beverage, and a vitamin complex, and includes compositions for health food in a conventional 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 ml of the composition of the present invention.

In addition to the above, the composition for a health food of the present invention may further 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, , Alcohols, carbonating agents 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 Rhododendron alumbaum extract or its fraction of the present invention exhibits a DPPH free radical scavenging effect and an ABTS free radical scavenging effect and has an excellent antioxidative effect. Therefore, the composition for health food for antioxidation or the composition for health prevention for aging prevention It can be seen that it can be usefully used as a component.

In addition, the present invention provides an antioxidative cosmetic composition comprising Rhodendron aroma extract or a fraction thereof as an active ingredient.

The present invention also provides an anti-aging cosmetic composition comprising Rhodendron aroma extract or a fraction thereof as an active ingredient.

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.

The cosmetic composition may further contain, in addition to the roodendron aroma extract of the present invention or its fractions, a lipid, an organic solvent, a solubilizer, a thickening agent and a gelling agent, a softener, an antioxidant, a suspending agent, a stabilizer, , A perfume, a surfactant, water, an ionic or nonionic emulsifier, a filler, a sequestering and chelating agent, a preservative, a vitamin, a barrier, a wetting agent, a essential oil, a dye, a pigment, a hydrophilic or lipophilic active agent, Or any other ingredient conventionally used in cosmetics. The cosmetic compositions of the present invention may also contain adjuvants commonly used in the cosmetics field.

In the cosmetic composition containing the Rhododendron alumb extract or a fraction thereof of the present invention, the extract of the present invention or its fraction 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 roodendron aroma 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, a stabilizer, ), Perfumes, surfactants, water, ionic or nonionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, blocking agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipids Or any other ingredient conventionally used in external preparations for skin. The present invention also relates to a cosmetic composition, In addition, the components can be introduced in amounts commonly used in the dermatology field.

The Rhododendron alumbaum extract or its fractions of the present invention exhibit DPPH free radical scavenging effect and ABTS free radical scavenging effect and have an excellent antioxidative effect. Therefore, it is useful as an active ingredient of an antioxidant cosmetic composition or an antioxidant cosmetic composition Can be used.

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 content of the present invention is not limited by the following Examples, Experimental Examples and Production Examples.

Example 1: Rhododendron Album (Rhododendron album Blume ) extract

Rhododendron alumb extract was purchased from the Overseas Plant Extract Bank of the International Biomaterials Hub Center, Korea Research Institute of Bioscience and Biotechnology (KRIB 0019989, Korea Research Institute of Bioscience and Biotechnology) ). In the following experiment, a Rhododendron alumbaum methanol extract was dissolved in dimethyl sulfoxide (DMSO) to a concentration of 20 mg / ml (hereinafter referred to as Rhodendron alumbum extract) and diluted by concentration.

< Example  2> Rhododendron Album  menstruum Fraction  Produce

To 360 mg of the Rhododendron alumbaum extract obtained in Example 1, distilled water was added to suspend the suspension, and the same amount of n-hexane was added to the mixture. The n-hexane-soluble fraction and the water-soluble fraction were separated three times Filtration and concentration under reduced pressure gave 17.2 mg of n-hexane fraction. Then, the n-hexane fraction was removed and chloroform was equally added to the remaining water layer to obtain 21.0 mg of a chloroform fraction. To the remaining water layer, ethyl acetate was added in the same amount to obtain 14.0 mg of an ethyl acetate fraction, An equal amount of butanol was added to the remaining water layer, and 47.3 mg of a butanol fraction was obtained in the same manner. The remaining water layer was concentrated to obtain 235.5 mg of a water fraction (FIG. 1).

< Example  3> Rhododendron Alom  The extract and Fraction TLC  Research

The Rhodendron alumbum extract or its fractions obtained in Examples 1 and 2 were dissolved in methanol at a concentration of 10 mg / ml and purified by silica gel reverse phase chromatography (TLC Silica gel 60 RP-18 F ₂₅₄ S, Merck) were used to investigate ethyl acetate and butanol fraction materials with DPPH and ABTS free radical scavenging activity. The developing solvent was 50% methanol (Fig. 2).

< Example  4> Rhododendron Album  extract, Fraction UPLC  analysis

The Rhododendron alumbaum extract or fractions thereof obtained in Example 1 and Example 2 were dissolved in methanol at 10 mg / ml and filtered through a membrane filter of 0.2 μm. UPLC was carried out using a Waters Acquity UPLC system and the column was equipped with ACQUITY UPLC ^™ BEHC ₁₈ (10 mm × 2.1 mm, id, 1.7 μm, waters, USA) at 35 ° C., mobile phase A was composed of water and formic acid, (100: 0.1, v / v), mobile phase B was initially maintained at 10% for 1 minute with acetonitrile and formic acid (100: 0.1, v / v) 45%, increased to 100% to 7.5%, analyzed by increasing the mobile phase B, and then maintained at 100% for 9 minutes, until the mobile phase B was reduced to 10% for 9.5 minutes and stabilized at 10% for 11 minutes Were used for analysis. The flow rate of the mobile phase was 0.4 ml / min and the sample injection volume was 3 μl.

CAD (Charge Aerosol Detector) is a detector that measures charge by attaching a charge to materials separated from UPLC. The relative content and the degree of separation of materials can be checked by chromatographic method (FIG. 3) .

Experimental Example 1: Rhododendron DPPH of alum extract Check free radical scavenging effect

The free radical scavenging effect of the Rhododendron alumbaum extract of Example 1 was measured by the method of 1,1-diphenyl-2-picryl-hydrazyl (DPPH, Sigma) .

Specifically, to measure the free radical scavenging effect, 90 μl of a 0.15 mM DPPH solution diluted in ethanol was placed in a 96-well plate, and the Rhododendron alumbaum extract was diluted 2.5, 5, 10, 20, 40 And 80 占 퐂 / ml, respectively, and left at room temperature for 20 minutes. The absorbance of the reaction was then measured at 517 nm with a microplate 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, the group treated with BHA, a synthetic antioxidant, and tocopherol, a natural antioxidant, were used as positive control.

A; Absorbance of the negative control not treated with the Rhododendron alumbaum extract of the present invention; And

B; The absorbance of the experimental group treated with the Rhododendron alumb extract of the present invention

As a result, as shown in FIG. 4, it was confirmed that the DPPH free radical scavenging effect was dependent on the concentration of Rhododendron alum extract, and the DPDH free radical scavenging ability of Rhododendron alum extract was much superior to the positive control tocopherol , And the activity was similar to that of the synthetic control group BHA (Fig. 4).

Experimental Example 2: Rhododendron ABTS of alum extract Check free radical scavenging effect

The ABTS free radical scavenging activity was confirmed using the Rhododendron alum extract of Example 1 above.

Specifically, ABTS and potassium persulfate were prepared to be 7 mM and 2.45 mM, respectively, and mixed and left in a dark place for 12 hours to form ABTS free radicals. This was diluted with distilled water to an absorbance of 0.7 at 734 nm, and the mixture was dispensed into a 96-well plate. The Rhododendron alumbaum extract was dispensed at concentrations of 2.5, 5, 10, 20, 40 and 80 / / The reaction was carried out at room temperature 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 tocopherol and BHA was used as a positive control.

As a result, as shown in FIG. 5, it was confirmed that the ABTS free radical scavenging effect was dependent on the concentration of Rhododendron alumbum extract, and the ABTS radical scavenging effect of Rhododendron albaum extract was superior to that of the positive control tocopherol (Fig. 5).

< Experimental Example  3> Rhododendron Alom  menstruum Fraction DPPH  And ABTS Free radical Scatters  Confirm

In the same manner as Experimental Example 1 and Experimental Example 2, n-hexane, chloroform, ethyl acetate, butanol, and water fractions of Rhodendron alum solvent fractioned in Example 2 were treated with DPPH and ABTS free radicals The scavenging ability was measured.

Specifically, each fraction was fractionated at a concentration of 40 μg / 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 a group treated with a natural antioxidant a-tocopherol and ascorbic acid or a synthetic antioxidant BHA was used as a positive control.

As a result, as shown in Table 1, the Rhodendron albumin solvent fraction was found to be excellent in DPPH and ABTS free radical scavenging effects, and in particular, Rhodendron alum bromo ethyl acetate and butanol fraction showed significant DPPH and ABTS free radical scavenging effects (Fig. 1). In addition, it was confirmed that even when compared with the group treated with a-tocopherol and ascorbic acid, which are positive control groups derived from natural products, the activity is remarkably excellent (Table 1).

< Manufacturing example  1> Rhododendron Alom  Extract or its The fraction  Active ingredient Preparation of pharmaceutical composition for antioxidation

<1-1> Sanje  Produce

2 g of the Rhododendron alumb extract of the present invention or a fraction thereof

Lactose 1 g

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

<1-2> Preparation of tablets

100 mg of the Rhododendron alumb extract of the present invention or a fraction thereof

Corn starch 100 mg

100 mg of milk

2 mg of magnesium stearate

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 of the Rhododendron alumb extract of the present invention or a fraction thereof

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 Rhododendron alumb 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 Rhododendron alumb extract of the present invention or a fraction thereof

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  2> Rhododendron Alom  Extract or its The fraction  Production of antioxidant health food containing active ingredient

<2-1> Production of flour food

0.5 to 5.0 parts by weight of the Rhododendron alumbaum extract or fraction thereof of the present invention was added to wheat flour, and the mixture was used to prepare breads, cakes, cookies, crackers and noodles.

<2-2> soup  And juicy ( gravies )

0.1-5.0 parts by weight of the Rhododendron alumb extract of the present invention or its fractions were added to the soup and the juice to prepare health promotion meat products, noodle soup and juice.

<2-3> Ground Beef  Produce

10 parts by weight of the Rhododendron alumb extract of the present invention or a fraction thereof was added to ground beef to prepare a health beverage ground beef.

<2-4> Dairy products ( dairy products )

5 to 10 parts by weight of the Rhododendron alumb extract of the present invention or its fractions were added to milk, and the milk was used to make various dairy products such as butter and ice cream.

<2-5> 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 Rhododendron alumbaum extract or its fraction of the present invention was concentrated under reduced pressure in a vacuum concentrator, sprayed and dried with a hot-air drier, and the resulting dried material was pulverized to a particle size of 60 mesh with a pulverizer to obtain a dried powder.

The grains, seeds, and Rhodendron alumb extract of the present invention prepared above were blended 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)

The Rhododendron alumbaum extract or fraction thereof (3 parts by weight) of the present invention,

(0.5 part by weight),

Rhubarb (0.5 parts by weight).

< Manufacturing example  3> Rhododendron Alom  Extract or its The fraction  Preparation of Antioxidant Drinks Containing Active Ingredients

<3-1> Health drink  Produce

The raw materials such as liquid fructose (0.5%), oligosaccharide (2%), sugar (2%), salt (0.5%) and water (75%) and 5 g of the Rhododendron alumb extract or its fraction After sterilization, they were packaged in small containers such as glass bottles and plastic bottles.

<3-2> Preparation of vegetable juice

Vegetable juice was prepared by adding 5 g of the Rhododendron alumb extract of the present invention or its fraction to 1,000 ml of tomato or carrot juice.

<3-3> Production of fruit juice

Fruit juice was prepared by adding 1 g of the Rhododendron alumb extract of the present invention or a fraction thereof to 1,000 ml of apple or grape juice.

< Manufacturing example  4> Rhododendron Alom  Extract or its The fraction  Antioxidant containing active ingredient Cosmetic  Produce

<4-1> Production of flexible lotion

The softened lotion containing rhodendron alumb extract or its fractions as the active ingredient of Example 1 or Example 2 was prepared as shown in Table 2 below.

Raw material Content (parts by weight) The Rhododendron alumb 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

<4-2> Preparation of nutritional cream

Nutritive creams containing the Rhododendron alumbaum extract or its fractions as the active ingredient of Example 1 or Example 2 were prepared according to the composition shown in Table 3 below.

Raw material Content (parts by weight) The Rhododendron alumb 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

Claims (8)

delete delete delete delete A composition for health food for antioxidation characterized by containing Rhodendron alumb extract or a fraction thereof as an active ingredient, wherein the fraction is any one of ethyl acetate fraction, normal-butane fraction and water fraction.
6. The composition for health food for antioxidation according to claim 5, wherein the Rhododendron alum extract is extracted using water, ethanol or a mixture thereof as a solvent.
delete A cosmetic composition for antioxidation characterized by containing Rhodendron aroma extract or a fraction thereof as an active ingredient, wherein the fraction is any one of ethyl acetate fraction, n-butane fraction and water fraction.

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