KR20130009649A - Composition for antioxidant comprising extract or fractions of rhododendron album blume as an active ingredient - Google Patents

Abstract

PURPOSE: A composition containing a Rhododendron album extract or a fraction thereof for antioxidation is provided to remove concentration-dependent DPPH free radical and ABTS free radical and to ensure excellent antioxidation effect. CONSTITUTION: A pharmaceutical composition for antioxidation contains a Rhododendron album extract or a fraction thereof as an active ingredient. The extract is prepared using water, alcohol of C1-C2, or a mixture thereof. The fraction is a normal-hexane fraction, a chloroform fraction, an ethyl acetate fraction, a normal-butanol fraction, or a water fraction. A composition for health foods for antioxidation contains the extract or the fraction as an active ingredient. [Reference numerals] (AA) DPPH radical scavenging ability(%); (BB) Rhododendron album; (CC) Alpha-tocopherol

Description

Composition for antioxidant comprising extract or fractions of Rhododendron album Blume as an active ingredient}

The present invention is Rhododendron album Blume) relates to a pharmaceutical composition for antioxidant, a health food composition and a cosmetic composition containing an extract or a fraction thereof as an active ingredient.

Free radicals that cause aging and disease in the human body are formed by biological reactions during normal metabolic processes in the human body, and are known to cause various diseases by causing harmful toxicity to cells and tissues (Folin O & Denis W, 1912, J Biol Chem 12: 239-249). The physiological action of inhibiting such harmful free radicals is similar to that of an electron donating ability and a superoxide dismutase which inhibits oxidation by donating electrons to oxidative free radicals. It converts superoxide anion radicals into normal oxygen.

Many bioactive ingredients with powerful antioxidant effects contribute greatly to the prevention of ulcers and spasms caused by oxidative stress, control of gastric juice secretion and diarrhea, prevention of diabetes, prevention 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 recently increased, various plant resources have been selected to develop resources containing a large amount of functional substances, such as anti-cancer, anti-allergy, anti-obesity, antioxidant, and antibacterial, and to develop them as raw materials for pharmaceuticals, food additives, and cosmetics. Research is underway worldwide (Shin DB, et al., 2006, Food Sci Biotechnol., 15: 357-362, Park et 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 (SOD), an antioxidant, breaks down free radicals (Superoxide radicals, O ^2- ), which are issued during cell respiration, into hydrogen peroxide and oxygen. Superoxide radicals themselves have relatively low reactivity, but are easily converted to H ₂ O ₂ to form highly reactive hydroxyl radicals or react with nitric oxides (NO) to form highly reactive peroxy nitrites. nitrite, ONOO ^- generate) to causes such as the oxidation of SH- groups, protein tyrosine nitride (nitration), lipid peroxidation, DNA damage. When hydrogen peroxide, a product of decomposition by superoxide dismutase, reacts with metal ions such as iron or copper ions, a more powerful active oxygen called a hydroxy radical is generated. The antioxidants catalase and glutathione peroxidase, which remove hydrogen peroxide before it reacts with metal ions, decompose hydrogen peroxide into water and carbon dioxide. 5% of the total hydrogen peroxide that antioxidants can't remove turns into hydroxy radicals in response to metal ions in the body, destroying cells and causing disease. Frequent exposure to radiation or ultraviolet light converts hydrogen peroxide into hydroxy radicals to promote aging, which can be delayed by taking antioxidants.

As a method of measuring free radical scavenging ability, DPPH receives an electron or hydrogen from an antioxidant and forms an irreversibly stable molecule. Therefore, it can measure antioxidant activity from an electron donating ability. At this time, there is a characteristic that the indigo blue color inherent in DPPH is thinned, and the color difference is measured to measure the electron donating ability (Jeon et al., J. Kor. Soc. Food sic. Nutr. 38 (1)). : 1-8, 2009). In addition, the ABTS method was designed in 1993 to measure the physiological activity of a sample, but has since been widely applied to test foods and naturally soluble phenolic substances. The cyan radical cation (ABTS ^{· +} ) produced by the mixture of ABTS [2,2′-azinobis (3-ethyl-benzo-thiaziline-6-sulfonate)] and potassium persulfate is the same as the phenolic compound. It reacts violently with a hydrogen donor (H-donor) to become colorless, and quantitatively measures the electron donating ability (Miller et al., Clin. Sci . 26: 265-277, 1993).

 Antioxidants are widely distributed in animals and plants. Antioxidants such as phenolic compounds, flavonoids, tocopherols, vitamin C, and selenium in fruits and vegetables delay or prevent the oxidation of fats, and cancer, cardiovascular diseases, etc. By preventing and delaying, it plays an important role in preventing aging. Tocopherols and vitamin C, which are natural antioxidants present in the plant kingdom, are widely used in food, medicine, and cosmetics. The rancidity of fats and oils or foods containing oils and fats is mainly caused by the combination with oxygen in the air. To prevent this, many synthetic or natural antioxidants have been developed.

Synthetic antioxidants include Butylated Hydroxy Toluene (BHT), Butylated Hydroxy Anisole (BHA), Propyl Gallate, and the like. Synthetic antioxidants are widely used in commercial foods because of their excellent antioxidant power and low price, but their usage is legally regulated due to concerns about stability, such as human side effects. Natural antioxidants, such as tocopherol, are safe for the human body, but have the disadvantage of low oxidative chain reaction inhibition ability and high price. Therefore, in recent years, researches are being actively conducted to develop safer and more natural antioxidants.

Rhododendron Alboom is a native plant of Indonesia that belongs to the rhododendron. The plant is white and in some cases light purple. Surgery was orange or pink (Bay Science Foundation, 2008). The habitats of these plants are threatened with species due to habitat loss for reasons of logging, crop cultivation and expansion of infrastructure (Whitten et al. 1996). In addition, this plant has a variety of varieties worldwide, especially in the United States, because of its high aesthetic value, typically Rhododendron. cawtawbiencse var. album The same plant is being developed and used in the landscape industry (Paghat, 2008). However, no antioxidant effect has been reported on Rhododendron album Blume extract at present.

Accordingly, the inventors of the present invention, while screening plants with excellent antioxidant effects, Rhododendron alboom extract or its fractions are excellent DPPH (1,1-diphenyl-2-picryl-hydrazyl) free radical scavenging effect and ABTS (2,2 ' -azinobis- (3-ethylbenzothiazoline-6-sulfonate)) Since the free radical scavenging effect was confirmed to have an excellent antioxidant effect, the present invention was completed by revealing that it can be usefully used as an active ingredient of the antioxidant composition.

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

The present invention is Rhododendron album The present invention relates to an antioxidant composition containing Blume) extract or a fraction thereof as an active ingredient. More specifically, the Rhododendron alboom extract of the present invention or a fraction thereof exhibits significant DPPH free radical scavenging effect and ABTS free radical scavenging effect. By having an antioxidant effect, it can be usefully used as an active ingredient of the pharmaceutical composition for antioxidant, the composition for health food and the cosmetic composition.

1 is a Rhododendron album Blume) is a diagram showing the manufacturing process of extracts and fractions.
2 is a diagram showing Thin Layer Chromatography (TLC) of Rhododendron Alboom extract and fractions:
T: rhododendron alboom extract;
H: rhododendron alboom n-hexane fraction;
C: rhododendron alboom chloroform fraction;
E: rhododendron alboom ethyl acetate fraction;
B: rhododendron alboom butanol fraction; And
W: Rhododendron Alboom water fraction.
Figure 3 is a diagram showing a CAD (Charge Aerosol Detector) of the rhododendron alboom extract or fractions thereof.
Figure 4 is a graph showing the DPPH free radical scavenging effect of the rhododendron alboom extract.
5 is a graph showing the ABTS free radical scavenging effect of the rhododendron alboom extract.

Hereinafter, the present invention will be described in detail.

The present invention is Rhododendron album Blume) provides an antioxidant pharmaceutical composition containing an extract or a fraction thereof as an active ingredient.

In addition, the present invention Rhododendron alboom extract ( Rhododendron album Blume) or a fraction thereof is provided as an anti-aging pharmaceutical composition.

Rhododendron alboom extract ( Rhododendron album Blume) is preferably prepared by a manufacturing method comprising the following steps, but not limited to:

1) extracting by adding an extraction solvent to the Rhododendron alboom;

2) filtering the extract of step 1); And

3) concentrating the filtered extract of step 2) under reduced pressure and drying to prepare an extract of rhododendron alboom.

In the above method, the Rhododendron alboom of step 1) can be used without limitation, such as grown or commercially available. The Rhododendron alboom is available in both leaves, stems or roots.

In the above method, the extraction solvent of step 1) is preferably water, alcohol or a mixture thereof. As the alcohol, C ₁ to C ₂ lower alcohols are preferably used, and as lower alcohols, 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 is not limited thereto. The extraction solvent is preferably extracted by adding 1 to 10 times to the dried Rhododendron alboom, and more preferably by adding 2 to 3 times, but not always limited thereto. The extraction temperature is preferably 20 to 100 ° C, 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. In addition, the number of extraction is preferably 1 to 5 times, more preferably 3 to 4 times repeated extraction 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. In addition, the drying is preferably reduced pressure drying, vacuum drying, boiling drying, spray drying or freeze drying, but is not limited thereto.

In addition, the fraction of the rhododendron alboom extract is preferably prepared as follows, but is not limited thereto.

1) suspending the rhododendron alboom extract in water, and then separating n-hexane layer by adding n-hexane;

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

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

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

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

In the above method, the organic solvent is preferably, but not limited to, normal-hexane, chloroform, ethyl acetate, or normal-butanol. The fraction may be obtained by repeating the fractionation process 1 to 5 times, preferably 3 times from the Rhododendron alboom extract, preferably concentrated under reduced pressure after the fraction, but is not limited thereto.

The organic solvent is more preferably ethyl acetate or butanol.

In a specific embodiment of the present invention, the present inventors purchased the Rhododendron alboom extract from the Overseas Plant Extract Bank of the Korea Center for Biomaterials, Korea Research Institute of Bioscience and Biotechnology, and uses the Rhododendron alboom extract to extract the rhododendron alboom fraction. Prepared (see FIG. 1).

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

In addition, in another specific embodiment of the present invention, in order to confirm the antioxidant effect of the rhododendron alboom extract, DPPH free radical scavenging effect of 1,1-diphenyl-2-picryl-hydrazil (1,1- As measured by diphenyl-2-picryl-hydrazyl, DPPH, Sigma) method, it was confirmed that the rhododendron alboom extract showed the scavenging effect of DPPH free radicals in a concentration-dependent manner (see FIG. 4).

In addition, in another specific embodiment of the present invention, in order to confirm the antioxidant effect of the rhododendron alboom extract, as a result of measuring the ABTS free radical scavenging effect according to various concentrations of the rhododendron alboom extract, the rhododendron alboom extract It was confirmed that this concentration-dependent scavenging effect of the ABTS free radicals (see Fig. 5).

In addition, in another specific embodiment of the present invention, as a result of confirming the DPPH and ABTS free radical scavenging ability of the rhododendron alboom fraction, it was confirmed that the rhododendron alboom solvent fraction was excellent in DPPH and ABTS free radical scavenging effect, in particular Rhodo The dendron alboom ethyl acetate and butanol fractions were found to have significantly higher DPPH and ABTS free radical scavenging effects, and also showed significantly better activity compared to the a-tocopherol and ascorbic acid treatment groups, which were derived from natural products. It was confirmed (see Table 1).

Therefore, the rhododendron alboom extract or fractions thereof of the present invention exhibit DPPH free radical scavenging effect and ABTS free radical scavenging effect in a concentration-dependent manner, and thus have an excellent antioxidant effect, and thus are effective for antioxidant or anti-aging pharmaceutical compositions. It can be seen that it can be usefully used as a component.

The composition containing the rhododendron alboom extract of the present invention or a fraction thereof may further contain one or more active ingredients exhibiting 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 the pharmaceutically acceptable additive may include starch, gelatinized starch, microcrystalline cellulose, lactose, povidone, colloidal silicon dioxide, calcium hydrogen phosphate, lactose , Mannitol, malt, gum arabic, pregelatinized starch, corn starch, powdered cellulose, hydroxypropyl cellulose, opiodry, sodium starch glycolate, lead carnauba, synthetic aluminum silicate, stearic acid, magnesium stearate, aluminum stearate, calcium stearate , Sucrose, dextrose, sorbitol, talc and the like can be used. The pharmaceutically acceptable additive according to the present invention is preferably included 0.1 to 90 parts by weight based on the composition, but is not limited thereto.

That is, the composition of the present invention can be administered in various oral and parenteral formulations during actual clinical administration, and when formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, surfactants, etc., which are commonly used It can be prepared using. Solid preparations for oral administration include tablets, pills, powders, granules, capsules and the like, and such solid preparations may contain at least one excipient such as starch, calcium carbonate (Rhododendron Alboom Extract or fractions thereof). ), Sucrose (Sucrose), lactose (Lactose) or gelatin can be prepared by mixing. In addition to simple excipients, lubricants such as magnesium styrate talc may also be used. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents, water and liquid paraffin. . Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

The composition of the present invention may be administered orally or parenterally according to a desired method, and when administered parenterally, external skin or intraperitoneal injection, rectal injection, subcutaneous injection, intravenous injection, intramuscular injection or intrathoracic injection injection method It is preferable to select. Dosage ranges depending on the patient's weight, age, sex, health condition, diet, time of administration, method of administration, rate of excretion and the severity of the disease.

The dosage of the composition of the present invention varies depending on the weight, age, sex, health status, diet, time of administration, administration method, excretion rate and severity of the disease of the patient, the daily dosage is Rhododendron alboom extract Or 0.0001 to 100 mg / kg, preferably 0.001 to 10 mg / kg, based on the amount of fractions thereof, and may 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 for inhibiting aging comprising administering to a subject a pharmaceutically effective amount of rhododendron alboom extract or a fraction thereof.

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

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

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 an anti-aging health food composition containing the rhododendron alboom extract or fractions thereof as an active ingredient.

The health food composition of the present invention may be added to the rhododendron alboom extract or fractions thereof as it is, or used with other food or food ingredients, and may be suitably used according to a conventional method.

There is no particular limitation on the kind of the composition for health food. Examples of foods to which the Rhododendron alboom extract or fractions thereof may be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, dairy products including gum, ice cream, and various soups. , Beverages, tea, drinks, alcoholic beverages and vitamin complexes, etc., and includes all of the composition for health food in the 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 proportion 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 health food of the present invention includes various nutrients, vitamins, electrolytes, flavors, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH regulators, stabilizers, preservatives, glycerin , 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.

Rhododendron alboom extract or fractions thereof of the present invention exhibits significant DPPH free radical scavenging effect and ABTS free radical scavenging effect, and thus has an excellent antioxidant effect, and thus is effective for antioxidant health food compositions or anti-aging health food compositions. It can be seen that it can be usefully used as a component.

In addition, the present invention provides a cosmetic composition for antioxidant containing Rhododendron alboom extract or fractions thereof as an active ingredient.

The present invention also provides an anti-aging cosmetic composition containing rhododendron alboom extract or fractions 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 may also be prepared in the form of a foam or in the form of an aerosol composition further containing a compressed propellant.

In addition, the cosmetic composition, in addition to the rhododendron alboom extract or fractions thereof of the present invention, fatty substances, organic solvents, solubilizers, thickening and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents (foaming agents) , Fragrances, surfactants, water, ionic or nonionic emulsifiers, fillers, metal ion sequestrants and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles Or any other ingredient commonly used in cosmetics, such as auxiliaries commonly used in the cosmetic field.

In the cosmetic composition containing the rhododendron alboom extract of the present invention or a fraction thereof, the amount of the extract or the fraction of the present invention is 0.1 to 50% by weight, preferably 1 to 10% by weight in the cosmetic composition usually contained Can be added.

When the Rhododendron alboom extract of the present invention or a fraction thereof is used as an external preparation for skin, it is further added to fatty substances, organic solvents, solubilizers, thickeners and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents ), Fragrances, surfactants, water, ionic or nonionic emulsifiers, fillers, metal ion sequestrants and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipids It may contain adjuvants commonly used in the field of dermatology, such as vesicles or any other ingredients conventionally used in external preparations for skin. In addition, the ingredients may be introduced in amounts generally used in the field of dermatology.

Rhododendron alboom extract or fractions thereof of the present invention exhibits significant anti-DPPH free radical scavenging effect and ABTS free radical scavenging effect, and thus has an excellent antioxidant effect, which is useful as an active ingredient of an antioxidant cosmetic composition or an anti-aging cosmetic composition. It can be seen that it 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 in detail, and the content of the present invention is not limited to the following Examples, Experimental Examples, and Preparation Examples.

Example 1 Rhododendron Alboom ( Rhododendron album Blume ) Extract Preparation

The Rhododendron Alboom Extract was purchased from the Overseas Plant Extract Bank of the Korea Center for Biomaterials, Korea Research Institute of Bioscience and Biotechnology, and was provided at the Herbarium of the Korea Research Institute of Bioscience and Biotechnology (KRIB 0019989). ) Is stored. In addition, the following experiment was used to dissolve the Rhododendron alboom methanol extract to a concentration of 20 mg / ㎖ in DMSO (dimethyl sulfoxide) (hereinafter referred to as Rhododendron alboom extract), it was used by diluting by concentration.

< Example  2> Rhododendron Alboom  menstruum Fraction  Produce

Suspended by adding distilled water to 360 mg of the Rhododendron alboom extract obtained in Example 1, and mixed with an equal amount of n-hexane, the n-hexane-soluble fraction and the water-soluble fraction were separated, and this was repeated three times. Filtration and concentration under reduced pressure gave 17.2 mg of n-hexane fractions. Then, by removing the n-hexane fraction and adding the same amount of chloroform to the remaining water layer to obtain 21.0 mg of the chloroform fraction in the same manner, to the remaining water layer was added to the same amount of ethyl acetate to give 14.0 mg of ethyl acetate fraction in the same manner. The same amount of butanol was added to the remaining water layer to obtain 47.3 mg of butanol fraction in the same manner, and the remaining water layer was concentrated to obtain 235.5 mg of water fraction (FIG. 1).

< Example  3> Rhododendron Alboom  Extract and Fraction TLC  Research

Rhododendron alboom extract or fractions thereof obtained in <Example 1> and <Example 2> were dissolved in methanol at a concentration of 10 mg / ml, and then silica gel reversed phase chromatography (TLC Silica gel 60 RP-18 F ₂₅₄ S, Merck) was performed to investigate the ethyl acetate and butanol fractions with DPPH and ABTS free radical scavenging activity. The developing solvent was 50% methanol (Fig. 2).

< Example  4> Rhododendron Alboom  extract, Fraction UPLC  analysis

The rhododendron alboom extract obtained in <Example 1> and <Example 2> or a fraction thereof was dissolved in methanol at 10 mg / ml and filtered through a 0.2 um membrane filter. UPLC was used Waters Acquity UPLC system, column was ACQUITY UPLC ^TM BEHC ₁₈ (10 mm x 2.1 mm, id, 1.7 μm, waters, USA) at 35 ℃, mobile phase A is water and formic acid Mobile phase B with (100: 0.1, v / v) was acetonitrile and formic acid (100: 0.1, v / v) and initially maintained mobile phase B at 10% for 1 minute and then until 5 minutes. After analyzing by increasing mobile phase B to 45%, increasing mobile phase B to 100% by 7.5 minutes, maintaining mobile phase B to 100% for 9 minutes, lowering mobile phase B to 10% by 9.5 minutes and stabilizing to 10% by 11 minutes. Used for analysis. The flow rate of the mobile phase was 0.4 ml / min and the sample injection amount was 3 μl.

CAD (Charge Aerosol Detector) is a detector that measures the charge by attaching a charge to the materials separated from the UPLC, and can check the relative content and the degree of separation in the chromatographic form (FIG. 3). .

Experimental Example 1 Rhododendron DPPH of Alboom Extract Check free radical scavenging effect

1,1-diphenyl-2-picryl-hydrazyl (1,1-diphenyl-2-picryl-hydrazyl, DPPH, Sigma) method of the free radical scavenging effect of the Rhododendron alboom extract of <Example 1> Measured by.

Specifically, in order to measure the free radical scavenging effect, 90 μl of 0.15 mM DPPH solution diluted in ethanol was put in a 96 well plate, and the Rhododendron alboom extract was 2.5, 5, 10, 20, 40 And 80 µg / ml each was dispensed and left for 20 minutes at room temperature. The absorbance of the reactants was then measured at 517 nm with a microplate reader. The free radical scavenging effect (%) was calculated by Equation 1 below. At this time, the DMSO-treated group was used as a negative control group, the group treated with a synthetic antioxidant BHA and a natural antioxidant tocopherol was used as a positive control group.

A; Absorbance of the negative control group not treated Rhododendron alboom extract of the present invention; And

B; Absorbance of Experimental Group Treated with Rhododendron Alboom Extract of the Present Invention

As a result, as shown in Figure 4, it was confirmed that the DPPH free radical scavenging effect is dependent on the concentration of the rhododendron alboom extract, and also the DPPH free radical scavenging ability of the rhododendron alboom extract is significantly superior to the tocopherol positive control group. , It was confirmed that the activity similar to the synthetic control BHA (Fig. 4).

Experimental Example 2 Rhododendron ABTS of Alboom Extract Check free radical scavenging effect

ABTS free radical scavenging activity was confirmed using the Rhododendron Alboom extract of <Example 1>.

Specifically, ABTS and potassium persulfate were prepared to be 7 mM and 2.45 mM, respectively, mixed, and then left in the dark for 12 hours to form ABTS free radicals. This was diluted with distilled water so that the absorbance was 0.7 at 734 nm, and the plate was dispensed into 96-well plates, and the rhododendron alboom extract was dispensed at concentrations of 2.5, 5, 10, 20, 40, and 80 μg / ml. The reaction was carried out for 7 minutes in a dark room temperature. Absorbance was measured using a microplate meter at 734 nm. At this time, the DMSO-treated group was used as a negative control group, the tocopherol and BHA-treated group was used as a positive control group.

As a result, as shown in Figure 5, it was confirmed that the ABTS free radical scavenging effect is dependent on the concentration of the rhododendron alboom extract, the ABTS radical scavenging effect of the rhododendron alboom extract is excellent when compared to the positive control tocopherol It was confirmed (FIG. 5).

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

For n-hexane, chloroform, ethyl acetate, butanol, and water fractions of the rhododendron alboom solvents fractionated in <Example 2>, DPPH and ABTS free radicals were prepared in the same manner as in <Experimental Example 1> and <Experimental Example 2>. Scavenging ability was measured.

Specifically, each fraction was fractionated at a concentration of 40 ㎍ / ㎖, respectively, and reacted at room temperature, the absorbance was measured by a microplate absorber. At this time, the DMSO-treated group was used as a negative control group, a-tocopherol as a natural antioxidant and ascorbic acid or a group treated with BHA as a synthetic antioxidant was used as a positive control group.

As a result, as shown in Table 1, it was confirmed that the rhododendron alboom solvent fractions were excellent in DPPH and ABTS free radical scavenging effects, and in particular, the rhododendron alboom ethyl acetate and butanol fractions were remarkably effective in DPPH and ABTS free radical scavenging effects. It was confirmed that the high, and also compared to the group treated with a-tocopherol and ascorbic acid, which is a positive control group derived from natural products, it was confirmed to show an excellent activity (Table 1).

< Manufacturing example  1> Rhododendron Alboom  Extract or its The fraction  Contains as an active ingredient Preparation of Antioxidant Pharmaceutical Compositions

<1-1> Sanje  Produce

2 g of rhododendron alboom extract of the present invention or a fraction thereof

1 g lactose

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

<1-2> Preparation of tablets

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

Corn starch 100 mg

Lactose 100 mg

2 mg 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

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

Corn starch 100 mg

Lactose 100 mg

2 mg 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 >

Rhododendron alboom extract of the present invention or fraction thereof 1 g

Lactose 1.5 g

1 g of glycerin

Xylitol 0.5 g

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

Rhododendron alboom extract of the present invention or fraction thereof 150 mg

Soy extract 50 mg

Glucose 200 mg

Starch 600 mg

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 Alboom  Extract or its The fraction  Preparation of Antioxidant Health Foods Containing as an Active Ingredient

<2-1> Production of flour food

0.5 to 5.0 parts by weight of the rhododendron alboom extract or fractions thereof of the present invention was added to flour, and bread, cake, cookies, crackers and noodles were prepared using this mixture.

<2-2> soup  And juicy ( gravies Manufacturing

0.1 to 5.0 parts by weight of the rhododendron alboom extract or fractions thereof of the present invention was added to soups and broths to prepare meat products for health promotion, soups and noodles for noodles.

<2-3> Ground Beef  Produce

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

<2-4> Dairy products ( dairy products Manufacturing

5-10 parts by weight of the rhododendron alboom extract or fractions thereof of the present invention was added to milk, and various dairy products such as butter and ice cream were prepared using the milk.

<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 alboom extract of the present invention or a fraction thereof was concentrated under reduced pressure in a vacuum concentrator, and the dried product obtained by spraying and drying with a hot air dryer was pulverized with a particle size of 60 mesh to obtain a dry powder.

The grains, seeds and the Rhododendron alboom extract of the present invention prepared above were formulated 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)

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

(0.5 part by weight),

Rhubarb (0.5 parts by weight).

< Manufacturing example  3> Rhododendron Alboom  Extract or its The fraction  Preparation of antioxidant drink containing as an active ingredient

<3-1> Health drink  Produce

Homogeneously subcomponents such as liquid fructose (0.5%), oligosaccharides (2%), sugar (2%), salt (0.5%), water (75%) and 5 g of rhododendron alboom extract or fractions thereof of the present invention. After sterilization by mixing, it was prepared by packaging in a small packaging container such as glass bottles, plastic bottles.

<3-2> Preparation of vegetable juice

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

<3-3> Production of fruit juice

1 g of rhododendron alboom extract of the present invention or a fraction thereof was added to 1,000 ml of apple or grape juice to prepare a fruit juice.

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

<4-1> Preparation of flexible lotion

The flexible lotion containing the rhododendron alboom extract of <Example 1> or <Example 2> or a fraction thereof as an active ingredient was prepared as shown in [Table 2].

Raw material Content (parts by weight) Rhododendron alboom extract of the present invention or fractions thereof 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 nourishing cream

The nourishing cream containing Rhododendron alboom extract of <Example 1> or <Example 2> or a fraction thereof as an active ingredient was prepared as shown in [Table 3].

Raw material Content (parts by weight) Rhododendron alboom extract of the present invention or fractions thereof 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)

Rhododendron album ) Pharmaceutical composition for antioxidant containing extract or fractions thereof as an active ingredient.
The method of claim 1, wherein the rhododendron alboom extract is water, C ₁ Pharmaceutical composition for antioxidant, characterized in that the extraction using a lower alcohol or a mixture of C ₂ ~ as a solvent.
The pharmaceutical composition for antioxidant according to claim 2, wherein the lower alcohol is ethanol or methanol.
The method of claim 1, wherein the fraction is obtained by suspending the rhododendron alboom extract in water, and then sequentially fractionated with normal-hexane, chloroform, ethyl acetate, normal-butanol and water. Antioxidant pharmaceutical composition, characterized in that any one of the acetate fraction, normal-butanol fraction or water fraction.
Rhododendron alboom extract or a composition for antioxidant health food containing the fraction thereof as an active ingredient.
The composition of claim 5, wherein the Rhododendron alboom extract is extracted using water, ethanol or a mixture thereof as a solvent.
The method of claim 5, wherein the fraction is obtained by suspending the rhododendron alboom extract in water, and then sequentially fractionated with normal-hexane, chloroform, ethyl acetate, normal-butanol and water. Antioxidant health food composition, characterized in that any one of the acetate fraction, normal-butanol fraction or water fraction.
Rhododendron alboom extract or cosmetic composition for antioxidant containing the fraction as an active ingredient.

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