KR20100121352A - A composition comprising complex crude drug extract for anti-aging and improvement of skin - Google Patents

Abstract

PURPOSE: A functional composition containing complex herb extract is provided to ensure skin elasticity, moisturizing, anti-wrinkle and whitening without side effects. CONSTITUTION: A complex herb composition with anti-aging and skin care contains herb extract of guava leaves, green tea, Artemisia princeps, aloe or grape seed as an active ingredient. A health food for preventing and treating skin aging contains the herb extract as an active ingredient. The health food is manufactured in the form of tablet, capsule, pill, or liquid.

Description

A composite composition complex crude drug extract for anti-aging and improvement of skin}

The present invention relates to a food composition having a skin aging prevention and cosmetic effect, and more particularly, containing as an active ingredient at least one compound herbal mixed extract selected from the group consisting of guava leaf, green tea, mugwort, aloe vera and grape seed. It is a food composition having excellent skin elasticity, moisturizing, anti-wrinkle and whitening effect through skin cell proliferation, while having no side effects and stable to the human body, thus preventing skin aging and skin beauty.

With the recent increase in living standards, modern people are paying more attention to maintaining healthy skin in addition to maintaining a healthy body. Therefore, interest in enhancing skin beauty and preventing skin aging is increasing.

As the largest organ of the human body, which occupies about 16% of the total body volume, the skin is in direct contact with the external environment and is important to protect the body from many deadly harmful factors such as temperature, humidity, and ultraviolet radiation. Act as a shield. However, as they age, skin cells are damaged due to various contaminants, strong ultraviolet rays, stress, and malnutrition, and cell proliferation is not performed properly, resulting in wrinkles, loss of elasticity, and keratinization.

Skin aging is largely divided into natural aging (or endogenous aging) and external aging (Doris EB, Cosmet. Toiletries, 1996, 111, 31), and natural aging (endogenous aging) is influenced by genetic factors, so artificial regulation On the other hand, external aging is relatively easy to artificially adjust because of environmental factors. Representative external aging factors include ultraviolet light, reactive oxygen species and stress, and the most prominent external aging phenomenon is wrinkle formation (Daniell HW, Ann. Intern. Med., 1971, 75 (6), 873; Grove GL et al., J. Am. Acad. Dermatol., 1989, 21 (3), 631; Griffiths CE et al., Arch. Dermatol., 1992, 128, 347). As a result, skin elasticity is significantly reduced.

Various studies have been conducted to investigate the mechanism of aging of the skin, revealing various causes of skin aging. When the skin is dried by a change in temperature, a decrease in humidity or wind, the skin physiological activity as a protective barrier against the outside is lowered to promote aging. In addition, exposure of the skin to harmful reactive oxygen species or free radicals causes oxidative action in the body to produce lipid peroxide and cause modification of the various proteins that make up the skin. One of the photoaging mechanisms caused by ultraviolet light is via free radical pathways (Harman D, J. Gerontol., 1956, 11 (3), 298). Free radicals are known to cause breakdown of connective tissue formation, such as cutaneous collagen, cell membrane dysfunction, facilitating DNA mutations, altering protein function, intercellular energy transfer, and metabolism related molecules (Lavker RM and Kligman AM, J. Invest.Drmatol., 1988, 90, 325). The fact that free radicals are involved in aging means that antioxidants or various chemical scavengers that inactivate the free radicals can be used to delay the aging process.

In addition, collagen and elastine related to the elasticity of the skin form a network structure in the dermal tissue of the skin. As the network structure is broken, that is, elastin is decomposed by elastin degrading enzyme (elastase). And wrinkling causes endogenous skin aging. Therefore, skin aging can be suppressed by lowering the activity of elastinase, one of the main causes of skin aging.

Conventionally, it has been relied on cosmetics that are almost applied to the skin surface to prevent skin aging. Korean Unexamined Patent Publication No. 2001-65755 discloses a beta-1,3-glucan and ginseng saponin-containing external preparation composition having skin wrinkle improvement and elasticity enhancing effects such as skin cell proliferation and collagen fiber synthesis. No. 423,521 discloses a cosmetic composition containing ginseng polysaccharides that exhibits anti-wrinkle effect, collagen synthesis promoting effect, damaged skin recovery effect by optical contact, and moisturizing effect. However, the external preparations and cosmetics applied directly to the skin surface are not absorbed up to the dermal layer, and act only on the epidermis, so that the skin aging prevention effect is temporary, and there is a problem that the skin is damaged as a side effect. In addition, the active oxygen in the body can not be removed, there was a limit to prevent the aging action by the active oxygen.

In addition, the cosmetic food compositions made of chemicals have a lot of side effects that cause a lot of discontinuation, the situation is urgently required to develop a highly stable skin aging prevention and improvement material even when using for a long time using natural materials. to be.

The present invention was created in order to solve the problems of the prior art as described above, the object of the present invention is a result of focusing on the research of foods with the prevention and improvement of skin aging derived from natural products, guava leaf, green tea, mugwort, aloe and The composition containing the extract of the complex herbal extract of grape seed is useful for skin beauty and health, and it is excellent for skin elasticity, moisturizing, anti-wrinkle and whitening effect due to the cell activity effect through skin cell proliferation, and is stable to human body because of no side effect. It is to provide a complex herbal composition having a skin aging prevention and cosmetic effect that can be applied to a functional food formulation having a skin aging prevention and improvement effect.

Composite herbal composition having a skin aging prevention and cosmetic effect according to an embodiment of the present invention for achieving the above object is an active ingredient extract of at least one herbal extract selected from the group consisting of guava leaf, green tea, mugwort, aloe and grape seed It is characterized by containing it.

As a preferred feature of the present invention, the extract is to include at least one or more than one of the herbal extract of guava leaf, green tea, mugwort, aloe vera and grape seed.

As another preferred feature of the present invention, the composition ratio of the complex herbal extract is guava leaf: green tea: mugwort: aloe vera: grape seed (1-3: 1-3: 1-2: 1-2: 1-2) It is in (w / w).

Health functional food for preventing and improving skin aging according to an embodiment of the present invention for achieving the above object is at least one herbal extract selected from the group consisting of guava leaf, green tea, mugwort, aloe vera and grape seed as an active ingredient It is characterized by including.

As a preferred feature of the present invention, the extract is to include at least one or more than one of the herbal extract of guava leaf, green tea, mugwort, aloe vera and grape seed.

As another preferable feature of the present invention, the compound herbal composition ratio is guava leaf: green tea: mugwort: aloe vera: grape seed (1-3: 1-3: 1-2: 1-2: 1-2) weight ratio (w / is in w).

Another preferable feature of the present invention is any one of tablets, capsules, pills, and liquids.

The features and advantages of the present invention will become more apparent from the following detailed description based on the accompanying drawings. Prior to this, the terms or words used in this specification and claims are not to be interpreted in a conventional and dictionary sense, and the inventors may appropriately define the concept of terms in order to best explain their invention in the best way possible. It should be interpreted as meaning and concept corresponding to the technical idea of the present invention based on the principle that the present invention.

The complex herbal composition having the skin aging prevention and cosmetic effect according to the present invention is composed of water-soluble extracts of guava leaf, green tea, mugwort, aloe and grape seed, and thus can be used as a functional composition and health functional food because it has excellent skin aging prevention and improvement effect. .

Hereinafter, with reference to the accompanying drawings illustrating a composite herbal composition having a skin aging prevention and cosmetic effects according to the present invention.

First, it should be noted that like elements or parts in the drawings are denoted by the same reference numerals as much as possible. In the following description of the present invention, detailed descriptions of well-known functions or configurations will be omitted in order not to obscure the subject matter of the present invention.

1 is a view showing the results of tyrosinase inhibitory activity of the functional composition of Example 2 of the present invention, Figure 2 is a view showing the changes in condition and constitution when ingesting the functional composition of Example 2 of the present invention.

And, Figure 3 is a view showing a change in skin condition appearing when ingesting the functional composition of Example 2 of the present invention.

The present invention provides a functional food composition having skin aging prevention and cosmetic effects containing at least one herbal extract selected from the group consisting of guava leaf, green tea, mugwort, aloe and grape seed.

In addition, the functional food composition of the present invention as the active ingredient, the collagen peptide, soybean extract, dietary fiber, pomegranate extract, vitamin C, vitamin B2 in order to enhance skin beauty and prevent skin aging as an auxiliary ingredient It may include vitamin B6 and the like, and may further include ingredients that are commonly added during food production. Commonly added ingredients in food manufacturing include various nutrients (vitamins, amino acids, antioxidants, etc.), pH adjusters (citric acid, sodium citrate), sweeteners (liquid fructose, erythritol, parachinose, mortgage, etc.), synthetic flavors, etc. Flavors, colorants, excipients (lactose, glucose, starch, lacchurose, etc.), stabilizers, glidants (magnesium stearate), fats and oils (wheat germ oil, olive oil, beeswax, wax, tocopherol, etc.). These components can be used independently or in combination.

Meanwhile, the functional food composition of the present invention may be prepared in various forms of food according to a conventionally known method, and may be prepared as food of various formulations such as pills, beverages, tea, granules, tablets, hard capsules, and soft capsules.

Specifically, the composition of the present invention is a guava leaf water-soluble extract 10-40% by weight, green tea water-soluble extract 10-40% by weight, mugwort water-soluble extract 20-70% by weight and aloe water-soluble extract 10-40% by weight, and grape seed water-soluble extract It consists of a food extract having a composition ratio of 10-50% by weight.

The present invention extracts guava leaf, green tea, mugwort, aloe and grape seeds from 80 to 110 ℃ hot water for 1 to 5 hours, concentrated to 10 to 40% solids, and then mixed with dry dry concentrate to prevent skin aging and beauty Functional food formulations and cosmetic formulations are provided.

The present invention also provides a food additive having a skin aging prevention and cosmetic effect containing one or more complex herbal extracts selected from the group consisting of guava leaf, green tea, mugwort, aloe and grape seed.

The food additive may be included in an amount of 0.01 wt% to 10 wt% based on the total weight of the food.

On the other hand, among the elements constituting the extract used in the present invention, guadium ( psidium guajava L.) is a genus of the genus Myrtaceae (Psidium) genus (Psidium) is a short tree originated in tropical America, leaves, trees Peel, fruit, etc. can be used for health food and medicinal. It is known that it has strong fertility and can be cultivated without using pesticides, and is generally effective for dysentery and diarrhea. Its content is high in tannins, and tannins are known to have a formal, gastrointestinal function, anti-aging and anticancer effects. Other ingredients include fatty oils, essential oils, gasoline, insulin, vitamin B group, vitamin C, magnesium, potassium and other vitamins and minerals, and antibacterial, astringent, branch, anti-inflammatory, hemostatic, histamine free, hypoglycemic, Pharmacological activities, such as antioxidant and vascular permeability inhibition, are known (Anduk Kyun, Korea Herb Book, Kyohaksa, 1999).

Green tea is a dried leaf of Camellia sinensis and belongs to Theaceae. The young and soft leaves of tea tree mainly contain purine alkaloid and caffeine, and its content is 1 to 5%. In addition to the trace amount, theobromine (theobromine), theophylline (theophylline) and xanthine (xanthine) is contained. Tannin in tea leaves is mainly galloyl - l - is going to epi catechol (galloyl- l -epigallocatechol), which is contained together with caffeine. The pharmacological action of tea leaves is mainly caused by xanthine derivatives (caffeine and theophylline), and also contains a large amount of tannic acid, thus acting as antibacterial and vitamin P. Pharmacological action is to excite the central nervous system, promote mental function, increase thinking ability, eliminate fatigue, expand coronary blood vessels, and diuretic (Jung Bo-seop and Shin Min-kyo, Ph.D., pp403-405, Younglimsa, 1998) . The young leaves of the tea tree can produce various kinds of teas with different flavors depending on the processing method. For example, the tea leaves, black tea, oolong tea, Iron ore, cannon tea, green tea, etc. can be manufactured.

The wormwood ( A. princeps var . Orientalis (PAMPAN.) HARA) is a perennial herb that is dried on the leaves of Yellow Sea Mugwort and its equivalents. Yellow sea wormwood contains essential oils and the highest content of essential oils is cineole, accounting for 25-30%. In addition, β-carophyllene (β-carophyllene), linalool (linalool), camphor (camphor), borneol (borneol) and the like is contained. Leaves are tetra Kosa play (tetracosanol), β- sitosterol (β-sitosterol), l - contains inositol (l -inositol) - swebul Rocky Tall (l -chebulachitol), l. Roots and stems contain polysaccharide artemis similar to inulin. The root contains a variety of polyin compounds, including heptadec-1, 7, 9, -triene-11, 13, 15-trine (heptadec-1, 7, 9, -trien-11, 13, 15-triyne), tetradeca-8, 10, 12, -tri-6-6-ene-3-one and methyl 2-decane- 4, 6, 8-triynate (methyl 2-decen-4, 6, 8-triynate). Small branches also contain substances that act like oxytoxins. In addition, numerous plants in the genus contain sedentin pene lactone (ridentin). In addition, it governs blood donation, drives out humidity and has the effects of warmth, haemostasis, and safety. Treatment of cold pain in the abdomen, diarrhea, muscle diarrhea, chronic bleeding, hemorrhage, nasal bleeding, bleeding, dysmenorrhea, abortion, crayfish, infertility, ongyang, improvement pp 1014-1016, 1998).

Aloe vera (Aloe barbadensis Miller) is a peeled aloe and using only jelly, which is a highly moisturizing polysaccharide (Polysaccharids) that enhances the body's immunity and wound healing, and glycoprotein that lowers blood sugar. (Glycoprotein) contains a large amount of active ingredients (Duk-kyun Ahn, Korean Herbal Painting, pp280, Kyohaksa, 2000: Changmin Kim, Primary Color Herbal Painting, pp306, Academy Book, 2001).

Grape seed in the main sichuan of Ishijin (珍 草 綱), the grapes control the strength and tenderness of the tendons and bones to strengthen the strength, strengthen the will, keep healthy, and also endure hunger and endure cold It is said that it is effective in strengthening, so if you eat grapes for a long time, your body will become lighter and not old. Since the extract of grape seed has excellent physiological efficacy, an attempt has been actively made to develop it as a functional food or a therapeutic agent. In Korean Patent Publication No. 98-51189, the grape seed extract as a tyrosinase inhibitor, Korean Patent No. 00-18117 Grape seed extract from natural anti-cavities, Korean Patent Publication No. 01-12238 has disclosed a functional food containing the grape seed extract, such as rice coated with grape seed extract, antioxidants in Korea Patent Publication No. 00-63265 Grape seed oil containing the same and a method for manufacturing the same, a method for producing grape products having improved antioxidant effect in Korean Patent Publication No. 01-04553, a grape seed extract such as a medicinal agent using prosanidine as an active ingredient in Korean Patent Publication No. 99-28877 The use of the antioxidant component of proanthocyanidin, which is a major component of the present invention, has been disclosed.

Hereinafter, a method of preparing a composite herbal composition having a skin aging prevention and cosmetic effect and a health functional food for preventing and improving skin aging according to the present invention and an embodiment thereof will be described in detail.

To prepare extracts of guava leaf, green tea, mugwort, aloe and grape seed, used in the present invention, specifically, after washing guava leaf, green tea, mugwort, aloe and grape seed, they are sufficiently dried in the shade and then chopped or crushed. , About 1 to 30 times the weight (kg), lower alcohols such as ethanol, or a mixed solvent thereof, preferably water, 20 to 120 ° C., preferably 100 to 110 ° C. at an extraction temperature of about 1 hour to 2 One, preferably about 2 to 12 hours, 1 to 10 times, preferably 2 to 5 times repeatedly extraction method, such as hot water extraction, ultrasonic extraction, reflux cooling extraction, more preferably 80 to 110 The crude extract, which is a soluble extract according to water, lower alcohol, or a mixed solvent thereof, was filtered by hot water extraction at 1 ° C. for 5 hours, and concentrated under reduced pressure or concentrated in vacuo to extract the extract concentrate. It may deukhal and lyophilized additionally is obtained in powder form.

The herbal plants used as the material of the composition of the present invention may be prepared directly or purchased commercially, the extracts of the herbal plants of the present invention may be used in the food composition in the form of powder or powder of 20 to 90% high concentration. have.

Functional food composition of the present invention can be used as a food, beverage, or beverage additives for health, the functional composition is mixed with water in 0.01% to 10% by weight relative to the total weight of food for the purpose of preventing and improving skin aging Applied in the field of functional foods in the form of soft or hard capsules, tablets, granules, drops and tea bags filled with dry powders of the functional beverages or functional ingredients and in the field of functional cosmetics in the form of skins, creams, lotions and soaps. It is preferable.

The composition containing the active ingredient of the present invention can be used in various foods and cosmetics for preventing and improving skin aging. Examples of the food to which the composition can be added include various foods, beverages, gums, candy, confectionery, tea, vitamin complexes, health functional foods, and the like, and cosmetics include skins, creams, lotions and soaps. have.

The water-soluble extracts of guava leaf, green tea, mugwort, aloe and grape seed of the present invention have been used for a long time as the extract of the present invention has little toxicity and side effects, so it can be used with confidence even for long-term use for prophylactic purposes. have.

Hereinafter, preferred examples and experimental examples are presented to help understand the present invention. However, the following Examples and Experimental Examples are provided only to more easily understand the present invention, and the present invention is not limited to the following Examples and Experimental Examples.

Example 1 Preparation of Essential Plant Extracts

30 g each of green tea, mugwort, aloe, grape seed, and guava purchased from Agriculture & Forestry Co., Ltd. (Baebang-myeon, Asan-si, Chungnam) and guava purchased from Jinyong Natural (Paldal-gu, Suwon-si, Gyeonggi-do) were put in 600 mL of water for 3 hours. Hot water is extracted and concentrated using a vacuum concentrator (Buchi R-114, Buchi, Switzerland) until the solid content reaches 30%, and the concentrate is freeze-dried with a freeze dryer (FD 5512, Ilsin, Korea) and guava. 3.1, 2.5, 2.3, 2.8 and 1.0 g of dry powder were prepared from leaves, green tea, mugwort, aloe and grape seed, respectively.

Example 2 Preparation of Functional Composition

Functional composition of the present invention by combining the guava leaf, green tea, mugwort, aloe and grape seed water-soluble extract prepared in Example 1 in a blending ratio of 25%, 15%, 15%, 18% and 27%, respectively, based on the solid content% Was prepared.

Experimental Example 1 Determination of Antioxidant Effects of Individual Plant Extracts

In order to confirm the antioxidant efficacy of each of the food extract prepared in Example 1 of the present invention and the functional composition prepared in Example 2, the free radical 1,1-diphenyl-2-picrylhydrazil (1,1 Experiments were performed by modifying the experimental method described in the literature using -diphenyl-2-picrylhydrazyl (hereinafter DPPH) (Yasushi, S et al., Chem. Pharm. Bull. 47, pp1369, 1999). After reacting at room temperature for 30 minutes by adding 1.5 mL of DPPH, 0.15 mL of sample (1000 ppm-10 ppm), and 1.35 mL of distilled water prepared by dissolving in ethanol to 2 × 10-4 M concentration, absorbance was UV at 520 nm. It was measured using a VIS spectrophotometer (Spectronic genesys 5, Milton Roy, USA). The DPPH capture effect of the samples of the present invention was compared by calculating DPPH radical capture activity calculated using Equation 1 as follows (Table 1-Table 2).

DPPH radical scavenging activity (%)

   = ((Acontrol-Bcontrol)-(Asample-Bsample)) / (Acontrol-Bcontrol) × 100

Acontrol: Absorbance of the control group without sample

Asample: Absorbance of reaction group with sample

Bcontrol: absorbance of control group without DPPH

Bsample: absorbance of reaction group without DPPH

As a result, as shown in Table 1 below, the water-soluble extracts of guava leaf, green tea, mugwort, aloe and grape seed all exhibited 60% or more of DPPH radical capture potency at 50 ppm, and the green tea, guava leaf, mugwort, grape seed, The low SC50 values in aloe were found to be excellent antioxidants.

Experimental Example 2 DPPH capture effect of the functional composition of the present invention

Functional composition of the present invention prepared in Example 2, and vitamin E (Sigma, USA, T-3251), (±) -catechin ((±) -Catechin; Sigma, USA, C-1788) as a positive control , Β-Carotene (β-Carotene; Sigma, USA, C-4582), Astaxanthin (Sigma, USA, A-9335), the same as above, free radical capture effect using DPPH as SC50 value Obtained and compared.

Experimental results showed that the functional composition prepared in Example 2 of the present invention exhibited lower antioxidant efficacy than (±) -catechin, but had better antioxidant power than vitamin E, beta-carotene, and astaxanthin used as other positive controls. It could be confirmed (Table 3).

Experimental Example 3. Superoxide Anion Radical Capture Effect of Functional Composition of the Present Invention

Superoxide anion radical scavenging effect measurement was used by modifying the method of Superoxide dismutase (SOD) activity detection kit (435-70601, Wako, Japan).

0.05 mL (500 ppm-10 ppm) of the functional composition of the Example 2 and the sample of the positive control, the coloring solution (0.4 mM xanthin + 0.1 M phosphate buffer (pH 8.0) + 0.24 mM nitroblue tetra Add 0.5 mL of nitroblue tetrazolium (NO2-TB), 0.5 mL of enzyme solution (0.048 unit / mL xanthine oxidase, from Butter milk solution + 0.1 mL of 0.1 M phosphate buffer (pH 8.0) and mix well The reaction was stopped at 37 ° C. for 20 minutes, and then 1 mL of reaction stop solution (709 mM sodium dodecyl sulfate) was added to stop the enzymatic reaction and the absorbance at 560 nm was measured by UV-VIS spectrophotometer (spectronic genesys 5). , Milton Roy, USA). As a positive control used in this experiment, vitamin E, (±) -catechin, beta-carotene, astaxanthin was used, the detailed experimental method is shown in Table 4 below, the superoxide anion radical capture effect is represented by Equation 2 It was calculated as shown in Table 5 below by comparing the SC50 value.

Superoxide Anion Radical Scavenging Activity (%)

= ((Acontrol-Bcontrol)-(Asample-Bsample)) / (Acontrol-Bcontrol) × 100

As can be seen in Table 5 below, the functional composition of Example 2 showed the best capture effect on the superoxide anion radical, in particular 1.63 times than the (±) -catechin used as a positive control, about 10.34 than astaxanthin. The effect was twice as high, and vitamin E and beta-carotene showed little effect of trapping superoxide anion radicals.

Experimental Example 4 Measurement of Lipid Peroxidation Inhibitory Effect of Functional Composition of the Present Invention

In order to confirm the lipid peroxidation inhibitory ability of the functional composition of the present invention, a study on the effect of inhibiting lipid peroxidation using ferric-thiocyanate (FTC) method (Takeshi et al, Food Chemistry , 75 , pp237-40, 2001 The experimental method described in () was applied and used.

0.2 mL of the functional composition prepared in Example 2 and each positive control sample (250 ppm), 0.5 mL of 0.2 M sodium phosphate buffer (pH 7.0), 0.5 mL of 2.5% linoleic acid dissolved in ethanol 50 μl of 0.1 M 2,2′-azobis-2-amidinopropane-dihydrochloride (AAPH), followed by reaction in the dark at 37 ° C. After 200 minutes, 0.1 mL of 20 mM ferric chloride (FeCl2) dissolved in 9.7 mL of 75% ethanol, 0.1 mL of 30% ammonium thiocyanate, and 3.5% hydrochloric acid solution (HCl) was added to 0.1 mL of the reaction sample solution. After mixing well for 3 minutes, absorbance at 500 nm was measured using a UV-VIS spectrophotometer (spectronic genesys 5, Milton Roy, USA). As a positive control, vitamin E, (±) -catechin, beta-carotene and astaxanthin were used. Lipid peroxidation inhibitory activity (%) was calculated using the following equation (3).

Lipid peroxidation inhibitory activity (%)

= ((Acontrol-Bcontrol)-(Asample-Bsample)) / (Acontrol-Bcontrol) × 100

Acontrol: Absorbance of the control group without sample

Asample: Absorbance of reaction group with sample

Bcontrol: absorbance of control group without linoleic acid

Bsample: absorbance of reaction group without linoleic acid added

As a result, the functional composition of Example 2 was 16%, 29%, 56%, 76% higher inhibitory effect than the inhibitory effects of vitamin E, (±) -catechin, beta-carotene and astaxanthin, respectively, used as the positive control. (See Table 6).

Experimental Example 5. Determination of the hydroxy radical trapping effect of the functional composition of the present invention

In order to confirm the hydroxy radical scavenging effect of the functional composition of Example 2 of the present invention, an experimental document for measuring the oxidation inhibition rate of 2-deoxyribose due to hydroxy radicals (Takeshi et al., Food Chemistry , 80 , pp29-33, 2003).

First, 150 μl of 10 mM ferrous sulfate (FeSO4) -EDTA, 150 μl of 10 mM 2-deoxyribose, 300 μl of the functional composition or positive control sample solution of Example 2 (10,000 ppm or 1,000 ppm), 0.2 M sodium phosphate 450 μl of salt phosphate buffer (pH 7.0), 150 μl of 10 mM hydrogen peroxide (H 2 O 2), and 300 μl of distilled water were added. The reaction is initiated by adding H 2 O 2. After 4 hours of reaction at 37 ° C., 750 μl of 2.8% trichloroacetic acid and 750 μl of 1.0% tribarbituric acid were added to stop the reaction, and heated for 10 minutes. After standing for 5 minutes in the state, the absorbance at 520 nm was measured using a UV-VIS spectrophotometer (spectronic genesys 5, Milton Roy, USA). At this time, vitamin E, (±) -catechin, beta-carotene, astaxanthin was used as a positive control, and the hydroxy radical trapping effect was calculated using Equation 4 below.

Hydroxy radical scavenging activity (%)

= ((Acontrol-Bcontrol)-(Asample-Bsample)) / (Acontrol-Bcontrol) × 100

Acontrol: Absorbance of the control group without sample

Asample: Absorbance of reaction group with sample

Bcontrol: absorbance of control group without hydrogen peroxide

Bsample: absorbance of reaction group without hydrogen peroxide

As a result, as can be seen in Table 7, the functional composition of Example 2 showed a higher inhibitory effect than the positive control at 2,000 ppm and 200 ppm.

As a result of measuring the antioxidant power of the four active oxygen, it was found that each positive control group selectively inhibited several kinds of active oxygen, but did not simultaneously inhibit several types of active oxygen. However, the functional composition of Example 2 not only retains high antioxidant power but also simultaneously inhibits various forms of free radicals, and particularly with respect to the hydroxy radical, which is the strongest form of free radicals in active oxygen metabolism mechanisms and the main culprit of aging and cancer. It is considered to have a strong inhibitory ability to protect the human body against various diseases caused by free radicals. However, the functional composition of Example 2 not only retains high antioxidant power but also simultaneously inhibits various forms of free radicals, and is particularly strong against the hydroxy radicals, which are the strongest forms of free radicals in the free radical metabolism mechanism and the main culprit of skin aging. It is thought to be able to prevent and improve skin aging caused by free radicals due to its inhibitory ability.

Experimental Example 6. Measurement of inhibitory ability of basophilic cytokine Interleukin-8 production of the functional composition of the present invention

In order to confirm the hydroxy radical scavenging effect of the functional composition of the present invention, the anti-inflammatory effect of the functional composition of Example 2 was measured. In general, THP-1 cells, which are human monocytes, secrete out of the cell the inter-inflammatory cytokines Interleukin-8 by LPS (Lipopolysaccharide). IL-8 is known to be an inflammation-inducing cytokine, and it can be judged whether it is anti-inflammatory effect by increasing or decreasing IL-8 production by THP-1 cells. To judge.

First, the cells cultured in a P35 dish are replaced with a medium mixed with a test sample (titration) after 24 hours, and then the medium used after the culture for 24 hours is used as a sample. After diluting the sample to the appropriate concentration, put it into each well of IL-8 microplate, and then reacting. After adding Conjugate and reacting with the substrate after a certain time, absorbance (A550 nm) was measured and the concentration of IL-8 according to the standard curve was measured. Calculate

As a result, as can be seen in Table 8, the functional composition of Example 2 reduced the production of IL-8 by about 50-60% in Experimental Group 1 using the highest concentration of 5% compared to the control containing no test substance. As a result, the functional composition was found to have an excellent anti-inflammatory effect.

Experimental Example 7 Measurement of Collagen Formation Capacity and Collagenase Synthesis Capacity of Functional Compositions of the Present Invention

Type I procollagen production ability was measured using fibroblasts for the functional composition of Example 2 to confirm the collagen production ability of the functional composition of the present invention. For comparison of collagen production ability, the prepared functional composition was treated at a concentration of 10 to 100 ppm in a state in which fibroblasts were placed at 5 x 10 4 / well in a 48-well plate. After incubation for 24 hours, the type I procollagen production in the culture supernatant was measured using a type I pN collagen EIA kit. After measuring the type I procollagen production by dividing the total protein mass, the results are shown in Table 9 compared with the group without the addition of a separate sample, the vitamin C treated group and the TGF-b treated group.

In order to measure collagenase synthesis inhibitory activity, 15 mJ / cm2 of UVB was irradiated with fibroblasts attached to a 48-well plate at 5 × 10 4 / well, and then the functional composition of Example 2 was concentrated at 500 ppm. After 24 hours of incubation, the amount of production of collagenase matrix metalloproteinases (MMP-1) in the culture supernatant was measured using an MMP-1 ELISA kit. The measured MMP-1 expression amount was divided by the total protein amount of the cell, and then corrected. The results are shown in Table 9 compared with the retinoic acid and TGF-b treated groups used as controls. .

As a result, as can be seen in Table 9, it can be seen that the collagen synthesis increases and collagenase production that degrades collagen decreases when fibroblasts are treated with the functional composition of Example 2. In other words, it is possible to maintain a corresponding effect to the unstable vitamin C or retinoic acid, which is a denatured and readily available herbal extract.

Experimental Example 8. Measurement test of the anti-aging effect of the functional composition of the present invention

In order to measure the anti-aging effect of the functional composition of the present invention, the proliferative effect of human normal fibroblasts (Korea Cell Line Bank) was measured by MTT method. This MTT method uses MTT dye (3- (4,5) -dimethylthiazol-2-yl) -2,5-diphenyl-tetrazolium bromide) to form blue formazan by mitochondrial dehydrogenase in mitochondria of living cells. This blue amount represents the amount of mitochondria and metabolic activity. Fibroblasts were placed in a 96-well plate at 1 × 10 4 / well and attached to the prepared functional composition at a concentration of 100 to 10,000 ppm. After 24 hours of incubation, MTT solution (5 μg / mL) dissolved in PBS was added, left in a CO2 incubator for 2 hours, and then the supernatant was removed. After shaking for 20 minutes, the absorbance was measured at 570 nm with an ELISA reader (VersaMax, Molecular device, USA). The control composition was compared with the group that did not add a separate sample as a control group, and as a result, the cell growth rate of the functional composition treatment group was 1.3 times higher than that of the control group. Therefore, it can be seen that the cell proliferation efficacy is excellent when the functional composition of Example 2 is treated.

The measured values were substituted into Equation 1 below to calculate the cell proliferation effect, and the results are shown in Table 10 below.

Cell proliferation effect (%) = [(absorbance of experimental group-absorbance of control group) / absorbance of control group] x 100

As can be seen in Table 10, compared to the control group not treated with the functional composition, the experimental group treated with the functional composition shows a very high fibroblast proliferation effect.

Experimental Example 9. Determination of the effect on tyrosinase activity of the functional composition of the present invention

In order to measure the whitening effect of the functional composition of the present invention, the degree of inhibitory effect of tyrosinase activity was measured. Tyrosinase is an enzyme that accelerates the process of oxidation of a substance called tyrosine in the body, helping to produce melanin. The whitening effect was measured using a method of inhibiting the function of this enzyme to inhibit the oxidation of tyrosine to form a black polymer called melanin. Inhibitory activity against tyrosinase was determined by adding 15 μL of sample to 96-well, 150 μL of 0.1 M phosphate buffer (pH 6.86), 25 μL of 1.5 mM L-tyrosine solution, and then 2,380 units / mL mushroom tyrosinase (0.05 M phosphate buffer). Solution, pH 6.86), 7 μL was added and reacted at 30 ° C. for 10 minutes, and then the absorbance was measured at 475 nm. Inhibition rate for tyrosinase (%) was calculated by the equation (6).

% Inhibition of tyrosinase activity = (1- (A-B) / C) x 100

A: Absorbance of the sample to which the sample, the enzyme, and the substrate are added

B: absorbance of the sample and the sample added only the substrate

C: absorbance of the sample added with enzyme and substrate only

As shown in FIG. 1, as a result of measuring the inhibitory effect of tyrosinase activity of the functional composition of the present invention, the functional composition of Example 2 showed higher inhibitory activity than arbutin currently used as a functional whitening agent.

Experimental Example 10 Determination of the Effect on the Elastase Activity of the Functional Compositions of the Present Invention

In order to measure the anti-wrinkle effect of the functional composition of the present invention, the effect of inhibiting the activity of elastase that inhibits the elasticity of the skin was tested. The test method is an in vitro measure of skin aging prevention and inhibits the enzyme elastase in the body that degrades elastin, which plays an important role in maintaining skin elasticity.

When elastase decomposes the synthetic substrate N-Suc- (Ala) 3-ρ-nitroanilide, it produces ρ-nitroanilide, which gives it a yellow color. This degree of color development was measured with an absorbance photometer to calculate the elastase inhibition rate. First, 1 unit / 100 μL elastase (porcine pancretic elastase) and the functional composition of Example 2 were added to a test tube in 0.2 M Tris-HCl buffer (pH 8.0), followed by preincubation at 25 ° C. for 15 minutes. 20 μL of 0.85 mM N-Suc- (Ala) 3-ρ-nitroanilide was added and reacted at 25 ° C. for 20 minutes, and the absorbance was measured at 405 nm to quantify the amount of ρ-nitroanilide generated from the substrate. One unit of elastase inhibitory activity was used to generate 1 μmol of ρ-nitroanilide per minute, and the inhibitory activity (%) was calculated from Equation 7.

Elastase inhibitory activity (%) = [1- (A-B) / (C-D)] x 100

A: absorbance of the sample reacted with the addition of enzyme

B: absorbance of the sample reacted without adding enzyme

C: absorbance of the control group reacted without adding sample

D: Absorbance of the control group reacted without adding sample and enzyme

As a result of measuring the inhibitory effect of elastase activity of the functional composition of the present invention, as shown in Table 11, it showed a concentration-dependent inhibition pattern showed more than 50% of the inhibitory effect at 5,000 ppm or more. As a result, it can be seen that the effect of inhibiting elastase activity, which is one of the causes of wrinkles, is excellent, and it can be given a function to suppress the occurrence of skin wrinkles.

Experimental Example 11. Toxicity test through animal experiment

In order to determine the toxicity of the functional composition of Example 2 of the present invention 5-week-old SD rats (male (250 g, 5), female (210 g, 5) received from Samtako (Seorang-dong, Osan-si, Gyeonggi-do) 15 rats were administered orally at a concentration of 3 g / kg with 5 control groups and 10 experimental groups, and survived for 2 weeks for a total health condition including survival, weight, hairiness, bristle, bowel movement, and overall movement. It was confirmed.

As a result of the above experiment, all 10 experimental groups orally administered the functional composition of Example 2 survived and were in good health and no specificity was found (see Table 12).

Experimental Example 12. Clinical Trials

In order to examine the changes in the condition, constitution and skin condition of the functional composition of Example 2 of the present invention, the normal type of women in their 40s, dry skin, 23 people who care about dry skin or cold constitution About 500 mg (two tablets once a day, twice a day) were administered for 8 weeks for a simple clinical trial (see FIG. 2-3).

As a result of the above experiments, most of the subjects showed improvement in ingestion of the functional composition of Example 2 in 13 symptoms such as appetite, drowsiness, and eye fatigue in relation to physical condition and constitution, and a survey related to skin elasticity and wrinkles, etc. Also showed an improvement effect.

It describes an example of the formulation of the functional composition comprising the extract of the complex herbal of the present invention, the present invention is not intended to limit it, but is intended to explain in detail only.

Formulation Example 1 Preparation of Powder

300 mg of the functional composition of Example 2

Lactose 100 mg

Talc 10 mg

The above ingredients are mixed and filled in an airtight cloth to prepare a powder.

Formulation Example 2 Preparation of Tablet

50 mg of the functional composition of Example 2

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

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

Formulation Example 3 Preparation of Capsule

50 mg of the functional composition of Example 2

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare capsules.

Formulation Example 4 Preparation of Healthy Food

1000 mg of functional composition of Example 2

Vitamin mixture proper amount

70 μg of Vitamin A Acetate

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

Vitamin B6 0.5 mg

0.2 μg of vitamin B12

Vitamin C 10 mg

10 μg biotin

Nicotinic Acid 1.7 mg

50 μg folic acid

Calcium Pantothenate 0.5mg

Mineral mixture

Ferrous Sulfate 1.75 mg

Zinc Oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Dibasic calcium phosphate 55 mg

Potassium Citrate 90 mg

Calcium Carbonate 100 mg

Magnesium chloride 24.8 mg

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

Formulation Example 7 Preparation of Healthy Drink

1000 mg of functional composition of Example 2

Citric acid 1000 mg

100 g oligosaccharides

Plum concentrate 2 g

1 g of taurine

Add 900 ml of purified water

The above components were mixed according to a conventional health drink manufacturing method, and the mixture was heated at 85 DEG C for about 1 hour with stirring, and the solution thus prepared was filtered to obtain a sterilized 2-liter container, which was sealed and sterilized, ≪ / RTI >

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

On the other hand, the present invention is not limited to the described embodiments, it is obvious to those skilled in the art that various modifications and variations can be made without departing from the spirit and scope of the present invention. Therefore, such modifications or variations will have to belong to the claims of the present invention.

1 is a view showing the tyrosinase inhibitory activity results of the functional composition of Example 2 of the present invention,

Figure 2 is a view showing the condition and constitution changes appearing when ingesting the functional composition of Example 2 of the present invention,

3 is a view showing a change in skin condition appearing when ingesting the functional composition of Example 2 of the present invention.

Claims (7)

A composite herbal composition having skin aging prevention and cosmetic effects, comprising at least one herbal extract selected from the group consisting of guava leaf, green tea, mugwort, aloe and grape seed. The complex herbal composition of claim 1, wherein the extract comprises at least one or one or more of the complex herbal extracts of guava leaf, green tea, mugwort, aloe vera, and grape seed. According to claim 2, wherein the compositional ratio of the herbal extract extract guava leaf: green tea: mugwort: aloe vera: grape seed (1-3: 1-3: 1-2: 1-2: 1-2) weight ratio (w) / w) complex herbal composition having a skin aging prevention and cosmetic effect, characterized in that. Health functional food for preventing and improving skin aging, comprising at least one herbal extract selected from the group consisting of guava leaf, green tea, mugwort, aloe vera and grape seed. [5] The health functional food for preventing and improving skin aging according to claim 4, wherein the extract comprises at least one or one or more of the complex herbal extracts of guava leaf, green tea, mugwort, aloe vera and grape seed. The composition of claim 5, wherein the compound herbal composition ratio is guava leaf: green tea: mugwort: aloe vera: grape seed (1-3: 1-3: 1-2: 1: 2) weight ratio (w / w) Functional food for preventing and improving skin aging, characterized in that. The health functional food for preventing and improving skin aging according to claim 4, wherein the substance containing the extract as an active ingredient is any one of tablets, capsules, pills, and liquids.

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