KR101429117B1 - Composition for skin-whitening comprising extract of calyx of peach blossom - Google Patents

Abstract

The present invention relates to a skin-whitening cosmetic composition and a skin-whitening health functional food containing an extract of a peach blossom calyx (Prunus persica calyx) as an active ingredient. According to the present invention, when a skin-whitening composition containing an extract of a peach blossom calyx is produced, a non-toxic peach blossom calyx does not have to be treated in a toxicity removing step. Accordingly, time and cost are reduced, and not only excellent skin-whitening activities but also melanin secretion controlling activities can be exhibited.

Description

TECHNICAL FIELD The present invention relates to a composition for skin whitening comprising peach calyx extract,

) 추출물을 함유하는 피부 미백용 조성물에 관한 것으로, 더욱 구체적으로 복숭아 꽃받침(도화악,

) 추출물을 유효성분으로 함유하는 피부 미백용 화장료 조성물 및 피부 미백용 건강기능식품에 관한 것이다.
The present invention relates to a peach calyx (Fig.

The present invention relates to a skin whitening composition containing an extract of a peach,

) Extract as an active ingredient, and a health functional food for skin whitening.

Whitening is one of the three functions (whitening, UV protection, wrinkle improvement) of the functional cosmetics currently allowed by the Food and Drug Administration. When skin is exposed to sunlight, melanin secreted from the dermal melanocyte protects the skin It is a phenomenon that the color of the epidermis appears to be black between the epidermis and the dermis. The color of human skin is determined by the amount of melanin, carotene and hemoglobin, among which melanin is the most crucial factor. Melanin is synthesized in melanocytes, a pigmented cell in the basal layer of the epidermis, and transformed into peripheral keratinocytes to determine skin color. Tyrosinase (dihydroxy-L-phenylalanine: oxygen oxidoreductase, EC 1.14.18.1) is a copper-containing enzyme that uses a wide range of phenolic compounds as a substrate. Tyrosine is used as a substrate for L- (L-3,4-dihydroxyphenylalanine) and then transferring it to L-DOPAquinone. The resulting product is subjected to continuous enzymatic oxidation (hydroxylation and oxidation) Melanin is biosynthesized. The melanocyte is activated by the mitosis of the melanocyte due to ultraviolet rays, and the synthesis of tyrosinase is promoted in the activated melanocyte to promote the production of melanin. The melanin produced in an excessive amount is eventually transported and discharged out of the epidermis, Which is a cause of pigmentation. When melanin is abnormally overproduced, it produces spots or freckles. On the other hand, when it is produced in less amount, it causes skin lesions such as albumin. Pigmentation is caused by various stimuli such as ultraviolet rays and inflammation from outside, and the extent of this pigment deposition is the amount of melanocytes in the epidermal basal layer reacted. Central to the mechanism of pigmentation is tyrosinase.

On the other hand, the biochemical mechanism for inhibiting the secretion of melanin is known to be relatively detailed. The first initiator of melanin production starts from tyrosine and is proportional to the increase in the number of melanocytes and is determined by an increase in tyrosinase activity. Tyrosinase catalyzes the oxidation of 3,4-dihydroxyphenylalanine (L-DOPA) to produce DOPA quinone (L-tyrosine) (red), eumelanin (brown), and pheomelanin (brown) by a series of enzymatic reactions such as tyrosinase, tyrosinase related protein-1 (TRP-1) and tyrosinase related protein- Are synthesized. The factors involved in melanin biosynthesis include tyrosinase, dopachrome conversion factor, prostaglandin (PG), interferon (IFN), melanocyte-stimulating hormone hormone, MSH), vitamin D3 (vitamin D3), and histamine (histamine). Arbutin, kojic acid, and hydroquinone are known to be the most effective inhibitors of tyrosinase, but they are known to have powerful whitening effects, but due to some adverse effects, Of a substance having a whitening activity.

Peach (Prunus persica L. Batsch) is a fruit of a peach tree belonging to the rose and cherry tree. It is also called a "copy tree" by its origin. It is a small tree which grows about 3 ~ 5 meters. It is an apple tree, tangerine tree, persimmon tree, It is known to grow a lot later. Peaches are broadly classified into Prunus persica for. Alba Schneider and Prunus persica for. Rubro - plena Schneider. Studies on the activity of inhibiting cholinesterase related to aging and Alzheimer 's disease, Reduction of toxic side effects is a typical effect. Peach contains allergens such as allergens, which are composed of lipid transfer protein, whereas 70% ethanol extract of fruits contains NF-κB, IL-6, TNF-α and histamine in the inflammation reaction of HMC-1 cells Inhibitory anti-inflammatory effects. As a study on the whitening effect of peaches, there is a study on the inhibitory effect of tyrosinase on fruit, seed and leaf extracts, but other studies are insufficient.

On the other hand, Korean Patent Laid-Open No. 10-2007-0032520 (composition for external application for skin containing oriental flower extract), Korean patent for cosmetics, functional health-oriented food, or fancy material using peach (Prunus persica L. Batsch) Korean Patent Laid-Open No. 10-2004-0078499 (cosmetic composition for preventing ultraviolet rays and anti-aging skin containing perillaic acid and extracts of peanuts), Korean Patent Laid- -0041927 (a composition containing an extract of pepper showing antioxidant, anti-cancer and whitening activity) and the like are known. Its functionalities include whitening activity, anti-aging activity, especially wrinkle-improving activity, antimicrobial activity, antioxidation and anticancer activity There is no activity on the support of peach blossom provided by the patent.

The peach blossoms (drawing) used for the existing whitening activity contain specific toxic components. This unique toxic component is cyanoglycoside, which is hydrolyzed by external factors and converted to cyanide. The resulting cyanide inhibits cytochrome oxidase and causes respiratory problems. For this reason, the use of peach flower (drawing) was accompanied by restriction. In order to solve this problem, it is necessary to make effort to extract the shade, extract it in hot water or to detoxify it through microbial fermentation process, JO Yong Jin, Journal of Toxicology and Public Health Vol.23 No.1, 2007.3, 47-53). Therefore, the calyx portion of these flesh or flower parts is relatively less likely to generate such cyanide compounds, and there is an advantage that the cost is low even when raw materials are supplied and received.

)을 무독화시킬 필요가 없기 때문에 시간과 경비를 줄이면서 우수한 피부 미백 활성은 물론 멜라닌 분비 억제 활성을 나타낼 수 있음을 확인하고, 본　발명을 완성하게 되었다.
Accordingly, the present inventors have made extensive efforts to overcome the problems of the prior art. As a result, in the case of a skin whitening composition containing an extract of a peach calyx, the peach calyx

), It is possible to exhibit an excellent skin whitening activity as well as a melanin secretion inhibiting activity while reducing time and expense, and completed the present invention.

Accordingly, it is a main object of the present invention to provide a skin whitening composition containing a peach calyx extract having no toxicity and no restrictions on its use, and having an activity of inhibiting melanin secretion as well as an excellent skin whitening activity.

) 추출물을 유효성분으로 함유하는 피부 미백용 화장료 조성물 및 피부 미백용 건강기능식품을 제공하는데 있다.
Another object of the present invention is to provide a skin whitening composition containing peach calyx extract,

) Extract as an active ingredient, and a health functional food for skin whitening.

) 추출물을 유효성분으로 함유하는 피부 미백용 화장료 조성물을 제공한다.According to one aspect of the present invention,

) Extract as an active ingredient.

)이라고도 하며 심피(心皮)·수술·꽃잎과 함께 꽃을 구성하는 4요소 중의 하나다. 흔히 녹색이나 갈색이다. 꽃받침은 대체적으로 꽃이 시들면 화관과 더불어 건조되어 열매가 커지면 자연적으로 소실 되어진다.In the present invention, the 'peach calyx' is a protection organ of the flower supporting the petals at the outermost of the components of the flower,

) And is one of the four elements that constitute flowers together with heart, skin, and petals. It is often green or brown. Calyx is generally dried together with a corolla when the flower is worn, and it is naturally lost when the fruit grows.

In the present invention, the extract can be extracted with any extraction solvent known in the art, and is preferably extracted with a solvent selected from the group consisting of water, a lower alcohol of C 3-6 and a lower organic acid of C-6 .

In accordance with the present invention, there is provided a cosmetic composition for skin whitening characterized in that the extract has tyrosinase inhibitory activity.

In the examples of the present invention, the extract of calyx peach showed higher tyrosinase inhibitory activity than the extract of peach flower, and as a result, the extract of the peach calyx showed tyrosinase (tyrosinase), which is a factor involved in melanin biosynthesis ), Thereby inhibiting melanin production and exhibiting skin whitening activity (Fig. 3).

In accordance with the present invention, there is provided a cosmetic composition for skin whitening characterized in that the extract has an effect of inhibiting melanin biosynthesis.

In the examples of the present invention, the melanin inhibitory effect was higher in the extract of the peach flower than in the peach flower extract, especially in the ethanol extract of the peach calyx. As a result, the extract of the peach calyx inhibited the melanin biosynthesis, (Fig. 4). The results are shown in Fig.

In the present invention, the composition may be used as a skin lotion, a skin softener, a skin toner, an astringent, a lotion, a milk lotion, a moisturizing lotion, a nutrition lotion, a massage cream, a nutritional cream, a moisturizer cream, a hand cream, Essences, packs, soaps, cleansing foams, cleansing lotions, cleansing creams, body lotions and body cleanser formulations.

The ingredients contained in the cosmetic composition of the present invention may contain, in addition to the above-mentioned extracts, the components conventionally used in cosmetic compositions, such as stabilizers, solubilizers, vitamins, pigments and fragrances, And a carrier.

) 추출물을 유효성분으로 함유하는 피부 미백용 건강기능식품을 제공한다. According to another aspect of the present invention,

) Extract as an active ingredient for skin whitening.

In the present invention, the extract can be extracted with any extraction solvent known in the art and preferably extracted with a solvent selected from the group consisting of water, a lower alcohol of C3-6 and a lower organic acid of C-6 .

In the examples of the present invention, the extract of calyx peach showed higher tyrosinase inhibitory activity than the extract of peach flower, and as a result, the extract of the peach calyx showed tyrosinase (tyrosinase), which is a factor involved in melanin biosynthesis ), Thereby inhibiting melanin production and exhibiting skin whitening activity (Fig. 3).

In the present invention, the above-described extract has a melanin biosynthesis inhibitory effect.

In the examples of the present invention, the melanin inhibitory effect was higher in the extract of the peach flower than in the peach flower extract, especially in the ethanol extract of the peach calyx. As a result, the extract of the peach calyx inhibited the melanin biosynthesis, (Fig. 4). The results are shown in Fig.

In the present invention, the extract may be contained in any one selected from a beverage, a circle, a tablet, a capsule, and a powder and a powder. to provide.

The components contained in the health functional food composition of the present invention can be selected and formulated by those skilled in the art according to the formulations other than the above extract as an active ingredient and can be formulated as a main ingredient of the food or added to other foods . Food-acceptable food additives may be used at this time.

According to another aspect of the present invention, the present invention provides a method for extracting a peach calyx extract comprising the steps of:

(a) removing the flower (fig) from the peach flower and separating only calyx (fig);

(b) drying the sample of step (a);

(c) pulverizing the sample of step (b) with a homogenizer;

(d) extracting the sample of step (c) with a solvent selected from the group consisting of water, a lower alcohol of C3-6 and a lower organic acid of C3-6; And

(e) filtering the extract of step (d) with a filter paper followed by lyophilization.

In the production method of the present invention, after extracting the peach calyx extract, ethanol is added thereto and extracted using an extraction method such as hot water extraction, cold extraction, reflux cooling extraction, or ultrasonic extraction.

In the production method of the present invention, in (a), the peach blossom is obtained by blooming in a peach farm, and obtained in the process of blooming overflowing flower for the removal of superior fruit.

The peach calyx used in the present invention can be obtained by harvesting peach blossom which is obtained by overflowing flowers as mentioned above, and obtaining the peach calyx, so that the present invention can be completed without difficulty in terms of time and expense.

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

Since the extract of the peach flower of the present invention is a natural substance, there is no toxicity, and thus it can be continuously used in large quantities as a health functional raw material and a medicine. B16F10 melanoma cells or RAW264.7 cells treated with water and ethanol extract of the peach flower of the present invention obtained above were treated with a concentration of 10 mg / ml of each extract, assay, it was confirmed that the number of B16F10 melanoma cells and RAW264.7 cells did not show any decrease in number, and that these extracts showed no toxicity to these cells. Therefore, in the long term, It is applicable to skin beauty.

The peach flower bud of the present invention is isolated, its whitening activity is found from its extract, its whitening and melanin inhibitory activity is confirmed, and examples of the preparation of a flavor or cosmetic food composition using the same are described below. I would like to explain more specifically.

Since the peach calyx extract of the present invention inhibits tyrosinase and melanin biosynthesis, it is effective for skin whitening, has no toxicity, and can be used as a skin whitening cosmetic composition for skin care and a health functional food composition for skin whitening .

Fig. 1 is a picture illustrating the shape of a peach flower and a peach calyx (figurine).
Fig. 2 shows the antioxidant activity (ORAC) of the extracts of peach blossoms and peach calyx
Fig. 3 is a picture showing tyrosinase inhibitory activity of an extract of peach blossom and a calyx peach (peach flower).
Fig. 4 is a graph showing the activity of inhibiting melanogenesis of peach blossoms and peach calyx extracts.

Hereinafter, the present invention will be described in more detail with reference to Examples. These embodiments are only for illustrating the present invention, and thus the scope of the present invention is not construed as being limited by these embodiments.

Example  1: calyx peach Dohwa ) ≪ / RTI >

)을 각각 분리하였다. 이 시료를 건조기(MOV-112F, Sanyo, Osaka, Japan)에서 열풍건조(60℃, 12h) 및 동결건조(lyophilizer, 48h, ilShinBioBase Co., Dongducheon, Korea)한 후, 균질기(homogenizer, Sinil, Seoul , Korea)를 이용하여 완전히 분쇄(80 메쉬, 400메쉬)한 다음, 각 시료에 20배(w/v)의 DW(60℃)와 10배(w/v)의 100% 에탄올(45℃)을 이용하여 12시간 동안 추출하였으며, 여과지(filter paper No. 1, Whatman, Buckinghamshire, UK)를 이용하여 불순물을 제외한 용액을 동결건조시켜 분말(100μg/mL)로 만들어 실험에 사용하였다. 본 분말의 일부는 보존 시료로 보존하였다.
(Baekhyang, Gyeongsang) harvested from farms in Qingdao Peach Experiment Station were used as the disclosure material, and flowers (竹 桃, 桃花) and calyx

Respectively. This sample was subjected to hot air drying (60 ° C, 12h) and lyophilizer (48h, ilShinBioBase Co., Dongducheon, Korea) in a drier (MOV-112F, Sanyo, Osaka, Japan) (W / v) of DW (60 ° C) and 10 times (w / v) of 100% ethanol (45 ° C) were added to each sample ) And extracted for 12 hours. The filtrate (filter paper No. 1, Whatman, Buckinghamshire, UK) was freeze-dried except for the impurities to prepare powder (100 μg / mL). A portion of this powder was preserved as a preservation sample.

Example  2: Antioxidant activity measurement

Ferric ion reducing antioxidant power (FRAP) and oxygen radical antioxidant capacity (ORAC) assay were used to measure the antioxidant capacity of the samples. 20 μl of sample or positive control trolox [(±) -6-Hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid, 238813, Sigma] was inoculated into 96-well black plate 160 ng of fluorescein (F2456, Sigma) was added to each well, and the mixture was allowed to stand at 37 占 폚 for 10 minutes. 20 μl of 20 mM AAPH [2-methylpropionamidine) dihydrochloride, 440914, Sigma] was added to the plate and the plate was irradiated with a 490 nm excitation wavelength and a 535 nm emission using a spectrophotometer (Wallac Victor 3 PerkinElmer, MA, USA) Were measured for 70 minutes at 5 minute intervals under the wavelength condition. The area under the curve (AUC) and ORAC value were calculated to compare the antioxidant capacity numerically. The ORAC value was determined based on the values measured in 1 μM trolox to compare the values between the samples.

[Formula 1]

[Formula 2]

The antioxidant activities of water extract (DW) and ethanol extract (EtOH) of peach petal and calyx were measured and the results showed that the reducing power was dependent on the concentration. As a result of measuring FRAP activity using a standard curve of ascorbic acid, a first-order equation of y = 0.176x × 0.084 (y value: absorbance, x value: ascorbic acid μg / mL) And the absorbance was increased. In order to compare the antioxidant effect of the samples numerically, the absorbance of each sample was substituted into the standard curve of ascorbic acid to compare the activity. Based on the concentration of 100 mg / mL of sample, x value of ascorbic acid concentration, which is equivalent to the equivalent antioxidative effect, was calculated by substituting the absorbance y value in the standard curve equation. As a result, the ethanol extract of peach petal (Pf-EtOH), 4.63% of ethanol extract (Pc-EtOH) of peach calyx (Pfu-EtOH), 3.63% of water extract (Pf-DW) of peach petal Pc-DW) in the order of 0.34%. Peach flower extracts were more active than base extracts, and ethanol extracts were more active than DW extracts. Again, ORAC (oxygen radical antioxidant capacity) activity was measured from this data. The ORAC assay is a standardized method for measuring the antioxidant activity. Pf-DW: 5.074, Pf-EtOH: 8.849, and Pc-DW: 1.217 were obtained at the concentrations of 10 μg / ml of the extracts, based on the values measured at 1 μM of the positive control, trolox , And Pc-EtOH: 5.723 (Fig. 2). The highest activity was Pf-EtOH, and the ethanol extracts showed higher activity than the DW extracts and the flower extracts had higher activity than the calyx extracts.

Example  3: Tyrosinase  Measurement of inhibitory activity

The substrate used was 3,4-dihydroxy-L-phenylalanine (D9628, Sigma), and the substrate used was mushroom tyrosinase (T3824, Sigma) Was used. The reaction solution was 0.1 M sodium phosphate buffer (pH 6.4). The enzyme and substrate except the sample were dissolved in sodium phosphate buffer and reacted with the e-tube. All reagents used in the experiment were prepared just before the experiment and preliminarily reacted at 37 ℃ in a water bath. The total amount of each e-tube was 200 μL, and the concentrations of the reagents were 5 units / mL of tyrosinase and 10 mM of 3,4-dihydroxy-L-phenylalanine, respectively. A sample, substrate, and enzyme mixture were prepared in an e-tube, reacted at 37 ° C for 1 hour, and centrifuged at 13,200 rpm for 10 minutes. The solution, in which 200 μL of 1 N NaOH was added to the precipitated product and dissolved, was measured for absorbance at 490 nm with a spectrophotometer. Each group was repeatedly measured three times. The positive control group (N-phenylthiourea, PTU) was used, and the degree of activity of tyrosinase was calculated as a percentage of the absorbance of the untreated control group.

[Formula 3]

On the other hand, we confirmed whether the results of antioxidant activity are related to whitening activity. The amount of L-dopaquinone (L-dopaquinone) produced by mushroom tyrosinase was measured with a spectrophotometer using L-DOPA as a substrate. As a result, the positive control PTU was 3 mM: 84%, 1 mM: 31% , 0.3 mM: 40%, and 0.1 mM: 24%. The inhibitory activity of tyrosinase inhibition by the sample was 40% for Pf-DW, 32% for Pf-EtOH, 53% for Pc-DW and 43% for Pc-EtOH based on the concentration of 10 mg / Respectively. As a result, the highest activity was observed in Pc-DW, and the inhibitory activity was stronger in the DW extract than in the ethanol extract, and stronger in the calyx than in the flower (Fig. 3).

Example  4: Inhibition of melanin biosynthesis

B16F10 melanoma cells (ATCC, CRL-6475, mouse melanoma cells) were incubated in a 6-well plate (1 ml) with 1 mM 3-isobutyl-1-methylxanthine (IBMX, I5879, Sigma) The cells were cultured for 48 hours at a concentration of 2 × 10 ⁵ cells / well in a plate (SPL Lifesciences Co., Ltd., Pocheon, Korea). When cells were grown to about 80%, 1, 3 and 10 μg / mL. After 72 hours of culture, the medium was removed and washed with phosphate buffered saline (PBS). The adherent cells were transferred to an e-tube and centrifuged to obtain a pellet. The shape of the precipitated B16F10 cells was photographed, and values obtained by measuring red (R) -green-blue (RGB) were respectively added to the relative intensity. All analyzes were expressed as means and standard deviations. The significance was tested by Duncan's multiple range test by SAS Statistical Analysis System (NC, USA) (p <0.05).

As a result, the degree of inhibition of melanin biosynthesis in B16F10 cells was compared. The melanin produced in the cells can be easily distinguished visually as a black precipitate as shown in FIG. The value in the group in which melanin production was induced by IBMX was set to 1, the value in the group not induced (IBMX-free group) was set to 0, and the degree of melanin production was calculated on the basis of this value. relative intensity). As a result, the melanin inhibitory effect was not observed at the cell level in the DW extract of the peach flower (Pf) and the calyx peach (Pc), but the ethanol extract showed a high inhibitory activity. (Pc-EtOH) inhibited 60% and 100%, respectively, of 82% inhibition at 10 μg / mL of peach flower ethanol extract (Pf-EtOH) 50% inhibition of cell growth was observed (data not shown) and 100 μg / mL inhibition was observed to be due to this (Fig. 4).

While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will appreciate that such specific embodiments are merely preferred embodiments and that the scope of the present invention is not limited thereby. something to do. It is therefore intended that the scope of the invention be defined by the claims appended hereto and their equivalents.

Claims (9)

Peach calyx

) Extract as an active ingredient.
The cosmetic composition for skin whitening according to claim 1, wherein the extract is extracted with a solvent selected from the group consisting of water, a lower alcohol of C3-6 and a lower organic acid of C-6.
The cosmetic composition for skin whitening according to claim 1, wherein the extract has a tyrosinase inhibitory activity or a melanin biosynthesis inhibitory effect.
The composition according to claim 1, wherein the composition is at least one selected from the group consisting of a lotion (skin lotion), a skin softener, a skin toner, an astringent, a lotion, a milk lotion, a moisturizing lotion, a nutrition lotion, a massage cream, Wherein the composition has a formulation selected from the group consisting of a nutritional essence, a pack, a soap, a cleansing foam, a cleansing lotion, a cleansing cream, a body lotion and a body cleanser formulation.
Peach calyx

) Extracts as active ingredients for skin whitening health food.
6. The skin care whitening health food according to claim 5, wherein the extract is extracted with a solvent selected from the group consisting of water, C3-6 lower alcohol and C-6 lower organic acid.
[Claim 5] The skin health whitening food product according to claim 5, wherein the extract has an inhibitory activity against tyrosinase or melanin biosynthesis.
6. The skin whitening health functional food according to claim 5, wherein the extract is contained in any one selected from a beverage, a circle, a tablet, a capsule, and a powder and a powder. .
A method for extracting a peach calyx extract comprising the steps of:
(a) removing the flower (fig) from the peach flower and separating only calyx (fig);
(b) drying the sample of step (a);
(c) pulverizing the sample of step (b) with a homogenizer;
(d) extracting the sample of step (c) with a solvent selected from the group consisting of water, a lower alcohol of C3-6 and a lower organic acid of C3-6; And
(e) filtering the extract of step (d) with filter paper followed by freeze-drying.

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