KR20210050727A - Composition for skin whitening comprising Lepisorus thunbergianus extract - Google Patents

Abstract

The present invention provides a composition for skin whitening comprising a Lepisorus thunbergianus extract or fractions thereof, and a method for preparing the same. The composition comprising the Lepisorus thunbergianus extract or fractions thereof (ethyl acetate fraction, butanol fraction, hexane fraction, and methylene chloride fraction) of the present invention exhibits excellent tyrosinase inhibitory activity, and is useful as a natural material exhibiting whitening efficacy in functional cosmetics and functional food fields.

Description

Composition for skin whitening comprising Lepisorus thunbergianus extract}

The present invention relates to a composition for skin whitening, and more particularly, to a composition for skin whitening comprising an extract of a leaf herb.

Weeping fern is an evergreen perennial plant of the fern family Goranaceae, and grows attached to the bark of old trees or rocks in the forests of the southern region. The rhizome extends to the side and is covered with scale fragments, and the leaves are leathery, linear, and have smooth edges. The sporangia group has a round shape and runs side by side in a row on both sides, centering on the middle leaf line above the back of the leaf, and there is no vesicle. It has been reported that it is effective in improving blood circulation, anti-cancer activity, and kidney function in relation to the plant, but there has not been any report on the skin whitening function.

On the other hand, tyrosinase is an enzyme containing copper that exists in nature such as mammals, plants, and fungi and accepts substrates such as phenol and catechol. This enzyme is involved in the first two steps of melanin biosynthesis, hydroxylation of L-tyrosine to L-DOPA (L-3,4-dihydroxyphenylalanine), and o-quinone (o-quinone). dopaquinone). Dopaquinone is converted into melanin through a reaction in several steps and acts as a causative agent of skin pigmentation. In the food industry, tyrosinase also serves as a target enzyme that controls the stability of fruits and vegetables. Accordingly, developing a safer and more effective tyrosinase inhibitor is recognized as a very important challenge in the food and cosmetics industry.

Korean Patent Registration No. 10-1690012 (2016.12.21.) Korean Patent Publication No. 10-2011-0086894 (2011.08.02.) Korean Patent Publication No. 10-2010-0080972 (2010.07.14.)

The inventors of the present invention found that while researching on a natural material having excellent skin whitening activity, it can be used as a whitening composition by confirming that the extract and its fractions inhibit tyrosinase activity.

Accordingly, it is an object of the present invention to provide a composition for skin whitening and a method for preparing the same, including an extract or a fraction thereof.

According to an aspect of the present invention, there is provided a cosmetic composition for skin whitening comprising an extract of a single leaf herb.

In one embodiment, the monoleaflet extract _{may be a mixed solvent extract of C 1} to C ₃ alcohol and water, preferably an extract of 50 to 80% (v/v) ethanol.

According to another aspect of the present invention, a cosmetic composition for skin whitening comprising one fraction selected from the group consisting of an ethyl acetate fraction, a butanol fraction, a hexane fraction, a methylene chloride fraction and a mixture thereof Is provided.

According to another aspect of the present invention, (a) _{extracting the monoleaflet with a mixed solvent of C 1} to C ₃ alcohol and water to obtain a monoleaflet extract; (b) suspending the monoleaflet extract obtained in step (a) in water to obtain a monoleaflet extract suspension; (c) mixing the suspension obtained in step (b) with hexane to separate an aqueous layer and a hexane layer, and obtaining a hexane fraction from the hexane layer; (d) the water layer in step (c) is sequentially mixed with methylene chloride, ethyl acetate, and butanol to obtain a methylene chloride fraction, an ethyl acetate fraction, and a butanol fraction.

In one embodiment, the C ₁ to C ₃ alcohol of step (a) may be 50 to 80% (v/v) ethanol.

According to another aspect of the present invention, there is provided a food composition for skin whitening comprising an extract of a single leaf herb.

In one embodiment, the monoleaflet extract _{may be a mixed solvent extract of C 1} to C ₃ alcohol and water, preferably an extract of 50 to 80% (v/v) ethanol.

According to another aspect of the present invention, a food for skin whitening comprising a fraction selected from the group consisting of an ethyl acetate fraction, a butanol fraction, a hexane fraction, a methylene chloride fraction, and a mixture thereof A composition is provided.

According to the present invention, it was found that a composition comprising a monoleaflet extract or its fractions (ethyl acetate fraction, butanol fraction, hexane fraction, methylene chloride fraction) exhibits excellent tyrosinase inhibitory activity, and thus can be used as a whitening composition.

Therefore, the composition comprising the monoleaflet extract of the present invention or its fractions (ethyl acetate fraction, butanol fraction, hexane fraction, methylene chloride fraction) can be usefully used as a natural material exhibiting whitening efficacy in the field of functional cosmetics and functional foods.

1 is a graph measuring _{IC 50} for inhibition of tyrosinase activity of a crude extract.
FIG. 2 is a graph measuring _{IC 50} for inhibition of tyrosinase activity of ethyl acetate fraction of an extract of Japanese herbaceous plants.
FIG. 3 is a graph measuring _{IC 50} for inhibition of tyrosinase activity of a butanol fraction of a Japanese herbaceous extract.
FIG. 4 is a graph measuring _{IC 50} for inhibition of tyrosinase activity of the hexane fraction of an extract of Japanese herbaceous plants.
FIG. 5 is a graph measuring _{IC 50} for inhibition of tyrosinase activity of the methylene chloride fraction of an extract of Japanese herbaceous plants.

The present invention provides a cosmetic composition for skin whitening comprising an extract of a single leaf herb.

In the evaluation of inhibition of tyrosinase enzyme activity, the extract of the present invention showed an IC ₅₀ = 0.461, showing excellent tyrosinase inhibitory efficacy.

In one embodiment, the monoleaflet extract _{may be a mixed solvent extract of C 1} to C ₃ alcohol and water, preferably an extract of 50 to 80% (v/v) ethanol.

The present invention also provides a cosmetic composition for skin whitening comprising one fraction selected from the group consisting of an ethyl acetate fraction, a butanol fraction, a hexane fraction, a methylene chloride fraction, and a mixture thereof as a fraction fractionated from a leaf extract. .

The present invention also provides a cosmetic comprising the cosmetic composition. The cosmetic composition for whitening of the present invention may be used in basic cosmetics, makeup cosmetics, and body care cosmetics, but is not limited thereto.

The basic cosmetics refer to face wash, makeup remover, lotion, emulsion, beauty liquid, face cream, pack, eye care, and other cosmetics for facial skin care. Face wash, such as a solid soap, a face wash foam, and a face wash powder, for example; Makeup removers such as cleansing foam, cleansing cream, cleansing milk, cleansing lotion, cleansing gel, cleansing oil and cleansing mask; Lotions such as liposomal lotion, flexible lotion, astringent lotion, washing lotion, and multilayer lotion; Emulsions such as emollient lotion, moisture lotion, milky lotion, nourishing lotion, nourishing milk, skin moisture, moisture emulsion, massage lotion, and keratin smoother for the face; Cosmetic liquids such as liposomal lotion, moisturizing cosmetic liquid, whitening cosmetic liquid, and ultraviolet ray-preventing cosmetic liquid; Creams such as emollient cream, nourishing cream, moisture cream, night cream, massage cream, cleansing cream, makeup cream, base cream, shaving cream, and keratin softening cream for face; Packs such as peel-off pack, powder pack, washing pack, oil pack, and cleansing mask; Eye care such as eye serum, eye gel, and eye cream; Facial UV protection emulsion, sun protection, sun protector, UV care milk, sunscreen, UV care such as sunscreen cream and suntan cream, gels such as moisture gel, face peeling, and other basic cosmetics such as face slimming agent. I can.

As makeup cosmetics, makeup bases, such as a makeup base, a base cream, a color control base, and a UV-blocking base; Powder concealer and concealer such as cream and liquid concealer; Foundations such as a powder foundation, a UV blocking foundation, a cream foundation, and a UV blocking cream foundation; And face powders such as loose powder, presto powder, face color, and white powder.

Examples of body care cosmetics include body lotions such as body lotion, body oil, and body mist; Body creams such as body cream, body milk, body gel and body mousse; Lip creams such as moisturizing lip cream, UV care lip cream, and color lip cream; Hand creams such as hand cream and hand gel; UV care such as body UV protection emulsion, sun protector, sun protector, UV care milk, sunscreen, sunscreen cream and suntan cream; Fine hair treatment such as depilatory cream, depilatory mousse, depilatory wax, fine hair bleaching agent and shaving cream for body hair; Foot care such as a foot massage agent, a foot slimming agent, a foot peeling agent, an exfoliating agent other than the face such as a heel, and an emollient agent; Limited deodorants such as deodorant lotion, deodorant powder, deodorant spray, and deodorant stick; Insect repellers, such as insect repellent spray, are mentioned.

The cosmetic may be preferably any one selected from the group consisting of skin, mist, serum, lotion, mask pack, eye cream, and nutrition cream.

Each of the fractions (a) _{extracting the monoleaflet with a mixed solvent of C 1} to C ₃ alcohol and water to obtain a monoleaflet extract; (b) suspending the monoleaflet extract obtained in step (a) in water to obtain a monoleaflet extract suspension; (c) mixing the suspension obtained in step (b) with hexane to separate an aqueous layer and a hexane layer, and obtaining a hexane fraction from the hexane layer; (d) sequentially mixing the water layer in step (c) with methylene chloride, ethyl acetate, and butanol to obtain a methylene chloride fraction, an ethyl acetate fraction, and a butanol fraction. I can.

In one embodiment, the C ₁ to C ₃ alcohol of step (a) may be 50 to 80% (v/v) ethanol.

The present invention also provides a food composition for skin whitening comprising an extract of uniweed.

In one embodiment, the monoleaflet extract _{may be a mixed solvent extract of C 1} to C ₃ alcohol and water, preferably an extract of 50 to 80% (v/v) ethanol.

The present invention also provides a food composition for skin whitening comprising one fraction selected from the group consisting of an ethyl acetate fraction, a butanol fraction, a hexane fraction, a methylene chloride fraction, and a mixture thereof as a fraction fractionated from the monoleaflet extract. .

Hereinafter, the present invention will be described in more detail through examples. However, the following examples are for illustrating the present invention, and the scope of the present invention is not limited thereto.

<Example>

1. Preparation of Japanese Herb Extract

30 g of dried single leaf vinegar was pulverized finely using a grinder. 300 ml of 70% (v/v) ethanol was added and extracted under reflux for 12 hours at 80°C. After the extract was immersed, the supernatant was collected. As for the residue, 300 ml of 70% ethanol was added again, and a supernatant was obtained through repeated extraction twice. All of the supernatant was collected and filtration under reduced pressure was performed twice using filter papers (ADVANTEC, No. 2 and No. 1), respectively. The extract was sufficiently concentrated using a vacuum concentrator, placed in a freeze dryer, and completely dried.

2. Preparation of solvent-specific fractions of Japanese herbaceous extract

3 g of ethanol extract obtained in the above extraction process and 30 ml of distilled water were added to a separatory funnel and sufficiently mixed, 70 ml of hexane was added and shaken, and then left until the layers were divided. The water layer was passed from the separatory funnel, collected, transferred to another separatory funnel, and then 70 ml of hexane was added again and the same process was repeated two more times to obtain a hexane fraction.

70 ml of methylene chloride (MC) was added to the remaining water layer, mixed, waited until the layers were divided, and then the MC fraction was collected. The above process is repeated two more times to obtain an MC fraction.

Add 70 ml of ethyl acetate (EA) to the water layer, mix and wait until the layers are divided, then run the water layer to collect it, transfer it to another separatory funnel, add 70 ml of EA again, and repeat the same process twice more. EA fraction was obtained.

Mix water and butanol 1:1, add 70 ml of butanol sufficiently saturated in the water layer, shake and mix, wait until the layers are divided, flow the water layer, collect, transfer to another separatory funnel, and then saturated butanol 70 ml was added and the same process was repeated two more times to obtain a butanol fraction.

Each of the obtained fractions was sufficiently concentrated using a vacuum concentrator, put in a freeze dryer, and dried completely.

3. Evaluation of inhibition of tyrosinase activity

(1) One leaf herb extract and each fraction sample were dissolved in DMSO to make 100 mg/ml, and ascorbic acid, a positive control, was prepared using PBS buffer (pH 6.5). Then, by diluting the ascorbic acid solution using PBS buffer (pH 6.5), prepared by concentration, dispensed by 50 μl, and then prepared a tyrosinase solution [mushroom tyrosinase, 200 U/ml (in PBS buffer, pH 6.5)] to prepare 50 Each μl was added and reacted at 37° C. for 10 minutes.

(2) A 4 mM L-DOPA solution (in PBS buffer) was prepared, 50 μl each was added to the reaction solution, and reacted at 37° C. for 10 minutes.

(3) Dissolve FITC (Fluorescein isothiocyanate) in DMSO to prepare 10 mg/ml, dilute with PBS buffer to prepare a 1.6 μM solution, add 50 μl each, and excitation wavelength using a fluorescence microplate reader. Fluorescence intensity was measured at 480 nm and emission wavelength 540 nm.

4. Results

The results obtained by the above measurement are shown in FIGS. 1 to 5. In the evaluation of the inhibition of tyrosinase activity, the extract of Japanese herbaceous plant itself showed IC ₅₀ = 0.461 mg/ml (FIG. 1). In addition, the ethylacetate fraction of the Japanese herbaceous extract showed IC ₅₀ = 0.217 mg/ml (FIG. 2), the butanol fraction of the Japanese leaf herb extract was IC ₅₀ = 0.125 mg/ml (FIG. 3), and the hexane fraction of the Japanese leaf herb extract was IC ₅₀ = 0.397 mg/ml (FIG. 4), the methylene chloride fraction of the monoleaflet extract showed IC ₅₀ = 0.451 mg/ml (FIG. 5). From the above results, the monoleaflet extract itself exhibited tyrosinase inhibitory activity, and all fractions further fractionated showed superior or similar tyrosinase inhibitory activity than the monoleaflet extract itself, and in particular, the ethyl acetate fraction, the butanol fraction, and the hexane fraction It was found to exhibit excellent tyrosinase inhibitory activity.

Claims (10)

A cosmetic composition for skin whitening comprising a leaf extract. The cosmetic composition according to claim 1, wherein the monoleaflet extract is a mixed solvent extract of _{C 1} to C _{3 alcohol and water.} The cosmetic composition according to claim 1, wherein the monoleaflet extract is an extract of 50 to 80% (v/v) ethanol. A cosmetic composition for skin whitening comprising one fraction selected from the group consisting of an ethyl acetate fraction, a butanol fraction, a hexane fraction, a methylene chloride fraction, and a mixture thereof as a fraction fractionated from a leaf extract. (a) _{extracting the monoleaflet with a mixed solvent of C 1} to C ₃ alcohol and water to obtain a monoleaflet extract;
(b) suspending the monoleaflet extract obtained in step (a) in water to obtain a monoleaflet extract suspension;
(c) mixing the suspension obtained in step (b) with hexane to separate an aqueous layer and a hexane layer, and obtaining a hexane fraction from the hexane layer;
(d) sequentially mixing the water layer in step (c) with methylene chloride, ethyl acetate, and butanol to obtain a methylene chloride fraction, an ethyl acetate fraction, and a butanol fraction.
A method for producing a fraction of a leaf herb extract comprising a. The method of claim 5, wherein the C ₁ to C ₃ alcohol of step (a) is 50 to 80% (v/v) ethanol. A food composition for skin whitening comprising a leaf extract. The food composition according to claim 7, wherein the monoleaflet extract is a mixed solvent extract of _{C 1} to C _{3 alcohol and water.} The food composition according to claim 7, wherein the monoleaflet extract is an extract of 50 to 80% (v/v) ethanol. A food composition for skin whitening comprising one fraction selected from the group consisting of an ethyl acetate fraction, a butanol fraction, a hexane fraction, a methylene chloride fraction, and a mixture thereof as a fraction fractionated from a leaf extract.

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