KR20170123824A

KR20170123824A - Cosmetic composition comprising paprika extract for whitening and moisturizing effect on the skin and the manufacturing method thereof

Info

Publication number: KR20170123824A
Application number: KR1020160052900A
Authority: KR
Inventors: 윤길영; 김봉우; 장재선; 김희대; 김미경
Original assignee: (주)또르르; 서원대학교산학협력단
Priority date: 2016-04-29
Filing date: 2016-04-29
Publication date: 2017-11-09

Abstract

The present invention relates to a skin whitening and moisturizing composition containing a paprika extract and a method of preparing the same, and a skin whitening cosmetic composition containing the paprika extract of the present invention as an active ingredient has antioxidative effect, inhibits the production of melanin , It can be usefully used for the production of functional cosmetics for improving skin whitening.

Description

TECHNICAL FIELD The present invention relates to a cosmetic composition for skin whitening and moisturizing containing a paprika extract and a preparation method thereof,

The present invention relates to a cosmetic composition for skin whitening and moisturizing containing a paprika extract and a process for producing the same.

Domestic whitening cosmetics is a field that is actively researched and developed compared to other functional cosmetics fields, and it is a field that focuses on the preference of the market, technology for natural extracts, differentiation strategy to survive in global competition, to be.

It contains various natural raw materials and new concept ingredients such as arbutin which is a representative whitening raw material, kojic acid, lucinol, vitamin C, resveratrol, phytoglue EL-1 and white flower combedless, licorice bark bark, It is recognized as a functional cosmetic material that has been recognized as effective.

The whitening products are step by step, line-by-line system, forming a high-end premium brand market. With the elimination of keratin and wastes from the skin, the skin cleansing line that increases the transparency of the skin, intensive skin tone improvement system, In recent years, the system is being developed in a step-by-step system. In recent years, a combined system that simultaneously attempts to improve whitening and wrinkle, protects and maintains the skin is in the limelight, and gives whitening effect to color cosmetics such as eyeshadow.

As a representative whitening ingredient using natural products, most of them use albutin (whitening) and licorice extract (whitening), which are notified raw materials, and there are also increasingly avoiding differentiation by introducing their own developed raw materials and new technology. In Korea, it is used as a whitening cosmetic composition by using ginseng adventitious roots, mahwang, donghwa, bokhwa, top, celestial god, yew tree,

Paprika ( Capsicum annuum L., Bell pepper, Paprika) is an improvement of green pepper. Origin is Central America. It is a low tree type crop with woody stem. In addition, it has a sweet taste, crisp texture, rich in vitamins A and C, and carotene, vitamin C, B6 and p-coumaric acid have been reported as major components of paprika extract.

Accordingly, the inventors of the present invention have found that paprika callus extract is effective for whitening and moisturizing while trying to find a natural product effective for whitening and moisturizing, and completed the present invention.

Korean Registered Patent KR10-0923141 Korean Registered Patent KR10-0967018

Accordingly, an object of the present invention is to provide a cosmetic composition for skin whitening and moisturizing which contains paprika callus extract as an active ingredient.

In order to achieve the above object, the present invention provides a cosmetic composition for skin whitening and moisturizing which contains an extract of paprika callus as an active ingredient.

Since the skin whitening and moisturizing cosmetic composition containing the paprika callus extract of the present invention as an active ingredient has an antioxidative effect, inhibits the production of melanin and has no toxicity, it is useful for the production of functional cosmetics for skin whitening and moisturizing You can use it.

FIG. 1 is a graph showing the results of confirming antioxidative effects of paprika extract (WE, RE) at different concentrations.
FIG. 2 is a graph showing the results of confirming melanocyte cytotoxicity of the hot-water extract of paprika and the ethanol extract. FIG.
FIG. 3 is a diagram showing results of culturing paprika callus and line selection (A) and genotyping (B) using a tissue culture method.
FIG. 4 is a graph showing melanin inhibitory effects on each of paprika callus lines (# 1, # 2, # 3, # 4, # 5, # 6 and # 8).
FIG. 5 is a graph showing melanin production inhibitory effects of the extracts of paprika callus (# 2 and # 6).
FIG. 6 is a diagram showing the result of confirming stability for 6 months in the results of the severe condition test (from the left to the right and 6 months to the right) for the stability test of the whitening product formulation.
Fig. 7 is a view showing the result of analyzing the moisture resistance of a formulation containing paprika callus extract. Fig.

Hereinafter, the present invention will be described in detail.

The present invention provides a skin whitening and moisturizing cosmetic composition comprising an extract of paprika callus as an active ingredient.

In one embodiment of the present invention, the paprika callus extract is preferably, but not exclusively, extracted with hot water and ethanol.

The dried plant material to produce the paprika extract has no problem in the composition study even if it is left for a long time, and the accurate extraction method is determined according to the part of the plant, the condition, the moisture content and the characteristics of the target ingredient. In the case of flowers or leaves, alcohol or methanol is most effective as an extraction solvent. Methanol is boiled on a water bath for 1-2 hours and then filtered through a humidifier. The number of times is determined by referring to the color of the eluent after eluting all components with methanol. When the dried material is extracted, the material is cut to a suitable size or powdered, and then extracted sequentially from a solvent having a low dielectric constant. Generally, the systematic extraction method used is extraction with petroleum ether, Et ₂ O, CHCl ₃ , MeOH, EtOH and H ₂ O in that order. Then organic compounds contained in the plant material were extracted from nonpolar materials in order of increasing polarity.

In one embodiment of the present invention, the paprika callus extract may contain from 0.01 to 30% by weight, preferably from 0.1 to 10% by weight, based on the total weight, of the composition containing from 0.1 to 30% by weight based on the total weight of the composition . If it is contained in an amount of less than 0.1% by weight, it is difficult to exert a desired skin improving effect, and if it is contained in an amount exceeding 10%

In the present invention, the expression "comprising as an active ingredient" means that the composition for external application for skin is capable of exhibiting skin improving effect, for example, improving skin cell proliferation activity and activating do.

In the present invention, paprika ( Capsicum annuum L., Bell pepper, Paprika) is a crop of improved bell pepper origin, which is a Central American tree with a tree-necked stem and a strong group-ignition characteristic. In addition, it has a sweet taste, crisp texture, rich in vitamins A and C, and carotene, vitamin C, B6 and p-coumaric acid have been reported as major components of paprika extract.

In one embodiment of the invention, the paprika callus is

1) culturing paprika callus; And

2) It is preferable to perform genotyping by observing polymorphism through RAPD (Random Amplification of Polymorphic DNA) method which selects callus having excellent antioxidative and whitening function in cultured callus.

In the present invention, the callus is a specific tissue or tissue which is produced when a tissue cut from a plant is cultured in a medium containing plant hormones (auxin and cytokinin), a wound is injured to some kind of plant, It is a cell mass. Usually callus is an amorphous tissue or cell mass that has lost its ability to cause normal organogenesis or tissue differentiation, and is mostly flowcytosed. In a broad sense, it also includes plant tumor tissue caused by infection such as agrobacterium.

When a callus, a undifferentiated cell mass, is formed through tissue culture, that is, the cells in the plant body are immediately directed when they multiply, and are regularly assembled into specific tissues and further into organs. However, when various plant growth hormones are given from the outside, it is interpreted that the control that has been maintained until that time is lifted and the cells grow wild. When the callus obtained by this dedifferentiation is put into a liquid medium having the same composition and shake-cultured, the cells are kept apart from each other and continue to multiply. This callus or cultured cell can be permanently cleaved and propagated by subculture in a new medium, which is essentially different from the differentiated cells in which the whole plant ages and dies. After several generations of subcultured callus or cultured cells are applied to a solid medium from which various plant growth hormones have been removed, eyes and roots come out and individual plants are restored. In other words, plants are restored from one somatic cell that constitutes the plant. Plant tissue cells isolated from the parent plant have the potential to be reproduced as a complete plant, that is, typical performance when cultured in a suitable culture environment. This typical performance is not only of academic importance such as plant embryology, but it is also highly useful for practical use of cosmetic raw materials.

In the present invention, the tissue of the plant for inducing callus may be any part of seed or plant. For example, disinfected or sterilized seeds may be germinated to cut off part of the plant, or some part of the plant may be cut and sterilized. In this way, callus is induced through aseptic tissue culture method. Preferably, a piece of plant is cultured in a tissue culture medium. As the above medium, a medium such as MS (Murashige & Skoog, 1962), N6 (Chu et al. 1975) medium, B5 (Gamborg et al., 1968), NN (Nitsch and Nitsch, 1967) . Plant growth regulators such as 2,4-D (2,4-dichlorophenoxyacetic acid), 2,4,5-T (2, 2, 3, , 4,5-trichlorophenoxyacetic acid, Dicamba (2-methoxy-3,6-dichlorobenzoic acid), Indole-3-acetic acid, IBA (Indole-3-butyric acid)

2-methyl-4-chlorophenoxyacetic acid (MCPA), 1-naphthylacetic acid (NAA), 2-naphthyloxyacetic acid (NOA), and 4-amino- 2,5,6-trichloropicolinic acid (Picora), and Cytokinin (6-benzylaminopurine), 2iP (N6- (2-isopentyl) adenine), Kinetin (6-furfurylaminopurine), Thidiazuron (1-phenyl- 3- (1,2,3-thiadiazol- , And zeatin (4-hydroxy-3-methyl-trans-2-butenylaminopurine) are used at plant concentrations appropriate for plant species, stage, area and ripening. In addition, in order to further enhance various physiological activities of callus induced by growth regulators, secondary metabolites such as alkaloids, terpenoids, prebonoids, carotenoids, phenol compounds and glycosides may be added to plant species May additionally be added.

A preferred method of inducing and culturing callus is summarized as follows. A part of the plant tissue is disinfected and cultured in a medium containing auxin or cytokinin growth regulator in the plant basal medium. After culturing for 2 ~ 3 weeks, if the cell of the tissue divides into cells and the callus, which is a mass of cells, is formed, the callus alone is separated and placed on a new medium, and then subcultured by observing the callus state at a certain time interval.

A skin whitening and moisturizing cosmetic composition containing the paprika callus extract according to the present invention can be prepared.

The cosmetic composition of the present invention may contain an acceptable carrier in cosmetic preparations. Herein, "an acceptable carrier for a cosmetic preparation" is a known or used compound or composition that can be contained in a cosmetic preparation, or a compound or composition to be developed in the future, which is toxic, instable or irritating It says nothing.

The carrier may be included in the skin topical composition of the present invention in an amount of from about 1% by weight to about 99.99% by weight, preferably from about 90% by weight to about 99.99% by weight of the composition, based on the total weight thereof. However, since the ratio varies depending on the above-described formulation in which the composition for external application for skin of the present invention is prepared, and its specific application site (face, neck, etc.) or its preferable application amount, And should not be construed as limiting the scope of the invention.

Examples of the carrier include alcohols, oils, surfactants, fatty acids, silicone oils, humectants, moisturizers, viscosifiers, emulsifiers, stabilizers, sunscreens, coloring agents and perfumes. The compounds / compositions that can be used as the above-mentioned alcohol, oil, surfactant, fatty acid, silicone oil, humectant, humectant, viscosifier, emulsion, stabilizer, ultraviolet screening agent, coloring agent and fragrance are already known in the art The appropriate substance / composition can be selected and used.

When the cosmetic composition is manufactured from a cosmetic composition, the cosmetic composition of the emulsified formulation includes nutritional lotion, cream, essence and the like. It can also be prepared in the form of adjuvants which can be applied topically or systemically, which are conventionally used in the field of dermatology by containing a dermatologically acceptable medium or base.

In addition, suitable cosmetic formulations include, for example, solutions, gels, solid or kneaded anhydrous products, emulsions obtained by dispersing an oil phase in water, suspensions, microemulsions, microcapsules, microgranules or ionic forms (liposomes) May be provided in the form of a follicular dispersing agent of the type, cream, skin, lotion, powder, ointment, spray or conical stick. It can also be prepared in the form of a foam or an aerosol composition further containing a compressed propellant.

Hereinafter, And The present invention will be described in more detail by way of examples. These preparation examples And The examples are intended to illustrate the present invention in more detail, And It is not limited to the embodiment.

< Preparation Example > Preparation of paprika extract

It was classified into light yellow (WE) and red (RE) according to the color of paprika. 30 g of each of the dried outposts is added to 200 ml of distilled water and 70% (v / v) ethanol, and the mixture is stirred for 24 hours. The extracted paprika juice is filtered with 4 layers of miracloth, then filtered with filter paper and concentrated to a final concentration of 30% using a vacuum concentrator. The protein content of the paprika extract is measured and standardized.

< Example 1> paprika Antioxidant efficacy of extracts

DPPH auto-oxidation inhibitory effect of paprika extract was confirmed.

Specifically, the free radical scavenging ability of DPPH was confirmed by treating the paprika extract extracted by the method described in the above preparation example with concentrations (0, 10, 20, 30, 40, 50 and 60 ug / ml) Vitamin C (ascorbic acid) was used.

As a result, it was confirmed that the extracts (WE, RE) according to the paprika radish color had antioxidative effects in a concentration-dependent manner (see FIG. 1).

< Example 2> Cytotoxicity analysis of paprika extract

(B16-F10) was subjected to concentration-dependent treatment in order to confirm whether or not the hot water extract of paprika (DW) and the ethanol extract (EtOH) of the red hull extracted by the method described in the above preparation example showed toxicity in melanocytes (B16-F10) The degree of cell death was confirmed.

As a result, it was confirmed that the paprika hydrothermal extract and the ethanol extract showed toxicity to melanocytes at a concentration of 200 ug / ml or higher (see FIG. 2).

< Example 3> Selection of paprika cell line and paprika Callus Extract preparation

<3-1> Paprika Callus Culture and cell line selection

In order to select paprika callus and cell line using tissue culture technique, we investigated callus growth rate according to culture conditions and established excellent culture conditions.

The paprika was immersed in 70% ethanol for 2 minutes, then washed with sterilized water, disinfected with disinfectant (10% lactose) for 10 minutes, and then washed again with sterilized water three times. The culture medium was prepared by adding 8 g / L agar and 30 g / L sucrose to MS base medium (Murashige and Skoog, Duchefa), adding solid medium (pH 5.8) supplemented with plant hormone 2.0 mg / L BA and 0.1 mg / (5 x 5 mm). Petri dishes with paprika sections were cultured in an incubator (25 ° C, RH 65%) and induction of calli. Slices and induced calli were subcultured in the same medium every 4 weeks. The induced calli were classified according to the color of paprika and callus induction period and used for cell line selection.

Genotyping was performed using the RAPD method to analyze the genetic characteristics of the selected cell lines. The random primers used for RAPD analysis were Operon primers (Operon, USA) The solutions and conditions are shown in Table 1 below.

Reagent Amount (ul) 10X reaction buffer 2 50 uM dNTP 4 15 mM MgCl ₂ 2 Primer 2 Taq polymerase One Genomic DNA 4 Total 20

PCR was carried out by predenaturation at 95 ° C for 10 min, denaturation at 94 ° C for 1 min, annealing at 43 ° C for 1 min, extension at 72 ° C for 1 min, followed by final extension at 72 ° C for 10 min, using ASTEC (PC320) % agarose gel electrophoresis. To confirm the genetic relationship between the cell lines, RAPD using the first sample was used to identify the polymorphism pattern according to the cell line using primers # 01 to # 04 using 5 samples. Seconds were performed using 10 selected green and 10 red and 9 red specimens and primer # 07, and the third was performed using 14 cell lines with primer # 09 (see FIG. 3A).

As a result of RAPD analysis using the selected cell line, possibility of confirming polymorphism according to the cell line was observed, and a line with high whitening function activity was first selected and the genotype of the highly active line was confirmed (see FIG. 3B).

<3-2> Paprika Callus Extract preparation

① High pressure extraction and freeze drying

In the above Example <3-1>, the efficacy related to whitening was examined using the first selected sample based on the growth rate and morphological characteristics of the cells, and the selected cells were extracted with a high pressure extractor, The relevant efficacy was tested.

② Organic solvent Extraction and concentration

During the tissue culture, the cell line was cultured according to the growth and morphological characteristics of the cells.

Specifically, 70% EtOH was adjusted to 10% with paprika cell tissue and then shaken at room temperature for 24 hours (Vision VS-250, Korea). The supernatant was collected by centrifugation, and the supernatant was concentrated under reduced pressure (EYEA N-1000, Japan), and then subjected to whitening function and related tests.

< Example 4> paprika Callus Analysis of inhibitory effect of extract on melanogenesis

Melanocytes (B16-F10 cell) Since B16-F10 melanoma is derived from melanoma of mouse skin and is used for inhibition of melanin production, B16-F10 cells were used as growth medium supplemented with 10% FBS and antibiotic antimycotic in DMEM medium .

Cell lines producing melanin pigments were cultured in a humidified incubator at 37 ° C, 5% CO ₂ , and subcultured when grown to 80% of culture dishes every 2-3 days. B16-F10 cells were cultured and the cells per well were plated on a 24-well plate at 1 × 10 ⁵ cells / well. The paprika cell lines (# 1, # 2, # 3, # 4, # 5, # 8), and cultured for 48 hours. Vitamin C (ascorbic acid) was used as a control reagent (con). After 48 hours of incubation, each well was washed with PBS, and 400 ul of 0.2N NaOH solution was added and dissolved at 60 ° C for 1 hour. Absorbance was measured at 405 nm using a mocroplatereader to determine the amount of melanin dissociated into the solution.

As a result, it was confirmed that the melanin inhibitory effect of each line of the paprika cell lines (# 1, # 2, # 3, # 4, # 5, # 6 and # 8) (See Fig. 4).

< Example 5> paprika Callus Analysis of inhibitory effect of melanin on the concentration of extract

In Example 4, melamine production inhibitory effect was confirmed according to the concentration of paprika cultured cell lines # 2 and # 6 having the best melanin production inhibitory effect.

Specifically, cell lines producing melanin pigments were cultured in a humidified incubator at 37 ° C, 5% CO ₂ , and subcultured when grown up to 80% of culture dishes every 2-3 days. B16-F10 cells were plated on a 24-well plate at 1 × 10 ⁵ cells / well in a 24-well plate, and the extracts of paprika cell lines (# 2 and # 6) %) And cultured for 48 hours. Vitamin C (ascorbic acid) was used as a control reagent (con). After 48 hours of incubation, each well was washed with PBS, and 400 ul of 0.2N NaOH solution was added and dissolved at 60 ° C for 1 hour. Absorbance was measured at 405 nm using a mocroplatereader to determine the amount of melanin dissociated into the solution.

As a result, it was confirmed that the melanin inhibitory effect was excellent according to the concentration of paprika cultured cell lines # 2 and # 6 (see FIG. 5).

< Example 6> paprika Callus Formulation analysis containing extracts

<6-1> Paprika Callus Stability analysis of formulations containing extracts

In the case of natural materials, it is known as an ideal material that can satisfy the application feeling requirements, but chemical stability is pointed out as a disadvantage and harmony with the basic composition (proper concentration and ratio) In order to maintain the chemical stability and functionality of the material, experiments were conducted to evaluate the stability and functionality of the materials using the basic formulation to evaluate the mixing ratio of the materials having proper concentration and complementary function.

Specifically, for the stability test of the formulation, the layer separation phenomenon and the browning were periodically observed under the severe condition. In the severe condition (37 ° C, RH 60%), (See Fig. 6).

Therefore, the product was manufactured using excellent formulations by checking the spreadability and the like.

<6-2> Paprika Callus The composition mixing ratio of the formulation containing the extract and Application feeling analysis

The stability, safety, stability and sensory evaluation of the functional ingredients and mixing ratios were investigated in order to maintain the stability of each formulation and smoothness of application.

As part of the process of deciding products that can satisfy the product's superiority and consumer's preferences, we performed skin sensitization experiments to confirm the application feeling. After the skin sensitization experiment, no skin specific phenomenon was found and the result was consistent with the toxicity test and the safety of the formulation was proved.

In addition, the final formulation was selected as the essence by the sensory test such as the color and flavor of the formulation. As a prototype for the sensory test was developed, the improvement and complementary items were evaluated through the internal evaluation. As a result, The higher the product, the lower the degree of oiliness, but both formulations were rated higher than the basic formulations in overall satisfaction. However, the BLAND TEST performed slightly different depending on the individual (skin type).

Therefore, even when considering the fact that it is evaluated according to the subjective standard as shown in Table 2 below, it is confirmed that a good level of formulation and composition can be obtained by integrating skin sensitization and sensory evaluation results of the prototype product.

Satisfaction of sensory evaluation result of prototype Evaluation items * Overall satisfaction Moisture responsiveness Oil level transparency Absorption degree Formulation 1
(essence) ◎◎◎ ○○○ Xx Xx ○○○ ○○○ Formulation 2
(cream) ◎ ◎ Xx Xx ○○○ ○ Xx Formulation 3
(Basic formulation) ◎ ◎ Xx Xx Xx Xx Xx

* Subjective evaluation results (n = 21)

Results of skin sensitization after skin sensitization experiment division Erythema edema itch Burning sensation Stiffness Tingling Other Mild X X X X X X X

< Example 7> paprika Callus Of the formulation containing the extract Moisture power analysis

In order to analyze the moisturizing power of the formulation containing paprika callus extract, a formulation containing paprika callus extract was applied to the subject to analyze the skin moisturizing ability.

Specifically, 22 subjects who met the selection criteria and did not fall under the exclusion criteria were selected, and the formulation containing paprika callus extract ² mg / cm ² Was applied once to the face of the subject. The instrument was evaluated using a corneometer and the stability of use by a dermatologist was evaluated.

As a result of the test of 22 subjects, the test site coated with the formulation containing the extract of porkpurica callus was statistically significantly higher (P <0.05) than the control site coated with the comparative product [moisturizing cream 2] It showed an effect of improving the moisture content of the stratum corneum and having a relatively superior skin moisturizing effect (see FIG. 7).

The present invention has been described with reference to the preferred embodiments.

Those skilled in the art will appreciate that the present invention is not limited to the embodiments described herein,

It is understood that the present invention can be implemented in a modified form without departing from the qualitative characteristics.

It will be possible. It is therefore to be understood that the disclosed embodiments are not to be considered in a limiting sense,

Should be taken into account. The scope of the present invention is not limited to the above description,

And all differences within the scope of equivalents are intended to be embraced by the present invention

.

Claims

A cosmetic composition for skin whitening and moisturizing containing paprika callus extract as an active ingredient.

The cosmetic composition for skin whitening and moisturizing according to claim 1, wherein the paprika callus extract is extracted with hot water and ethanol.

The cosmetic composition for skin whitening and moisturizing according to claim 1, wherein the paprika callus extract contains 0.1 to 10% by weight based on the total weight of the composition.

The method of claim 1, wherein the paprika callus is
1) culturing paprika callus; And
2) genotyping is performed by RAPD (Random Amplification of Polymorphic DNA) method which selects callus having excellent antioxidative and whitening function in cultured callus, and polymorphism is observed and selected A cosmetic composition for whitening and moisturizing.

The method according to claim 1,
The cosmetic composition may be in the form of an emulsion, cream, lotion, essence, pack, gel, powder, lipstick, makeup base, foundation, lotion, ointment, gel, patch, essence, cleansing foam, cleansing cream, cleansing water, Wherein the cosmetic composition is at least one selected from the group consisting of a cosmetic and a cosmetic composition.