KR20110047824A - Cosmetic composition containing citron extract - Google Patents

Abstract

Embodiments of the present invention relates to a cosmetic composition comprising a citron extract.

Embodiments of the present invention are directed to a method of preparing a cosmetic composition comprising the citron extract.

Cosmetic composition comprising citron extract according to the embodiments of the present invention has a whitening, anti-aging, antioxidant and anti-inflammatory effect.

Citron Extract, Flavonoids

Description

Cosmetic composition containing citron extract {Cosmetic Composition Comprising Citron Extracts}

Embodiments of the present invention relates to a cosmetic composition comprising a citron extract.

Citron is a kind of citrus produced in Northeast Asia including Korea, China and Japan. Korea has the strongest fragrance and thickest skin. Korea's major producing regions are Goheung, Wando, Jangheung, Jindo, and Jeju Island, Jeollanam-do.

As the main ingredient of citron, vitamin C contains three times more than lemon, which is good for colds and skin care, and contains many organic acids that prevent aging and fatigue. In addition, vitamin B, sugars, and protein are more than other citrus fruits and contain flavonoids that protect capillaries, thereby preventing cerebrovascular disorders and wind. It also drains and excretes the waste that builds up in the body.

Flavonoid and limonoid compounds present in citrus fruits have a very high bioactive effect. Flavonoids such as naringenin, hesperidin, and nobiletin in citron seeds have been reported to be effective in treating colorectal cancer. It has also been reported to inhibit expression and inhibit lymph node or lung metastasis of these tumors. In addition, it has been reported that citrus flavonoids inhibit the activity of tumor growth promoters, and also inhibit the activity of anticancer drug resistance genes of cancer cells that pump out therapeutic drugs entering cancer tissues.

There are also studies showing that flavonoid compounds lower blood cholesterol levels and that citrus flavonoids have antiviral, antibacterial and antifungal effects by acting like vitamins. Citrus limonoid compounds, in particular, limonin (limonin) and nomilin (nomilin) has been reported to suppress stomach cancer, lung cancer, skin cancer, etc. Recently, studies have been reported to effectively inhibit breast cancer.

According to the domestic functional cosmetics law, functional cosmetics are defined as having an effect on anti-aging wrinkle prevention, whitening, and skin protection against ultraviolet rays. Skin aging can be divided into photoaging observed in skin exposed to ultraviolet rays for a long time and intrinsic aging (chronological aging) observed in non-exposed areas, and more than 70% of the causes of skin aging are caused by photoaging. have. Ultraviolet rays are known to have the greatest effect on skin aging, among other energy sources from the sun. The effect of ultraviolet rays is that wrinkles are formed when UV rays are irradiated to hairless mice. Also, when UV rays are irradiated to cultured fibroblasts, expression of collagen (procollagen), which is a skin protein, decreases and MMP- The importance of 1 expression is increased.

Basically, all functional cosmetics use functional physiologically active materials that can prevent skin damage to ultraviolet rays. Recently, the global trend of technological development is actively developing antioxidant materials obtained from natural products. Therefore, it is possible to develop ultraviolet light products using known components of citron hesperidin, carotenoids and vitamin C. In particular, hesperidin has a great potential as a novel flavonoid.

Numerous studies have been published in addition to those illustrated above that the flavonoid and limonoid compounds present in citrus fruits have a very high physiological activity. Seeds in particular are known to contain much more of these substances than rinds and tissues. However, in the country, the use of citron is mainly used for the production of citron, and the seeds which occupy about 10-15% of the whole fruit are disposed of all 1,800 tons per year.

Therefore, the development of a new functional cosmetic material using citron seed extract, which is a citron by-product, is likely to be converted into a high value-added product from industrial waste to a new functional cosmetic, and thus, it is very necessary to develop a new functional cosmetic material through citron seed extract. Situation.

Embodiments of the present invention to provide a cosmetic composition comprising a citron extract.

In order to solve the above problems, embodiments of the present invention provides a cosmetic composition comprising a citron extract.

Embodiments of the present invention also provide a method of extracting the citron extract from the citron.

The cosmetic composition according to the embodiments of the present invention has a skin aging inhibitory effect, a photoaging inhibitory effect and a whitening effect through skin irritation relief by ultraviolet rays.

Embodiments of the present invention provide a cosmetic composition comprising a citron extract.

In another embodiment of the present invention, the citron may use only citron rind or citron seed, and the citron extract of the present invention may include flavonoids such as naringenin, hesperidin, nobiletin and the like. In addition, the citron extract of the present invention may include limonin (limonin) and nomilin (nomilin).

Embodiments of the present invention provide a cosmetic composition comprising a citron extract.

As can be seen through the following examples, the cosmetic composition comprising the citron extract according to an embodiment of the present invention is excellent in anti-aging, anti-inflammatory and whitening effect. Therefore, it can be usefully used in the manufacture of functional cosmetics for preventing skin aging or whitening.

In one embodiment of the invention, in addition to the cosmetic composition comprising the citron extract of the present invention, fatty substances, organic solvents, solubilizers, thickening and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents ), Fragrances, surfactants, water, ionic or non-ionic emulsifiers, fillers, metal ion sequestrants and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles Or additives commonly used in the field of dermatology, such as any other ingredients commonly used in external preparations for the skin. The ingredients may also be introduced in amounts generally used in the field of dermatology.

According to one embodiment of the present invention, the cosmetic composition containing the citron extract is used, but the cosmetic composition is not limited thereto. For example, supple cosmetics, nourishing cosmetics, massage creams, nourishing creams, packs, gels, essences, Lipstick, makeup base, foundation, lotion, ointment, gel, cream, cleansing, face wash, soap, shampoo, rinse, treatment and essence. Such cosmetics may include conventional ingredients such as aqueous vitamins, oily vitamins, polymer peptides, polymer polysaccharides, sphingolipids, and the like, and may be readily prepared according to techniques well known to those skilled in the art.

According to one embodiment of the present invention, the content of the cosmetic including the citron extract may be 0.001 to 30% by weight, 0.001 to 15% by weight, or 0.001 to 5% by weight based on the total weight of the cosmetic composition. Preferably from 0.02 to 0.2% by weight is used.

Embodiments of the present invention provide a method for preparing citron extract.

Citron is classified by skin, genus seeds and seed shells and lyophilized by classified parts. Thereafter, stored at low temperature in a dry powder state. As for the temperature to store, -50 degreeC --5 degreeC is suitable, Preferably -30 degreeC --10 degreeC is good.

Each powder is hydrothermally extracted with an organic solvent. At this time, the amount of the organic solvent is preferably used 500 parts by weight to 1500 parts by weight relative to 100 parts by weight of powder. The temperature of the hot water is 40 ° C to 80 ° C, preferably 50 ° C to 70 ° C.

The organic solvent may be an organic solvent commonly used in the art, and examples thereof include alcohols such as phenoxyethanol, propiparaben, oxyphenol, benzophenone, and ethanol, and preferably ethanol. do.

The extract is concentrated 7 times to 13 times, preferably 8 times to 12 times under reduced pressure to prepare a sample, and then stored in an ultra-cold refrigerator having a temperature of -50 ° C or less, preferably -60 ° C to -80 ° C.

Hereinafter, examples will be described in detail to help understand the present invention. However, the following examples are merely to illustrate the content of the present invention is not limited to the scope of the present invention. The embodiments of the present invention are provided to more completely explain the present invention to those skilled in the art.

< Example  1> Citron Extract Manufacturer

1 is a schematic diagram of a process for preparing a sample containing citron extract.

After the production of Goheungsan Citron, the frozen by-products were dissolved and classified into skins, soybean seeds and seed shells. Freeze-drying for each classified site and stored at -20 ℃ for each site in dry powder form. At the time of extraction, 20 g of each powder was extracted with hot water at 60 ° C. with 200 ml ethanol. Extract was concentrated 10 times under reduced pressure to prepare a sample, and then stored in -70 ℃ ultra low temperature freezer.

Since Examples 2 to 6 are cell experiments (in vitro), the samples were further freeze-dried to obtain a dried powder, which was then dissolved in dimethyl sulfoxide. At this time, 1 part by weight of dry powder relative to 100 parts by weight of dimethyl sulfoxide was used.

< Example  2> Evaluation of cytotoxicity and cell growth promoting effect MTT assay )

HaCaT Keratinocytes were placed in DMEM (Dulbecco's Modified Eagle's Medium) medium containing penicillin (100 IU / mL), streptomycin (100 g / mL) and 10% FBS (fetal bovine serum) and maintained at 37 ° C. And cultured in an incubator containing 5% carbon dioxide. The obtained keratinocytes were dispensed into 12-well plates at 1 × 10 ⁵ per well and incubated for 24 hours under the same cell culture conditions. After incubation, the extract was added to the cells at a concentration of 0.0025% and 0.005%, and then cultured for 24 hours. After that, the number of cells was measured to measure the degree of cell death and cell growth promoting effect. The results are shown in FIG.

The experiment was divided into UV treated experimental group (Fig. 2a) and untreated experimental group (Fig. 2b). There was no cytotoxicity at concentrations of 0.0025% and 0.005% of citron extract and showed cell growth promoting effect on UV.

< Example  3> Inhibition of inflammatory reactions caused by ultraviolet rays

In general, when skin cells, especially keratinocytes, which are epidermal cells, are irradiated with ultraviolet light, reactive oxygen species are formed. In addition, the inflammatory cytokines such as TNF-a and IL-1a are increased, and the action of AP- It is known that a series of skin aging processes proceed as transcription factors are activated. The keratinocytes in culture were irradiated with UV light and simultaneously treated with citron extracts to reduce these inflammatory cytokines.

HaCaT Keratinocytes were placed in DMEM (Dulbecco's Modified Eagle's Medium) medium containing penicillin (100 IU / mL), streptomycin (100 g / mL) and 10% FBS (fetal bovine serum) and maintained at 37 ° C. And cultured in an incubator containing 5% carbon dioxide. The obtained keratinocytes were dispensed into 12-well plates at 1 × 10 ⁵ per well and incubated for 24 hours under the same cell culture conditions. After culturing, the cells were irradiated with 12.5mJ / Cm ² of ultraviolet rays, and then citron extract was added at a concentration of 0.0025% and 0.005%, and further cultured for 24 hours. After culturing, the culture supernatant was recovered and the amount of inflammatory cytokine TNF-α was measured. The measurement results are shown in FIG. 3.

3, the secretion of TNF-α, an inflammation-induced cytokine (pro-inflammatory cytokine) in which the secretion of citron extracts is increased by ultraviolet rays at 0.0025, 0.005% concentration, is reduced.

< Example  4> Collagen Degrading Enzyme Determination Anti-aging Experiment

Normal human fibroblasts are placed in DMEM (Dulbecco's Modified Eagle's Medium) medium containing penicillin (100 IU / mL), streptomycin (100 g / mL) and 10% FBS (fetal bovine serum). The culture was maintained in an incubator containing 5% carbon dioxide. The obtained fibroblasts were dispensed into 6-well plates at 1 × 10 ⁵ per well and incubated for 24 hours under the same cell culture conditions. After incubation for 24 hours in a medium not containing FBS, and irradiated with 15mJ / Cm ² of ultraviolet rays to the cells after the addition of the citron extract at each concentration was further cultured for 72 hours. After the culture, the culture supernatant was collected and the amount of collagen degrading enzyme (MMP) was measured by Western blot. The results are shown in FIG.

Referring to Figure 4, it can be seen that the secretion of collagen degrading enzymes (MMPs), which are increased by the ultraviolet rays at concentrations of 0.0025 and 0.005% of the citron extract is reduced.

< Example  5> Review of whitening effect by measuring melanin secretion

Mouse-derived malignant melanin tumor cell line B16F1 (KCLB 8007) was placed in DMEM (Dulbecco's Modified Eagle's Medium) medium containing penicillin (100 IU / mL), streptomycin (100 g / mL) and 10% FBS (fetal bovine serum) Culture was maintained in an incubator containing 5% carbon dioxide while maintaining 37 ℃. The obtained melanin tumor cells were seeded in 1 × 10 ⁴ cells / well in a 24-well plate and cultured for 1 day. After incubation, the medium was removed, and in order to induce melanin synthesis, melanin synthesis was induced by adding 5 nM of alpha-MSH (melanocyte stimulating hormone) except for the untreated group. (Positive control) DMEM medium to which 800 μM was added was added and cultured for 3 days. Cells of each treatment group were harvested, 10% DMSO (dimethyl sulfoxide) and 100 μl of 1N NaOH were added to dissolve melanin, and the melanin content was measured by measuring absorbance at 405 nm. The results are shown in FIG.

Referring to Figure 5, it can be seen that the amount of secreted melanin secreted by the α-MSH at the concentration of 0.02% citron extract decreased in the skin and skin. In particular, at 0.02% concentration, it showed similar activity to Kojic acid as a standard.

< Example  6> Tyrosinase  Inhibitory activity measurement effect

Mouse-derived malignant melanin tumor cell line B16F1 (KCLB 8007) was placed in DMEM (Dulbecco's Modified Eagle's Medium) medium containing penicillin (100 IU / mL), streptomycin (100 g / mL) and 10% FBS (fetal bovine serum) Culture was maintained in an incubator containing 5% carbon dioxide while maintaining 37 ℃. The obtained melanin tumor cells were seeded in 1 × 10 ⁴ cells / well in a 24-well plate and cultured for 1 day. After incubation, the medium was removed, and in order to induce melanin synthesis, melanin synthesis was induced by adding 5 nM of alpha-MSH (melanocyte stimulating hormone) except for the non-treated group, and at the same time, citron extract and kojic acid (Positive control group) 800 μM was added to the DMEM medium and cultured for 48 hours, and then cells were lysed using RIPA buffer. The amount of protein was equally corrected by using a Laemmli sample buffer to which β-mercaptoethanol was not added through the Bradford assay. After heating the protein and skipping 8% SDS-PAGE (Sodium Dodecyl Sulfate Poly Acrylamide Gel Electrophoresis), the gel was equilibrated while shaking with 0.1M SPB (Sodium phosphate buffer, pH 6.8) for 30 minutes. After one additional round of equilibration, staining was dissolved in 5 mM L-DOPA in SPB and allowed to react at 37 ° C for about 1 hour until color development appeared. The results are shown in FIG.

Referring to Figure 6 it can be seen that the amount of tyrosinase involved in melanogenesis by α-MSH at the concentration of 0.02% citron extract is reduced in the skin and peel. In particular, at 0.02% concentration, it showed better activity than the standard kojic acid.

Having described the specific parts of the present invention in detail, it will be apparent to those skilled in the art that such specific descriptions are merely preferred embodiments, and thus the scope of the present invention is not limited thereto. will be. Thus, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

1 is a schematic diagram of a process for preparing a sample containing citron extract.

2 is a cytotoxicity and cell growth promoting effect evaluation (MTT assay), a is a group treated with ultraviolet rays, b is a group not treated with ultraviolet rays.

Figure 3 is a graph of the results of the inflammatory response inhibition test induced by ultraviolet light.

4 is a result of measuring the amount of collagenase enzyme secretion.

5 is a graph of Secreted Melanin Assay results.

6 shows the results of measuring tyrosinase inhibitory activity.

Claims (12)

Cosmetic composition containing citron extract. The method of claim 1, Citron extract is a cosmetic composition derived from citron shell and citron seed. The method of claim 1, Fatty substances, organic solvents, solubilizers, thickening and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water, ionic or non-ionic emulsifiers, fillers, metal ion containment And a chelating agent, preservative, vitamin, blocker, wetting agent, essential oils, dyes, pigments, hydrophilic or lipophilic actives and a cosmetic composition further comprising at least one additive selected from the group consisting of lipid vesicles. Classifying and crushing the citron by portions and lyophilizing; Hydrothermal extraction of the lyophilized powder with an organic solvent, and Method for producing a citron seed extract comprising the step of concentrating the extract under reduced pressure. The method of claim 4, wherein The site of citron is seed, skin, and pulp. The method of claim 4, wherein Lyophilization temperature is -50 ℃ to -5 ℃ manufacturing method of citron seed extract. The method of claim 4, wherein The organic solvent is at least one selected from the group consisting of phenoxyethanol, propiparaben, oxyphenol, benzophenone and ethanol. The method of claim 7, wherein The amount of the organic solvent is 500 to 1500 parts by weight based on 100 parts by weight of citron powder, the method of producing a citron seed extract. The method of claim 4, wherein The temperature of hot water is 40 ℃ to 80 ℃ manufacturing method of citron seed extract. The method of claim 4, wherein Concentration under reduced pressure is carried out 7 times to 13 times the preparation method of the citron seed extract. Whitening cosmetic composition containing citron extract Anti-aging cosmetic composition containing citron extract

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