KR20040101709A - Cosmetic composition containing camellia japonica extract stabilized with nanoliposome - Google Patents

Abstract

PURPOSE: Provided is a cosmetic composition which comprises the extract of Camellia japonica stabilized in nanoliposome for preventing skin-aging. The cosmetic composition whitens skin and improves moisturization and skin elasticity to prevent skin-aging. CONSTITUTION: The cosmetic composition comprises, as an active ingredient, 0.1-20 wt.% of the extract of Camellia japonica which is obtained by supercritical extraction and stabilized in nanoliposome, wherein the extract of Camellia japonica is contained in the nanoliposome in the amount of 10-40 wt.%.

Description

Cosmetic composition containing nanoliposomes stabilized camellia extract {COSMETIC COMPOSITION CONTAINING CAMELLIA JAPONICA EXTRACT STABILIZED WITH NANOLIPOSOME}

The present invention relates to a cosmetic composition containing a camellia extract, and more particularly to a cosmetic composition containing a nanoliposome stabilized camellia extract obtained by the supercritical extraction method.

Recently, as women's desire for the function of cosmetics has increased, high-functional products having anti-aging effects such as whitening effects, wrinkle improvement and suppression of skin, as well as clean and aesthetic functions, which are basic functions of cosmetics, have emerged. .

The mechanism of skin aging is not explained by any one theory. According to the findings and ongoing research, it is almost impossible to regenerate the skin of cigars with growing growth by rebuilding the skin in a healthy state, but it is possible to make more active improvement on the already created wrinkles, making it more elastic and It turns out that you can get smooth skin.

In this regard, US Pat. No. 5,340,568 contains 2-Deoxy-2-halo-lysophosphatidic acid, which functions similar to growth factors and induces cell proliferation. One composition is described as inhibiting the production of skin wrinkles, WO 94/04184 describes a composition in which protein growth factors are stabilized to prevent skin aging such as skin wrinkles, sagging of the skin, epidermal growth as a protein growth factor Factor (Epidermal growth factor, EGF), insulin-like growth factor (IGF), platelet-derived growth factor (PDGF), fibroblast growth factor (FGF) It is described as being.

However, the skin wrinkle improvement active ingredients, which can not be used as a cosmetic raw material or very unstable, it is not easy to transfer to the skin requires a special stabilization system and delivery system, such as the improvement effect of the skin wrinkles is not visible There is a problem.

Accordingly, attention has recently been focused on skin protection agents containing retinoids. Currently, retinoids are used as a means to solve photoaging phenomena such as wrinkles, skin thickening, sagging and elasticity, which are the result of daylight accumulation. have. However, retinoids are very unstable compounds, which are susceptible to light (ultraviolet light), moisture, heat, air (oxygen) and easily cause chemical changes.

In addition, materials with high antiwrinkle effects, such as retinol and retinoic acid, are difficult to use as raw materials for cosmetics due to chemical instability, difficulty in use, complexity of formulation technology, and skin irritation, such as retinyl palmitate or retinyl acetate. Β-carotene, a precursor of retinyl ester and vitamin A, has a low anti-wrinkle effect, but is widely used as a raw material for cosmetics due to its ease of use and high safety.

Vitamin C is also recognized as the most powerful antioxidant in relation to the aging of the human body, but due to its stability, it is difficult to use as a cosmetic because of the fact that many derivatives are used instead of vitamin C itself.

Camellia Japonica is an evergreen broad-leaved arboreous tree, with many twigs on the stem of long roots, long oval or semi-spherical, with bark smooth gray, leaves alternate, oval or long oval. It is 5-12cm long and 3-7cm wide, and its surface is dark green. There is 66% oil in the seed and its main ingredient is glyceride of oleic acid. In addition, glycosides camellia (C18H34O7) and camellia saponin (C58H92O25) are present in the seeds (components and use of herbs). Camellia has a strong action, Camellia saponin is known to be known for hemolytic action, camellia oil is used as a base oil-based ointment, camellia flowers are used in the intestinal bleeding, uterine bleeding, hemorrhage, etc. as a medicine.

Supercritical fluid extraction technology is a fluid state when a certain material exceeds a certain high pressure and high temperature (critical point), using the specific material as a solvent to isolate only the active ingredients with high purity, which are beneficial to humans, without toxic or residual substances. The supercritical fluid as a solvent is used at or near the critical temperature (Tc) and the critical pressure (Pc), and as a commonly used supercritical fluid, the critical Carbon dioxide which is easy to isolate | separate and refine | purify a material which is low in temperature with low temperature is mentioned. By using carbon dioxide, a supercritical fluid solvent, it is non-toxic to skin because it uses carbon dioxide that is harmless to the human body instead of the general solvent, and can be easily removed from the product, thereby preventing environmental pollution, which is a problem when extracting an organic solvent. This inexpensive and simplifies the extraction process as much as possible to achieve biochemical stability (oxygen blocking, microbial contamination).

An object of the present invention is to obtain a cosmetic composition containing nanoliposomes stabilized camellia extract, excellent anti-aging effect, and excellent in bioavailability through nanoliposome stabilization by extracting the anti-aging active ingredient as much as possible.

1 is a graph showing the cell proliferation ability according to Test Example 2,

2 is a graph showing collagen biosynthesis according to Test Example 3.

Therefore, according to the present invention, the nanoliposomes comprising the Camellia extract by the supercritical extraction method, characterized in that it contains 0.1 to 20% by weight based on the total weight of the cosmetic composition, containing the nanoliposomes stabilized Camellia extract A cosmetic composition is provided.

The present invention is described in detail as follows.

The usable part of the camellia according to the present invention is bark, seed, leaf, fruit, and the use of bark is preferable in view of effect.

In order to prepare a camellia extract, first the camellia is dried and then pulverized to produce a camellia powder.

As the extraction solvent, 1,3-butylene glycol and ethanol are mixed in the same mass ratio, followed by stirring to form a single phase, which is used as a cosolvent.

Supercritical fluid extraction used in the present invention can be carried out by a conventional method using carbon dioxide, specifically, it is preferably carried out as follows.

The pulverized camellia and cosolvent are mixed and stirred. The mixing ratio is preferably a mixture of camellia and cosolvent in a mass ratio of 1: 1 to 1: 3, and mixing in a mass ratio of 1: 2 is the best in terms of yield. Subsequently, the mixture is introduced into a supercritical extraction tank, and carbon dioxide is supplied to the extraction tank using a high pressure gas pump to increase the pressure. Then, the temperature inside the extraction tank is raised to 40 ° C. and maintained therein. Then, while continuously supplying carbon dioxide to the extraction tank, the extract is released using a pressure regulator attached to the outlet side. By controlling the temperature of the pressure regulator, the pressure and temperature drop of the discharge prevent carbon dioxide from transferring to solid dry ice and clogging the tube at the final outlet. The released extract is decompressed to atmospheric pressure to separate the carbon dioxide and Camellia extract, the carbon dioxide is released into the atmosphere and the Camellia extract is recovered.

The Camellia extract thus obtained is contained in the nanoliposomes in order to stabilize the camellia.

The nanoliposome in the present invention uses a liposome consisting of phospholipid, glutathione, polysorbate 20, and ethanol seven times through a high pressure emulsifying device to reduce its size.

At this time, the Camellia extract is added in 10 to 40% by weight based on the total weight of the extract-containing nanoliposomes. If it is added less than 10% by weight, the effect is hard to expect, and if it exceeds 40% by weight, no significant increase in efficacy effect is seen.

The nanoliposomes obtained by stabilizing the obtained Camellia extract can be added to moisturizing cosmetics such as nourishing lotion, nourishing cream and essence, and in particular can be added to liquid or transparent cosmetics.

Nanoliposomes stabilized camellia extract is preferably added in an amount of 0.1 to 20% by weight, more preferably 1 to 10% by weight based on the total weight of the composition. If it is less than 0.1% by weight, it is difficult to expect an effective effect. If it exceeds 20% by weight, there is no apparent increase in efficacy even if the content is increased.

The following Examples, Comparative Examples, Preparation Examples, and Test Examples are provided to explain the present invention in more detail, and the scope of the present invention is not limited thereto.

(Example 1)

Preparation of Camellia Extract

Camellia bark was dried until there was no change in weight, and ground to a size of 0.3 cm or less using a grinder. In addition, 1,3-butylene glycol and ethanol were mixed in the same mass ratio and stirred to prepare a single-phase cosolvent.

The crushed camellia bark and the cosolvent were mixed at a mass ratio of 1: 2, stirred for 30 minutes, and then the mixture was put into a supercritical extraction tank having an internal volume of 1.5 L and the extraction tank was sealed. The high pressure gas pump was used to supply carbon dioxide to the extraction tank and increase the pressure, thereby increasing the pressure of the extraction tank.

The temperature inside the extraction tank was raised to 40 ° C. at a rate of 0.5 / min and maintained for 50 minutes using a heating wire wrapped outside the extraction tank using a PID temperature controller.

The extract was released while continuously supplying carbon dioxide to the extraction tank. The extracted extract was decompressed to atmospheric pressure to separate carbon dioxide and bark extract and carbon dioxide was released into the atmosphere. The separated bark extract was recovered using a glass collector.

(Test Example 1)

Fibroblast Proliferation Capacity Comparison Experiment

Using the Camellia extract of Example 1, cell proliferation using human skin-derived fibroblasts was tested. Fibroblasts were used to proliferate cells stored in liquid nitrogen. Fibroblasts cultured in T-75 flasks were washed 2-3 times with PBS, and the adherent cells were dropped using trypsin, followed by centrifugation at 1500 rpm for 5 minutes (Heraeus, Labofuge 400R) to collect the cells. After centrifugation, the supernatant was discarded, and 10 ml of DMEM (10% FBS) was added to the precipitated cells and mixed evenly. By measuring the number of cells using a blood cell counter ⁴ to 10 ea / well degree of cell to each well of a 96 well it was dispensed by 200㎕. When the cells of about 50% of the well area were grown while incubating at 37 ° C. and 5% CO ₂ , the camellia extract of Example 1 was changed by concentration (0.00001%, 0.0001) while replacing the medium with fresh DMEM (FBS10%). %, 0.001%, 0.01%, 0.1%, 1%), cells were incubated for 48 hours.

MTT {(3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl tetrazolium bromide (Sigma)} was dissolved in PBS to 2 mg / ㎖ was stored in the refrigerator and used Each well was treated with 20 μl of MTT reagent (5 mg / ml in PBS), incubated for 2 more hours, discarded the supernatant, and mixed with 100 μl of 0.04 N-HCl, followed by ELISA reader (μQuant, Absorbance was measured at 560 nm using Bio-Tek instrument INC. (Reference: 650 nm).

Cell proliferation was calculated according to the following equation, and the results are shown in FIG. 1.

In Fig. 1, the Camellia extract of Example 1 showed a cell proliferation effect of about 85% after 48 hours at 0.01% of the treatment, while the treatment-free treatment was about 40%.

(Test Example 2)

Collagen Biosynthesis Effect

Fibroblasts incubated in T-75 flasks were washed 2-3 times with PBS, the adhered cells were dropped using trypsin, and the cells were collected by centrifugation (Heraeus; Labofuge 400R) at 1500 rpm for 5 minutes. After centrifugation, the supernatant was discarded, and 10 ml of DMEM (10% FBS) was added to the collected cells and mixed evenly. By measuring the number of cells using a hemocytometer, cells of about 10 ⁴ ea / well were dispensed into each 200-well well of 96 wells. When incubating at 5% CO ₂ at 37 ° C and growing about 50% of the well area, the Camellia extract of Example 1 was changed by concentration (0.00001%, 0.0001%) while exchanging with fresh DMEM (FBS 10%). , 0.001%, 0.01%, 0.1%, 1%), and the cells were incubated for 48 hours.

Collagen synthesis performance measurement kit (Procollagen Type-I C-peptide EIA kit (Takara, MK 101)) was used. For determination using immunological methods, 100 μl of Ab-POD conjugate solution was transferred to one well and aliquoted, and 20 μl of cell culture or standard solution was added. A 96 well plate was wrapped in foil and incubated at 37 ° C. for about 3 hours. After 3 hours, all the solutions in each well were removed and washed 4 times with PBS. After the washing step, 100 μl of substrate solution was added to each well and incubated at 20-30 ° C. for 15 minutes. In the final step, 100 µl of 1N H ₂ SO ₄ , the reaction termination solution, was added and mixed gently. The absorbance was measured with an ELISA reader and the result was used. After the reaction solution was added, the plate was stored at room temperature for about 1 hour and absorbance was measured at 450 nm.

Synthesis (%) was calculated according to the following equation, and the results are shown in FIG. 2.

In Figure 2, it can be seen that as shown in the results of cell proliferation, the enhancement effect is large.

(Test Example 3)

Melanin production inhibitory effect

The inhibitory effect of melanin production was tested using the Camellia bark extract of Example 1.

Melanocytes isolated from human tissues were primaryly cultured and passaged two times, and then cultured in 60 mm culture dishes at a concentration of 2 × 10 ⁴ cells / culture plate using RPMI 1640 medium. After confirming that the cells attached to the dish, camellia extract was treated by concentration up to 0.001, 0.01, 0.1, 1%. After 3 days of treatment, cells were detached with trypsin-EDTA and the 2 × 10 ⁵ cells were evenly distributed. 100 μl of 1N NaOH solution was added thereto and treated at 37 ° C. for 12 hours to completely lyse the cells. Absorbance was measured at 490 nm for 500 μl of cells in which cells were completely lysed. In addition, absorbance was measured using the extract-free treatment as a control.

As a standard indicator, 10 mg of synthetic melanin (M8601, manufactured by Sigma Co., Ltd.) was dissolved in 10 ml of 1N NaOH to prepare a mother solution (1 mg / ml), followed by 700 µg / ml, 300 µg / ml, 100 µg / ml, 70 Dilutions of μg / ml, 30 μg / ml, 10 μg / ml, 7 μg / ml, 3 μg / ml, 1 μg / ml, 0.1 μg / ml and 0 μg / ml were prepared and the absorbance was measured at 490 nm. The amount of melanin reduction was calculated according to the following equation.

The results are shown in Table 1 below.

Melanin reduction Camellia Extract (%) Melanin Reduction (%) One 32.35 0.1 46.06 0.01 53.12 0.001 16.42

According to Table 1, camellia extract can be seen that the effect of whitening by inhibiting the action of biosynthesis melanin.

(Comparative Example 1)

The bark of camellia was dried at room temperature until there was no change in weight. It was ground by pulverization to powder. 50 g of camellia bark dry powder was added to 50 g of ethanol, and the mixture was heated at about 60 ° C. for about 48 hours. The extract removed from the debris was first filtered using a 300 mesh filter paper, and the filtered extract was filtered again and again with 0.40 μm filter paper to obtain an extract.

(Test Example 4)

Free radical scavenging ability measurement

For the supercritical camellia extract obtained in Example 1 and the ethanol camellia extract of Comparative Example 1, the content of catechin as a substance having active oxygen scavenging ability was measured. Since reactive oxygen species cause various adult diseases and aging, in particular, skin aging and hyperpigmentation, its scavenging ability is essential in functional anti-wrinkle and whitening cosmetics.

Specifically, the Camellia extract of Example 1 and Comparative Example 1, catechin standard (manufactured by Sigma) were precisely weighed by about 5.0 mg each, dissolved in 50 ml of a mobile phase, and then added to the mobile phase to make 100 ml. It was.

Test solution, and by using liquid chromatography (1100Series, AT, Inc.) as the standard solution was determined 20㎕ category cue acid, peak area ratios of the parcel retinoic {Q _T (sample area) / Qs (standard size) × 100}.

The operating conditions of the liquid chromatography for the test are as follows.

Mobile phase: 1.0 g of phosphoric acid is dissolved in 50% methanol solution to make 2 liters.

Detector: UV absorbance photometer (wavelength 280nm)

Column: C ₁₈

Flow rate: 1.0 ml / min

The test results are shown in Table 2 below.

sample Number of tests Catechin content (%) Average Supercritical Extract Camellia Bark Extract One 95.0 94.93 2 95.2 3 94.6 Ethanol Camellia Extract One 31.5 30.77 2 30.2 3 30.6

(Test Example 5)

Skin moisturizing effect

In order to measure the skin moisturizing effect of simple raw materials before nanoliposomalization, Na-PCA (Sodium Pyrolidone Carboxylic acid), Chondroitin, which is a component of Natural Moisturizing Factor (NMF) as a comparison target with Camellia extract of Example 1 Na-Condroitin Sulfate and 1% aqueous solution of Na-hyaluronate were measured to reduce the epidermal moisture loss (TEWL) after barrier damage, which is a standard measure of skin barrier function.

The specific method is as follows.

Acetone was instilled into the back of 20 hairless mice aged 8-12 weeks to damage the skin barrier. When transepidermal water loss (TEWL) reached 4.0 g / m ² / h, the sample was applied to an area of 5 cm ² . TEWL was measured with Tewameter, an evaporimeter from CK (Courage + Khazaka, Cologne, Germany). After 6 hours of application, TEWL was measured to evaluate the extent to which TEWL was reduced to evaluate the extent to which skin barrier function was restored.

The recovery rate used for the efficacy evaluation was calculated according to the following equation.

(Bt = 6: TEWL measured value 6 hours after skin barrier injury)

Bt = 0: TEWL measurement before skin barrier injury

Bt = d: TEWL measured immediately after skin barrier damage)

The results are shown in Table 3 (unit: mg / m 2 / hr).

Hairless Mouse Number Bark Extract (1%) Na-PCA aqueous solution (1%) Na-CS aqueous solution (1%) Na-hyaluronate aqueous solution (1%) One 14 18.5 17.4 18.4 2 16 19 18 19 3 15.8 20.1 19 19.2 4 17.6 21 19 19.6 5 13.5 20.5 20 19.7 6 13 18.5 21 20.2 7 15.4 18.9 18 20.3 8 15.2 18.3 19.3 21.4 9 17.1 18.8 19 19.7 10 14.5 20.3 18 18.9 11 13.9 17.4 18.2 18.6 12 13.4 17.5 17.9 19.3 13 15.9 19.8 19 20.1 14 15.1 18.8 17.6 21.1 15 15.8 18.5 20 19.8 16 16.1 21.5 18 18.6 17 16.8 19.3 18 19 18 17.0 19.1 19.5 19.2 19 16.2 20.7 19.8 20.1 20 16.6 19.5 20.2 20.1 Average measurement value 15.445 19.3 18.845 19.615

The TEWL measurement results, 19.97% compared to Na-PCA (Sodium Pyrolidone Carboxylic acid) used in the present invention, 18.04%, Na- compared to Na-Condroitin Sulfate 21.26% better than hyaluronate.

(Example 2)

Preparation of Nanoliposomes with Stabilized Camellia Extract

Phase A in Table 4 was heated to 95 ° C., uniformly mixed and dissolved. It was cooled to 40 ° C and phase B was mixed. This was passed through a high pressure emulsifier (Microfludizer M210EH, Microfluidies, USA) seven times under a pressure of 1200 bar and a flow rate of 500 m / s to obtain a nanoliposome stabilized with Camellia extract. The size of the liposome obtained through one pass was about 133 nm, but the size of the seventh was about 75 nm.

Raw material name Content (% by weight) A Phospholipids 4 Glutathione 0.1 95% modified ethanol 20 glycerin 30 Polysorbate 20 4 B Camellia Extract of Example 1 20 Purified water Remaining amount system 100

(Comparative Example 2)

Nanoliposomes were prepared in the same manner as in Example 2, except that Camellia extract was not added.

(Manufacture example 1)

Preparation of Cosmetics Containing Nanoliposomes Stabilizing Camellia Extract

Using the raw materials shown in Table 5 below, an emulsion was prepared according to a conventional cosmetic preparation method.

Raw material name Content (unit: weight%) Polysorbate-60 1.5 Sorbitan stearaytan 0.5 Stearic acid One Squalane 5 Cetostearyl alcohol 2 Cetyl octanoate 5 Carbomer 0.15 Triethanol amine 0.2 glycerin 5 Nanoliposomes Stabilizing Camellia Extract of Example 2 0.1 incense A reasonable amount antiseptic A reasonable amount Purified water Remaining amount total 100

(Comparative Production Examples 1 to 3)

Emulsion in the same manner as in Preparation Example 1, except that 5% by weight of Na-PCA, sodium chondroitin sulfate, and Na-hyaluronate were used instead of the nanoliposomes stabilized with the Camellia extract of Example 2. Were prepared, and each was made into Comparative Production Examples 1-3.

(Comparative Production Example 4)

Instead of using the nanoliposomes stabilized Camellia extract of Example 2, except that 1% by weight of Camellia extract of Example 1 and 4% by weight of nanoliposomes of Comparative Example 2 is used, the same as in Preparation Example 1 The emulsion was prepared by the method.

(Comparative Production Example 5)

An emulsion was prepared in the same manner as in Preparation Example 1, except that 1 wt% of the Camellia extract of Example 1 was used instead of the nanoliposomes stabilized Camellia extract of Example 2.

(Production Examples 2 to 5)

An emulsion was prepared in the same manner as in Preparation Example 1, except that 1, 5, 10, and 20 wt% of the liposomes stabilized the Camellia extract of Example 2 were added, respectively, to Preparation Examples 2 to 5.

(Comparative Production Examples 6 and 7)

An emulsion was prepared in the same manner as in Preparation Example 1, except that 0.05 and 25 wt% of the liposomes stabilized the Camellia extract of Example 2 were added, respectively, to Comparative Preparation Examples 6 and 7.

(Test Example 6)

Skin elasticity effect

60 healthy women (average age 29.5 years old) of 20 years or older (mean age 29.5 years) were divided into 6 groups at the temperature of 22-24 degreeC and 55% of the relative humidity, and the cosmetic composition of manufacture example 3 and comparative manufacture examples 1-5 was centered around the eyes After application for 12 weeks (2 times / day), skin elasticity was measured using a skin elasticity measuring instrument (Cutometer SEM 575, C + K Electronic Co., Germany). The test results are described in Table 5 below as ΔR5 values (R5 (12 weeks) -R5 (0 weeks)), which are average values for each test group of the Cutometer SEM 575. The closer it is, the better the elasticity is.

In addition, the improvement degree was computed by the following formula.

Improvement (%) = (△ R5 / Before Application) X 100

Skin elasticity test result (12 weeks) Before application After application Skin elasticity enhancing effect (△ R5) % Improvement Preparation Example 3 0.74 1.12 0.38 51.35 Comparative Production Example 1 0.71 0.72 0.01 1.40 Comparative Production Example 2 0.71 0.75 0.04 5.63 Comparative Production Example 3 0.73 0.76 0.03 4.11 Comparative Production Example 4 0.72 0.75 0.03 4.16 Comparative Production Example 5 0.73 0.84 0.11 15.07

According to Table 6, when using Preparation Example 3 containing the nano-liposomes stabilized Camellia extract, skin elasticity is 49.95% compared to Comparative Preparation Example 1, 45.72% compared to Comparative Preparation Example 2, Comparative Preparation Example 3 47.24%, 47.19% compared to Comparative Preparation Example 4, 36.28% compared to Comparative Preparation Example 5. Therefore, cosmetics containing nanoliposomes stabilized camellia extract according to the present invention, skin elasticity than when using the components of the natural moisturizing factor (NMF) or camellia extract that is not stabilized with nanoliposomes Can be effectively promoted.

(Test Example 7)

Skin moisture measurement

Seventy healthy women (mean age 29.5 years) were divided into seven groups of 70 healthy women (22 ~ 24 ℃, 55% relative humidity, no air flow). The cosmetics of Preparation Examples 1 to 5 and Comparative Preparation Examples 6 and 7 were applied to the entire face twice a day for 6 weeks, mainly around the eyes of each group. By using Skicon-200 (IBS Co. Japan), the moisture content of the cheek area was measured before and after application to measure the persistence of the moisturizing effect. The results are shown in Table 7 as an average value for each test group.

The improvement was calculated according to the following equation.

Before the test 6 weeks after application Improvement (%) Preparation Example 1 630 ± 10 645 ± 10 2.33 Preparation Example 2 630 ± 10 690 ± 10 8.70 Preparation Example 3 630 ± 10 850 ± 10 25.88 Preparation Example 4 630 ± 10 663 ± 10 4.98 Preparation Example 5 630 ± 10 650 ± 10 3.08 Comparative Production Example 6 630 ± 10 640 ± 10 1.56 Comparative Production Example 7 630 ± 10 645 ± 10 2.33

According to the table, it is preferable to contain 0.1 to 20% by weight of stabilized nano-liposomes in cosmetics.

(Test Example 8)

Stability experiment

Stability of the formulation for the cosmetic containing nanoliposomes stabilized Camellia extract according to the present invention was measured by the following method.

Samples stored in an opaque vitreous container in a constant temperature bath maintained at 45 ° C. in Preparation Example 3, Comparative Examples 4 and 5 were stored for 12 weeks in an opaque glass container in a completely shaded refrigerator maintained at 4 ° C. The samples stored for 12 weeks, and samples stored for 12 weeks in a circulation chamber (once / day) circulating at 37 ° C. at −5 ° C. were compared and measured for the degree of discoloration and precipitation.

The results of the evaluation of product separation and discoloration graded into the following six grades are shown in Table 8.

Product discoloration evaluation criteria:

0: no change 1: very little separation (discoloration)

2: Slightly separated (discolored) 3: Slightly separated (discolored)

4: Extremely separated (discolored) 5: Extremely separated (discolored)

Separation and discoloration degree of test substance Temperature Preparation Example 3 Comparative Production Example 4 Comparative Production Example 5 45 ℃ 0 3 3 4 ℃ 0 2 2 cycle 0 2 3

According to the Table 8, Preparation Example 3 stabilized Camellia extract through nanoliposomes is stable without discoloration or separation symptoms in 4 ℃, 45 ℃ and circulation test, Comparative Preparation Example 4 and used directly in the formulation without stabilization In case of 5, separation (discoloration) occurred and it was found to be unstable.

In addition, the results obtained by classifying the product precipitation degree into the following six grades are shown in Table 9 below.

Product discoloration evaluation criteria:

0: no change 1: very slight precipitation

2: little precipitate 3: little precipitation

4: settling heavily 5: settling extremely severe

Precipitation degree of test substance Temperature Preparation Example 3 Comparative Production Example 4 Comparative Production Example 5 45 ℃ 0 3 3 4 ℃ 0 3 2 cycle 0 3 2

According to Table 9, it was found that the cosmetics stabilized camellia extract according to the present invention with nanoliposomes did not precipitate, it was stabilized.

As such, the Camellia extract is added without stabilization, causing easy reaction with other weak ionic polymers in the formulation, causing precipitation and discoloration. However, when the camellia extract is stabilized with nanoliposomes as in the present invention and applied to the formulation, it can be seen that precipitation and discoloration reaction are minimized even when the Camellia extract is added in excess.

(Test Example 9)

Measurement of Transdermal Absorption by Size of Nanoparticles

Nanoparticles having sizes of 50 nm, 80 nm, 120 nm and 500 nm were prepared using PMMA polymers to which fluorescent substances were added by chemical bonding, and applied to the skin of female hairless guinea pigs. The cross section was cut to measure the fluorescence emitted from the percutaneously absorbed nanoparticles.

Preparation of fluorescent PMMA polymer nanoparticles allowed the fluorescent molecules to be covalently bonded to the carboxyl group of methacrylic acid of polymethylmethacrylate-co-methacrylic acid (molar ratio of methylmethacrylate and methacrylic acid is 84:16).

The preparation method is briefly described as follows.

After 34 g of polymethylmethacrylate-co-methacrylic acid was dissolved in 150 ml of anhydrous methylene chloride, 132 mg of dicyclohexylcarbodiimide and 74 mg of N-hydroxysuccinimide were added to activate the carboxyl group. After stirring for 1 hour, 222 mg of fluoresceamine was added to react the primary amine of the fluorescein amine with the activated carboxyl group to form an amide bond, thereby allowing the fluorescent molecules to be covalently bonded to the polymer chain. The reaction proceeded for 5 hours at room temperature and dark conditions. Dicyclohexyl urea, a by-product formed after the reaction, was removed through a 0.45 μm nylon filter and then precipitated in diethyl ether to remove the reaction reagents and unreacted residues. Subsequently, it was left in excess tertiary distilled water for one day to remove the remaining residues, and then dried using a vacuum oven at 40 degrees. As a result, PMMA nanoparticles having sizes of 50 nm, 80 nm, 120 nm, and 500 nm to which fluorescent materials were added were prepared.

Eight-week-old female hairless guinea pigs (strain IAF / HA-hrBR) were used for the nanoparticle transdermal absorption test by size. The skin of the abdominal region of the guinea pig was cut and mounted in Franz-type diffusion cells (Lab Fine Instruments, Korea). A 50 mM phosphate buffer (pH 7.4, 0.1 M NaCl) was added to the receiving vessel (5 ml) of the plasma-type diffusion cell. The diffusion cell was mixed and dispersed at 600 rpm while maintaining 32 ° C. 50 μl of a solution containing 10% (w / v) of fluorescent PMMA nanoparticles prepared for each size was placed in each donor container. gave. Absorption and diffusion were performed according to a predetermined time (average 12 hours), and the area of skin where absorption and diffusion occurred was 0.64 cm ² . After the absorption and diffusion of nanoparticles was finished, the non-absorbed nanoparticles remaining on the skin were washed with dried Kimwipes, and the cross-section of the skin was cut to measure the distribution of fluorescent PMMA nanoparticles absorbed on the skin section.

The intradermal distribution measurement results of the nanoparticles are shown in Table 10 below.

Average size of nanoparticles Percutaneous absorption area 50 nm 35 μm 80 nm 17 μm 120 nm 12㎛ 500 nm 7㎛

From the above experiments, it was confirmed that the percutaneous absorption of the nanoparticles largely depends on the average particle diameter. Nanoparticles having an average diameter of about 50nm size was passed through the epidermis, it was confirmed that the absorption and diffusion to the top of the dermal layer. When the particle size exceeds 500 nm, the nanoparticles stay on the skin surface and no longer absorb into the skin. Skin absorption of nanoparticles with an average diameter of about 50 nm is diffused into the intercellular lipids of the skin, and the hydrophobicity of the polymer is thought to have an effect of promoting such transdermal absorption. In addition, when the active ingredient is poorly or unstable, and the active ingredient is trapped in the nanoparticles having an average particle diameter of about 50 nm, it may be possible to transmit the percutaneous absorption of the active ingredient through delivery into the skin by a polymer.

According to the present invention, by containing a camellia extract maximized in the content of active ingredients such as catechins in the nanoliposomes by the supercritical extraction method, while having excellent anti-aging effects such as whitening, moisturizing, elasticity, stability, skin transmittance and feeling of use This excellent cosmetic composition can be obtained.

Claims (3)

A cosmetic composition comprising a nanoliposomal stabilized Camellia extract, characterized in that containing 0.1 to 20% by weight of the nanoliposome comprising the Camellia extract by the supercritical extraction method with respect to the total weight of the cosmetic composition. The method of claim 1, The nanoliposomes are obtained by passing a liposome consisting of phospholipid, glutathione, polysorbate 20, ethanol seven times through a high-pressure emulsifying apparatus, cosmetic composition containing the nano-liposomes stabilized camellia extract. The method of claim 1, The Camellia extract is characterized in that it is contained in 10 to 40% by weight relative to the nanoliposomes, cosmetic composition containing the nano-liposomes stabilized Camellia extract.