KR102058445B1 - Cosmetics composition with Extract of Sparassis crispa Wulf. ex Fr. - Google Patents

Abstract

The present invention relates to a cosmetic composition using nanoliposome containing an extract of peach blossoms. The cosmetic composition using nanoliposome containing an extract of peach blossoms, contains 0.1-20 wt% of nanoliposome comprising an extract of peach blossoms extracted by supercritical and ultrasound extraction methods with respect to the total weight of the cosmetic composition. The cosmetic composition comprising the extract of peach blossoms according to the present invention has excellent moisturizing and tightening effects, has a whitening effect, has excellent stability and skin permeability, and improves strength and thicknesses of hair, by obtaining the extract of peach blooms based on extraction of supercritical and ultrasound extraction methods, maximizing content of a catechin component to be contained in the nanoliposome.

Description

Cosmetic composition using nanoliposomes containing peach blossom extract {Cosmetics composition with Extract of Sparassis crispa Wulf. ex Fr.}

The present invention relates to a cosmetic composition, and more specifically, by extracting the peach blossom extract by supercritical method and ultrasonic wave to maximize the content of catechin component in the nanoliposomes, while moisturizing, elasticity effect, whitening effect It relates to a cosmetic composition using a nano liposome containing peach blossom extract, having stability and excellent skin permeability.

Recently, as women's desire for the function of cosmetics has increased, high-functional products have appeared, which are not only clean and aesthetic functions, which are the basic functions of cosmetics, but also whitening to bright and white skin, improving and suppressing wrinkles on the skin. In particular, wrinkle improvement products are of interest to many women who want to stay young, and efforts are being made to improve wrinkles on the skin.

The mechanism of skin aging is not explained by any one theory. According to the research results or ongoing research, it is almost impossible to regenerate the skin when the growth phase is stopped by rebuilding the active skin structure, but it is possible to improve the already created wrinkles more aggressively and smoothly. It can be found that it can be applied to the skin.

In general, active oxygen is a superoxide anion radical in which the oxygen molecule is reduced by one electron, and hydrogen peroxide and hydroxyl radical in which it is reduced again. In addition, there is singlet oxygen caused by energy transfer, not electron transfer. Although there are four of the above-mentioned active oxygens in a narrow sense, they broadly include their oxidation products, lipid peroxides and nitric oxides (Hitoshi Masaki. Et al., Fargrance j., 19-20 (2000)).

On the other hand, US Patent No. 5,340,568 (Patent Document 1) is a 2-deoxy-2-halo- lysophosphatidic acid (2Deoxy-2-halo-lysophosphatidic acid) that functions similar to the growth factor and induces cell proliferation The composition containing is also described as inhibiting the production of skin wrinkles, and WO 94/04184 also discloses that a composition which stabilizes protein growth factors prevents skin aging such as skin wrinkles and sagging skin. Protein growth factors used in this technique include epidermal growth factor (EGF), insulin-like growth factor (IGF), platelet-derived growth factor (PDGF), and fibroblasts. Growth factor (Fibroblast growth factor (FGF)).

However, the above described skin wrinkle improvement active ingredients cannot be used as some cosmetic ingredients or are very unstable, and it is not easy to transfer to the skin, so a special stabilization system and delivery system are needed, and the effect of improving wrinkles is not visible. Recently, attention has been gradually 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 decreased elasticity, which are the result of daylight accumulation. It is becoming. However, these retinoids are very unstable compounds that are sensitive to light (ultraviolet light), moisture, heat, and air (oxygen), and easily cause chemical changes.

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, as mentioned above. Retinyl palmitate or reti Retinyl esters such as nile acetate and β-carotene, which are precursors of vitamin A, have low wrinkle improvement effects, but are widely used as raw materials for cosmetics due to their ease of use and high safety.

Vitamin C is regarded as the most powerful antioxidant in relation to the aging of the human body. However, due to its stability, the use of cosmetics is difficult. Currently, the cosmetics industry has made a lot of efforts to use vitamin C itself, but has not overcome the destabilizing properties, the reality is to eliminate the effect using a number of derivatives other than vitamin C itself. The most important issue with regard to skin aging is collagen. Many researchers have shown that aging is progressed and repaired by various complexes such as collagen and elastin.

US 5,340,568 (1994.08.23)

The present invention has been made to solve the above problems, the object of the present invention is a peach that has an excellent moisturizing, elasticity effect, whitening effect and excellent stability and skin permeability, effective in improving the strength and thickness of hair It is to provide a cosmetic composition comprising a flower extract.

In order to solve the above problems, the present invention is characterized in that it contains 0.1 to 20% by weight of the nanoliposome comprising the peach blossom extract extracted by supercritical and ultrasonic extraction method with respect to the total weight of the cosmetic composition.

In the above configuration, the nanoliposomes are characterized in that the liposome consisting of phospholipid, glutathione, polysorbate 20, ethanol through a high pressure emulsifying device.

In addition, the peach blossom extract extracted by the supercritical and ultrasonic extraction method is characterized in that it is contained in 10 to 40% by weight relative to the total weight of the nanoliposomes comprising the peach blossom extract.

As described above, the cosmetic composition comprising the peach blossom extract according to the present invention by extracting the peach blossom extract by supercritical and ultrasonic extraction method to maximize the content of the catechin component in the nanoliposomes, moisturizing, elasticity While excellent in effect, it has an effect of having a whitening effect, excellent stability and skin permeability, and obtaining a cosmetic composition effective for improving the strength and thickness of hair.

1 is a graph comparing the fibroblast proliferation capacity using the peach blossom extract according to the present invention.
Figure 2 is a graph of collagen biosynthesis using peach blossom extract according to the present invention.
Figure 3 is a graph of melanin biosynthesis inhibition using peach blossom extract according to the present invention.

Cosmetic composition comprising a peach blossom extract according to the present invention for achieving the above object is a cosmetic composition stabilized with nanoliposomes, the total weight of the cosmetic composition of nanoliposomes comprising peach blossom extract extracted by supercritical and ultrasonic extraction method It is characterized by containing 0.1 to 20% by weight relative to.

Preferably, the nanoliposomes are obtained by passing a liposome consisting of phospholipid, glutathione, polysorbate 20, ethanol through a high pressure emulsifier.

Preferably, the peach blossom extract extracted by the supercritical and ultrasonic extraction method is characterized in that it is included in 10 to 40% by weight based on the total weight of the nano liposomes containing the peach blossom extract.

Hereinafter, the present invention will be described in detail with reference to embodiments and drawings of the present invention. These examples are only for illustrating the present invention in more detail, it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples in accordance with the gist of the present invention. .

The present invention extracts the peach blossoms by supercritical and ultrasonic extraction method, extracts the active ingredient related to skin aging prevention as much as possible, and then stabilizes with nanoliposomes to apply to cosmetics to overcome skin aging and to improve the existing functional ingredients for wrinkles To obtain a new raw material of the reinforced function, and to use it to obtain a cosmetic for improving wrinkles of new materials.

Nanoliposomes used in the present invention are made to reduce the size by passing the liposome consisting of phospholipid, glutathione, polysorbate 20, ethanol seven times through a high-pressure emulsion device. The peach blossom extract is added in 10 to 40% by weight based on the total weight of the peach blossom extract-containing nanoliposomes. It is because it is hard to expect an effect when it adds less than 10 weight%, and even if it adds more than 40 weight%, it does not show a clear increase of an efficacy effect.

Nanoliposomes stabilized peach blossom extract obtained by supercritical and ultrasonic extraction can be added to moisturizing and elastic cosmetics such as nourishing lotion, nourishing cream, essence, etc., in particular can be added to liquid or transparent cosmetics.

In addition, the cosmetic composition comprising the peach blossom extract according to the present invention preferably contains 0.1 to 20% by weight of the nanoliposome stabilized peach blossom extract relative to the total weight of the cosmetic composition. If it is less than 0.1% by weight, it is difficult to expect an effective effect, and if it exceeds 20% by weight, there is no apparent increase in efficacy. More preferably, it is contained in 0.1 to 10% by weight.

First, 1,3BG (Butylene Glycol) and ethanol were mixed in the same mass ratio and stirred to form a single phase, which was used as a cosolvent to be used in the extraction process using supercritical carbon dioxide. The peach blossoms were dried until no change in weight and ground to a size of 0.3 cm or less using a grinder. The pulverized peach blossom and the cosolvent were mixed at a mass ratio of 1: 2, stirred for 30 minutes, and subjected to primary extraction for about 3 hours using ultrasonic waves at 60 ° C. The co-solvent mixture was introduced into a supercritical extraction tank having an internal volume of 1.5 L and sealed to increase the pressure by supplying carbon dioxide using a high pressure gas pump. The temperature inside the extraction tank is raised to 40 ° C. at a rate of 0.5 ° C./min, and maintained for 50 minutes. The carbon dioxide is released while supplying carbon dioxide and decompressed at atmospheric pressure to separate carbon dioxide and peach blossom extract, and carbon dioxide is released into the atmosphere. Peach blossom extracts were collected with a collector.

As a comparison target of the peach blossom extract extracted by the supercritical extraction and ultrasonic method, the peach blossom was dried and pulverized to a size of 0.3 cm or less using a pulverizer, and peach blossom extract by ethanol extraction was used in the present invention. .

Nanoliposomes were prepared using the peach blossom extracts obtained in Example 1.

The nanoliposomes were made small by passing the liposomes consisting of phospholipid, glutathione, polysorbate 20, and ethanol seven times through a high pressure emulsifier. First, phospholipid, glutathione, 95% modified ethanol, glycerin, and polysorbate 20 were emulsified under high pressure to make a closed bilayer nanoliposome. The phase A of Table 1 was heated up to 95 degreeC, and it mixed and dissolved uniformly. It was cooled to 40 ° C and phase B was mixed. Then, the resultant was passed through a high pressure emulsifier (Microfludizer M210EH, Microfluidies, USA) seven times under pressure of 1200 bar and a flow rate of 500 m / s to obtain nanoliposomes in which the peach blossom extract obtained in Example 1 was stabilized. The size of the nanoliposome obtained through one pass was about 133 nm, but the size of the seventh was about 75 nm.

Division (top) Raw material name Content (% by weight)

A Phospholipids 4 Glutathione 0.1 95% modified ethanol 20 glycerin 30 Polysorbate 20 4 B Peach Blossom Extract 20 Purified water Remaining amount system 100

Experimental Example 1 Comparative Experiment of Fibroblast Proliferation

A fibroblast proliferation comparison experiment was performed using the peach blossom extract obtained in Example 1. Cell proliferation was tested using human skin derived fibroblasts. Fibroblasts were used by proliferating cells stored in -70 ° C or liquid nitrogen. Fibroblasts cultured in T-75 flasks were washed 2-3 times with PBS, and the adherent cells were dropped using trypsin, and then cells were collected by centrifugation (Heraeus, Labofuge 400R) at 1500 rpm for 5 minutes. After centrifugation the supernatant was discarded to give uniform mix by the addition of DMEM (10% FBS) of the 10㎖ precipitated cells, ⁴ to 10 ea / well degree of cells using a blood cell counter measuring the number of cells in each well of the 96well 200 μl were dispensed.

Subsequently, when about 50% of the well area of the cells grew while incubating at 37 ° C. and 5% CO ₂ , the medium was exchanged with fresh DMEM (FBS10%), and each well for each treatment group of Example 1 was prepared by concentration. The cells were incubated at 37 ° C. and 5% CO ₂ at different times. MTT (3- (4,5-dimethylthiazol-2-yl) 2,5-diphenyl tetrazolium bromide, Sigma) was dissolved in PBS to 2 mg / ml and stored in the refrigerator. MTT reagent (5 mg) was used in each well. / Ml in PBS) was treated with 20ul each and incubated for 2 hours, and then the supernatant was discarded and mixed with 150µl of DMSO (DMSO) or mixed with 100µl of 0.04N-HCl. After all steps were measured at 560nm using ELISA leader (μQuant, Bio-Tek instrument INC.) (Reference: 650nm).

The measurement results are shown in Figure 1 and the effect of the peach blossom extract showed a cell proliferation effect of about 85% compared to when the control was 0 after 48 hours at 0.01% treatment.

Experimental Example 2 Collagen Biosynthesis Effect

In Experimental Example 2, fibroblasts cultured in T-75 flasks were washed 2-3 times with PBS using collagen biosynthesis effect using the peach blossom extract obtained in Example 1, and trypsin was tested. After dropping the attached cells by using, centrifugation (Heraeus; Labofuge 400R) for 5 minutes at 1500rpm to collect the cells. After centrifugation the supernatant was discarded to give the collected mixture even by the addition of DMEM (10% FBS) of 10㎖ the cells, the 10 ⁴ ea / well degree of cells using a blood cell counter measuring the number of cells in each well of the 96well Dispense 200 μl each. After culturing at 37 ° C. and 5% CO ₂ , about 50% of the cells grew well and exchanged with fresh DMEM (FBS 10%).

Collagen synthesis performance measurement kit (Procollagen Type-I C-peptide EIA kit (Takara, MK 101)) was used. For measurement using immunological methods, transfer 100 μl of Ab-POD conjugate solution into one well and dispense, and add 20 μl of cell culture or standard solution. The 96 well plate was wrapped with foil or the like and incubated at 37 ° C. for about 3 hours. After 3 hours, the solution contained in each well was 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, add 100 µl of 1N H ₂ SO ₄ , the reaction termination solution, mix gently, and measure the absorbance using an ELISA leader.The resultant is used to store the plate containing the reaction solution at room temperature for about 1 hour. After measuring at 450nm.

The results are shown in Figure 2 and can be seen that the enhancement effect is shown as well as the results of cell proliferation ability.

Experimental Example 3 Measurement of Active Oxygen Scavenging Ability

Experimental Example 3 measured the active oxygen scavenging ability of the peach blossom extract active ingredient using the peach blossom extract obtained in Example 1. With respect to the peach blossom extract obtained by supercritical and ultrasonic extraction obtained in Example 1 and the ethanol peach blossom extract to be compared, active oxygen scavenging ability (active oxygen species causes various adult diseases and aging, in particular, skin aging , Its scavenging ability is essential in functional anti-wrinkle cosmetics) was measured the content of catechin (Catechin).

Each peach flower extract, about 5 mg of catechin standard (manufactured by Sigma) was precisely weighed, and each was dissolved in 50 ml of a mobile phase. Then, 100 ml of the mobile phase was added to prepare a sample solution and a standard solution.

With the sample solution and the standard solution 20㎕ using liquid chromatography (1100Series, AT G) is obtained, and _T Q Qs of catechins for the peak area of material.

The amount of catechin as polyphenol was determined by the following formula (1).

[Equation 1]

Amount of catechin (mg.) As polyphenol = amount of catechin standard (mg) × (Q _{T /} Qs)

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

Mobile phase: Dissolve 1.0 g of phosphoric acid in 50% methanol solution and make 2 L.

Detector: UV absorbance photometer (wavelength 280nm)

Column: C ₁₈

Flow rate: 1.0ml / min

The results of Experimental Example 3 are shown in Table 2 below.

sample Number of tests As polyenol
Content of catechin (% by weight)) Average Peach Blossom Extract Obtained by Supercritical and Ultrasonic Extraction One 95.0 94.93 2 95.2 3 94.6 Ethanol Peach Blossom
extract One 31.5 30.77 2 30.2 3 30.6

Experimental Example 4 Skin Moisturizing Effect Experiment

This experiment tested the skin moisturizing effect. In order to measure the skin moisturizing effect of simple raw material before nanoliposomalization, Na-PCA (Sodium Pyrolidone Carboxylic acid), Chondroitin, which is a component of NMF (Natural Moisturizing Factor) as a comparison target with the peach blossom extract of Example 1 Na-Condroitin Sulfate and Na-hyaluronate 1% aqueous solution was measured to reduce the TEWL after barrier damage, which is a standard measure of skin barrier function. The specific method is as follows.

Acetone was instilled in the back of 8-12 week old hairless mice 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 by 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 in the efficacy evaluation was calculated as shown in Equation 2 below.

[Formula 2]

(However, Bt = 6: TEWL measurement value after 6 hours after skin barrier damage, Bt = 0 ': TEWL measurement value before skin barrier damage, Bt = d: TEWL measurement value immediately after skin barrier damage)

The results of Experimental Example 4 are shown in Table 3 (unit: mg / m 2 / hr).

Peach Blossom Extract Na-PCA
Aqueous solution (1%) Na-CS
Aqueous solution (1%) Na-Hyaluronate
Aqueous solution (1%) One 12 18.5 17.4 18.4 2 16.5 19 18 19 3 16.1 20.1 19 19.2 4 18.5 21 19 19.6 5 14.3 20.5 20 19.7 6 13 18.5 21 20.2 7 14.9 18.9 18 20.3 8 15 18.3 19.3 21.4 9 16.5 18.8 19 19.7 10 14.6 20.3 18 18.9 11 13.8 17.4 18.2 18.6 12 13.5 17.5 17.9 19.3 13 16.8 19.8 19 20.1 14 15.2 18.8 17.6 21.1 15 16.4 18.5 20 19.8 16 15.3 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 16.8 19.5 20.2 20.1 Average measurement value 14.75 19.3 18.845 19.615

As a result of TEWL measurement, the peach blossom extract used in the present invention was 26.8%, 23.58%, and 25.13%, respectively, compared to Na-PCA (Sodium Pyrolidone Carboxylic acid), which was used as a comparative study. 21.73% and Na-Hyaluronate showed 28.07% and 24.8% better effects, respectively.

Comparative Example 1

Nanoliposomes were prepared in the same manner as in Example 2, except that no peach blossom extract was added.

A cosmetic (Example 3) containing nanoliposomes stabilized peach blossom extract of Example 2 was prepared.

Using the raw materials shown in Table 4 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 with Stabilized Peach Blossom Extracts 0.1 incense A reasonable amount antiseptic A reasonable amount Purified water Remaining amount total 100

The emulsion of Example 4 was prepared in the same manner as in Example 3, except that 1 wt% of the nanoliposome stabilized with the peach blossom extract of Example 2 was added.

An emulsion of Example 5 was prepared in the same manner as in Example 3, except that 5% by weight of the nanoliposome stabilized in the peach blossom extract of Example 2 was added.

An emulsion of Example 6 was prepared in the same manner as in Example 3, except that 10 wt% of the nanoliposome stabilized with the peach blossom extract of Example 2 was added.

An emulsion of Example 7 was prepared in the same manner as in Example 3, except that 20 wt% of the nanoliposome stabilized in the peach blossom extract of Example 2 was added.

Comparative Example 2

The emulsion of Comparative Example 2 was prepared in the same manner as in Example 3, except that 0.05 liposomes stabilized with the peach blossom extract of Example 2 were added.

Comparative Example 3

An emulsion of Comparative Example 3 was prepared in the same manner as in Example 3, except that 0.25 of the lipophilic stabilized peach blossom extract of Example 2 was added.

Experimental Example 5 Measurement of Skin Moisture Content

Experimental Example 5 measured the moisture content of the skin using Examples 3 to 7 and Comparative Preparation Examples 2 and 3.

Eighty healthy women (mean age 29.5 years old) were divided into eight groups of 22 healthy women (22 ~ 24 ℃, 55% relative humidity, no air flow). The cosmetics of Examples 3 to 7 and Comparative Production Examples 2 and 3 were applied to the entire face twice a day for 6 weeks, mainly around the eyes of each group. Using the 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 5 as an average value for each test group.

Improvement was calculated according to the following equation 3 and the results are shown in Table 5.

[Equation 3]

Before the test 6 weeks after application Improvement (%) Example 3 630 ± 10 650 ± 10 3.17 Example 4 625 ± 10 685 ± 10 9.6 Example 5 630 ± 10 890 ± 10 41.27 Example 6 630 ± 10 675 ± 10 7.14 Example 7 630 ± 10 650 ± 10 3.17 Comparative Example 2 630 ± 10 635 ± 10 0.78 Comparative Example 3 630 ± 10 630 ± 10 0

According to Table 5, it can be confirmed that it is preferable to contain 0.1 to 20% by weight of the nano-liposomes stabilized peach blossom extract in the cosmetic.

[Comparative Examples 4 to 6]

Emulsion in the same manner as in Example 3, except that 5% by weight of Na-PCA, sodium chondroitin sulfate, and Na-hyaluronate were used instead of the nanoliposomes stabilized with the peach blossom extract of Example 2. Were prepared, and each was made into Comparative Examples 4-6.

Comparative Example 7

Instead of using the nanoliposomes stabilized peach blossom extract of Example 2, except that 0.5% by weight of the peach blossom extract of Example 1 and 4% by weight of nanoliposomes of Comparative Example 1 are used. The emulsion was prepared by the method.

Comparative Example 8

Emulsion was prepared in the same manner as in Preparation Example 1, except that 0.5 wt% of the peach blossom extract of Example 1 was used instead of the nanoliposomes stabilized with the peach blossom extract of Example 2.

Experimental Example 6 Skin Elasticity Effect Experiment

In this example, the skin elasticity effect was tested. 60 healthy women (average age 29.5 years) over 20 years old were divided into 6 groups under the conditions of temperature of 22-24 ° C and 55% relative humidity, and the cosmetic compositions of Example 5 and Comparative Examples 6 to 8 were respectively centered around the eyes. After weekly application (2 times / day), skin elasticity was measured using a skin elasticity analyzer (Cutometer SEM 575, C + K Electronic Co., Germany). The test results are described in Table 5 as ΔR5 values (R5 (12 weeks) -R5 (0 weeks)), which are average values of the respective test groups of the Cutometer SEM 575. The closer it is, the better the elasticity is.

In addition, the degree of improvement was calculated by the following equation 4 and the results are shown in Table 6 below.

[Equation 4]

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

Skin elasticity test result (12 weeks) Before application After application Skin elasticity enhancing effect (△ R5) % Improvement Example 5 0.74 1.16 0.42 56.76 Comparative Example 4 0.71 0.80 0.09 12.68 Comparative Example 5 0.71 0.81 0.1 14.08 Comparative Example 6 0.73 0.75 0.02 2.74 Comparative Example 7 0.72 0.80 0.08 10.00 Comparative Example 8 0.73 0.77 0.04 5.19

According to Table 6, when using Example 5 containing the stabilized nanoliposome, the skin elasticity is 44.08% compared to Comparative Example 4, 42.68% compared to Comparative Example 5, 54.02% compared to Comparative Example 6, Comparative Example 7 46.76% and 51.57% compared to Comparative Example 8. As can be seen from these results, it can be seen that the cosmetic composition comprising the peach blossom extract according to the present invention effectively enhances skin elasticity.

Experimental Example 7 Melanin Synthesis Amount Inhibitory Effect

In this experimental example, the melanin synthesis amount inhibitory effect was tested using the peach blossom extract of Example 1. Melanocytes isolated from human tissues were primaryly cultured and passaged two times, followed by incubation at a concentration of 2 × 10 ⁴ / dish using RPMI 1640 medium in a 60 mm culture dish. After confirming that the cells attached to the plate, the peach blossom extract was treated by concentration up to 0.001 ~ 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 the sample in which the cells were completely lysed. 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.

Melanin reduction was calculated according to Equation 5 below and the results are shown in Table 7 and FIG. 3.

[Equation 5]

% Melanin reduction = {(control of absorbance-absorbance at each concentration) / control of absorbance} X 100

Peach Blossom Extract (%) Melanin Reduction (%) One 26.35 0.1 34.26 0.01 48.26 0.001 21.10

As can be seen in Table 7, it can be seen that the peach blossom extract also shows the efficacy of whitening by inhibiting the action of biosynthesis of melanin.

Experimental Example 8 Formulation Stability Measurement

Stability of the formulation for the cosmetic composition comprising the peach blossom extract according to the present invention was measured by the following method.

Samples 5 and Comparative Examples 7 and 8 were placed in an opaque glass container in a constant temperature bath maintained at 45 ° C. and stored for 12 weeks in an opaque glass container in a completely shaded refrigerator kept at 4 ° C. For samples stored for 12 weeks and samples stored for 12 weeks in a circulation chamber (once / day) circulating at -5 ° C to 37 ° C, separation, discoloration, and precipitation were compared.

The results of the evaluation of the product separation and discoloration degree classified 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 of test substance Temperature Example 5 Comparative Example 7 Comparative Example 8 45 ℃ 0 3 3 4 ℃ 0 2 2 cycle 0 2 3

Through Table 8, Example 5 stabilized through nanoliposomes was stable without discoloration or separation symptoms at 4 ° C., 45 ° C. and circulating tests, but separated in the case of Comparative Examples 7 and 8 used for direct formulation without stabilization. (Discoloration) 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 sedimentation evaluation criteria:

0: no change 1: very little precipitation

2: settle slightly 3: settle slightly

4: Severe precipitation 5: Extremely precipitation

Precipitation degree of test substance Temperature Example 5 Comparative Example 7 Comparative Example 8 45 ℃ 0 3 3 4 ℃ 0 3 2 cycle 0 3 2

Through Table 9, it was found that the cosmetic composition comprising the peach blossom extract according to the present invention was not precipitated and stabilized.

As such, when the peach blossom extract is added without stabilization, precipitation and discoloration occur easily by reacting with other weak ionic polymers in the formulation. However, when the peach blossom extract is stabilized with nanoliposomes as in the present invention and applied to the formulation, the precipitation and discoloration reactions are minimized even when the peach blossom extract is added in excess.

Experimental Example 9 Transdermal Absorption Measurement Experiment

Percutaneous absorption measurement experiment according to the size of the nanoparticles for the cosmetic composition comprising a peach blossom extract according to the present invention was performed by the following method.

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 were 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 the 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. 34 g of polymethylmethacrylate-co-methacrylic acid is dissolved in 150 ml of anhydrous methylene chloride, and then 132 mg of dicyclohexylcarbodiimide and 74 mg of N-hydroxysuccinimide are 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, is removed through a 0.45 μm nylon filter, and then precipitated in diethyl ether to remove reaction reagents and unreacted residues first, and to stand in excess distilled water for one day. The residue was removed and 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 weeks of female hairless guinea pigs (strain IAF / HA-hrBR) were used for percutaneous absorption testing of nanoparticles by size. The skin of the abdomen of the guinea pig was cut and mounted in a Franz-type diffusion cell (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 10% (w / v) solution of fluorescent PMMA nanoparticles prepared for each size was placed in each donor container. 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 diffusion of nanoparticles was finished, the non-absorbed nanoparticles remaining on the skin were washed with dried Kimwipes or ethanol 10ml, 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 45㎛ 80 nm 16 μm 120 nm 14 μm 500 nm 10 μm

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 upper dermis 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 believed 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.

Experimental Example 10

The nanoliposome prepared in Example 2 was mixed with water to 1 wt%, and the hair was immersed in this solution for 20 minutes, and the tensile strength of the hair was measured. At this time, the tensile strength measurement of the hair was used a digital force gauge (DPS-5R). The measurement was carried out 10 times, and compared with the hair immersed in pure water for 20 minutes for comparison.

The average value of the measured results was that the tensile strength of the hair immersed in the solution containing the nanoliposomes was 0.125 kg, whereas the tensile strength of the hair soaked in pure water was 0.095 kg.

Therefore, the nanoliposomes according to the present invention have been shown to be effective in improving hair strength.

Experimental Example 11

The hair was soaked for 20 minutes in the same manner as in Experimental Example 10, and then the average thickness was calculated by measuring the thickness of the hair 10 times using a hair thickness gauge (ID-C112BS).

At this time, the thickness measurement position measured two points 5cm, 7cm from the lowermost part of the hair.

As a result, the hair immersed in the water containing the nanoliposomes according to the present invention showed that the average thickness of the 5cm point was increased from 0.075mm to 0.083mm, and the increase rate was about 5% from 0.080mm to 0.084mm at the 7cm point.

On the other hand, there was no change in the comparison group.

Therefore, the nanoliposomes according to the present invention have been shown to be effective in improving hair thickness.

In the present specification, the inventors describe only a few examples of various manufacturing and analysis experiments performed according to the manufacturing method according to the present invention, but the technical idea of the present invention is not limited thereto and is variously modified by those skilled in the art. Of course it can be implemented.

Claims (3)

4% by weight of phospholipids, 0.1% by weight of glutathione and 20% by weight of modified ethanol and 30% by weight of glycerin and 4% by weight of polysorbate 20 were heated to 95 ° C., uniformly mixed and then cooled to 40 ° C. next,
After drying the peach blossoms and pulverizing to less than 0.3cm in diameter, mixed with the same mass ratio of butylene glycol and ethanol and mixed in a mass ratio of 1: 2 and stirred at 60 ° C for 3 hours using ultrasonic waves After extraction, the co-solvent mixture was added to a supercritical extraction tank, and the temperature was raised to 40 ° C. in the extraction tank while carbon dioxide was supplied to the extraction tank. After adding water to the raw material cooled to 40 ℃ and mixed,
Nanoliposomes which stabilized the peach blossom extract obtained by passing seven times through a high pressure emulsifier (Microfludizer M210EH, Microfluidies, USA) under a pressure of 1200 bar and a flow rate of 500 m / s,
Characterized in that 1% by weight of the entire composition,
Cosmetic composition for improving hair strength and thickness. delete delete

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