KR102487634B1 - A cosmetic composition containing eisenia bicyclis extract for skin whitening and wrinkle treatment, and manufacturing method thereof - Google Patents

Abstract

The present invention relates to a cosmetic composition for skin whitening and wrinkle improvement containing a rhubarb enzyme extract as an active ingredient, and specifically, by adding an enzyme to a rhubarb mixture prepared by mixing rhubarb and water, a rhubarb enzyme mixture is prepared and extracted under pressure It relates to a cosmetic composition for skin whitening and wrinkle improvement containing rhubarb enzyme extract as an active ingredient, which not only improves the extraction yield and antioxidant ability, but also has skin whitening and wrinkle improvement effects by preparing the rhubarb enzyme extract.

Description

A COSMETIC COMPOSITION CONTAINING EISENIA BICYCLIS EXTRACT FOR SKIN WHITENING AND 　WRINKLE TREATMENT, AND MANUFACTURING METHOD THEREOF}

The present invention relates to a cosmetic composition for skin whitening and wrinkle improvement containing rhubarb enzyme extract as an active ingredient and a method for manufacturing the same, and specifically, to prepare a rhubarb enzyme mixture by adding an enzyme to a rhubarb mixture prepared by mixing rhubarb and water. and pressurized extraction to prepare a rhubarb enzyme extract, which not only improves the extraction yield and antioxidant capacity, but also improves skin whitening and wrinkles. A cosmetic composition for skin whitening and wrinkle improvement containing rhubarb enzyme extract as an active ingredient and a method for manufacturing the same it's about

As the standard of living and income rises and improves, people are escaping from the daily lifestyle of food, clothing and shelter, and interest in personal aesthetic life is increasing (Study on purchasing patterns and preferences of functional cosmetics and functional foods. Pyeon-suk Pyeon, Graduate School of Seokyeong University). .Master's thesis. 2010.). In particular, as the average lifespan has increased and become more relaxed due to the development of modern medicine, there is a lot of interest in anti-aging and skin health (Kim, S.H. Yong, H.J. Shin, C. and Ko, S.G. Research of traditional herbal medicines for anti-aging, inhibition effect of wrinkle and whitening effect in the skin.Korean J. Ori. Physiol. Pathol. 2008, 22, 691-698.).

Skin aging can be divided into intrinsic aging and extrinsic aging according to the factors. Intrinsic aging is the decline of the structure and physiological functions of the skin with age, and extrinsic aging is the aging phenomenon caused by accumulated external stress such as sunlight. In particular, extrinsic aging such as exposure to various pollutants and ultraviolet rays makes the skin thinner, wrinkles increase, elasticity decreases, and spots, freckles, and age spots increase (Gilchrest BA. Skin aging and photoaging. Dermatol Nurs. 1990, 2, 79-82., Kang, K.S. Kim, I.D. Kwon, R.H. Heo, Y.Y. Oh, S.H. Kim, M.A. Jung, H.J. Kang, H.Y. Ha, B.J. The evaluation of anti-wrinkle effects in oriental herb extract.J. Life Sci, 2007, 17, 1147-1151.). As skin aging progresses, the ability to produce structural proteins such as collagen, elastin, and hyaluronic acid, which constitute the skin, decreases, and the biosynthesis of type-1 collagenase increases, resulting in increased expression of matrix metalloproteinases (MMPs), which leads to increased collagen fibers in the dermis. , elasticity fibers, induces degradation of matrix proteins such as fibronectin, which reduces skin elasticity and causes skin wrinkles (Brenneisen P, Sies H, Scharffettter-Kochannek K. “Ultraviolet-B irradiation and matrix metalloproteinases: from induction via signaling to initial events” Ann. N. Y. Acad. Sci, 2002, 973, 31-43.).

Accordingly, studies on cosmetics, foods, and medicines using various natural substances are being actively conducted in order to maintain and improve overall health of the body, such as skin whitening and wrinkle improvement effects.

Korea is made up of the sea on three sides, so there are many kinds of seaweed that live in the sea area. These seaweeds have long been widely used as food, health food, medicine, raw materials for seaweed industry, and fertilizers. Recently, as seaweed contains various functional components as much as plants, active research is being conducted on the separation of these functional components and the verification and utilization of their efficacy (Choi, D.M., Kim, D.S., Lee, D.S., Kim, H.R., and Pyeun, J.H. 1995. Trace components and functional sacchsrides in seaweed-1. J. Korean Fish. Soc. 28: 49-59.).

The large brown algae rhubarb (Eisenia bicyclis) is a perennial seaweed belonging to the genus Rhubarb in the genus Gamtaeidae and forms large colonies in Ulleungdo and Dokdo in Korea, and in the east coast, colonies are developed in Yeongdeok, Gyeongsangbuk-do. Effects of temperature, light intensity, and photoperiod Lee Min-jung Lee, Graduate School of Gyeongsang National University. Master's thesis. 2018). Rhubarb contains a large amount of iodine and potassium, and has been used as a substitute for kelp since ancient times due to its unique taste. Master's thesis. 2009.).

Research on the functionality of rhubarb is also being actively conducted. 43, 117-124.), anti-inflammatory (Jung, HA, Jin SE, Ahn BR, Lee CM and Choi JS. 2013. Anti-inflammatory activity of edible brown alga Eisenia bicyclis and its constituents fucosterol and phlorotannins in LPS-stimulated RAW264 .7 macrophages. Food Chem Toxicol 59, 199-206.), antihyperlipidemia (Jang YH, Choi SW and Cho SH. 2008. Effect of Eisenia bicyclis and its pill on serum lipid status in rats fed high fat diet. Korean J Food Nutri 41, 5-12.), antidiabetic (Okada Y, Ishimaru A, Suzuki R and Okuyama T. 2004. A new phloroglucinol derivative from the brown alga Eisenia bicyclis: potential for the effective treatment of diabetic complications. J Nat Prod 67 , 103-105.) have been reported to have various efficacies. In addition, rhubarb contains a large amount of physiologically active substances such as phlorotannin, and phlorotannin isolated from rhubarb includes eckol, phlorofucofuroeckol-A, dieckol, and 8,8'-bieckol, and is reported to have antioxidant activity. (Okada Y, Ishimaru A, Suzuki R and Okuyama T. 2004. A new phloroglucinol derivative from the brown alga Eisenia bicyclis: potential for the effective treatment of diabetic complications. J Nat Prod 67, 103-105.).

On the other hand, Korean Patent Publication No. 10-2013-0141874 relates to an antimicrobial composition containing a rhubarb-derived phlorotannin compound as an active ingredient, and the antimicrobial composition has excellent antibacterial activity against MRSA (Methicillin-Resistant Staphylococcus aureus). Disclosed is an antibacterial composition comprising a phlorotannin compound having as an active ingredient.

Korean Patent Registration No. 10-0966128 discloses a rhubarb enzyme extract having a therapeutic effect on hyperlipidemia and diabetes prepared by manufacturing rhubarb powder from seaweed rhubarb, extrusion molding and enzymatic hydrolysis, and adding rhubarb powder, kelp powder and excipients to the rhubarb enzyme extract. It discloses a rhubarb tablet pill prepared by mixing and a manufacturing method thereof.

Korean Patent Publication No. 10-2011-0057586 discloses a pharmaceutical composition for preventing or treating diabetes and a health functional food composition for improving diabetes consisting of brown algae rhubarb powder, rhubarb enzyme extract, kelp powder, and excipients.

As described above, various studies on rhubarb are being conducted and its functionality is known, but its utilization is still insufficient, and in particular, functional verification for skin improvement and industrial application to cosmetics are very low.

Korean Patent Publication No. 10-2013-0141874 (2013.12.27.) Korean Patent Registration No. 10-0966128 (2010.06.17.) Korean Patent Publication No. 10-2011-0057586 (2011.06.01.)

The present invention has been made to solve the problems described above and provide necessary technology,

An object of the present invention is to provide a cosmetic composition for skin whitening and wrinkle improvement containing as an active ingredient a rhubarb enzyme extract having skin whitening and wrinkle improvement effects as well as improving extraction yield and antioxidant activity, and a manufacturing method thereof.

As one embodiment of the present invention for achieving the above object,

One embodiment of the present invention is a rhubarb mixture preparation step of preparing a rhubarb mixture by mixing 2000 to 4000 parts by weight of water with respect to 100 parts by weight of dried and pulverized rhubarb powder; 2 to 5 parts by weight of any one enzyme selected from the group consisting of neutrase, viscozyme and celluclast with respect to 100 parts by weight of rhubarb powder in the rhubarb mixture prepared in the rhubarb mixture preparation step A stirring step of preparing a rhubarb enzyme mixture by adding part and stirring at a temperature of 50 to 70 ° C. at 200 rpm for 15 to 20 hours; And pressurized extraction of the rhubarb enzyme mixture prepared in the stirring step at 1.2 atm and a temperature of 100 to 130 ° C. for 20 to 70 minutes, followed by centrifugation at 2000 to 4000 rpm for 10 to 30 minutes to prepare a rhubarb enzyme extract by taking only the supernatant It provides a method for producing a cosmetic composition for skin whitening and wrinkle improvement using the rhubarb enzyme extract as an active ingredient, characterized in that it is prepared by including; rhubarb enzyme extract preparation step.

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Another embodiment of the present invention provides a cosmetic composition for skin whitening and wrinkle improvement comprising the rhubarb enzyme extract, characterized in that prepared by the above method, as an active ingredient.

In the present invention, the cosmetic composition for skin whitening and wrinkle improvement is from the group consisting of lotion, lotion, serum, massage cream, nutrient cream, eye cream, moisture cream, hand cream, sunscreen, essence, nutrient essence, body lotion, and face wash It may be characterized in that it is prepared in any one selected formulation.

A cosmetic composition for skin whitening and wrinkle improvement comprising a rhubarb enzyme extract prepared according to an embodiment of the present invention as an active ingredient is prepared by adding an enzyme to a rhubarb mixture prepared by mixing rhubarb and water to prepare a rhubarb enzyme mixture and pressurize it. As the rhubarb enzyme extract is prepared by extraction, the extraction yield and antioxidant capacity are improved, as well as skin whitening and wrinkle improvement effects, so it can be used as a cosmetic composition of various formulations.

1 is a graph showing the results of measuring the tyrosinase inhibitory activity of rhubarb enzyme extract.
Figure 2 is a graph showing the results of measuring the elastase inhibitory activity for the rhubarb enzyme extract.
Figure 3 is a graph showing the results of measuring the viscosity when the cream containing the rhubarb enzyme extract was stored at 4 ℃.
Figure 4 is a graph showing the results of measuring the viscosity when the cream containing the rhubarb enzyme extract was stored at 25 ℃.
Figure 5 is a graph showing the results of measuring the viscosity when the cream containing the rhubarb enzyme extract was stored at 40 ℃.
Figure 6 is a photograph showing the results of observation of discoloration and odor when the cream containing the rhubarb enzyme extract was stored at 4 ° C.
Figure 7 is a photograph showing the results of observation of discoloration and odor when the cream containing the rhubarb enzyme extract was stored at 25 ° C.
Figure 8 is a photograph showing the results of observation of discoloration and odor when the cream containing the rhubarb enzyme extract was stored at 40 ° C.

Hereinafter, embodiments of the present invention will be described in detail so that those skilled in the art can easily practice the present invention. Embodiments of the present invention are provided to more completely explain the present invention to those skilled in the art. Accordingly, the embodiments of the present invention can be modified in many different forms, and the scope of the present invention is not limited to the embodiments described below.

Throughout the specification of the present invention, when a part "includes" a certain component, it means that it may further include other components without excluding other components unless otherwise stated.

The terms "about," "substantially," and the like used throughout the specification of the present invention are used at or close to that value when manufacturing and material tolerances inherent in the stated meaning are given, and the present invention Accurate or absolute figures are used to prevent unfair use by unscrupulous infringers of the disclosed disclosures mentioned for the sake of understanding. The term “step of (doing)” or “step of” used throughout the present specification does not mean “step for”.

The present invention provides a cosmetic composition for skin whitening and wrinkle improvement (hereinafter also referred to as 'cosmetic composition') containing rhubarb enzyme extract as an active ingredient.

In the cosmetic composition according to the present invention, a rhubarb enzyme mixture is prepared by adding an enzyme to a rhubarb mixture prepared by mixing rhubarb and water, and a rhubarb enzyme extract is prepared by pressurized extraction, so that the extraction yield and antioxidant activity are not only improved, but also the skin It is expected to have high whitening and wrinkle improvement effects.

According to one embodiment of the present invention, a rhubarb mixture preparation step of preparing a rhubarb mixture by mixing rhubarb powder and water, stirring to prepare a rhubarb enzyme mixture by adding an enzyme to the rhubarb mixture prepared in the rhubarb mixture preparation step and stirring It may be characterized in that it is prepared by including a rhubarb enzyme extract production step of preparing a rhubarb enzyme extract by extracting the rhubarb enzyme mixture prepared in the steps and the stirring step under pressure and then centrifuging to obtain only the supernatant.

In addition, according to one embodiment of the present invention, the rhubarb mixture may be characterized in that it is prepared by mixing 2000 to 4000 parts by weight of water with respect to 100 parts by weight of dried and pulverized rhubarb powder.

In addition, according to one embodiment of the present invention, the rhubarb enzyme mixture is added with 2 to 5 parts by weight of enzyme based on 100 parts by weight of rhubarb powder and stirred for 15 to 20 hours at a temperature of 50 to 70 ° C. at 100 to 200 rpm. It can be characterized in that the rhubarb enzyme mixture is prepared by doing.

In addition, according to one embodiment of the present invention, the enzyme used to prepare the rhubarb enzyme mixture is added to any one selected from the group consisting of neutrase, viscozyme and celluclast. that can be characterized.

In addition, according to one embodiment of the present invention, the rhubarb enzyme extract is extracted under pressure from the rhubarb enzyme mixture at 1.2 atm and a temperature of 100 to 130 ° C. for 20 to 70 minutes, and then centrifuged at 2000 to 4000 rpm for 10 to 30 minutes. It can be characterized in that it is prepared by taking only the supernatant.

If the rhubarb enzyme extract is out of the limited range as described above, antioxidant, skin whitening and wrinkle improvement effects cannot be expected due to the low content or dilute concentration of useful ingredients in the extract. Although anti-inflammatory and anti-wrinkle effects may have an enhancing effect, there is a concern that manufacturing cost may increase and skin toxicity may also occur in cosmetic manufacturing.

At this time, when using the rhubarb enzyme extract as a cosmetic composition, the cosmetic composition may be characterized in that the rhubarb enzyme extract is included in a ratio of 0.1 to 90% by weight based on the total weight% of the cosmetic composition.

This is because when the rhubarb enzyme extract is included in an amount of less than 0.1% by weight with respect to the total weight% of the cosmetic composition, the antioxidant, skin whitening and wrinkle improvement effects may not be properly expressed, and the rhubarb extract is 90% by weight relative to the total weight% of the cosmetic composition. If it is included in a ratio exceeding weight %, a problem in that the manufacturing cost increases in manufacturing cosmetics may occur.

In addition, the cosmetic composition according to the present invention is selected from the group consisting of lotion, lotion, serum, massage cream, nutrient cream, eye cream, moisture cream, hand cream, sunscreen, essence, nutrient essence, body lotion, and face wash in a conventional method. It may be formulated into any one formulation and used.

Hereinafter, the present invention will be described in detail according to specific examples. The cosmetic composition prepared according to an embodiment of the present invention has a rhubarb enzyme extract yield measurement experiment, a total polyphenol content measurement experiment, a total sugar and total protein content measurement experiment, an antioxidant activity measurement experiment, and a tyrosinase inhibitory activity measurement test. An experiment, an experiment to measure elastase inhibitory activity, and an experiment to evaluate the storage stability of a cream containing a rhubarb enzyme extract were conducted. The cosmetic composition according to an embodiment of the present invention can be more clearly understood by experiments described later.

Preparation of rhubarb enzyme extract

1. Rhubarb mixture preparation step: A rhubarb mixture is prepared by mixing 3000 parts by weight of water with respect to 100 parts by weight of dried and pulverized rhubarb.

2. Stirring step: Add 3 parts by weight of any one selected from the group consisting of neutrase, viscozyme and celluclast based on 100 parts by weight of rhubarb powder to the above rhubarb mixture and stir at 150 rpm A rhubarb enzyme mixture was prepared by stirring at a temperature of 60 ° C. for 18 hours.

3. Rhubarb enzyme extract preparation step: The rhubarb enzyme mixture is pressure-extracted at 121 ° C. and 1.2 atm for 30 to 60 minutes, and then centrifuged at 3000 rpm for 20 minutes to prepare a rhubarb enzyme extract by taking only the supernatant.

Rhubarb enzyme extract yield, total polyphenol content, total sugar and total protein content measurement experiment

The yield of the rhubarb enzyme extract was measured by the solid content of the raw material dry matter used in the dried sample after drying using a freeze dryer.

Analysis of the total polyphenol content was performed according to the Folin-Denis method by adding 1 mL of 1 N Folin Ciocalteu reagent to 1 mL of the sample, mixing thoroughly, adding 1 mL of 20% Na ₂ CO ₃ and reacting for 30 minutes in the dark at room temperature. Absorbance was measured at 725 nm using a spectrophotometer. The total polyphenol content was calculated from a standard curve prepared using gallic acid.

Total sugar content was analyzed by the phenol-sulfuric acid method according to the method of saha and brewer. That is, after reacting 1 mL of 5% phenol (w/v) and sulfuric acid with 1 mL of sample, the absorbance was measured at 525 nm using a spectrophotometer and calculated using glucose as a standard curve.

Total protein content was measured by the method of Lowry et al. and calculated using bovine serum albumin (BSA) as a standard curve.

Experiment result

The yield of the rhubarb enzyme extract was the highest at 34.0% in the experimental group (N60), which was extracted under pressure for 60 minutes after treatment with neutrase.

The total polyphenol content was 35.05 ± 0.35 mg / 100 g and 32.50 ± 0.17 mg / 100 g, respectively, in the N30 and N60 sections extracted after treatment with neutrase. On the other hand, the total sugar content was the highest at 38.62g/100g in V30 using viscozyme. The total protein content was the highest at 28.33 g/100 g and 26.53 g/100 g, respectively, in N30 and N60 using neutrase. The measurement results for the yield, total polyphenol content, total sugar and total protein content of the rhubarb enzyme extract are shown in Table 1.

Sample ¹⁾ Yield
(%) Total polyphenols
(mg/g) Total sugar
(g/100g) Total protein
(g/100g) WE 28.2±0.5 ^cd2) 128.23±0.76 ^d 33.36± ^0.43c 17.49± ^0.34d AC 19.9±1.2 ^f 120.23± ^1.04e 34.46± ^0.31b 17.09±0.55 ^d C30 27.2±1.6 ^de 128.90±1.73 ^d 32.95±0.45 ^cd 16.95±0.17 ^d C60 29.5± ^0.6bc 114.57±1.15 ^f 34.96± ^0.56b 15.41± ^0.08f V30 26.5± ^0.5e 141.57±0.58 ^c 38.62±0.25 ^a 18.70± ^0.25c V60 28.2±0.3 ^cd 119.07± ^1.44e 32.55±0.53 ^cd 16.22± ^0.50e N30 29.8± ^0.8b 180.07±1.76 ^a 30.59± ^0.49e 28.33±0.08 ^a N60 34.0± ^0.6a 167.40± ^0.87b 32.11± ^1.21d 26.53± ^{0.34b 1)} WE: rhubarb hot water extract,
AC: pressurized rhubarb extract extracted by autoclave,
C30: pressurized rhubarb extract extracted by autoclave for 30 minutes after celluclast treatment,
C60: pressurized rhubarb extract extracted by autoclave for 30 minutes after celluclast treatment,
V30: pressurized rhubarb extract extracted by autoclave for 30 minutes after viscozyme treatment,
V60: pressurized rhubarb extract extracted by autoclave for 30 minutes after viscozyme treatment,
N30: pressurized rhubarb extract extracted by autoclave for 30 minutes after neutrase treatment,
N60: Pressurized rhubarb extract extracted by autoclave for 30 minutes after neutrase treatment.

²⁾ Means±SD (n=3) with different letters (af) within column are significantly different by Duncan's multiple range test (p<0.05).

Antioxidant activity measurement experiment of rhubarb enzyme extract

The reducing power by the Ferric reducing antioxidant potential (FRAP) method was measured according to the method of Benzie and Strain in 300 mM acetate buffer (pH 3.6) and 10 mM TPTZ (2,4,6-tripyridyl-s-triazine) solution dissolved in 40 mM HCl. and 20 mM FeCl _{3.6H 2} O were mixed at a ratio of 10:1:1 (v:v:v) and warmed on a water bath at 37° C., and then used as a FRAP substrate solution. 25 μL of sample solution diluted to 5 mg/mL concentration and 175 μL of FRAP substrate solution were sequentially mixed in a 96-well plate, reacted at 37 ° C for 4 minutes, and then absorbance was measured at 590 nm using a microplate reader.

2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activity was determined by mixing 7.4 mM ABTS and 2.45 mM potassium persulfate to a final concentration and leaving it in the dark at room temperature for 24 hours to obtain ABTS ⁺ After forming, it was diluted with phosphate buffer saline (PBS, pH 7.4) so that the absorbance was 0.70±0.02 at 732 nm. 100 μL of the sample was mixed with 1900 μL of the diluted solution, reacted for 1 minute, and then the absorbance was measured at 732 nm using a spectrophotometer. The difference in ABTS radical scavenging activity before and after the addition of the extract was calculated.

Experiment result

As a result of measuring the antioxidant activity by the FRAP method, the rhubarb enzyme extract showed a higher effect than the rhubarb hot water extract, but it was confirmed that the 30-minute experimental group showed higher antioxidant activity than the 60-minute experimental group subjected to pressure extraction. Among the enzymes, the antioxidant activity was high in the experimental group using neutrase, which is considered to be excellent in antioxidant activity because the total polyphenol content was high. Similar experimental results were shown in ABTS radical scavenging activity, and both experimental groups showed a concentration-dependent effect on the sample. The measurement results for antioxidant capacity are shown in Table 2.

Sample FRAP (μM) ABTS radical scavenging activity (%) 5,000 µg/mL 10,000 µg/mL 5,000 µg/mL 10,000 µg/mL WE 372.46±2.34 ^l 569.25± ^6.33g 67.87±2.66 ⁱ 91.49± ^0.45e AC 410.61± ^2.50k 645.14±3.79 ^d 72.82± ^4.65h 96.58±0.79 ^c C30 370.60±5.61 ^l 625.90± ^7.42e 77.67± ^2.23g 98.17± ^0.91b C60 349.36± ^5.19m 590.92±8.64 ^f 70.94±2.08 ^hi 94.75±0.87 ^d V30 426.08±11.00 ^j 704.84±5.99 ^{c 79.65} ±1.22g 99.06± ^0.31ab V60 361.53±8.92 ^l 654.57±3.55 ^d 71.44±1.30 ^hi 95.99± ^0.93c N30 469.60±2.86 ^h 785±53± ^4.59a 87.03±0.84 ^f 99.65± ^0.09a N60 444.64±8.45 ⁱ 749.97± ^2.48b 84.75±1.40 ^f 98.91± ^{0.48ab 2)} Means±SD (n=3) with different letters (ai) within column are significantly different by Duncan's multiple range test (p<0.05).

Tyrosinase inhibitory activity measurement experiment

The tyrosinase inhibitory activity was measured by adding 20 μL of mushroom tyrosinase (100 U/mL) to a mixture of 40 μL of 0.175 M phosphate buffer solution (pH 6.8), 40 μL of 5 mM L-DOPA solution, and 100 μL of sample. After reacting for 5 minutes in a ℃ incubator, the absorbance was measured at 475 nm, and the tyrosinase inhibitory activity showed the absorbance decrease rate of the group with and without the addition of the sample solution.

Tyrosinase inhibition activity (%) =

S : absorbance of sample at 475 nm

C: absorbance of control at 475 nm

Experiment result

As a result of analyzing tyrosinase inhibitory activity, at a concentration of 10,000 ug/mL, depending on the type of enzyme, neutrase 54.21% and 55.12%, viscozyme 48.45% and 48.65, celluclast Inhibition rates of 43.93% and 46.83% were shown, and the lowest tyrosinase inhibitory activity was shown in the hot water extract of rhubarb without enzyme treatment with WE 39.44% and AC 43.53%, and the pressurized extract had higher inhibitory activity than the hot water extract. It was confirmed that the most effective tyrosinase inhibitory activity appeared in the rhubarb enzyme extract. The results of measuring tyrosinase inhibitory activity are shown in FIG. 1 below.

Elastase inhibitory activity measurement experiment

To measure elastase inhibitory activity, 200 μL of buffer was added to 20 μL of the diluted sample, and then 20 μL of 0.8 mM N-succinyl-(Ala) ₃ -ρ-nitroanilide was added. This was incubated at 25° C. for 10 minutes, and then 20 μL of 1.0 μg/mL porcine pancreatic elastase was added. The reaction mixture was incubated again at 25° C. for 20 minutes, and absorbance was measured at 405 nm. Elastase inhibitory activity showed the rate of decrease in absorbance of the sample solution added group and non-added group.

Elastase inhibition activity (%) =

S : absorbance of sample at 405 nm

C: absorbance of control at 405 nm

Experiment result

WE, AC, C30, C60, V30, and V60 were 14.67%, 16.39%, 17.79%, 20.09%, 17.41%, and 19.71%, respectively, showing no statistically significant difference, whereas N30 and N60 were 25.89% and 26.79%, respectively. It is thought that the rhubarb enzyme extract extracted after decomposition of the neutrase enzyme will have an effect on improving skin wrinkles. Elastase inhibitory activity measurement results are shown in Figure 2 below.

Evaluation of storage stability of cream containing rhubarb enzyme extract

1. Preparation of cream with rhubarb enzyme extract added

Cream preparation was prepared according to the recipe in Table 3. After completely dissolving Part A, a fat-soluble material, at 75°C, Part B, a water-soluble material adjusted to 70°C, was slowly added to dissolve completely, and emulsified with a blender. In addition, it was homogenized while adding Part C containing rhubarb extract according to the extraction method to the mixture of Parts A and B.

Part commercial name Control Sample A Avocado Butter 10 10 Camellia Oil 7 7 Babassu Oil 3 3 Sorbitan Olivate 3 3 Cetyl Alcohol 2 2 B Green Tea Water 42 42 D.W. 5 0 C Aloe Vera Gel 20 20 D-Panthenol 2 2 Sodium Hyaluronate 3 3 β-glucan 2 2 Vitamin E One One Enzyme extract of sea oak 0 5 Total 100 100

2. Chromaticity measurement

For color difference, lightness (L), redness (a-value), and yellowness (b-value) were measured using a CR-400 color difference meter (KONICA Minolta, Kenis, Japan). At this time, the standard color was measured three times with an L value of 93.53, a value of -0.28, and b value of 4.11, respectively, and was expressed as the average value.

3. pH measurement

After the cream to which the rhubarb enzyme extract was added was placed in a cosmetic storage container, the pH stability was measured at 0 ° C, 25 ° C, and 40 ° C for 4 weeks at 1-week intervals. The measurement method was to measure the pH after taking 1 g of the sample, filling it with distilled water to 9 mL, and sonication for 1 hour with a sonicator.

4. Viscosity measurement

Since the cream used in the viscosity measurement experiment was a fluid viscous liquid, the viscosity was measured with a Brookfield viscometer using a T-bar spindle. That is, the cream was measured using a device that measures the viscosity by detecting the torque value of the viscous resistance of the cream moving on the spindle rotating at a constant acceleration.

5. Observation of discoloration and odor

After putting the cream with rhubarb enzyme extract into a cosmetic storage container, visually inspect the layer separation and aggregation, discoloration and odor to evaluate storage stability at 0℃, 25℃, and 40℃ for 4 weeks at 1-week intervals. reviewed.

Experiment result

1. Chromaticity measurement result

Overall, no significant change was observed with the storage temperature and storage period of the sample under all conditions, but the value of b, which represents yellowness, was higher than that of a, because the color of the rhubarb extract was brown. The L value was the highest in the cream without the rhubarb extract, and when the rhubarb extract was added, the L value was lower than that of the non-added group due to the color of the extract. In the case of the a value representing red color, the lowest value was shown in the section where the rhubarb extract was not added, and the highest redness was shown in N30. Overall, it showed a tendency to increase as the storage temperature increased. The b value also showed a slight increase as the storage temperature increased, but did not show a significant difference. Therefore, the cream with the addition of rhubarb extract is judged to be stable in the future application of the product because the color change is constant or almost indistinguishable depending on the storage period and temperature. The chromaticity measurement results are shown in Table 4, Table 5 and Table 6 below.

Chromaticity L value measurement result Sample and storage temperature Storage Period (Day) 0 7 14 21 28 NO 4℃ 84.97±0.11 85.57±0.16 85.68±0.20 84.78±0.15 25℃ 85.10±0.19 84.82±0.12 84.47±0.13 84.48±0.18 83.96±0.13 40℃ 83.71±0.10 83.00±0.04 82.76±0.06 82.27±0.09 WE 4℃ 74.38±0.02 84.55±0.16 84.35±0.14 83.74±0.04 25℃ 83.74±0.09 83.44±0.05 83.05±0.14 83.17±0.02 82.72±0.09 40℃ 82.21±0.08 82.07±0.06 81.73±0.06 80.54±0.39 AC 4℃ 83.89±0.04 84.32±0.24 84.32±0.10 83.47±0.02 25℃ 83.87±0.13 83.62±0.04 83.35±0.05 83.36±0.04 82.92±0.06 40℃ 82.37±0.04 81.76±0.16 81.26±0.13 80.15±0.29 C30 4℃ 83.82±0.24 83.71±0.16 84.37±0.33 83.52±0.14 25℃ 83.72±0.07 84.27±0.12 83.92±0.11 83.91±0.09 83.48±0.15 40℃ 83.31±0.03 82.77±0.02 82.40±0.12 81.64±0.26 V30 4℃ 84.51±0.08 84.33±0.09 84.94±0.16 84.06±0.08 25℃ 84.43±0.02 84.09±0.01 83.70±0.05 83.75±0.04 83.43±0.05 40℃ 83.43±0.06 82.89±0.10 82.57±0.04 81.77±0.07 N30 4℃ 84.33±0.09 83.97±0.03 84.54±0.10 83.82±0.07 25℃ 83.99±0.15 83.82±0.06 83.49±0.15 83.49±0.04 83.13±0.06 40℃ 82.96±0.13 82.67±0.19 82.26±0.21 81.75±0.16

Chromaticity a value measurement result Sample and storage temperature Storage Period (Day) 0 7 14 21 28 NO 4℃ -0.69±0.01 -0.84±0.03 -0.36±0.04 -0.12±0.01 25℃ -0.65±0.03 -0.76±0.02 -0.87±0.03 -0.41±0.03 0.01±0.02 40℃ -0.68±0.00 -0.72±0.02 -0.23±0.01 0.03±0.03 WE 4℃ 0.11±0.01 -0.04±0.01 0.41±0.02 0.66±0.01 25℃ 0.23±0.01 0.23±0.01 0.16±0.02 0.56±0.02 0.86±0.01 40℃ 0.33±0.04 0.21±0.06 0.69±0.05 0.78±0.08 AC 4℃ -0.05±0.02 -0.25±0.02 0.25±0.01 0.51±0.02 25℃ 0.07±0.01 0.04±0.01 -0.05±0.02 0.40±0.01 0.71±0.02 40℃ 0.16±0.01 0.05±0.00 0.54±0.02 0.68±0.02 C30 4℃ -0.05±0.01 0.09±0.01 0.42±0.02 0.47±0.01 25℃ -0.03±0.02 -0.01±0.01 0.21±0.01 0.59±0.02 0.67±0.02 40℃ 0.12±0.02 0.37±0.02 0.74±0.01 0.71±0.02 V30 4℃ 0.00±0.02 0.14±0.01 0.45±0.01 0.49±0.01 25℃ 0.02±0.02 -0.02±0.01 0.20±0.02 0.57±0.01 0.63±0.01 40℃ 0.07±0.02 0.31±0.01 0.67±0.01 0.65±0.01 N30 4℃ 0.34±0.02 0.48±0.01 0.82±0.03 0.83±0.02 25℃ 0.42±0.02 0.37±0.02 0.59±0.03 0.93±0.01 0.98±0.01 40℃ 0.47±0.02 0.67±0.02 1.04±0.01 0.95±0.04

Chromaticity b value measurement result Sample and storage temperature Storage Period (Day) 0 7 14 21 28 NO 4℃ 2.65±0.02 2.38±0.09 2.22±0.07 2.28±0.04 25℃ 2.63±0.19 2.81±0.04 3.04±0.08 2.94±0.07 2.92±0.05 40℃ 2.95±0.03 3.50±0.02 3.40±0.01 3.48±0.05 WE 4℃ 4.17±0.02 4.34±0.04 4.24±0.02 4.25±0.04 25℃ 4.44±0.02 4.64±0.02 4.88±0.04 4.87±0.01 4.94±0.02 40℃ 5.18±0.06 5.66±0.04 5.72±0.03 6.04±0.07 AC 4℃ 4.45±0.02 4.28±0.01 4.36±0.01 4.44±0.02 25℃ 4.49±0.02 4.86±0.02 5.04±0.03 4.99±0.02 5.03±0.03 40℃ 5.11±0.02 5.62±0.02 5.64±0.06 6.11±0.08 C30 4℃ 4.64±0.03 4.52±0.03 4.32±0.03 4.48±0.03 25℃ 4.82±0.02 5.04±0.02 5.08±0.03 5.00±0.03 5.04±0.04 40℃ 5.28±0.02 5.54±0.06 5.62±0.05 5.84±0.05 V30 4℃ 4.88±0.03 4.77±0.03 4.68±0.04 4.80±0.05 25℃ 4.98±0.04 5.16±0.02 5.18±0.05 5.13±0.01 5.17±0.03 40℃ 5.44±0.02 5.71±0.05 5.80±0.02 6.05±0.05 N30 4℃ 4.99±0.04 4.99±0.03 4.85±0.05 4.89±0.06 25℃ 5.19±0.06 5.34±0.02 5.40±0.13 5.52±0.02 5.64±0.04 40℃ 5.72±0.07 5.96±0.12 6.16±0.07 6.29±0.07

2. pH measurement results

The initial pH of the cream without the addition of rhubarb extract was 7.29, which was higher than that of the cream with the addition of rhubarb enzyme extract (7.04~7.16). Overall, the pH tended to decrease slightly as the storage period increased, and the pH tended to decrease as the temperature decreased depending on the storage temperature, but the difference was not large, confirming stability. The pH measurement results are shown in Table 7.

pH measurement result Sample and storage temperature Storage Period (Day) 0 7 14 21 28 NO 4℃ 7.38±0.01 7.38±0.03 7.21±0.03 7.25±0.02 25℃ 7.29±0.01 7.31±0.00 7.24±0.02 7.18±0.00 7.16±0.01 40℃ 7.25±0.00 7.20±0.02 7.11±0.01 7.17±0.01 WE 4℃ 7.04±0.01 7.08±0.02 7.01±0.00 7.06±0.02 25℃ 7.13±0.01 7.05±0.01 7.04±0.02 6.99±0.01 6.97±0.00 40℃ 7.09±0.01 7.09±0.02 7.00±0.01 7.02±0.02 AC 4℃ 7.13±0.01 7.12±0.01 7.13±0.01 7.11±0.01 25℃ 7.16±0.01 7.15±0.01 7.06±0.02 7.06±0.01 7.03±0.01 40℃ 7.13±0.00 7.09±0.02 7.03±0.01 7.03±0.00 C30 4℃ 7.08±0.02 7.13±0.01 7.05±0.01 7.03±0.01 25℃ 7.13±0.01 7.13±0.01 7.02±0.01 7.03±0.01 7.05±0.01 40℃ 7.04±0.02 6.99±0.01 6.98±0.01 7.00±0.01 V30 4℃ 7.08±0.01 7.13±0.01 6.95±0.03 7.04±0.01 25℃ 7.11±0.02 7.11±0.01 7.08±0.01 6.93±0.03 7.01±0.01 40℃ 7.04±0.00 7.04±0.01 6.95±0.01 6.95±0.02 N30 4℃ 7.00±0.01 7.12±0.00 6.99±0.01 7.03±0.01 25℃ 7.04±0.01 7.07±0.02 7.11±0.01 7.04±0.01 7.07±0.01 40℃ 7.05±0.01 6.92±0.01 6.94±0.00 6.96±0.01

3. Viscosity measurement results

The stability of cosmetics during the storage period is very important because the quality of cosmetics should not deteriorate due to temperature changes. Viscosity of cosmetics may change due to period or temperature, which may affect quality. Therefore, a viscosity stability test was conducted to confirm whether the cream containing the rhubarb extract had an effect on the change in viscosity at temperature. As a result of measuring the viscosity of the cream containing and not containing the rhubarb enzyme extract, it was confirmed that the initial viscosity of the cream containing the rhubarb enzyme extract was 11,266 to 12,856 cP, lower than the initial NO viscosity of 13,400 cP. The viscosity according to temperature showed a decrease in viscosity after 4 weeks from the initial viscosity in all conditions of the cream with rhubarb added and the control cream without added rhubarb at 4℃, 25℃ and 40℃. In the case of 40 ℃, the viscosity did not decrease significantly compared to 4 ℃ and 25 ℃. Viscosity measurement results are shown in Figures 3, 4 and 5 below.

4. Observation of discoloration and odor

As a result of observing the color and odor at 4℃, 25℃ and 40℃ according to the storage period of creams containing and without rhubarb enzyme extract, there was no change in cream color and no specific odor. The same phenomenon was not observed either. The results of discoloration and odor change are shown in FIGS. 6, 7 and 8 below.

In conclusion, through the results of Examples 1 to 6, the cosmetic composition according to one embodiment of the present invention contains the active ingredient of the rhubarb enzyme extract prepared using enzymes, thereby exhibiting antioxidant activity, skin whitening and wrinkle improvement effects. confirmed that there is

Above, the present invention has been described in detail with examples, but the present invention is not limited to the above embodiments, and can be modified in various forms, and within the technical spirit of the present invention, those skilled in the art It is clear that many variations are possible by the ruler. In addition, various forms of substitution, modification and change will be possible by those skilled in the art within the scope of the technical spirit of the present invention described in the claims, which also falls within the scope of the present invention. something to do.

Claims (7)

A rhubarb mixture preparation step of preparing a rhubarb mixture by mixing 2000 to 4000 parts by weight of water with respect to 100 parts by weight of dried and pulverized rhubarb powder;
2 to 5 parts by weight of any one enzyme selected from the group consisting of neutrase, viscozyme and celluclast with respect to 100 parts by weight of rhubarb powder in the rhubarb mixture prepared in the rhubarb mixture preparation step A stirring step of preparing a rhubarb enzyme mixture by adding part and stirring at a temperature of 50 to 70 ° C. at 200 rpm for 15 to 20 hours; and
The rhubarb enzyme mixture prepared in the stirring step is pressure-extracted at 1.2 atm and a temperature of 100 to 130 ° C. for 20 to 70 minutes, and then centrifuged at 2000 to 4000 rpm for 10 to 30 minutes to prepare a rhubarb enzyme extract by taking only the supernatant. Enzyme extract preparation step; Method for producing a cosmetic composition for skin whitening and wrinkle improvement comprising the rhubarb enzyme extract as an active ingredient, characterized in that produced by including. delete delete delete delete A cosmetic composition for skin whitening and wrinkle improvement comprising rhubarb enzyme extract, characterized in that it is prepared by the method of claim 1, as an active ingredient. The method of claim 6,
The cosmetic composition for skin whitening and wrinkle improvement is any one selected from the group consisting of lotion, lotion, serum, massage cream, nutrient cream, eye cream, moisture cream, hand cream, sunscreen, essence, nutrient essence, body lotion, and face wash A cosmetic composition for skin whitening and wrinkle improvement comprising rhubarb enzyme extract as an active ingredient, characterized in that it is prepared in a dosage form.

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