KR20220075077A - 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 using a rhubarb enzyme extract as an active ingredient, specifically, by adding an enzyme to a rhubarb mixture prepared by mixing rhubarb and water to prepare a rhubarb enzyme mixture and extracting it under pressure It relates to a cosmetic composition for skin whitening and wrinkle improvement, which includes, as an active ingredient, a rhubarb enzyme extract, which not only improves extraction yield and antioxidant capacity, but also has skin whitening and wrinkle improvement effects.

Description

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 using a rhubarb enzyme extract as an active ingredient and a method for manufacturing the same, specifically, by adding an enzyme to a rhubarb mixture prepared by mixing rhubarb and water to prepare a rhubarb enzyme mixture and extracting it under pressure to prepare a rhubarb enzyme extract, which not only improves the extraction yield and antioxidant capacity, but also contains a rhubarb enzyme extract, which has skin whitening and wrinkle improvement effects, as an active ingredient. it's about

As living and income levels rise and improve, people are getting away from their daily lifestyles according to food, clothing and shelter, and interest in personal aesthetic life is also increasing. (Master's thesis, 2010.). In particular, as the average life expectancy is increased 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.).

Aging of the skin can be divided into intrinsic aging and extrinsic aging according to the factors. Intrinsic aging is the decline in 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, due to extrinsic aging such as exposure to various pollutants and UV rays, the skin becomes thinner, wrinkles increase and elasticity decreases, as well as 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 are substances constituting the skin, decreases, and the biosynthesis of type-1 collagenase increases and the expression of matrix metalloproteinases (MMPs) increases. , induces the breakdown of matrix proteins such as elastic fibers and fibronectin, which reduces skin elasticity and causes skin wrinkling (Brenneisen P, Sies H, Scharffetter-Kochannek K. “Ultraviolet-B irradiation and matrix metalloproteinases: from induction via signaling to initial events” Ann. N. Y. Acad. Sci, 2002, 973, 31-43.).

In order to maintain and improve overall health of the body, such as skin whitening and anti-wrinkle effects, research on cosmetics, foods, and pharmaceuticals using various natural products is being actively conducted.

Korea has three sides of the sea, so the types of seaweed inhabiting the sea area are very diverse and abundant. These seaweeds have long been widely used as food, spices, medicines, raw materials for seaweed industry, fertilizers, and the like. Recently, seaweed contains as many functional ingredients as plants, so the isolation of these functional ingredients and the verification and utilization of their efficacy are being actively studied (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.).

Large brown algae rhubarb (Eisenia bicyclis) is a perennial seaweed belonging to the genus Eisenia bicyclis, belonging to the genus Eisenia bicyclis. Effects of temperature, light intensity, and photoperiod. Lee Min-jung, Gyeongsang National University Graduate School. Master's thesis. 2018.). Rhubarb contains a large amount of iodine and potassium, and because of its unique taste, it has been eaten as a substitute for kelp since ancient times, and has recently been used as a raw material for alginic acid (physiological activity and application of rhubarb extract. Dae-yong Kim. Graduate School of Pukyong National University. Master's thesis. 2009.).

Research on the functionality of rhubarb is also being actively conducted. In particular, Eom SH, Lee BJ and Kim YM. 2010. Effect of yeast fermentation on the antioxidant and anti-inflammatory activity of sea tangle water extract. Korean J Fish Aquat Sci 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.) has been reported to have various effects. In addition, rhubarb contains a large amount of physiologically active substances such as phlorotannin, and phlorotannin isolated from rhubarb includes eckol, phlorofucofuroeckol-A, dieckol, 8,8'-bieckol, etc. It 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 Application Laid-Open No. 10-2013-0141874 relates to an antibacterial composition comprising a phlorotannin compound derived from rhubarb as an active ingredient, and the antimicrobial composition has particularly excellent antibacterial activity against MRSA (Methicillin-Resistant Staphylococcus aureus). Disclosed is an antibacterial composition comprising a phlorotannin compound having as an active ingredient.

Korea Patent No. 10-0966128 discloses a rhubarb enzyme extract having a hyperlipidemia and diabetes treatment effect produced by manufacturing rhubarb powder from seaweed rhubarb, extrusion molding and enzymatic hydrolysis, and rhubarb powder, kelp powder and excipients in the rhubarb enzyme extract Disclosed is a rhubarb tablet prepared by mixing and a manufacturing method thereof.

Korean Patent Application Laid-Open No. 10-2011-0057586 discloses a pharmaceutical composition for preventing or treating diabetes and a health functional food composition for improving diabetes comprising 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 utility is still insufficient, and in particular, functional verification for skin improvement and industrial application to cosmetics are very poor.

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 devised to solve the problems as described above and provide the necessary technology,

It is an object of the present invention to provide a cosmetic composition for skin whitening and wrinkle improvement, and a method for manufacturing the same, comprising, as an active ingredient, a rhubarb enzyme extract, which not only improves extraction yield and antioxidant ability but also has skin whitening and wrinkle improvement effects.

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

One embodiment of the present invention is a rhubarb mixture manufacturing step of mixing rhubarb powder and water to prepare a rhubarb mixture; A stirring step of adding an enzyme to the rhubarb mixture prepared in the rhubarb mixture preparation step and stirring to prepare a rhubarb enzyme mixture; And after pressure extraction of the rhubarb enzyme mixture prepared in the stirring step, centrifugation to prepare a rhubarb enzyme extract by taking only the supernatant to prepare a rhubarb enzyme extract; skin whitening and Provided is a method for preparing a cosmetic composition for improving wrinkles.

In the present invention, the rhubarb mixture may be 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, the rhubarb enzyme mixture is prepared by adding 2 to 5 parts by weight of an enzyme to 100 parts by weight of rhubarb powder and stirring at 100 to 200 rpm at a temperature of 50 to 70 ° C. for 15 to 20 hours. can be done with

In addition, the enzyme used to prepare the rhubarb enzyme mixture may be characterized by adding any one selected from the group consisting of neutrase, viscozyme and celluclast.

In addition, the rhubarb enzyme extract is prepared by extracting the rhubarb enzyme mixture under pressure 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 take only the supernatant. can do.

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

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

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

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

Hereinafter, embodiments of the present invention will be described in detail so that those of ordinary skill in the art to which the present invention pertains can easily carry out the present invention. The embodiments of the present invention are provided in order to more completely explain the present invention to those of ordinary skill in the art. Accordingly, the embodiment of the present invention may be modified in various other 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 other components may be further included, rather than excluding other components, unless otherwise stated.

The terms "about", "substantially", etc. to the extent used throughout the specification of the present invention are used in or close to the numerical value when manufacturing and material tolerances inherent in the stated meaning are presented, and the present invention It is used to prevent an unconscionable infringer from using the disclosure in which exact or absolute figures are mentioned to help the understanding of the As used throughout this specification, the term “step of (to)” or “step of” 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.

The cosmetic composition according to the present invention prepares 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. It is expected that the effect of whitening and wrinkle improvement will be high.

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

In addition, according to an 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 an embodiment of the present invention, the rhubarb enzyme mixture is stirred for 15 to 20 hours at a temperature of 50 to 70 ℃ by adding 2 to 5 parts by weight of the enzyme with respect to 100 parts by weight of rhubarb powder and 100 to 200 rpm. It may be characterized in that the rhubarb enzyme mixture is prepared.

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. can be characterized as

In addition, according to an embodiment of the present invention, the rhubarb enzyme extract is centrifuged for 10 to 30 minutes at 2000 to 4000 rpm after pressurized extraction of the rhubarb enzyme mixture at 1.2 atm and a temperature of 100 to 130 ° C. for 20 to 70 minutes Thus, it may be characterized in that it is prepared by taking only the supernatant.

When the rhubarb enzyme extract is produced outside the limited range as described above, antioxidant, skin whitening and wrinkle improvement effects cannot be expected due to the low or dilute concentration of useful components in the extract. Although the anti-inflammatory and wrinkle-improving effects may have an improvement effect, there is a concern that the manufacturing cost increases and skin toxicity may also occur in the manufacture of cosmetics.

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 with respect to the total weight% of the cosmetic composition.

This is that when the rhubarb enzyme extract is included in a ratio 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% with respect to the total weight of the cosmetic composition. When it is included in a proportion exceeding the weight %, there may be a problem in that the manufacturing cost is increased in manufacturing the cosmetic.

In addition, the cosmetic composition according to the present invention is selected from the group consisting of lotions, lotions, serums, massage creams, nourishing creams, eye creams, moisture creams, hand creams, sun creams, essences, nourishing essences, body lotions, and face washes by 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 is a rhubarb enzyme extract yield measurement experiment, total polyphenol content measurement experiment, total sugar and total protein content measurement experiment, antioxidant activity measurement experiment, tyrosinase inhibitory activity measurement Experiments, tests for measuring elastase inhibitory activity, and evaluation of storage stability of creams containing rhubarb enzyme extract were conducted. The cosmetic composition according to an embodiment of the present invention can be more clearly understood by the experiments described below.

Preparation of Rhubarb Enzyme Extract

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

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

3. Rhubarb enzyme extract preparation step: The rhubarb enzyme mixture is extracted under pressure 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 relative to the dry matter of the raw material used in the dried sample after drying using a freeze dryer.

To analyze the total polyphenol content, according to the Folin-Denis method, add 1 mL of 1 N Folin Ciocalteu reagent to 1 mL of sample, mix thoroughly, add 1 mL of 20% Na ₂ CO ₃ , and react for 30 minutes in a dark place 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 phenol-sulfuric acid method according to the method of saha and brewer. That is, 1 mL (w/v) of 5% phenol and sulfuric acid were reacted with 1 mL of sample, and 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 34.0% in the experimental group (N60), which was extracted under pressure for 60 minutes after neutrase treatment, showing the highest extraction yield.

The total polyphenol content was high as 35.05±0.35 mg/100 g and 32.50±0.17 mg/100 g, respectively, in the N30 and N60 sections extracted after neutrase treatment. 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 in N30 and N60 using neutrase at 28.33 g/100 g and 26.53 g/100 g, respectively. Table 1 shows the measurement results for the yield, total polyphenol content, total sugar and total protein content for the rhubarb enzyme extract.

Sample ¹⁾ Yield
(%) Total polyphenols
(mg/g) Total sugar
(g/100 g) total protein
(g/100 g) WE 28.2±0.5 ^cd2) 128.23±0.76 ^d 33.36±0.43 ^c 17.49±0.34 ^d AC 19.9±1.2 ^f 120.23±1.04 ^e 34.46±0.31 ^b 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±0.6 ^bc 114.57±1.15 ^f 34.96±0.56 ^b 15.41±0.08 ^f V30 26.5±0.5 ^e 141.57±0.58 ^c 38.62±0.25 ^a 18.70±0.25 ^c V60 28.2±0.3 ^cd 119.07±1.44 ^e 32.55±0.53 ^cd 16.22±0.50 ^e N30 29.8±0.8 ^b 180.07±1.76 ^a 30.59±0.49 ^e 28.33±0.08 ^a N60 34.0±0.6 ^a 167.40±0.87 ^b 32.11±1.21 ^d 26.53±0.34 ^{b 1)} WE: rhubarb hot water extract;
AC: autoclave-derived rhubarb pressurized extract,
C30: Rhubarb pressurized extract extracted with autoclave for 30 minutes after celluclast treatment,
C60: Rhubarb pressurized extract extracted with autoclave for 30 minutes after celluclast treatment,
V30: Rhubarb pressurized extract extracted with autoclave for 30 minutes after viscozyme treatment,
V60: Rhubarb pressurized extract extracted with autoclave for 30 minutes after viscozyme treatment,
N30: rhubarb pressurized extract extracted with autoclave for 30 minutes after neutrase treatment,
N60: Pressurized extract of rhubarb extracted with 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 determined according to the method of Benzie and Strain, with a solution of 10 mM TPTZ (2,4,6-tripyridyl-s-triazine) dissolved in 300 mM acetate buffer (pH 3.6) and 40 mM HCl. and 20 mM FeCl _3. 6H ₂ O were mixed in a ratio of 10:1:1 (v:v:v), respectively, and heated in a water bath at 37° C. was used as a FRAP substrate solution. In a 96-well plate, 25 μL of the sample solution diluted to a concentration of 5 mg/mL and 175 μL of the FRAP substrate solution were sequentially mixed and 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 obtained 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 , the test was conducted by diluting with phosphate buffer saline (PBS, pH 7.4) so that the absorbance becomes 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 absorbance was measured at 732 nm using a spectrophotometer. The difference in ABTS radical scavenging activity was calculated before and after addition of the extract.

Experiment result

As a result of measuring the antioxidant capacity by FRAP method, it was confirmed that the rhubarb enzyme extract showed a higher effect than the hot water extract of rhubarb, but higher antioxidant activity was shown in the experimental group that was subjected to pressure extraction for 60 minutes than in the experimental group that was extracted for 60 minutes. Among the enzymes, the antioxidant activity was high in the experimental group using neutrase, which was judged to have excellent antioxidant activity because the total polyphenol content was high. Similar experimental results were shown in ABTS radical scavenging ability, and all of the experimental groups were sample concentration-dependent. The measurement results for the 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±6.33 ^g 67.87±2.66 ⁱ 91.49±0.45 ^e AC 410.61±2.50 ^k 645.14±3.79 ^d 72.82±4.65 ^h 96.58±0.79 ^c C30 370.60±5.61 ^l 625.90±7.42 ^e 77.67±2.23 ^g 98.17±0.91 ^b C60 349.36±5.19 ^m 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±1.22 ^g 99.06±0.31 ^ab V60 361.53±8.92 ^l 654.57±3.55 ^d 71.44±1.30 ^hi 95.99±0.93 ^c N30 469.60±2.86 ^h 785±53±4.59 ^a 87.03±0.84 ^f 99.65±0.09 ^a N60 444.64±8.45 ⁱ 749.97±2.48 ^b 84.75±1.40 ^f 98.91±0.48 ^{ab 2)} Means±SD (n=3) with different letters (ai) within column are significantly different by Duncan's multiple range test (p<0.05).

Tyrosinase (tyrosinase) inhibitory activity measurement experiment

Tyrosinase inhibitory activity was obtained 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 in an incubator at ℃ for 5 minutes, absorbance was measured at 475 nm, and tyrosinase inhibitory activity showed a decrease in absorbance in 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, neutrase 54.21% and 55.12%, viscozyme 48.45% and 48.65, celluclast, depending on the enzyme type at a concentration of 10,000 ug/mL The inhibition rates were 43.93% and 46.83%, WE 39.44% and AC 43.53% showed the lowest tyrosinase inhibitory activity in the hot water extract of rhubarb without enzyme treatment, and the pressurized extract showed higher inhibitory activity than the hot water extract. It was confirmed that the most effective tyrosinase inhibitory activity was shown in the rhubarb enzyme extract. The measurement result of tyrosinase inhibitory activity is 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 diluted sample, and then 20 μL of 0.8 mM N-succinyl-(Ala) ₃ -ρ-nitroanilide was added. After incubating this at 25°C for 10 minutes, 20 μL of 1.0 μg/mL porcine pancreatic elastase was added. The reaction mixture was again incubated at 25° C. for 20 minutes, and then absorbance was measured at 405 nm. The elastase inhibitory activity showed a decrease in absorbance in the group with and without the sample solution.

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 showed no statistically significant difference at 14.67%, 16.39%, 17.79%, 20.09%, 17.41%, and 19.71%, respectively, 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 affect the improvement of skin wrinkles as it exhibits high elastase inhibitory activity. Elastase (Elastase) inhibitory activity measurement results are shown in Figure 2 below.

Storage stability evaluation of cream containing rhubarb enzyme extract

1. Preparation of cream with rhubarb enzyme extract

Cream preparation was prepared according to the recipe in Table 3. Part A, which is a fat-soluble substance, was completely dissolved at 75°C, and then part B, a water-soluble substance adjusted to 70°C, was slowly added to dissolve completely, and then emulsified with a blender. And 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 differences, brightness (L), redness (a-value), and yellowness (b-value) were measured using a CR-400 colorimeter (KONICA Minolta, Kenis, Japan). At this time, the standard color was measured repeatedly three times, with an L value of 93.53, a value of -0.28, and b value of 4.11, and expressed as the average value.

3. pH measurement

After putting the cream containing rhubarb enzyme extract into a cosmetic storage container, pH stability was measured at 0°C, 25°C and 40°C for 4 weeks at intervals of 1 week. As for the measurement method, 1 g of a sample was taken, filled with 9 mL of distilled water, sonicated for 1 hour with a sonicator, and pH was measured.

4. Viscosity Measurement

Since the cream used in the viscosity measurement experiment is 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 viscosity resistance of the cream moving on a spindle rotating with a constant acceleration.

5. Observation of discoloration and odor

After putting the cream with the rhubarb enzyme extract into the cosmetic storage container, visually inspect for layer separation and agglomeration, discoloration and odor, etc. to evaluate storage stability at 0°C, 25°C and 40°C for 4 weeks at intervals of 1 week. reviewed.

Experiment result

1. Chromaticity measurement result

Overall, no significant changes were observed depending on the storage temperature and storage period of the sample under all conditions. However, the value of b indicating yellowness was higher than that of a because the color of the rhubarb extract was brown. The L value was 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 itself. In the case of a value indicating red, the lowest value was shown in the section where rhubarb extract was not added, and the highest redness was shown in N30, and 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, it is judged that the cream containing rhubarb extract will be stable when applied in the future as the color change is constant or there is little difference depending on the storage period and temperature. The chromaticity measurement results are shown in Tables 4, 5 and 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 result

The initial pH of the cream without the addition of rhubarb extract was 7.29, which was higher than the pH of the cream with the addition of rhubarb enzyme extract, 7.04~7.16. In general, the longer the storage period, the lower the pH was, and the lower the temperature, the lower the pH. 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 result

The stability of cosmetics during storage period is very important because the quality of cosmetics should not change due to temperature change. The viscosity of cosmetics may change depending on the period or temperature, which may affect the quality. Therefore, a viscosity stability test was conducted to confirm whether the cream containing the rhubarb extract affects the viscosity change at temperature. As a result of measuring the viscosity of the cream containing the rhubarb enzyme extract and the cream without it, the initial viscosity of the cream containing the rhubarb enzyme extract was 11,266-12,856 cP, which was lower than the initial NO viscosity of 13,400 cP. Viscosity according to temperature showed a decrease in viscosity after 4 weeks from the initial viscosity in all conditions of the cream with and without the addition of rhubarb at 4°C, 25°C, and 40°C. In the case of 40℃, the viscosity did not decrease significantly compared to 4℃ and 25℃, which is thought to be due to loss of moisture and increased viscosity as cosmetics are stored at high temperatures. The viscosity measurement results are shown in FIGS. 3, 4 and 5 below.

4. Results of observation of discoloration and odor

According to the storage period of the cream with and without rhubarb enzyme extract, the color and smell were observed at 4℃, 25℃, and 40℃. As a result, there was no change in cream color and no specific odor. The same phenomenon was not observed. Discoloration and discoloration observation results are shown in FIGS. 6, 7 and 8 below.

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

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

Claims (7)

A rhubarb mixture manufacturing step of mixing rhubarb powder and water to prepare a rhubarb mixture;
A stirring step of adding an enzyme to the rhubarb mixture prepared in the rhubarb mixture preparation step and stirring to prepare a rhubarb enzyme mixture; and
Skin whitening using rhubarb enzyme extract as an active ingredient, characterized in that it is prepared including; pressurized extraction of the rhubarb enzyme mixture prepared in the stirring step, followed by centrifugation to prepare a rhubarb enzyme extract by taking only the supernatant and a method of manufacturing a cosmetic composition for wrinkle improvement. The method according to claim 1,
The rhubarb mixture is
A method of manufacturing a cosmetic composition for skin whitening and wrinkle improvement using a rhubarb enzyme extract as an active ingredient, 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. The method according to claim 1,
The rhubarb enzyme mixture is
Rhubarb enzyme extract, characterized in that by adding 2 to 5 parts by weight of the enzyme to 100 parts by weight of rhubarb powder and stirring at 100 to 200 rpm at a temperature of 50 to 70 ° C. for 15 to 20 hours to prepare a rhubarb enzyme mixture as an active ingredient A method of manufacturing a cosmetic composition for skin whitening and wrinkle improvement. The method according to claim 1,
The enzyme used to prepare the rhubarb enzyme mixture is,
Method for preparing a cosmetic composition for skin whitening and wrinkle improvement using a rhubarb enzyme extract as an active ingredient, characterized in that any one selected from the group consisting of neutrase, viscozyme and celluclast is added . The method according to claim 1,
The rhubarb enzyme extract,
Rhubarb enzyme extract, characterized in that it is prepared by extracting the rhubarb enzyme mixture under pressure at 1.2 atm and a temperature of 100 to 130 ° C. for 20 to 70 minutes, and then centrifuging at 2000 to 4000 rpm for 10 to 30 minutes, and taking only the supernatant as an active ingredient A method of manufacturing a cosmetic composition for skin whitening and wrinkle improvement. A cosmetic composition for skin whitening and wrinkle improvement comprising a rhubarb enzyme extract as an active ingredient, which is prepared by the method of any one of claims 1 to 5. 7. The method of claim 6,
The cosmetic composition for skin whitening and wrinkle improvement is selected from the group consisting of lotion, lotion, serum, massage cream, nourishing cream, eye cream, moisture cream, hand cream, sun cream, essence, nourishing essence, body lotion, and face wash. A cosmetic composition for skin whitening and wrinkle improvement comprising a rhubarb enzyme extract as an active ingredient, characterized in that it is prepared in a formulation.

Images