KR102062036B1 - Cosmetic composition for skin whitening and anti-wrinkle comprising extract of fruit and vegetables as effective component - Google Patents

Abstract

The present invention relates to a cosmetic composition for improving skin whitening and anti-wrinkle comprising fruit vegetables extract as an active ingredient, the cosmetic composition according to the present invention inhibits tyrosinase and elastase, and has an effect of lowering melanin index and erythema index have.
Therefore, the cosmetic composition according to the present invention can be usefully used for skin whitening and wrinkle improvement. In particular, it can be usefully used as a pack for skin whitening and wrinkle improvement.

Description

Cosmetic composition for skin whitening and anti-wrinkle comprising extract of fruit and vegetables as effective component}

The present invention relates to a cosmetic composition for improving skin whitening and wrinkles comprising fruit extract as an active ingredient.

In modern society, there is a growing recognition that appearance is one of social competitiveness, and as medical technology and income levels improve, the average life expectancy of humans increases, and the desire for healthy life and beauty is strengthened. Based on these changes and interests of the times, the market of cosmetics is expanding rapidly, and related researches are being conducted in various angles.

In particular, with increasing interest in healthy skin, there is a growing interest in the prevention and treatment of various skin problems such as inflammatory acne or skin diseases such as pigmented melasma (freckle) or freckles (freckle).

The research on anti-aging cosmetics of natural materials is being actively conducted, and it is widely used as a raw material for pharmaceuticals and functional cosmetic products while paying much attention to various physiological activity effects of natural materials, and due to the problems of stability and safety of compounds, The research on is getting more active.

Recently, there are many cases of using natural materials as functional ingredients. In the foodmetic fever combining food and cosmetics, cosmetics containing fruit have emerged as a trend, and aka fruitmetic is also attracting attention. There are a lot of fruit beauty products that are not only rich in nutrients but also color and aroma of fruit.

However, unlike conventional compounds, the skin whitening and wrinkle improvement effects of natural materials have not been verified yet. Therefore, cosmetics prepared using selected materials without sufficient verification of the skin whitening and wrinkle improvement effects of natural materials are used. The effect may be insignificant.

Aronia melanocarpa is a rose family belonging to the Rosaceae family and grows in northern America. Black chokeberry ( Aronia melanocarpa ), Red chokeberry ( Aronia) Arborutifolia ) and purple chokeberry (purple chokeberry, Aronia prunifolia ) are divided into, but generally included in Aronia ( Aronia melanocarpa ). The main useful components of Aaronia are polymeric proanthocyanidins, chlorogenic acid, neochlorogenic acid, and cyanidin-3-0-galactoside. 3-O-galactoside) and cyanidin-3-0-arabinoside have various physiological functions and are widely used as food and functional materials. Representative studies on radical scavenging activity and antioxidant effects on polyphenols, which are abundantly contained in aronia, have been actively conducted, and tumor suppression activity on cancer cell lines is also being studied.

Tomato ( Lycopersicon esculentum ) is an integral part of the human diet and major agricultural products. Lycopene, a lycopen found in tomatoes or tomato products, is the main component of a dark red color. It was first known as a preventive agent for prostate cancer. In addition, the most abundant carotinoids (80 to 90% of the total pigment content in tomatoes), recently attracted attention in various research fields, is a carotenoid-based antioxidant without the activity of provitamun-A (provitamun-A). It exhibits physiological activities such as cancer cell suppression, and mainly serves to prevent cardiovascular disease and skin cancer.

Citrus fruit is used as a raw material of herbal medicine and herbal medicine because it contains a lot of functional substances and active ingredients. Citrus fruits have a variety of flavonoids (flavonoids) and more than about 60 types of structures have been revealed to date, mainly hesperidin (nesperidin), neohesperidin (neohesperidin) and naringin (naringin) has been studied significantly. It is also important as a source of vitamins, dietary fiber, organic acids and free sugars, and contains flavonoids as a functional ingredient, which is highly valuable for cold resistance, skin cancer and atherosclerosis prevention.

Meanwhile, Korean Patent No. 10-1239361 discloses a fruit extract for preparing a cosmetic composition for whitening, a preparation method thereof, and a cosmetic composition comprising the same, and Korean Patent No. 10-1798026 as an active ingredient. Although a cosmetic composition for skin whitening is disclosed, there is no disclosure on skin whitening and wrinkle improvement cosmetic compositions prepared by the method of enzymatic decomposition and low temperature high pressure extraction of aronia, tomatoes and citrus fruits of the present invention.

The present invention is derived by the above requirements, the present invention provides a cosmetic composition for skin whitening and wrinkle improvement comprising the fruit vegetable extract as an active ingredient, the cosmetic composition according to the present invention is tyrosinase and elastase By inhibiting and lowering the melanin index and erythema index, the present invention was completed by confirming that the skin whitening and wrinkle improvement effect was obtained.

In order to achieve the above object, the present invention comprises the steps of (1) obtaining a hydrolyzate by treating the hydrolysing enzymes in fruit vegetables;

(2) low-temperature and high-pressure extraction of the enzymatic digest of fruit vegetables obtained in step (1); And

(3) provides a cosmetic composition for skin whitening and wrinkle improvement using the fruit vegetable extract extracted by the extraction method comprising the step of depressurizing the enzyme decomposition product of the fruit vegetable extracted at low temperature and high pressure in the step (2).

The present invention also provides a pack containing the cosmetic composition for improving skin whitening and wrinkles.

The present invention relates to a cosmetic composition for skin whitening and wrinkle improvement comprising fruit extract as an active ingredient, the cosmetic composition according to the present invention inhibits tyrosinase and elastase, and lowers the melanin index and erythema index It has a whitening and wrinkle improvement effect. Therefore, the cosmetic composition according to the present invention can be usefully used in cosmetics for skin whitening and wrinkle improvement, in particular, there is a useful value as a pack.

1 is a photograph of a dry powder of the Aaronia extract of the present invention.
Figure 2 is a photograph of the dry powdered tomato extract of the present invention.
Figure 3 is a dry powdered photograph of the tangerine extract of the present invention.
4 is a fruit vegetable extract (ELA: aronia enzymatic degradation and low temperature high pressure extract, ELL: tomato enzymatic degradation and low temperature high pressure extract, ELC: tangerine enzyme extracted by the method of enzymatic degradation and low temperature high pressure extraction of aronia, tomato and citrus fruits of the present invention) Degradation and low temperature and high pressure extract). As a comparative example, the enzymatic digest (EA: aronia enzymatic digestion, EL: tomato enzymatic digest, and EC: tangerine enzymatic digest) which performed only enzymatic digestion without low temperature and high pressure extraction was shown below.
Figure 5 shows the FRAP activity of the fruit vegetable extract extracted by the enzymatic decomposition and low temperature high pressure extraction method of aronia, tomato and citrus of the present invention.
Figure 6 confirms the tyrosinase inhibitory activity of the fruit vegetable extract extracted by the enzyme decomposition and low temperature high pressure extraction method of Aaronia, tomato and citrus of the present invention.
Figure 7 shows the elastase inhibitory activity of the fruit vegetable extract extracted by the enzyme decomposition and low temperature high pressure extraction method of Aaronia, tomato and citrus of the present invention.

The present invention relates to a cosmetic composition for improving skin whitening and wrinkles comprising fruit extract as an active ingredient.

In order to achieve the object of the present invention, the present invention comprises the steps of (1) processing the hydrolase to fruit vegetables to obtain an enzyme digest;

(2) low-temperature and high-pressure extraction of the enzymatic digest of fruit vegetables obtained in step (1); And

(3) provides a cosmetic composition for skin whitening and wrinkle improvement using the fruit vegetable extract extracted by the extraction method comprising the step of depressurizing the enzyme decomposition product of the fruit vegetable extracted at low temperature and high pressure in the step (2).

In step (1) of the present invention, the fruit is characterized in that any one selected from aronia, tomato and tangerine.

Also in the step (1), the hydrolase of the vegetable is cellulase (cellulase), pectinase (pectinase), hemifectinase (hemipectinase), hemicellulase (hemicellulase), arabinase, beta-glucanase ( One or more of β-glucanase) and xylanase is preferable, and may be more preferably cellulase, but is not limited thereto.

In addition, 0.5 to 5 parts by weight of purified water is mixed with respect to 1 part by weight of fruit vegetables, aronia, tomato or tangerine of step (1), and 0.1 to 0.4 parts by weight of enzyme is added to 40 to 60 parts by weight of 1 part by weight of the fruit vegetables. Although it is preferable to obtain the enzyme decomposition product of fruit vegetables by standing still at 1 to 8 hours, it is not limited to this.

In addition, in the step (2), the enzyme digested product may be extracted at low temperature and high pressure for 10 to 30 minutes at 30 to 40 ° C. and 500 to 2,000 bar (bar) pressure condition using a low temperature high pressure extractor, but is not limited thereto.

In addition, in the step (3), the step of filtration before and after the step of concentrated under reduced pressure to enzymatic digestion of low temperature high pressure extracted fruit vegetables can be carried out.

In addition, after the step (3), the step of spray drying may be further performed, the hot air inlet temperature of the spray dryer during the spray drying (inlet temperature) 100 ~ 200 ℃, outlet temperature (outlet temperature) 50 ~ 150 ℃, Feed rate (feed rate) may be set to 10 ~ 15mL / min, atomizer (rotator speed) 10,000 ~ 20,000rpm, but is not limited thereto.

Further, when spray drying, before the spray drying, 0.5 to 5 parts by weight of an emulsifier is added to 1 part by weight of the reduced pressure concentrate, preferably 1 part by weight of an emulsifier is added and spray dried to homogenize using a high pressure homogenizer. It may include the step of, the emulsifier is preferably Dextrin or pectin can be used, but is not limited thereto.

Cosmetic composition according to an embodiment of the present invention is an external skin ointment, cream, supple cosmetics, nourishing cosmetics, pack, essence, hair tonic, shampoo, conditioner, hair conditioner, hair treatment, gel, skin lotion, skin softener, skin toner , Astringent, lotion, milk lotion, moisture lotion, nutrition lotion, massage cream, nutrition cream, eye cream, moisture cream, hand cream, foundation, nutrition essence, sunscreen, soap, cleansing foam, cleansing lotion, cleansing cream, body lotion And it may have a formulation selected from the group consisting of a body cleanser, but is not limited thereto. The composition of each of these formulations may contain a variety of bases and additives necessary for the formulation of the formulation and are suitable, and the types and amounts of these components can be readily selected by those skilled in the art.

The cosmetic composition of the present invention may further contain one or more components for improving skin whitening or wrinkles, which exhibit the same or similar functions, in addition to the active ingredient. Skin whitening Skin whitening or anti-wrinkle ingredients include kojic acid and its derivatives, arbutin, ascorbic acid and its derivatives, hydroquinone and its derivatives, resorcinol, cycloalkanone, methylenedioxyphenyl alkanol, 2,7-di Plant extracts such as nitroindazole or vine tangerine extract, rice extract, licorice extract, and the like, but are not limited thereto.

When the formulation of the present invention is a paste, cream or gel, animal carriers, vegetable fibers, waxes, paraffins, starches, tracantes, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicas, talc or zinc oxide, etc. may be used as carrier components. Can be.

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used, in particular, in the case of spray, additionally chlorofluorohydrocarbon, Propellant such as propane / butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solvating or emulsifying agent is used as the carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 Fatty acid esters of, 3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan.

When the dosage form of the present invention is a suspension, liquid carrier diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline Cellulose, aluminum metahydroxy, bentonite, agar or tracant and the like can be used.

When the formulation of the present invention is a surfactant-containing cleansing, the carrier component is an aliphatic alcohol sulfate, an aliphatic alcohol ether sulfate, a sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide. Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, linolin derivatives or ethoxylated glycerol fatty acid esters and the like can be used.

When the cosmetic composition for skin whitening or wrinkle improvement as described above is formulated into a cosmetic, it may include a known excipient applicable to the skin to act as a carrier for the active ingredient. Formulations in cosmetics may refer to the information disclosed in the 67th edition (1995) of the International Cosmetic Ingredient Dictionary. Such documents are incorporated as part of this specification.

In the cosmetic composition according to an embodiment of the present invention, the cosmetic composition for skin whitening may be contained from 0.0001 to 10% by weight based on the total weight, preferably 0.005 to 1% by weight, but may be preferred It includes, but is not limited to, an effective amount that can provide skin whitening or wrinkle improvement.

The invention also relates to a pack comprising a cosmetic composition according to the invention.

The pack is preferably made of any one of a mask pack, a sleep pack, a cleansing pack, a wash off pack and a peel off pack, but is not limited thereto.

Hereinafter, the present invention will be described in detail by Examples, Comparative Examples and Experimental Examples. However, the following Examples, Comparative Examples and Experimental Examples are merely illustrative of the present invention, and the content of the present invention is not limited to the following Examples, Comparative Examples and Experimental Examples.

Example  One. Aaronia And enzymatic breakdown of tomatoes and tangerines and Low temperature and high pressure extraction  Preparation of Fruit Vegetable Extracts

Aronia, tomato and tangerine were each stored frozen at -20 ° C or lower after washing, and each frozen fruit was crushed using a crusher.

1 part by weight of purified water was mixed on the basis of 1 part by weight of each fruit sample, and 0.2% (w / w) cellulase was added based on the weight of each fruit sample to enzymatically decompose at 50 ± 5 ° C. for 2-6 hours.

The obtained enzyme digested product was extracted at low temperature and high pressure for 20 minutes at 35 ° C. and 1,000 bar pressure by a low temperature high pressure extractor (FT110, Benchtop rapid extractor, ARMFIELD, England), and then each low temperature high pressure extract was concentrated under reduced pressure (Model N-). 1N, Eyela Co., Tokyo, Japan) to prepare the respective Aaronia, tomato and tangerine extracts.

Comparative example  One. Aaronia , Tomatoes and tangerine enzymes Decomposition  Produce

Aronia, tomato and citrus fruits of the embodiment of the present invention were frozen and stored at -20 ° C. or lower after washing, and each frozen fruit was crushed using a crusher.

1 part by weight of purified water is mixed based on 1 part by weight of each fruit sample, and 0.2% (w / w) cellulase is added based on the weight of each fruit sample, followed by enzymatic reaction at 50 ± 5 ° C. for 2 to 6 hours. Enzyme digest was obtained and used as a comparative example.

Example  2. Aaronia , Tomatoes and tangerine Spray drying  Powder manufacturing

Spray dried powder was prepared by spray drying the low temperature and high pressure extract prepared in the process of Example 1.

The spray drying conditions were set at a hot air inlet temperature of 140 ° C., an outlet air temperature of 100 ° C., a feed rate of 12 ml / min, and an atomizer rotational speed of 15,000 rpm.

Experimental Example  One. Aaronia , Tomato and tangerine enzymes resolvent  And low temperature high pressure extract of  Extraction yield, total polyphenol content, total flavonoid content and Per total  content

Experimental Example 1 The experimental results of all the experimental examples below were analyzed by ANOVA using IBM SPSS Statistics (19.0, IBM Corp., Armonk, NY, USA), and the significance of each measured mean value (p <0.05) was Duncan's multiple range. The test was carried out and assayed.

Aaronia enzymatic digestion and low temperature high pressure extract, tomato enzymatic digestion and low temperature high pressure extract and tangerine enzymatic digestion prepared according to Example 1 and Changes in the active ingredient contents of the low-temperature and high-pressure extracts and the aronia enzymatic digest, tomato enzymatic digest and tangerine enzymatic digest prepared according to Comparative Example 1 were compared. As a control, aronia extract, tomato extract, and tangerine extract were performed.

The extraction yields of enzymatic digestion and low temperature high pressure extraction for Aaronia, tomato and tangerine were lyophilized (Free Zone 2.5, Labconco Co.) and the weight of the building was expressed as a percentage of the raw material dryness used to prepare the sample. .

Total polyphenol content was measured according to the Folin-Denis method. After mixing 1 ml of sample and 1 ml of 1N Folin Ciocalteu reagent, 1 ml of 20% Na ₂ CO ₃ was added and reacted for 30 minutes in the dark at room temperature, followed by spectrophotometer (Ultrospec 2100pro, Biochrom Ltd.). , Cambridge, UK) was used to measure the absorbance at 725 nm. Total polyphenol content was calculated by preparing a standard curve from tannic acid (Sigma-Aldrich Co., St. Louis, MO, USA).

Total flavonoid content was measured using the method of Jia et al. 1 ml of sample and 150 l of 5% NaNO ₂ were mixed and reacted at room temperature for 6 minutes, then mixed with 300 l of 10% AlCl _3, and then reacted at room temperature for 5 minutes, followed by mixing with 1 ml of 1N NaOH and then a spectrophotometer (Ultrospec 2100pro, Biochrom). Ltd.) and the absorbance was measured at 510 nm. Total flavonoid content was calculated by preparing a standard curve in rutin (Sigma-Aldrich Co.).

Total sugar content was measured by applying the phenol-sulfuric acid method. That is, 1 ml of sample, 1 ml of 5% phenol, and 5 ml of concentrated sulfuric acid (H ₂ SO ₄ ) were added and reacted at room temperature for 20 minutes, using an spectrophotometer (Ultrospec 2100pro, Biochrom Ltd.). Measured at 470 nm.

Extraction yield, total polyphenol content, total flavonoid content and total sugar content of each control, enzyme digest and low temperature high pressure extract are shown in Table 1.

sample Kinds Extraction yield (%) Total polyphenols
(g / 100g) Total Flavonoids
(g / 100g) Total equivalent
(g / 100g) Control
(extract) Aaronia 68.70 ± 0.12 ^d 17.82 ± 0.50 ^c 9.00 ± 0.37 ^bc 31.69 ± 0.45 ^g tomato 36.15 ± 0.26 ⁱ 2.45 ± 0.33 ^h 0.88 ± 0.09 ^f 34.01 ± 0.11 ^ef tangerine 51.21 ± 0.41 ^g 9.64 ± 1.04 ^e 1.13 ± 0.36 ^e 36.12 ± 0.18 ^d Comparative Example 1
(enzyme
resolvent) Aaronia 75.60 ± 0.06 ^b 21.32 ± 1.63 ^b 10.30 ± 0.27 ^b 33.74 ± 0.77 ^f tomato 45.20 ± 0.81 ^h 4.60 ± 0.44 ^g 1.04 ± 0.41 ^e 35.89 ± 0.30 ^e tangerine 66.93 ± 1.17 ^e 12.61 ± 0.68 ^d 3.40 ± 0.32 ^c 40.54 ± 0.48 ^b Example 1
(Enzymatic digestion + low temperature high pressure extract) Aaronia 82.87 ± 0.25 ^a 27.89 ± 0.39 ^a 12.46 ± 1.06 ^a 38.91 ± 0.12 ^c tomato 55.00 ± 1.05 ^f 5.63 ± 0.69 ^f 1.48 ± 0.19 ^e 37.47 ± 0.98 ^d tangerine 73.47 ± 0.14 ^c 17.38 ± 1.49 ^c 4.99 ± 0.36 ^d 48.72 ± 0.47 ^a

Comparing the extraction yield of each sample, in the control group the extraction yield was the lowest as 68.70% Aronia extract, 36.15% tomato extract, 51.21% tangerine extract.

In Comparative Example 1, the extraction yield was 75.60% of aronia enzyme digestion products, 66.93% of tangerine enzyme digest products, and 45.20% of tomato low-temperature and high-pressure extracts.

In Example 1, Aronia low-temperature and high-pressure extract 82.87% showed the highest extraction yield among all the samples, 73.47% of tangerine low-temperature and high-pressure extracts, tomato low-temperature and high-pressure extract 55.00%, the highest overall extraction yield.

It can be seen that the extraction yield was improved by performing both enzyme digestion and low temperature and high pressure extraction process in all samples.

Comparing the total polyphenol content of each sample, the total polyphenol content of the Aronia extract in the control group was 17.82 g / 100 g, and the lowest total polyphenol content was 2.45 g / 100 g tomato extract and 9.64 g / 100 g tangerine.

In Comparative Example 1, the total polyphenol content of the aronia degradant was 21.32 g / 100 g, and the total polyphenol content was relatively high as tomato enzyme degradant 4.60 g / 100 g and tangerine 12.61 g / 100 g.

In Example 1, the highest total polyphenol content of all samples was obtained with 27.89 g / 100 g of Aaronia low temperature and high pressure extract, and the total polyphenol content was the highest with 5.63 g / 100 g and 17.38 g / 100 g of tomato cold and high pressure extract. High.

Comparing the total flavonoid content of each sample, the total flavonoid content of the Aronia extract in the control group was 9.00g / 100g, the tomato extract 0.88g / 100g, the tangerine 1.13g / 100g, the lowest total polyphenol content.

In Comparative Example 1, the total flavonoid content of the Aaronia enzymatic degradation product was 10.30 g / 100 g, and the total flavonoid content was relatively high as 1.04 g / 100 g of tomato enzyme degradation product and 3.40 g / 100 g of tangerine.

In Example 1, 12.46 g / 100 g of Aronia low temperature and high pressure extract showed the highest total polyphenol content among all samples, and 1.48 g / 100 g of tomato low temperature and high pressure extract and 4.99 g / 100 g of tangerine showed the highest total polyphenol content. High.

Of each sample Comparing the total sugar content, the total sugar content of the Aronia extract in the control group was 31.69g / 100g, and the lowest total sugar content was 34.01g / 100g tomato extract and 36.12g / 100g tangerine.

In Comparative Example 1, the total sugar content was 33.74 g / 100 g, the tomato enzyme digest 35.89 g / 100 g, and the tangerine digest 40.54 g / 100 g, respectively.

In Example 1, the total sugar content of the Aaronia low temperature high pressure extract was 38.91 g / 100 g, the tomato low temperature high pressure extract 37.47 g / 100 g, and the orange low temperature high pressure extract 48.72 g / 100 g.

Although the active ingredient content of both the enzyme decomposition and the low-temperature high-pressure fruit vegetable extract according to Example 1 and the enzyme digested product according to Comparative Example 1 were improved, the overall active ingredient content of the enzyme decomposition and low-temperature and high pressure fruit vegetable extract according to Example 1 was further increased. It can be seen that the improvement.

Experimental Example  2. Aaronia , Tomato and tangerine enzymes resolvent  And low temperature high pressure extract of DPPH  Radical scavenging activity

Phenolic compounds contained in fruits have been reported to have excellent antioxidant effects, which are reported to be due to phenol rings that can stabilize free radicals.

DPPH radical scavenging activity was tested by dissolving DPPH solution in 100 mL of 99.9% ethanol in DPPH and then adding 100 mL of distilled water to adjust the absorbance to about 1.5 at 517 nm. 0.5 ml of the sample and 5 ml of the DPPH reagent were mixed and reacted at room temperature for 15 minutes, and then the absorbance was measured and calculated with a spectrophotometer (Ultrospec 2100pro, Biochrom Ltd.).

Experimental Example 2 The treatment concentrations of the samples of all the experimental examples below were the same at 100 µg / ml, 250 µg / ml, 500 µg / ml and 1,000 µg / ml, respectively.

DPPH radical scavenging activity showed a significant concentration-dependent result in all groups, ranging from 12.68% to 79.23%. The highest value of DPPH radical scavenging activity was obtained in Aronia low-temperature and high-pressure extract of 1,000㎍ / ㎖ (79.23%). Next, the DPPH radical scavenging activity was shown in the order of the tangerine low temperature high pressure extract, tomato low temperature high pressure extract, aronia enzymatic digestion, tangerine enzymatic digestion, tomato enzymatic digestion (Fig. 4).

The enzyme decomposition and low temperature and high pressure fruit vegetable extract according to Example 1 and the enzyme digested product according to Comparative Example 1 had DPPH radical scavenging activity, but the DPPH radical scavenging activity of the enzyme decomposition and low temperature and high pressure vegetable extract according to Example 1 It can be seen higher.

Experimental Example  3. Aaronia , Tomato and tangerine enzymes resolvent  And Low temperature  FRAP Activity of Extracts

Ferric tripyridyltriazine (Fe ^{3 +} -TPTZ) complex was ferrous tripytidyltriazine (Fe ^{2 +} ) by the reducing agent at low pH. -TPTZ), the activity of reducing power is represented by the absorbance value of reducing the metal ions by a chain reaction of free radicals providing hydrogen atoms, indicating that the higher the reducing power, the better the antioxidant activity.

FRAP was measured according to the method of Benzie and Strain. FRAP solution was prepared in 25 ml acetate buffer (300 mM, pH 3.6), 10 mM 2,4,6-tris (2-pyridyl) -s-triazine (2,4,6-tris) dissolved in 40 mM HCl. 2-pyridyl) -s-triazine} (TPTZ, Sigma-Aldrich Co.) 2.5ml and 20mM ferric chloride (FeCl ₃ ) was prepared by mixing, and then warmed at 37 ℃ for 10 minutes.

30 μl of the sample, 900 μl of the prepared FRAP solution and 90 μl of distilled water were added thereto, followed by reaction at room temperature for 30 minutes, and then absorbance was measured at 510 nm using a spectrophotometer (Ultraspec 2100pro, Biochrom Ltd.). FRAP was calculated by preparing a standard curve with FeSO ₄ · 7H ₂ O (Sigma-Aldrich Co.).

FRAP activity was found to be significant concentration-dependent in all groups, with a minimum of 140.66 μM and a maximum of 1,060.11 μM. Aronia low temperature high pressure extract showed the highest FRAP activity at 1,060.11 μM at 1,000 μg / ml concentration, and was highest in all groups at the other concentrations, followed by tangerine low pressure high pressure extract, aronia degradant and tangerine degradant , Tomato low temperature and high pressure extract, tomato enzyme decomposed products in order (Fig. 5).

Enzymatic degradation and low temperature and high pressure fruit vegetable extracts according to Example 1 and enzyme digests according to Comparative Example 1 both had FRAP activity, but overall, the enzyme decomposition and low temperature and high pressure vegetable extracts according to Example 1 had higher FRAP activity. It can be seen.

Experimental Example  4. Aaronia , Tomatoes and tangerine Tyrosinase ( Tyrosinase Inhibitory activity

In the continuous enzymatic oxidation reaction that forms melanin in tyrosine, tyrosinase is the first step in which tyrosine is oxidized to DOPA and the next step, DOPA is oxidized to dopaquinone. It is an enzyme that promotes the reaction of the step.

Tyrosinase inhibitory activity was measured according to the method of Yagi et al. The reaction group was treated with mushroom tyrosinase (110 U) in a mixed solution of 0.2 ml of substrate solution dissolved in 10 mM L-DOPA in 0.5 ml of 0.175 M sodium phosphate buffer (pH 6.8) and 0.1 ml of sample solution. / mL) 0.2ml was added and reacted for 2 minutes at 25 ℃ to measure the DOPA chrome produced in the reaction solution at 475nm.

[Formula 1] shows the tyrosinase inhibitory activity calculation formula.

[Equation 1]

Tyrosinase inhibitory activity (%) = (absorbance of 1-sample added group / absorbance of no additive group) × 100

Tyrosinase inhibitory activity showed a significant concentration-dependent result in all groups, ranging from 12.47% to 59.01%. Aronia low temperature high pressure extract showed the highest tyrosinase inhibitory activity at 1,000㎍ / ㎖ (59.01%) and highest in all other concentrations, followed by tangerine low temperature high pressure extract, aronia degrading product and tomato low temperature High pressure extract, tangerine enzyme digest and tomato enzyme digest were shown in this order (FIG. 6).

Enzymatic digestion and low temperature and high pressure fruit vegetable extract according to Example 1 and the enzymatic digestion product according to Comparative Example 1 had tyrosinase inhibitory activity, but the enzymatic digestion and low temperature and high pressure fruit vegetable extract according to Example 1 were tyrosinase. It can be seen that the activity of inhibitor is higher.

Experimental Example  5. Aaronia , Tomatoes and tangerine Elastase ( Elastase Inhibitory activity

Elastin (Elastin) is one of the components constituting the extracellular matrix (ECM), which is a skin elastic fiber, the reduction of elastin causes a decrease in the elasticity of the skin along with a decrease in collagen (collagen) and wrinkle formation.

Elastase is an enzyme that breaks down elastin, a substrate protein that is important for maintaining skin elasticity in the dermis, and is involved in the aging process and wrinkle formation caused by UV exposure. Therefore, elastase inhibitors are known to have an effect on improving skin wrinkles.

Elastase inhibitory activity measurements were performed according to the method of James et al. Aaronia, tomato and tangerine samples were diluted with 0.2 M Tris-HCl buffer (pH 8.0) at a concentration of 10-1,000 μg / ml, depending on the dilution factor, and 200 μl of buffer in 20 μl of diluted sample. then added, 0.8mM N- succinyl-was added the nitro ρ- not ride {N-succinyl- (Ala) 3 -ρ-nitroanilide} 20㎕ - ( Ala) 3.

After incubating the sample for 10 minutes at 25 ° C., 20 μl of 1.0 μg / ml porcine pancreatic elastase (PPE) was added to the sample.

After incubating the sample mixture again at 25 ° C. for 20 minutes, the reaction was stopped by cold aspiration and the absorbance was measured at 405 nm.

As a control, distilled water was added instead of the sample to measure enzyme activity.

[Formula 2] shows the inhibitory activity of elastase.

[Equation 2]

Elastase Inhibitory Activity (%) = [1- (B-C) / (A-D)] × 100

A: Absorbance after reaction by adding distilled water instead of sample and adding enzyme

B: absorbance of the sample after the addition of enzyme

C: absorbance of sample after reaction by adding distilled water instead of enzyme

D: Absorbance after reaction by adding distilled water instead of sample and enzyme

Elastase inhibitory activity of all groups showed significant concentration-dependent results, ranging from 6.74% up to 55.23%. Aronia low-temperature high-pressure extract showed the highest elastase inhibitory activity at 55.23% at 1,000 ㎍ / ㎖, and the highest in all groups at the other concentrations. Cold high-pressure extract, tangerine enzyme digest, tomato enzyme digest in the order of (Figure 7).

Enzyme digestion and low temperature and high pressure fruit vegetable extracts according to Example 1 and enzyme digests according to Comparative Example 1 both had an elastase inhibitory activity, but the enzyme digestion and low temperature and high pressure fruit vegetable extracts according to Example 1 were generally elastas. It can be seen that the activity of inhibitor is higher.

Experimental Example  6. Aaronia , Tomatoes and tangerine Spray drying  powder Fruit pack  Skin response measurement

The skin response of the fruit packs prepared using the spray-dried powders of aronia, tomato and tangerine prepared by the process of Example 2 was measured.

Table 2 shows the fruit pack manufacturing conditions.

Ingredient Name Content ratio (wt%) Aloe veragel 40 glycerin 10 water 20 Each fruit spray drying powder 30

The ingredients shown in Table 2 were mixed and applied to the facial skin from which moisture was removed, and left for 20 minutes, the face was massaged for 1 minute, washed with lukewarm water and secondly washed with cold water.

Skin measurement monitoring was conducted on 15 women in their 20s and 30s and 15 men in their 20s and 30s. The skin changes before and after 30 minutes of fruit pack using Aaronia, tomato, and tangerine spray-dried powder were checked. It was. Moisture was used as a corneometer, oil was used as a semeter, melanin and mexameter as a mexameter.

After a total of five skin measurements, the average value of the measured values was obtained, and a test site having a predetermined position and size was designated to set a value before using the pack as a control.

Table 3 is a result of comparing the skin moisture capacity, skin oil dose, melanin index, erythema index when using each fruit pack prepared by the process of Example 2.

sample Kinds Skin moisture
Capacity (AU) Skin oil
Dose (μg / cm ² ) Melanin index Erythema index Control Before use 18.70 3.00 169.08 201.40 Example 2
(Spray drying) Aaronia 34.22 8.45 117.36 149.06 tomato 32.92 6.32 122.06 155.21 tangerine 29.67 5.18 125.64 175.55

Before using the fruit pack using Aaronia, tomato and tangerine spray-dried powder, the skin condition was 18.70 AU for water, 3.00 µg / cm ^{2 for} oil, 169.08 for melanin and 201.40 for erythema.

After using three fruit packs made from Aaronia, tomato and tangerine spray-dried powder, the skin condition was 34.22 AU, 32.92 AU and 29.67 AU, respectively, and the oil doses were 8.45 ㎍ / cm ² and 6.32, respectively. Μg / cm ² , 5.18 μg / cm ² . All kinds of fruit packs have the effect of increasing the moisture and oil content of the skin, especially the fruit packs made of Aaronia spray-dried powder had the best effect.

The melanin index was 117.36 for Aronia spray dried fruit pack, 122.06 for tomato spray dried fruit pack, and 125.64 for tangerine spray dried fruit pack, and the erythema index was 149.06, 155.21, 175.55. It has the effect of reducing the erythema index, in particular, the effect of fruit packs made of Aaronia spray dried powder was the best.

Fruit packs made of Aaronia, tomato and tangerine spray-dried powder according to Example 2 can be seen that the effect of increasing the moisture, oil capacity of the skin, and both melanin index and erythema index.

Claims (8)

(1) treating cellulase (cellulase) hydrolase to any one vegetable selected from aronia, tomato and tangerine, and standing at 40 to 60 ° C. for 1 to 8 hours to obtain an enzymatic digest of the fruit vegetable;
(2) extracting the enzymatic digest of the fruit vegetable obtained in step (1) at low temperature and high pressure for 10 to 30 minutes under a pressure condition of 500 to 2,000 bar at 30 to 40 ° C; And
(3) a cosmetic composition for skin whitening and wrinkle improvement using the fruit vegetable extract extracted by the extraction method comprising the step of depressurizing the enzyme decomposition product of the fruit vegetable extracted at low temperature and high pressure in the step (2). delete delete delete The cosmetic composition for skin whitening and wrinkle improvement according to claim 1, further comprising spray drying after the step (3) of decomposing the enzyme digest under reduced pressure. The method of claim 5, wherein the spray drying is the skin whitening, characterized in that the spray dryer is performed in a spray dryer rotating inlet temperature of 100 ~ 200 ℃, outlet temperature is 50 ~ 150 ℃, 10,000 ~ 20,000rpm Cosmetic composition for improving wrinkles. A pack comprising the cosmetic composition according to any one of claims 1, 5 and 6. The method of claim 7, wherein the pack is made of any one of a mask pack, a sleep pack, a cleansing pack, a wash off pack, and a peel off pack. Pack made with.

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