KR100891294B1 - Cosmetic compositions containing complex plant extracts - Google Patents

Abstract

The present invention relates to a cosmetic raw material composition that helps antioxidant and whitening as a complex extract (Schisandra chinensis, Schisandra chinensis, Licorice) of the herbal medicines, and confirmed that the extract of the extract of Schizandra chinensis (Schisandra chinensis) was superior to each extract. This complex extract exhibits its effect by scavenging free radicals (free radicals) and inhibiting tyrosinase activity. In addition, it has antibacterial and anti-inflammatory effects.

Herbal Medicine, Cosmetics, Antioxidant, Whitening, Schisandra chinensis, Lettuce, Licorice

Description

Cosmetic raw materials using complex herbal extracts {Cosmetic compositions containing complex plant extracts}

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to cosmetic raw material compositions, and in particular to cosmetic raw material compositions comprising Schizandra chinensis, Morus bark, Licorice complex extract (OSG) as a main component. In addition, it relates to the use as a whitening agent by identifying the tyrosinase activity inhibitory ability of the composition, and relates to the use as an antioxidant of the above extract having the excellent free radical scavenging and active oxygen scavenging ability and having this effect.

When the ultraviolet light is irradiated, tyrosine, an intracellular substance, is oxidized by tyrosinase, an active enzyme, converted into dopakinone, and finally becomes melanin, causing pigmentation on the skin. Arbutin is a whitening ingredient widely used to prevent pigmentation of the skin by preventing the increase of melanin pigment.

Oxygen is required to breathe, but is directly or indirectly involved in various diseases such as aging, arthritis, and cancer. It refers to activated oxygen species, which are free radicals that are produced by cellular immune responses, energy-generating responses in the intracellular electron transport system, ultraviolet light, stress, and bacteria. In addition, when produced in the body is known to cause various problems such as cell destruction, wrinkle formation, skin cancer, atopic dermatitis, acne. Antioxidants are widely distributed in the copper and plant systems, and many phenolic compounds in fruits and vegetables, such as flavonoids, tocopherols, and vitamin C, prevent or slow down the oxidation of lipids.

An object of the present invention is to provide a cosmetic raw material composition having a whitening effect is superior to arbutin, and at the same time as a raw material with an antioxidant effect.

The object of the present invention is achieved by a cosmetic raw material composition containing Schizandra chinensis, Morus bark, Licorice complex extract (OSG) as a main component.

The extraction method uses a method of atmospheric pressure extraction or ultrahigh pressure extraction at a temperature between normal temperature and the boiling point of the solvent using water, ethanol or butylene glycol, or a mixed solvent thereof.

The ratio of Schisandra chinensis, Morus bark and Licorice is preferably 6-15 parts by weight: 8-20 parts: 14-35 parts by weight based on the weight of the dried raw material.

The extract of the present invention is a substance having high affinity with tyrosinase, and when penetrated into the skin, it binds to the tyrosinase enzyme and inhibits the reaction with tyrosine. Tyrosinase inhibitory effect is much higher than that of arbutin, a whitening ingredient widely used to prevent pigmentation of the skin by preventing the increase of melanin pigment.

Hereinafter, the present invention will be described in more detail with reference to Examples.

Example 1 Heat Extraction Method Using 50% Ethanol

15 g of Schizandra chinensis, 20 g of lettuce, and 35 g of licorice were put in 700 ml of 50% ethanol, and concentrated to 1/10.

Example 2. Room temperature immersion extraction using 50% ethanol

15 g of Schizandra chinensis, 20 g of lettuce, and 35 g of licorice were added to 700 ml of 50% ethanol and extracted by stirring at room temperature for 24 hours. The extract was concentrated to 1/10.

Example 3. Hydrothermal extraction

15 g of Schizandra chinensis, 20 g of lettuce, and 35 g of licorice were put in 700 ml of distilled water, and then boiled and concentrated to 1/10.

Example 4. Extraction Method of Room Temperature Using Butylene Glycol and Ethanol

15 g of Schizandra chinensis, 20 g of lettuce, and 35 g of licorice were added to 700 ml of a solvent having a ratio of butylene glycol and 95% ethanol at 3: 7, followed by extraction with stirring at room temperature for 24 hours. The extract was concentrated to the maximum.

Example 5. Ultra high pressure extraction method using 50% ethanol (cold isotatic press, CIP extraction method)

6 g of Schizandra chinensis, 8 g of lettuce, and 14 g of licorice were added to 280 ml of 50% ethanol, and extracted by applying a pressure of 4,000 bar (extracted by an ultrahigh pressure sterilizer) and the amount was concentrated to 1/10.

Example 6. Supercritical fluid system (SCF extraction) using butylene glycol and ethanol

6 g of Schisandra chinensis, 8 g of lettuce, and 14 g of licorice were added to 280 ml of a solvent having a ratio of butylene glycol and 95% ethanol at 3: 7, and extracted by applying a pressure of 4,000 bar (extracted by an ultrahigh pressure sterilizer) to maximize concentration.

Test Example

Test Methods

Extracts obtained by the six extraction methods select the most suitable extracts for cosmetics, taking into account the condition of color and flavor precipitates. Selected extracts are evaluated by confirming their physiological activity effects and applying the product.

Test Example 1 . DPPH On by Free radical  Reduction Activity by the Elimination Method

1,1-diphenyl-2-picryl-hydrazine (DPPH, D9132, Sigma, USA) is a water-soluble substance with chemically stabilized free radicals, which has a maximum absorbance at around 515 nm to 520 nm and has antioxidant activity. When it meets with, it releases electrons and the radical (DPPH) disappears and its color changes. Chemically stable DPPH can analyze the antioxidant effects of extracts, beverages and oils, and pure phenolic compounds inherent in various antioxidants. This experiment is to determine the activity of eliminating radicals by sample after initiating oxidation with DPPH. To 1 ml of 0.2 mM DPPH solution in which DPPH was dissolved in methanol, a composite extract sample and each individual extract sample of each concentration were added. After reacting at room temperature for 10 minutes, the absorbance was measured at 517 nm by UV (Varian, Germany). In the control group, methanol is added instead of the sample solution and the DPPH solution to obtain a correction value. Free radical scavenging rate was calculated according to the following equation.

% Clearance = (absorbance of control group-absorbance of test group) / absorbance of control group X 100

The control group used 6-hydroxy-2,5,6,8-tetramethylchroman-2-carboxylic acid (Trolox, 97%, Aldrich, USA). Trolox is a vitamin E analogue that is highly antioxidant. In the test, samples diluted with 0.1% concentration in ethanol and similar to or higher than Trolox based on 50% of activity were considered to have good activity.

The results are published in Table 1 below and in FIG. 1.

Extract of Example 1 was more active than the control at 1000ppm, 500ppm

Exam 2 . SOD  Active measurement

Reactive oxygen species SOD activity is the oxidation of NBT (nitrobluetetrazolium, Sigma, USA) by active oxygen using an active oxygen generation system by xanthine / xanthine oxidase (Sigma, USA) enzyme reaction. Absorbance change by the measurement is measured and evaluated. 0.05mM Na₂CO₃ (Mw 105.99, Yakuri Pure Chemicals Co., Ltd, Japan) 2.4 ml, 3 mM xanthine (X-0626, Sigma, USA) 0.1 ml, 3 mM EDTA (E-5134, Sigma, USA) 0.1 ml, BSA (bovine serum albumin , A-7906, Sigma, USA) 0.1 ml, 0.75 mM NBT (N-6875, Sigma, USA) 0.1 ml, 0.1 ml of each sample (same as Test Example 1) and mixed well, and left to stand at 25 ° C for 10 minutes. I was. Add 0.1 ml of xanthine oxidase (X-4376, Sigma, USA) and react for 20 minutes at 25 ° C.₂ (Mw 170.48, Daejung Chemicals & Metals Co., LTD, Korea) 0.1 ml was added to stop the reaction, and the absorbance was measured at 560 nm with a UV-VIS spectrophotometer (Cary 50 Bio, Varian, Germany). In the control group, purified water was added instead of the sample, and purified water was added instead of xanthine oxidase to obtain a color correction value. Scavenging rate was calculated according to the formula used for DPPH radical scavenging rate. Vitamin C (l-ascorbic acid, A-1417, Sigma, USA) was used as a control. The activity was based on 60% to 80% and was considered to be high when the value was similar or high. The results are published in Table 1 and FIG.

SOD-like activity was also higher than that of the control at 1000 and 500 ppm.

Exam 3 . Tyrosinase activity inhibition test ( In in vitro  Tyrosinase inhibition assay )

TyrosinaseIs an enzyme involved in the most important early rate determining step in the melanin biosynthesis pathway in human body. This test is a method of evaluating the degree of inhibition of the activity of a tyrosinase enzyme in a sample. 0.9 ml of each sample (same as Test Example 1), 1.0 ml of 0.1 M phosphate buffer (pH 6.5), and 1.0 ml of 1.5 mM L-tyrosine (T8566, Sigma, USA) were added and then maintained at 37 ° C. for 10 minutes. After 0.1 ml of mushroom tyrosinase (T3824, Sigma, USA) was added and reacted at 37 ° C. for 10 minutes, the absorbance was measured at 480 nm using a UV-VIS spectrophotometer (Cary 50 Bio, Varian, Germany). The tyrosinase activity inhibition rate was calculated according to the following formula.

% Inhibition of tyrosinase activity = (absorbance of control group-absorbance of test group) / absorbance of control group X 100

2% arbutin was used as a control. The activity was based on 60% to 80% and was considered to be high when the value was similar or high. The results are posted in Table 1 below and in FIG. 3.

The activity was higher than that of arbutin at 500 and 1,000 ppm. The activity was 50% or more even at 50 and 100 ppm.

Exam 4 . Lipoxygenase ( Lipoxigenase)  Anti-inflammatory test by inhibition of activity

The inhibitory effect of lipid peroxide production was confirmed by measuring the peroxide value, an intermediate product of oxidation of unsaturated fatty acids, using linoleic acid (L2376, Sigma, assay 98%, USA) as a substrate. 1 ml, linolenic acid, 3000, 5000, 7000, 10000ppm of the composite extract sample of Example 1 at each concentration, 0.1ml of each individual extract sample, 0.9ml of lipoxidase (L7395, Sigma, USA) After reacting for 10 minutes at 25 ° C, mix 20% w / v trichloroacetic acid (TCA, Junsei Chemicals Co., Ltd, Japan) 0.5ml and 60% w / v Thiobarbituric acid (TBA, T5500, Sigma, USA) After heating for 10 minutes in boiling water, the color was cooled. 2 mL of butanol was added to the cooled sample, mixed with a vortex mixer for 20 seconds, and centrifuged at 3,500 rpm for 5 minutes to obtain a supernatant. Absorbance was measured at 535 nm using a UV-VIS spectrophotometer (Cary 50 Bio, Varian, Germany). The inhibition rate was calculated according to the following equation.

% Inhibition of lipoxygenase activity = (absorbance of control group-absorbance of test group) / absorbance of control group X 100

The results are published in Table 1 and FIG. 4. The activity was 58.66% at 1000 ppm. The control group was based on a very good extract and shows 70 to 90% of its activity at 10,000 ppm.

Exam 5 . Antimicrobial effect

The fungus used Gram-positive bacteria Staphylococcus aureus KCTC3881, Gram-negative bacteria Pseudomonas aeruginosa KCTC 1750, yeast Candida albicans KCTC 7965, and fungus Aspergillus niger KCTC 6196. The potato dextrose agar (PDA, Difco, USA) was used and the diluent was 0.85% NaCl saline. Bacteria were incubated at 37 ° C for 24 hours, and fungi were observed at 25 ° C for 48 to 72 hours. The antibacterial / antifungal effect was confirmed by the medium disk diffusion method. Before the test, MIC (minimum inhibition concentration) was calculated and applied. The results are posted in Table 1 and FIG. 5.

It was not active against fungi and had antimicrobial activity against bacteria. It had antimicrobial activity against both Gram-negative and positive, and showed a little more antimicrobial activity on the positive side than negative.

Exam 6 . Photo - hemolysis of RBC assay  Of red blood cells Photohemolysis  exam)

RBC artificially caused hemolysis by irradiation with UV, and extract was divided into control and treatment groups to test the anti-oxidative effect by checking the inhibition rate of hemolysis.

At 0.05%, red blood cells were lysed and tested only at a concentration of 0.02%. The photohemolysis inhibition rate of 10-30% was confirmed. ROS defense activity was 0.02% to 28.5% and 0.01% to 10.4%. At the extract concentration below that, it had an activity of 10% or less.

Exam7 . Melanogenesis inhibition assay  ( Intracellular  Melanin production inhibition test)

It is a test method to compare the amount of melanin production in cells with a blank sample during cell culture. Murine melanoma (B-16 F1) cells were placed in DMEM medium containing 10% FBS (fetal bovine serum) 5> 10³10% CO after inoculation at cells / 96well concentration₂, And incubated at 37 ℃. After culturing for 24 hours, the medium was removed, and the sample was replaced with a medium containing 0.05%, 0.02%, 0.01%, 0.002%, and 0.001%, and then cultured for 24 hours. The concentration range of the sample was determined by the toxicity test using the MTT assay. After 4 days of culture, the cells from which the medium was removed were washed with PBS (phosphated buffer saline) and treated with trypsin to recover the cells. Recovered cells were counted using a hematocytometer. 5> 10⁶The cells were diluted in 1 ml of 1 M sodium hydroxide solution (NaOH solution), heated at 100 ° C for 30 minutes, centrifuged at 13,000 rpm for 20 minutes, and the supernatant was measured for absorbance at 405 nm. The maximum concentration which does not cause toxicity by MTT assay was calculated | required. The concentration was 0.05% at maximum, and the tested concentrations were 0.001, 0.002, 0.01, 0.02, and 0.05%. Arbutin was tested as a control, and arbutin concentrations were 0.05, 0.02, 0.01, 0.03, and 0.001%. The results are shown in FIGS. 6 and 7.

As a result of measuring melanin production inhibition rate, it was confirmed that 80% of melanin was decreased at the concentration of 0.05%. The effect was better than a 70% reduction in arbutin. At 0.01% concentration, it had almost twice the activity, and it was confirmed that the effect was very excellent at 0.002% or less (FIG. 6). 7 is a photograph of a state in which melanin synthesis is suppressed experimentally. The phenomenon that inhibits melanin formation can be visually observed. Arbutin and OSG have a large concentration-dependent activity that inhibits melanin formation (compared to FIGS. 7, extracts 1 and 2).

Exam 8 . In hyperpigmentation  Whitening effect evaluation test Efficacy evaluation on hyper melanosis )

Human skin melanin and erythema levels are measured using a Derma-spectrometer (Dermalab., Netherlands). Twenty 18 to 60-year-old adult male and female extracts were added twice a day to pigmented areas and the rate of pigment reduction was confirmed over time (double blind method).

The rate of decrease of melanin was measured every 7 days from the beginning, and it is considered to have whitening activity as the rate of decrease increases over time. The results are shown in FIG.

Between 1 and 2 weeks, subjects had prolonged UV exposure and therefore increased melanin levels. That is, it was confirmed that the reduction rate was lowered. In the case of the control group, it took longer to recover after exposure to UV rays than the rate of melanin reduction, and the decrease was less than 28 days after the initial decrease. On the other hand, in the test group containing the extract, the final reduction rate was higher than the initial reduction rate, and the recovery after UV exposure was better than the control group. Therefore, the melanin reduction effect of the OSG extract is thought to be superior to the control.

Table 1. Comparison of activity of complex and individual extracts. Schizandra chinensis Baillon : O, Morus alba L .: S, and Glycyrrhiza glabra L .: G. Pa : P. aeruginosa KCTC1750 , Sa : S. aureus KCTC3881 . (n = 5)

+: Degree of activity,? +: Faint activity

O: Schisandra chinensis extract S: Morus bark extract G: Licorice extract

OSG: Schisandra chinensis, Morus bark, Licorice Complex Extract (Extract of Example 1)

When using an active extract as a cosmetic raw material, the safety of the product must be considered. Whether the color of the extract affects the product or becomes discolored later, the smell of the extract, or the toxicity can be a problem when applied to the skin as well as the product. Therefore, OSG extracts by various extraction methods were compared in color, activity and toxicity, and the most suitable method was heat extraction using 50% ethanol (Example 1). Cytotoxicity was not less than 0.05%, activity was confirmed at less than 0.05% in vitro, in vivo test and the activity of the extract was excellent. The extracts of Schisandra chinensis, Morus bark and Licorice were more active than the test results, and they had various functions. In particular, the whitening activity was more effective than the arbutin control group, it was confirmed that the antioxidant effect, the ability to eliminate free radicals and free radicals. It also has antibacterial activity and anti-inflammatory activity, so it is thought to be effective for skin trouble improvement. This extract is more active when applied as a complex extract than the activity of the individual extract, and can have the maximum effect in a small amount. Therefore, it can use suitably as a raw material of composite functional cosmetics.

1 is a graph showing the free radical scavenging effect according to the concentration of the complex extract of the present invention.

Figure 2 is a graph showing the active oxygen scavenging effect according to the concentration of the complex extract of the present invention.

Figure 3 is a graph showing the inhibition rate of tyrosinase activity according to the concentration of the complex extract of the present invention.

Figure 4 is a graph showing the anti-inflammatory effect according to the concentration of the complex extract of the present invention.

Figure 5 is a photograph showing the antimicrobial activity test results of the composite extract and the individual extract of the present invention (O: Schisandra chinensis extract S: Morus extract G: Licorice extract OSG: Schisandra chinensis, Morus extract, licorice complex extract (extract of Example 1) )

Figure 6 is a graph comparing the melanin synthesis inhibition rate of arbutin and the complex extract of the present invention.

Figure 7 is a photograph of a state in which melanin synthesis of arbutin and the complex extract of the present invention is suppressed.

8 is a graph showing the melanin / erythema reduction rate of the complex extract of the present invention.

Claims (5)

In the cosmetic raw material composition consisting of a complex extract of Schisandra chinensis, Morus bark and Licorice, the mixing ratio of Schisandra chinensis, Morus bark, and Licorice is 6-15 parts by weight: 8-20 parts: 14-35 parts by weight based on the weight of dried raw materials Cosmetic raw material composition, characterized in that. The cosmetic raw material composition according to claim 1, wherein the extract is an extract extracted using water, ethanol, butylene glycol, or a mixed solvent thereof. The cosmetic raw material composition according to claim 2, wherein the extract is an extract extracted using a mixed solvent of water and ethanol. 4. The cosmetic raw material composition according to claim 3, wherein the ethanol concentration of the mixed solvent of water and ethanol is 50%. delete

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