Composition, application thereof and cosmetic

Abstract

The invention belongs to the field of daily chemicals, and discloses a composition which comprises tetrahydrocurcumin and pterostilbene, the weight ratio of the tetrahydrocurcumin to the pterostilbene is (0.5-10): 1. The composition is composed of tetrahydrocurcumin and pterostilbene. The core innovation of the invention lies in that: 1, the tetrahydrocurcumin and the pterostilbene exert strong compounding performance under a specific proportion; 2, other effects except for oxidation resistance, inflammation resistance and whitening are found; and 3, the product has four effects of whitening, aging resistance, inflammation resistance and oxidation resistance, and in the field of cosmetics, the purpose is realized only by using two raw materials, and the condition that the four effects are relatively excellent is not common. Meanwhile, the invention further provides a mixture containing the composition, application and cosmetics.

Description

Technical Field

[0001] The invention belongs to the field of daily chemicals, and in particular relates to a composition and an application thereof and cosmetics. Background Art

[0002] CN202080072290.4 discloses a method for protecting mammalian skin against the harmful effects of UV radiation and environmental pollutants, using one or more plant actives selected from a composition comprising: not less than 10% w / w oroxylin A, not less than 10% w / w baicalein, and not less than 2% w / w chrysin; 95% w / w oxidized resveratrol; 95% w / w tetrahydrocurcumin; 90% w / w pterostilbene, and a composition comprising at least 10% w / w β-gallic acid glucoside and at least 10% w / w total mucic acid gallates.

[0003] The research direction of this program is the anti-pollution ability of the composition, which is specifically characterized by the estimation of superoxide dismutase (SOD) activity, estimation of glutathione peroxidase (GPx) activity, DPPH free radical scavenging analysis, IL-8 inhibition and other properties.

[0004] It is demonstrated in Example 4 that tetrahydrocurcumin inhibits IL-8 and has antioxidant and anti-collagenase activities;

[0005] It is demonstrated in Example 5 that when tetrahydrocurcumin at a concentration of 5 μg / ml and pterostilbene at a concentration of 2.5 μg / ml are combined, the anti-pollution effect is almost non-existent.

[0006] During the further screening of the relevant compatible combinations of tetrahydrocurcumin by the applicant, we found a different phenomenon.

[0007] The technical problem to be solved in this case is: how to develop a composition with excellent whitening, anti-aging, anti-inflammatory and antioxidant capabilities. Summary of the invention

[0008] In view of the deficiencies in the prior art, the object of the present invention is to provide a composition, which consists of tetrahydrocurcumin and pterostilbene. The core innovations of the present invention are: 1. It is found that the two have a strong composite performance under a specific ratio; 2. It is found that the composition has other effects besides anti-oxidation, anti-inflammatory and whitening; 3. It has four effects of whitening, anti-aging, anti-inflammatory and anti-oxidation. In the field of cosmetics, it is not common to achieve this purpose with only two raw materials and all four effects are excellent.

[0009] Meanwhile, the present invention also provides a mixture, an application and a cosmetic containing the composition.

[0010] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:

[0011] A composition consisting of tetrahydrocurcumin and pterostilbene;

[0012] The weight ratio of tetrahydrocurcumin to pterostilbene is 0.5-10:1;

[0013] When the tetrahydrocurcumin and pterostilbene are used in cosmetics, the total concentration of the tetrahydrocurcumin and pterostilbene is not less than 300 μg / ml.

[0014] The realization of the purpose of the present invention requires that tetrahydrocurcumin and pterostilbene exist at a certain concentration at the same time, and the ratio is 0.5-10:1; when these two conditions are met, the synergistic effect is very significant.

[0015] In terms of the single performance of the raw material, tetrahydrocurcumin is a natural functional whitening raw material, which is hydrogenated from curcumin isolated from the rhizome of the ginger plant turmeric (curcuma longa). Tetrahydrocurcumin has a strong activity in inhibiting tyrosinase, and its whitening effect is better than arbutin; tetrahydrocurcumin can effectively inhibit the generation of oxygen free radicals and can remove the free radicals that have been formed, and has a significant antioxidant effect.

[0016] The most significant effect of pterostilbene is to reduce free radical damage. Some documents mention that its ability to inhibit DPPH free radicals is far superior to that of tetrahydrocurcumin, and it also has excellent anti-inflammatory effects. In some documents, it is recorded that it has the effects of inhibiting melanin and removing spots. In addition, there are documents that disclose that after it is encapsulated in liposomes, its penetration effect can be further improved.

[0017] As for the above two raw materials, there is no very clear literature record that they have obvious anti-aging effects;

[0018] To analyze the anti-aging properties of these two raw materials, we used UVA to irradiate fibroblasts and evaluated the anti-wrinkle effect of the samples by detecting the changes in the content of 8-hydroxydeoxyguanosine (8-OHdG), type I collagen (Collagen I), and γ-H2AX. It was found that when both of them exist at the same time, they have a very strong anti-aging effect. This is beyond expectations.

[0019] At the same time, through the verification of antioxidant, anti-inflammatory and whitening effects, it was also found that the two have strong synergy, which is completely different from the rules recorded in CN202080072290.4.

[0020] In the above composition, the weight ratio of tetrahydrocurcumin to pterostilbene is 8-3:1.

[0021] The above composition also contains a synergistic ingredient, which is one or more combinations of tranexamic acid, ectoine, and narcissus bulb extract.

[0022] In the above composition, the weight ratio of the synergistic component to tetrahydrocurcumin in the composition is 0.1-0.5:1.

[0023] At the same time, the present invention also discloses the use of any of the above combinations in preparing cosmetics.

[0024] In the above-mentioned use, the cosmetics are cosmetics having at least one of anti-aging effects, whitening effects, anti-inflammatory effects, and antioxidant effects.

[0025] In addition, the present invention also discloses a cosmetic comprising any of the above-mentioned compositions, wherein when the tetrahydrocurcumin and pterostilbene are applied to the cosmetic, the total concentration thereof is not less than 300 μg / ml.

[0026] In the above-mentioned cosmetics, the total concentration of tetrahydrocurcumin and pterostilbene in the cosmetics is not higher than 5000 μg / ml.

[0027] Compared with the prior art, the present invention has the following beneficial effects:

[0028] The combination of the present invention has been verified to have very excellent anti-aging, anti-oxidation, anti-inflammatory and whitening effects; by analyzing the anti-aging properties of the two raw materials, UVA was used to irradiate fibroblasts, and the anti-wrinkle effect of the sample to be tested was evaluated by detecting the changes in the content of 8-hydroxydeoxyguanosine (8-OHdG), type I collagen (Collagen I), and γ-H2AX. It was found that when the two exist at the same time, they have extremely strong anti-aging effects. At the same time, through the verification of the antioxidant, anti-inflammatory and whitening effects, it was also found that the two have strong synergy. DETAILED DESCRIPTION

[0029] The technical solution of the present invention is further described below by specific implementation methods. It should be understood by those skilled in the art that the embodiments are only to help understand the present invention and should not be regarded as specific limitations of the present invention.

[0030] Source of reagents:

[0031] Tetrahydrocurcumin: purchased from Sami-Sabinsa Group Limited, USA, product model: CURCUMALONGA ROOT EXTRACT MICRONIZED CH;

[0032] Pterostilbene: purchased from Sami-Sabinsa Group Limited, USA. The product model is PTEROCARPUSMARSUPIUM BARK EXTRACT (PTERO WHITE 90%) CH.

[0033] The above two raw materials are both oil-soluble and water-soluble products.

[0034] Part I

[0035] In the embodiments and comparative examples, the compositions were prepared according to the concentrations in Table 1 and stirred to form a uniform phase.

[0036] Table 1 Formula table Unit: g

[0037] Tetrahydrocurcumin μg / ml Pterostilbene μg / ml Water Example 1 455 45 100 Example 2 445 55 100 Example 3 425 75 100 Example 4 400 100 100 Example 5 375 125 100 Example 6 335 165 100 Example 7 300 200 100 Example 8 250 250 100 Example 9 200 300 100 Example 10 100 400 100 Embodiment 11 50 450 100 Example 12 225 75 100 Example 13 750 250 100 Embodiment 14 3750 1250 100

[0038] Part 2

[0039] In the embodiments and comparative examples, the compositions were prepared according to the concentrations in Table 2 and stirred to form a uniform phase.

[0040] Table 2 Formula table Unit: g

[0041]

[0042]

[0043] Part III Performance Test

[0044] The performance test is divided into 4 parts, namely whitening performance test, anti-aging performance test, antioxidant performance test, and human body experiment. The following describes them one by one.

[0045] Test 1 Whitening performance test

[0046] In the biosynthesis of melanin in the skin, tyrosinase is a key enzyme that acts on DOPA to form DOPAQUINONE, which spontaneously undergoes a series of reactions to finally form melanin. In a phosphoric acid solution at pH 6.8, tyrosinase can catalyze the conversion of DOPA into DOPAQUINONE, and the absorbance value can be measured at 475nm on a spectrophotometer. Cosmetics that inhibit tyrosinase activity can reduce the conversion of DOPA into DOPAQUINONE, thereby reducing the absorbance value. Based on the change in absorbance value, the inhibitory effect of cosmetics on tyrosinase activity can be evaluated.

[0047] Test Basis

[0048] The test was carried out according to the “Melanin content and tyrosinase activity detection method based on melanocytes”.

[0049] Purpose of the test

[0050] This test is based on melanocytes and evaluates the whitening effect of the sample by detecting changes in tyrosinase activity.

[0051] Test Materials

[0052] 1Test system

[0053] The cells used in this test are melanocytes, batch number: MC23122801

[0054] Main reagents

[0055] DF-12 culture medium (Gibco), PBS (Solabo), kojic acid (Yuanye Biotechnology), melanin standard (Sigma), DMSO (Sigma), sodium hydroxide (Sinopharm Group), anhydrous ethanol (Sinopharm Group), and ether (Sinopharm Group).

[0056] 2 Main Equipment

[0057] CO2 incubator (Thermo, 150I), clean bench (Sujing Antai, SW-CJ-1F), and microplate reader (BioTek, Epoch).

[0058] 4 Test Methods

[0059] Tyrosinase activity inhibition rate test method

[0060] 1) Cell inoculation: After the cells are revived, when the plating rate reaches about 60%, the cells are inoculated into a 6-well plate and incubated overnight in a CO2 incubator (37°C, 5% CO2).

[0061] (2) Treatment of control and test samples

[0062] Active composition: Dilute with 95% ethanol to a sample concentration of 10%;

[0063] Positive control (kojic acid, purity ≥96%): dilute with 95% ethanol to a positive control concentration of 0.1%;

[0064] Negative control: pure water

[0065] 3) Administration: According to the test grouping, when the cell plating rate in the 6-well plate reaches 40% to 60%, group administration is carried out, and 3 replicate wells are set up for each group. 2 mL of culture medium is added to each well of the blank control group, 2 mL of culture medium containing kojic acid is added to each well of the positive control group, and 2 mL of culture medium containing the corresponding concentration of the sample to be tested is added to each well of the sample group. After the administration is completed, the 6-well plate is placed in a CO2 incubator (37°C, 5% CO2) and cultured for 24 hours. After 24 hours, the culture medium is discarded and washed once with PBS.

[0066] 4) Collect cells: Add 500 μL PBS to each well, transfer the cells into a 1.5 mL centrifuge tube using a cell scraper, and centrifuge at 6000 rpm for 5 min.

[0067] 5) Lysing cells: discard PBS, add 500 μL of 0.5% sodium deoxycholate to each well to lyse cells, lyse at 0°C for 1 h, centrifuge at 12000 rpm, 4°C for 20 min, and transfer the supernatant to a 1.5 mL centrifuge tube.

[0068] 6) Protein quantification: Protein quantification was performed on all samples according to the BCA kit instructions.

[0069] 7) L-DOPA reaction: 90 μL of the sample was added to a centrifuge tube, and 10 μL of 0.1% L-DOPA was added to each tube.

[0070] 8) OD value reading: At 0 min and 60 min, read the absorbance of each sample at a wavelength of 475 nm using an ELISA reader.

[0071] 9) Tyrosinase inhibition rate calculation formula: inhibition rate (%) = 1-[(sample 60 min OD value-sample 0 min OD value) / (control group 60 min OD value-control group 0 min OD value)*100%.

[0072] The test results can be seen in Table 3;

[0073] Table 3 Whitening performance test results

[0074]

[0075]

[0076]

[0077] Results Analysis

[0078] 1. From Example 1 to Example 11, when the total amount of tetrahydrocurcumin and pterostilbene is controlled at 500 μg / ml, the performance reaches a peak when the ratio of the two is 3 to 4:1; the value of Example 10 fluctuates. In the early experiments, we believe that it is an experimental error. After repeated experiments and comparison of the results of subsequent free radical scavenging experiments, it is found that when the ratio of the two is 1:4, there is indeed a certain performance improvement.

[0079] 2. From Example 12 to Example 14, according to the ratio of 3:1, when the sum of the concentrations of the two is between 500 and 1000 μg / ml, the performance is optimal, that is, at 1000 μg / ml, the performance is close to the top.

[0080] 3. From Examples 15 to 17, it can be seen that the performance can be further improved when tranexamic acid, ectoine, narcissus bulb extract, lactobionic acid, ergothioneine, tremella extract, tetrahydrocurcumin and pterostilbene are used in combination.

[0081] From comparative examples 1 to 6, all six have certain effects in terms of tyrosinase inhibition rate.

[0082] From the overall performance results, the screened tranexamic acid, ectoine, and narcissus bulb extracts have stronger stimulatory effects on low-dose tetrahydrocurcumin and pterostilbene.

[0083] Test 2 Antioxidant performance test

[0084] Excessive production of free radicals can lead to natural aging and photoaging of the skin, thus causing wrinkles. Therefore, whether a product has the ability to scavenge free radicals is one of the important indicators for evaluating anti-aging and anti-wrinkle cosmetics (raw materials).

[0085] This test refers to the laboratory method (HMC-WI-030 DPPH free radical scavenging rate) and compares the DPPH free radical scavenging rate test results of the test sample with that of the negative control. If the scavenging rate of the test sample is higher than that of the negative control and there is a significant difference (P<0.05), it can be considered that the test sample has certain anti-wrinkle and antioxidant effects.

[0086] Test methods

[0087] (1) Treatment of control and test samples

[0088] Active composition: Dilute with 95% ethanol to a sample concentration of 0.1%;

[0089] Positive control (vitamin E, purity ≥96%): dilute with 95% ethanol to a positive control concentration of 0.1%;

[0090] Negative control: pure water.

[0091] (2) Test operation steps

[0092] Set up sample tubes, sample background tubes, DPPH tubes and solvent background tubes. Each group needs to set up 3 parallel tubes. Add different

[0093] Gently shake the reagent solution and let it stand at room temperature for 5 minutes. Transfer each group of reaction solutions into a 1 cm cuvette and measure the absorbance at 517 nm.

[0094] (3) Calculation formula

[0095] The calculation formula of free radical scavenging rate (%) is:

[0096]

[0097] Where: T—absorbance of sample tube, i.e. absorbance of solution after sample reacts with DPPH;

[0098] T0—sample background absorbance;

[0099] C—the average value of the absorbance of the DPPH tube three times, that is, the absorbance of the DPPH solution when no sample is added; C0—the background absorbance of the solvent;

[0100] The test results can be seen in Table 4;

[0101] Table 4 Antioxidant performance test results

[0102]

[0103]

[0104] Note: The DPPH free radical scavenging rate is rounded to three decimal places. P<0.05 indicates that there are significant differences between the sample and the positive control and the negative control.

[0105] Results Analysis

[0106] 1. From Example 1 to Example 11, when the total amount of tetrahydrocurcumin and pterostilbene is controlled at 500 μg / ml and the ratio of the two is 2 to 4:1, the performance reaches the peak.

[0107] 2. From Example 12 to Example 14, according to the ratio of 3:1, when the sum of the concentrations of the two is between 500 and 1000 μg / ml, the performance is optimal, that is, at 1000 μg / ml, the performance is close to the top.

[0108] 3. From Examples 15 to 20, it can be seen that when tranexamic acid, ectoine, narcissus bulb extract, lactobionic acid, ergothioneine, tremella extract, tetrahydrocurcumin and pterostilbene are used in combination, the performance is not significantly improved.

[0109] From Comparative Examples 1 to 6, it can be seen that tranexamic acid and lactobionic acid have certain effects in scavenging free radicals, while the other four do not show obvious free radical scavenging effects.

[0110] That is, there may be no obvious synergy when tranexamic acid, ectoine, narcissus bulb extract, lactobionic acid, ergothioneine, tremella extract, tetrahydrocurcumin, and pterostilbene are used together.

[0111] Test 3 Anti-aging performance test

[0112] Collagen is the most abundant extracellular matrix protein in the human body, and type I collagen is the most abundant protein in the skin. The distribution of type I collagen in zebrafish is the same as that in humans and shows a high degree of conservation with humans. Their innate expression decreases significantly at 6 days after fertilization and becomes very low between 10 and 12 days after fertilization. Testing the expression of zebrafish type I collagen genes (col1a1a, col1a1b and col1a2), comparing the relative expression changes of zebrafish type I collagen genes in the treatment group and the blank control group, and calculating the promotion rate of type I collagen gene expression can evaluate the anti-wrinkle and firming effects of raw materials or products.

[0113] 1. Instruments and equipment

[0114] 1Zebrafish breeding equipment

[0115] The equipment must be equipped with temperature control devices, water circulation and filtration devices, and the breeding container should be made of glass or common food-grade PC materials. You can use general zebrafish breeding equipment or equip the fish tank according to laboratory needs (such as 45cm×45cm×30cm in length, width and height).

[0116] 2 spawning boxes / tanks

[0117] Made of glass, stainless steel or other inert materials, equipped with a mesh screen (mesh size is 2mm×2mm, made of stainless steel or multiple materials to protect fish eggs).

[0118] 3. Water quality monitoring equipment: pH meter, dissolved oxygen meter, salinity meter (conductivity meter).

[0119] 4 Analytical balance: accuracy 0.1mg.

[0120] 5. Microscope: A stereo microscope equipped with a camera system and a maximum magnification greater than 80 times.

[0121] 6 Test container: glass or polystyrene test container (such as 6-well plate, culture dish). If the test substance may be adsorbed to the polystyrene container (such as non-polar chemicals), inert materials (such as glass) should be selected to reduce adsorption.

[0122] 7 Thermostat: Accuracy ±1°C.

[0123] 8. Brine shrimp incubator: The equipment needs to be equipped with a hatching bucket, air pipe, air volume regulating valve, air pump cut-off switch and check valve device.

[0124] 9 Ultra-micro UV spectrophotometer: The wavelength range includes 260nm and 280nm.

[0125] 10PCR amplifier.

[0126] 11Real-time PCR (qPCR) instrument.

[0127] 12Other conventional testing instruments and equipment

[0128] Pipettes, centrifuge tubes, glass containers (such as beakers, volumetric flasks, etc.), vortex mixers, ultrasonic water baths, centrifuges, low-temperature refrigerators, adjustable pipettes and tips.

[0129] 2 Main reagents and consumables

[0130] 1 Water complies with GB / T 6682.

[0131] 2. Methanol, dimethyl sulfoxide, ethanol and other co-solvents are of analytical grade.

[0132] 3. Water for broodstock breeding

[0133] Weigh 4g of sea salt and dissolve it in 10L of water to prepare a solution with a salinity of 0.25‰ to 0.50‰, a conductivity of 500μS / cm to 800μS / cm, a dissolved oxygen saturation of 80%, a pH value of 6.5 to 8.5, and a hardness of 30mg / L to 300mg / L (calculated as calcium carbonate).

[0134] 4 Brine shrimp (Artemia salina) eggs

[0135] The dormant eggs of brine shrimp need to be kept dry and dark. Weigh about 15g of sea salt and dissolve it in 1L of water to prepare the brine shrimp egg hatching water. The density should not exceed 4g / L brine shrimp eggs.

[0136] 5Fish embryo culture medium

[0137] Weigh 2940 mg of anhydrous calcium chloride, 1233 mg of magnesium sulfate heptahydrate, 630 mg of sodium bicarbonate, and 55 mg of potassium chloride and dissolve them in 10 L of water. The pH value is 6.5 to 8.5. All chemicals are of analytical grade.

[0138] 6 Phosphate buffered saline (PBS) solution

[0139] 7. Acetyl hexapeptide-8 solution: Prepare 0.8 g / L acetyl hexapeptide-8 solution with water.

[0140] 8RNAlater solution (Invitrogen; AM7020) was stored frozen.

[0141] 9TRIzol reagent (Invitrogen; 15596018) was stored in a refrigerator.

[0142] 10 Chloroform

[0143] 11 Isopropyl alcohol is stored frozen.

[0144] 12 Ethanol Weigh 37.5 mL of ethanol and dissolve it in 12.5 mL of ultrapure distilled water and store it frozen.

[0145] 13gDNA Eraser PrimerScript RT Kit (Takara; Cat no.RR047A)

[0146] 14 Real-time PCR kit: SYBR Premix Ex Taq (Takara; Cat no.RR420A)

[0147] 3 Experimental methods

[0148] The test needs to set up a blank control group (fish embryo culture medium), a positive control group (acetyl hexapeptide-8) and a test group (test substance). If an organic solvent is used to assist dissolution, the solvent concentration in each group of solutions must be the same.

[0149] 1.1 Blank control group setting

[0150] 36 zebrafish were randomly selected and evenly distributed into a 24-well plate with 3 wells, each well containing 12 zebrafish and 2.5 mL of fish embryo culture medium.

[0151] 1.2 Positive control group setting

[0152] 36 zebrafish were randomly selected and evenly distributed into a 24-well plate with 3 wells, each well containing 12 zebrafish and 2.5 mL of acetyl hexapeptide-8 working solution.

[0153] 1.3 Test article treatment

[0154] 36 zebrafish were randomly selected and evenly distributed into 24-well plates with 3 wells, each containing 12 zebrafish and 2.5 mL of the test solution. They were placed in a 28°C±1°C incubator for 24 h±1 h. The 12 zebrafish in each well were collected into a 1.5 mL tube, the solution was removed, 0.5 mL of RNAlater solution was added, and the tubes were frozen for storage.

[0155] 2 RNA extraction

[0156] Remove RNAlater solution and rinse with PBS solution 3 times. Place in ice bath, remove PBS solution, add 500μL TRIzol reagent. Homogenize embryos with a pellet pestle and place in ice bath for 10 minutes. Add 100μL chloroform, vortex for 1 minute, and place in ice bath for 5 minutes. Centrifuge sample for 20 minutes, transfer the upper solution to a 1.5mL centrifuge tube, add 250μL isopropanol, vortex for 1 minute, and place in ice bath.

[0157] Place the sample in an ice bath for 10 minutes and centrifuge the sample for 20 minutes. Remove all supernatant, add 500 μL ethanol, vortex for 1 minute, centrifuge the sample for 5 minutes, and place in an ice bath for 10 minutes. Repeat this step 3 times. Remove ethanol, centrifuge the sample for 5 minutes, and air-dry the sample in a fume hood with the lid open for 10 minutes. Add 10 μL DNase / RNase-free ultrapure distilled water and heat at 55°C for 15 minutes.

[0158] 3cDNA synthesis

[0159] The RNA sample was diluted to 1000 ng / 7 μL with DNase / RNase-free ultrapure distilled water. The RNA was synthesized into cDNA using the PrimerScript RT Kit with gDNA Eraser (Takara; Cat no.RR047A) and stored frozen.

[0160] 4. Real-time RT-PCR

[0161] Prepare primer mix and real-time PCR master mix for each primer. Dilute cDNA sample 10-fold with DNase / RNase-free ultrapure distilled water. Add 9 μL PCR master mix and 1 μL diluted cDNA sample to each well of 96-well PCR plate. Seal the 96-well PCR plate with optical film, centrifuge the plate for 5 minutes, and perform real-time PCR amplification.

[0162] 5 Data and result calculation

[0163] Real-time PCR data were collected. Ct was used as the amplification result, the amplification amount of β-actin gene was used as the housekeeping gene, and the relative expression of each gene col1a1a was calculated as the test result.

[0164] Note: If a co-solvent is used in the preparation of the test substance, the blank control group value in the formula shall be replaced by the solvent control group value.

[0165] The mean and standard error of each test group were calculated, and the statistical results were expressed as mean ± standard error. The data were analyzed by variance analysis using statistical software, and the relative expression of the target gene between the test group and the blank control group (or solvent control group) was tested by two-tailed T test to obtain the p value. p<0.05 indicated a significant difference.

[0166] Results Report

[0167] 1. Collagen gene expression promotion rate

[0168] The promotion rate of collagen gene expression by the test substance at the corresponding test concentration.

[0169] 2 reviews

[0170] The ability of the test substance to promote collagen gene expression was evaluated, and whether the detection values ​​between the test substance treatment group and the blank control group were statistically different (p<0.05).

[0171] 3. Interpretation of the correlation of results

[0172] If the type I collagen gene expression promotion rate is positive and statistically significant (p<0.05), the test substance is judged to have anti-wrinkle and firming effects and can be used to support anti-wrinkle and firming effect claims. The results of the type I collagen gene expression promotion rate can be referred to Table 5;

[0173] Table 5 Promotion rate of zebrafish type I collagen-based col1a1a expression

[0174]

[0175]

[0176] Results Analysis

[0177] 1. From Example 1 to Example 11, when the total amount of tetrahydrocurcumin and pterostilbene is controlled at 500 μg / ml and the ratio of the two is 2 to 5.6:1, the performance reaches the peak value;

[0178] 2. From Example 12 to Example 14, according to the ratio of 3:1, when the sum of the concentrations of the two is between 500 and 1000 μg / ml, the performance is optimal, that is, at 1000 μg / ml, the performance is close to the top, and this trend is consistent with the above-mentioned tyrosinase inhibition rate and free radical scavenging.

[0179] 3. From Examples 15 to 20, it can be seen that the performance can be further improved when tranexamic acid, ectoine, narcissus bulb extract, lactobionic acid, ergothioneine, tremella extract, tetrahydrocurcumin and pterostilbene are used in combination.

[0180] From comparative examples 1 to 6, the effects of the six on tyrosinase inhibition rate are not significant. When tranexamic acid, ectoine, narcissus bulb extract, lactobionic acid, ergothioneine, tremella extract, tetrahydrocurcumin and pterostilbene are used in combination, tranexamic acid, ectoine and narcissus bulb extract show more obvious synergistic effects.

[0181] From the above three performance test results, the use of tranexamic acid, ectoine, and narcissus bulb extract can improve the whitening and anti-aging functions of the low-dose tetrahydrocurcumin and pterostilbene combination, and has a more obvious antioxidant effect.

[0182] Safety Testing

[0183] Safety test was performed using Example 13 and Example 14

[0184] 1. Materials and Methods

[0185] 1. Test substances: Example 13 and Example 14

[0186] 2. Negative control: blank control.

[0187] 3. Subjects: 30 people, male 9 People, Female 21 People, Age 21 to 52 years old, meeting the volunteer inclusion criteria.

[0188] 4. Test site: flexed forearm

[0189] 5. Spot test method: Select qualified spot test equipment, weigh 0.020mL~0.025mL of the test substance and put it into the small chamber of the spot tester. Apply the spot tester with the test substance to the test area with low-allergenic tape, and gently press it with the palm of your hand to evenly apply it to the skin for 24 hours. Observe the skin reaction according to the standard 30 minutes (after the indentation disappears), 24 hours and 48 hours after removing the test substance spot tester, and record the observation results. The rating standards can be seen in Table 6;

[0190] Table 6 Human skin patch test skin reaction grading standard

[0191]

[0192] 2. Test results

[0193] The test results refer to Table 7;

[0194] Table 7 Summary of cosmetics human skin patch test results

[0195]

[0196]

[0197] Note: The skin reactions of 30 subjects at different observation times are shown in the attachment.

[0198] The results of human skin patch tests showed that among 30 people 0 cases Appear Level 1 and above Reaction. For details, please see Annex 1 and Annex 2;

[0199] Attachment 1:

[0200]

[0201]

[0202] Attachment 2:

[0203]

[0204]

[0205]

[0206] Through the above tests, it can be found that the composition of the present invention is safe and non-irritating.

[0207] Application formula

[0208] The product of the present invention can be applied to various application formulas. The following lists essence milk and essence cream as examples, and the formulas are as follows:

[0209] The formula of the essence milk can be referred to Table 8;

[0210] Table 8 Recipe

[0211]

[0212]

[0213] The formula of the essence cream can be found in Table 9;

[0214] Table 9 Recipe

[0215]

[0216]

[0217]

[0218] The applicant declares that the present invention illustrates the process method of the present invention through the above-mentioned embodiments, but the present invention is not limited to the above-mentioned process steps, and it does not mean that the present invention must rely on the above-mentioned process steps to be implemented. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of the raw materials selected by the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

Claims

1. A composition, characterized in that including tetrahydrocurcumin and pterostilbene; The weight ratio of tetrahydrocurcumin to pterostilbene is 0.5-10:1; When the tetrahydrocurcumin and pterostilbene are used in cosmetics, the total concentration of the tetrahydrocurcumin and pterostilbene is not less than 300 μg / ml.

2. The composition according to claim 1, characterized in that The weight ratio of tetrahydrocurcumin to pterostilbene is 8-3:

1.

3. The composition according to claim 1, characterized in that The invention also contains synergistic ingredients, which are one or more combinations of tranexamic acid, ectoine, and narcissus bulb extract.

4. The composition according to claim 3, characterized in that The weight ratio of the synergistic component to pterostilbene in the composition is 0.1-0.5:

1.

5. Use of the combination according to any one of claims 1 to 4 in preparing cosmetics.

6. The use according to claim 5, characterized in that The cosmetics are cosmetics having at least one of anti-aging effects, whitening effects, anti-inflammatory effects, and antioxidant effects.

7. A cosmetic, characterized in that: A composition according to any one of claims 1 to 4, wherein when the tetrahydrocurcumin and pterostilbene are applied to cosmetics, the total concentration thereof is not less than 300 μg / ml.

8. The cosmetic according to claim 7, characterized in that: In the cosmetic, the total concentration of tetrahydrocurcumin and pterostilbene is not higher than 5000 μg / ml.

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