A soothing, moisturizing and repairing composition and its application

Abstract

The present invention provides a soothing, moisturizing and repairing composition and its application, which relates to the technical field of daily chemicals. The present invention has a soothing, moisturizing and repairing composition with Artemisia annua extract, Centella asiatica extract and Gentiana yunnanensis extract as main components, and the raw materials are selected from natural ingredients such as Artemisia annua, Centella asiatica, Gentiana yunnanensis, etc., which is non-toxic and non-irritating to human skin and has high safety; further, the product of the present invention can be used to repair skin damage caused by various reasons, and has anti-inflammatory and moisturizing effects. It is applied to cosmetics and skin care products, such as gel, emulsion, cream, essence, etc., which is gentler to the human body, safe and non-irritating.

Description

Technical Field

[0001] The present invention relates to the field of cosmetics, and in particular to a soothing, moisturizing and repairing composition and its application and its application in cosmetics. Background Art

[0002] There are dry skin, oily skin, mixed skin, etc. on the human face, and many of them are sensitive skin. Sensitive skin or oily skin is more susceptible to environmental influences and symptoms such as acne and inflammation. Therefore, you should pay more attention to skin care and choose appropriate skin care products to make the skin more radiant, soothe the skin, and moisturize and repair damaged epidermis.

[0003] Skin care gels are easy to use and are facial skin cosmetics that can clean, care and nourish, and are very popular among consumers. Products that can soothe, moisturize and repair are more popular among consumers with sensitive skin. Therefore, a product that can soothe, moisturize and repair is particularly important.

[0004] Studies have shown that Artemisia annua and Gentiana yunnanensis can have a good moisturizing effect and a certain repairing effect. Centella asiatica extract can also have a soothing and repairing effect. Combining these three in a certain ratio may have a better soothing, moisturizing and repairing effect. The present invention is based on the above hypothesis. Summary of the invention

[0005] In order to achieve the above-mentioned purpose of the invention, the present invention provides a soothing, moisturizing and repairing composition, including Artemisia annua extract, Centella asiatica extract and Gentiana yunnanensis extract.

[0006] Artemisia carvifolia is an annual herbaceous plant of the genus Artemisia in the Asteraceae family. It is distributed in all parts of the north and south and coastal areas. Its aerial parts are rich in bioactive substances, including sesquiterpenes, diterpenes, flavonoids, phenylpropionic acid, coumarins, flavonoids and volatile oils. These substances have pharmacological activities such as anti-malarial, anti-tumor, antibacterial and insecticidal, antipyretic and anti-inflammatory, and immunomodulatory.

[0007] Centella asiatica is a perennial herbaceous plant of the genus Centella in the family Apiaceae, native to India. The whole plant contains a variety of bioactive substances, including asiaticoside, madecassoside, asiatic acid, madecassic acid, succinic acid, rutin, and salix succinoside. These ingredients have a variety of pharmacological effects, such as promoting skin growth, healing wounds, anti-inflammation, and anti-tumor. Therefore, it is widely used in soothing, moisturizing, and repairing products.

[0008] Gentiana rigescens Franch.Et Sav., a perennial herb of the Gentianaceae family, is widely distributed in Yunnan, Guizhou and Sichuan. The whole plant can be used as medicine. In particular, its roots and rhizomes contain a variety of biologically active substances, mainly including gentiopicroside, gentiopicrin, gentioside, gentianine, gentioflavin, gentiosaccharide, etc. These ingredients have multiple pharmacological effects such as liver protection, anti-inflammatory, antibacterial, antiviral, antioxidant, and anti-tumor, and are widely used in soothing, moisturizing, and repairing products. After fermentation with Saccharomyces cerevisiae and Lactobacillus plantarum, the glycosides and gentianines in Gentiana rigescens can be more effectively resolved, and their biological activity is stronger.

[0009] The present invention innovatively combines Artemisia annua extract, Centella asiatica extract and fermented Gentiana yunnanensis extract into a soothing, moisturizing and repairing composition. Experimental verification shows that Artemisia annua extract, Centella asiatica extract and fermented Gentiana yunnanensis extract interact with each other to achieve a synergistic effect, making the soothing, moisturizing and repairing composition more effective.

[0010] Preferably, the raw materials of the composition include, by mass fraction, 0.2-1.7 parts of Artemisia annua extract, 0.05-0.75 parts of Centella asiatica extract, and 0.1-1.7 parts of Gentiana yunnanensis extract. By mass fraction ratio, the Artemisia annua extract, Centella asiatica extract and Gentiana yunnanensis extract are in a ratio of 1:0.05-8:0.1-10.

[0011] The present invention has found through experiments that when the Artemisia annua extract, the Centella asiatica extract and the Gentiana yunnanensis extract are compounded in the mass ratio, the soothing, moisturizing and repairing effects are better.

[0012] More preferably, the ratio of the Artemisia annua extract, Centella asiatica extract and Gentiana yunnanensis extract is 1:0.07-1.25:0.1-1.35 by mass fraction. The present invention has found through experiments that when the Artemisia annua extract, Centella asiatica extract and Gentiana yunnanensis extract are compounded in this mass fraction ratio, their soothing, moisturizing and repairing effects are further improved.

[0013] Most preferably, the ratio of the Artemisia annua extract, Centella asiatica extract and Gentiana yunnanensis extract is 1:0.1:1 by mass fraction. The present invention has found through experiments that when the Artemisia annua extract, Centella asiatica extract and Gentiana yunnanensis extract are compounded in this mass fraction ratio, the soothing, moisturizing and repairing effects of the soothing, moisturizing and repairing composition are better.

[0014] The invention provides a method for preparing the gentiana yunnanensis extract, which comprises the steps of crushing the whole gentiana yunnanensis plant and mixing and fermenting the gentiana yunnanensis extract with fermentation strains.

[0015] Preferably, the method comprises the following steps:

[0016] S1. Dry and crush the whole herb of Gentiana yunnanensis, add it into a specific culture medium for sterilization, and the solid-liquid ratio is 1:0.1-5;

[0017] S2, adding fermentation bacteria to the product of S1, and fermenting at a specific oxygen content, fermentation temperature, and fermentation time;

[0018] S3, collecting the filtrate of the S2 product;

[0019] S4. Dry the product of S3 to prepare the Gentiana yunnanensis extract.

[0020] More preferably, the specific culture medium in S1 is yeast extract peptone dextrose medium (YPD).

[0021] More preferably, the material-liquid ratio in S1 is 1:3.

[0022] More preferably, the specific oxygen content in S2 is 8% to 12%, the specific fermentation temperature is 28 to 32° C., and the specific fermentation time is 24 to 72 hours.

[0023] Most preferably, the specific oxygen content in S2 is 11%, the specific fermentation temperature is 30° C., and the specific fermentation time is 68 h.

[0024] More preferably, the fermentation bacteria in S2 are any one or both of Saccharomyces cerevisiae and Lactobacillus plantarum.

[0025] More preferably, the fermentation bacteria in S2 are Saccharomyces cerevisiae and Lactobacillus plantarum, and the mass ratio thereof is 1:0.1-2.

[0026] Most preferably, the fermentation bacteria in S2 are Saccharomyces cerevisiae and Lactobacillus plantarum, and the mass ratio thereof is 1:1.

[0027] The present invention also provides application of the soothing, moisturizing and repairing composition in cosmetics.

[0028] The present invention also provides a cosmetic comprising the soothing, moisturizing and repairing composition. The cosmetic can be in the form of cream, lotion, essence, mask, gel, etc., but is not limited thereto.

[0029] Preferably, the soothing, moisturizing and repairing composition has a weight percentage of 2wt% to 10wt% in the cosmetic.

[0030] When the soothing, moisturizing and repairing composition is applied to cosmetics at the weight percentage, it can provide the soothing, moisturizing and repairing effects of the cosmetics.

[0031] Most preferably, the soothing, moisturizing and repairing composition has a weight percentage of 5wt% in the cosmetic. When added at this content, the soothing, moisturizing and repairing effects are better.

[0032] When the soothing, moisturizing and repairing composition is applied to cosmetics at this weight percentage, it can effectively enhance the soothing, moisturizing and repairing effects of the cosmetics.

[0033] The present invention also provides a soothing, moisturizing and repairing gel, which is composed of the following components in percentage by weight:

[0034] The soothing, moisturizing and repairing composition comprises 2% to 10%, 0.1% to 0.8% of preservative, 1% to 10% of emollient and 0.06% to 2% of viscosity regulator and the balance of water. The soothing, moisturizing and repairing gel of the present invention is mild and non-irritating to the skin, safe to use, has excellent soothing, moisturizing and repairing effects, and is suitable for all skin types.

[0035] Preferably, the preservative is at least one selected from phenoxyethanol, methylparaben, ethylparaben and butylparaben.

[0036] Preferably, the emollient is at least one of glycerin, dimethicone, cyclopentasiloxane, caprylyl methicone, grape seed oil, jojoba oil, olive oil, ethylhexyl stearate, caprylic / capric triglyceride, tridecyl trimellitate, and isononyl isononanoate.

[0037] Preferably, the viscosity regulator is at least one of acrylic acid (esters) / C10-30 alkyl acrylate cross-linked polymer, magnesium aluminum silicate, xanthan gum, and carbomer.

[0038] More preferably, the soothing, moisturizing and repairing composition gel is composed of the following weight percentages:

[0039] The soothing, moisturizing and repairing composition comprises 5%, methyl paraben 0.4%, glycerin 2%, carbomer 0.6% and deionized water as the balance. The skin care gel prepared by the formula is mild and non-irritating to the skin, safe to use, has excellent soothing, moisturizing and repairing effects, and is suitable for all skin types.

[0040] Preferably, the method for preparing the soothing, moisturizing and repairing gel comprises the following steps:

[0041] The soothing, moisturizing and repairing composition, preservative, emollient and viscosity regulator are added into a beaker, deionized water is added, stirred at 1000 r / min for 1 h at 50° C. until the mixture is uniformly mixed, and sterilized under high pressure to obtain the skin care gel.

[0042] Compared with the prior art, the beneficial effects of the present invention are as follows: the present invention innovatively uses a mixture of Saccharomyces cerevisiae and Lactobacillus plantarum to ferment Yunnan Gentiana, and after fermentation, the activity and richness of the schizocyclopentadiene terpenoids in Yunnan Gentiana are significantly improved, and the functionality and application pathways of natural products such as glycosides, terpenes and plant polysaccharides are significantly improved. By combining the glycosides and terpenoids in the Yunnan Gentiana fermentation extract with the active products such as artemisinin, artemisinic acid, artemisinin alcohol in the Artemisia annua extract and asiaticoside and asiatic acid in Centella asiatica, it has significant antioxidant, anti-inflammatory, soothing and other effects. It can also significantly help wound healing, promote cell migration rate, and improve the skin's own barrier repair function.

[0043] It has been found through experiments that the soothing, moisturizing and repairing composition can repair skin damage caused by various reasons, has anti-inflammatory, soothing, moisturizing and repairing effects, and is suitable for various skin types. The present invention has found that under the premise of the same usage amount, the specific proportion of Artemisia annua extract, Centella asiatica extract and Gentiana yunnanensis extract defined in the present invention has better effects than using them alone or any three of them together, producing a synergistic effect, so that its soothing, moisturizing and repairing effects are better. The present invention also provides a formula that enables the soothing, moisturizing and repairing composition to achieve better effects in soothing, moisturizing and skin repair, and also provides a preferred formula for the soothing, moisturizing and repairing composition to be applied to gel. BRIEF DESCRIPTION OF THE DRAWINGS

[0044] Figure 1 Examples 4-9 HCEC survival rate.

[0045] Figure 2 Hyaluronidase inhibition rate of Examples 4 to 9 and Comparative Examples 1 to 16.

[0046] Figure 3 The TNF-α inflammatory factor inhibition rate of Examples 5, 7 to 9 and Comparative Examples 1 to 16.

[0047] Figure 4 The inhibition rate of zebrafish tail area reduction in Examples 5, 7 to 9 and Comparative Examples 1 to 16.

[0048] Figure 5 Hacat cell migration rate of Examples 5, 7-9 and Comparative Examples 1-16.

[0049] Figure 6 Relative transepidermal water loss rates of Examples 10 to 14 and Comparative Example 17. DETAILED DESCRIPTION

[0050] In order to better illustrate the purpose, technical scheme and advantages of the present invention, the present invention is further described by the following examples. It should be understood that the examples of the present invention are only used to illustrate the technical effects of the present invention, but not to limit the scope of protection of the present invention. In the examples, the methods used are conventional methods unless otherwise specified.

[0051] Some of the raw materials in the present invention come from the following sources:

[0052] Artemisia annua extract: purchased from Xi'an Qiancao Biotechnology Co., Ltd.

[0053] Centella asiatica extract: purchased from Xi'an Qiancao Biotechnology Co., Ltd.

[0054] Gentiana extract: purchased from Xi'an Qiancao Biotechnology Co., Ltd.

[0055] Yunnan Gentian: purchased from the farmers' market.

[0056] Saccharomyces paradoxus: purchased from Ningbo Testo Biotechnology Co., Ltd.; 3000 CFU / mL.

[0057] Lactobacillus plantarum: purchased from Ningbo Testo Biotechnology Co., Ltd.; 3000 CFU / mL.

[0058] The preparation method of the gentiana yunnanensis of the present invention is as follows:

[0059] S1. Dry and crush the whole herb of Gentiana yunnanensis, add YPD for sterilization, and the solid-liquid ratio is 1:0.1-5;

[0060] S2, adding fermentation bacteria to the product of S1, and fermenting at a specific oxygen content, fermentation temperature, and fermentation time;

[0061] S3, collecting the filtrate of the S2 product;

[0062] S4. Dry the product of S3 to prepare the Gentiana yunnanensis extract.

[0063] The specific parameters are shown in Table 1.

[0064] Table 1 Yunnan Gentian fermentation process parameters

[0065] step Strange yeast Lactobacillus plantarum Oxygen content Fermentation temperature Fermentation time Material-liquid ratio Example 1 1wt% 0wt% 8％ 28℃ 24h 1：1 Example 2 1wt% 1wt% 11％ 30℃ 68h 1：3 Example 3 0wt% 1wt% 12％ 32℃ 72h 1；5

[0066] The preparation of the soothing, moisturizing and repairing composition comprises the following steps:

[0067] S1 Weigh a specific mass portion of plant extract in a specific ratio and place it in a beaker.

[0068] The specific parameters are shown in Table 2.

[0069] Table 2 Composition ratio

[0070]

[0071] Note: The preparation process parameters of the gentian extract described in the above table are consistent with those of the Yunnan gentian extract.

[0072] A preparation method of a skin care gel comprises the following steps:

[0073] The soothing, moisturizing and repairing composition, preservative, emollient and viscosity regulator are added into a beaker, deionized water is added, stirred at 1000 r / min for 1 h at 50° C. until the mixture is uniformly mixed, and sterilized under high pressure to obtain the skin care gel.

[0074] The specific parameters are shown in Table 3.

[0075] Table 3 Gel preparation process parameters

[0076] Experimental Group Composition Addition amount Example 10 Example 5 4wt% Embodiment 11 Example 5 5wt% Example 12 Example 5 2wt% Example 13 Example 5 8wt% Embodiment 14 Example 5 10wt% Comparative Example 17 Example 5 0wt%

[0077] 1. Safety Testing

[0078] With reference to the 2015 Technical Specification for Safety of Cosmetics, the soothing, moisturizing and repairing compositions of Examples 4 to 9 and the skin care gels of Examples 10 to 14 were evaluated for cosmetic irritation. The test method was a skin patch test, and 110 randomly distributed people aged 16 to 65 were tested.

[0079] Test method: Put the test substance into the patch tester, the dosage is 0.020-0.025g, cover the patch tester with the test substance on the back or forearm of the subject with non-irritating cloth-based tape, and gently press it with the palm to make it evenly adhere to the skin surface for 24 hours. After removing the test patch tester, observe the skin reaction after the indentation disappears 30 minutes later. If the result is negative, observe again 24 hours and 48 hours after the patch test.

[0080] Evaluation criteria:

[0081] Grade 0: negative reaction;

[0082] Grade 1: Suspicious reaction, only slight erythema;

[0083] Grade 2: weak positive reaction, erythema, infiltration, edema, and papules may be present;

[0084] Grade 3: Strong positive reaction, erythema, infiltration, edema, papules, and the reaction may extend beyond the test area;

[0085] Grade 4: Very strong positive reaction, obvious erythema, severe infiltration, edema, confluent herpes, and the reaction exceeds the test area.

[0086] Test results: Skin reactions were negative in all subjects.

[0087] The above test results show that the soothing, moisturizing and repairing composition and skin care gel provided by the present invention are mild and non-irritating to the skin and safe to use.

[0088] 2. Cytotoxicity Test

[0089] Take human corneal epithelial cells HCEC with stable growth state and adjust the cell concentration to 1×10 5 / mL, 100μL per well. Inoculate into 96-well cell culture plates (except the outermost wells and blank control wells). Add 200μL of PBS to the outermost wells of the cell culture plate to reduce the evaporation of the culture medium. Place in a carbon dioxide incubator and culture at 37℃±1℃, CO2 concentration 5.0%±1% until the cell fusion reaches more than 90%. Add samples, the sample concentration is 10mg / L, set up blank control and sample groups, 100μL per well. Start adding samples (sample group) and PBS (blank control) at the same time interval between each two columns to ensure that the intervention time of each test substance on the cells is 5min. After 5min, remove the test substances in turn, add PBS to each well to wash the test substances, and wash twice. When using a vacuum suction pump or a discharge gun to remove the liquid in the well, be careful to avoid sucking away the surviving cells and affecting the experimental results. Add 100μL of complete culture medium and 10μL of CCK-8 solution to each well, and place in an incubator to culture for about 2h. At a wavelength of 450 nm, the absorbance (OD) value was measured and the cell viability was calculated; the results are shown in Table 4.

[0090] Formula (1)

[0091] Table 4 Cytotoxicity test

[0092] Experimental Group HCEC survival rate Example 4 (95.55±1.03)％ Example 5 (98.69±1.36)％ Example 6 (96.64±3.09)％ Example 7 (97.02±6.76)％ Example 8 (97.42±5.23)％ Example 9 (98.12±1.17)％ blank (100)％

[0093] According to the experimental results in Table 4, the cell survival rate was maintained above 95%. The combination of the present experiment had little effect on the viability of HCEC cells.

[0094] 3. Hyaluronidase Activity Inhibition Experiment

[0095] Hyaluronic acid, also known as hyaluronic acid, is a powerful moisturizer. It can attract and lock in moisture, maintain skin moisture, and improve dry, rough and dehydrated skin conditions. Hyaluronic acid can also enhance the biological activity of cells and has a strong synergistic effect on repairing and protecting the skin. Therefore, by inhibiting hyaluronidase, the decomposition rate of hyaluronic acid in the skin is reduced, thereby achieving long-term moisture retention and repair of the skin, which is of great significance.

[0096] (1) Take four groups of clean test tubes, label them A, B, C, and D, add 50 μL of the sample (10 mg / L) to test tubes A and B, add 50 μL of distilled water to test tubes C and D, add 50 μL of 500 U / mL hyaluronidase solution to test tubes A and C, add 50 μL of pH 5.6 acetate buffer to test tubes B and D, and place the test tubes in a 37°C incubator for 20 min.

[0097] (2) Add 10 μL of 2.5 mol / L CaCl2 solution to each of the four test tubes and place them in an incubator at 37°C for 20 min.

[0098] (3) Add 50 μL of 0.5 mg / mL sodium hyaluronate solution to test tubes A and C, and add 50 μL of pH 5.6 acetate buffer to test tubes B and D. Place the test tubes in a 37°C incubator for 40 min and then take them out and place them at room temperature for 10 min.

[0099] (4) Add 50 μL of distilled water, 10 μL of 5 mol / L NaOH solution, and 50 μL of acetylacetone solution to the four test tubes respectively. Place the test tubes in a boiling water bath for 30 min, then in an ice bath for 10 min, and finally at room temperature for 10 min.

[0100] (5) Add 100 μL of P-DAB reagent to each test tube, and then measure the absorbance at 530 nm using an ELISA reader.

[0101] Formula (2)

[0102] Wherein: A is the OD value of the sample solution; B is the OD value of the sample blank; C is the OD value of the control sample solution; D is the OD value of the control sample blank.

[0103] Table 5 Hyaluronidase activity inhibition rate

[0104]

[0105]

[0106] Analysis of the results from Table 5: From the results of Examples 4 to 6, it can be seen that the composition provided by the embodiments of the present invention has good hyaluronidase inhibitory efficacy, especially the effect of Example 5 is better, indicating that using Example 2 as the fermentation process parameter of Gentiana yunnanensis can effectively improve the inhibitory effect of the composition on hyaluronidase.

[0107] From the results of Examples 5 and 7 to 9, the composition of Artemisia annua extract, Centella asiatica extract and Gentiana yunnanensis extract has a significant effect of inhibiting hyaluronidase. The effect of Example 5, that is, when the mass fraction ratio of the composition is 1:0.1:1, the hyaluronidase inhibition effect is the best.

[0108] Comparing Example 5 with Comparative Examples 1 to 12, Example 5 has the best effect, indicating that the composition of different components can have a certain degree of influence on the efficacy of inhibiting hyaluronidase. Comparing Example 5 with Comparative Examples 1 to 4, Comparative Examples 5 to 8, and Comparative Examples 9 to 12 respectively show that the Artemisia annua extract, Centella asiatica extract, and Gentiana yunnanensis extract in the composition have a certain synergistic effect.

[0109] By comparing the results of Example 5 with Comparative Examples 13 to 16, it can be seen that the combination of Artemisia annua extract, Centella asiatica extract and Gentiana yunnanensis extract has a synergistic effect, and Gentiana yunnanensis produced by the same fermentation process cannot achieve the inhibitory effect of Gentiana yunnanensis on hyaluronidase as defined in the present invention.

[0110] IV. Inflammatory factor inhibition test

[0111] TNF-α is the most common inflammatory factor in inflammatory reactions. It can induce the phenotype of vascular endothelial cells to change to an inflammatory type, leading to changes in the body's metabolism and hemodynamics, and promoting the production of inflammatory mediators. Inflammatory mediators can increase vascular permeability, causing local congestion and edema, and may stimulate nerve endings to cause itching and pain. Therefore, the inhibition of TNF-α helps soothe and repair the skin.

[0112] RAW264.7 cells grown to the logarithmic phase were cultured at 1.5×10 5 Cells were inoculated at a density of 100 μg / mL in a 24-well plate, with 800 μL of cell suspension per well, and cultured in a 37°C, 5% CO2 incubator for 24 hours. When the cells grew to near fusion, 50 μL of the sample concentration of 10 mg / L was administered, with 3 replicates per group. The positive control was 50 μL of 10 μg / mL dexamethasone, and 1 μg / mL lipopolysaccharide (LPS) was used for stimulation 1 hour later. After incubation in a cell culture incubator for 24 hours, the cell culture fluid was transferred to a centrifuge tube, centrifuged at 5000 r / min for 10 minutes, and the cell supernatant was collected and tested according to the instructions of the mouse TNF-α enzyme-linked immunosorbent assay kit.

[0113] Formula (3)

[0114] Table 6 Results of RAW secretion of TNF-α

[0115]

[0116]

[0117] From the results of Example 5 and Examples 7 to 9 in Table 6, it can be seen that the composition of Artemisia annua extract, Centella asiatica extract and Gentiana yunnanensis extract has a significant effect of inhibiting TNF-α inflammatory factors. The effect of Example 5, that is, when the mass fraction ratio of the composition is 1:0.1:1, the effect of inhibiting TNF-α inflammatory factors is the best.

[0118] Comparing Example 5 with Comparative Examples 1 to 12, Example 5 has the best effect, indicating that the combination of different components can have a certain degree of influence on the efficacy of inhibiting TNF-α inflammatory factors. Comparing Example 5 with Comparative Examples 1 to 4, Comparative Examples 5 to 8, and Comparative Examples 9 to 12 respectively show that the Artemisia annua extract, Centella asiatica extract, and Gentiana yunnanensis extract in the composition have a certain synergistic effect.

[0119] By comparing the results of Example 5 with Comparative Examples 13 to 16, it can be seen that the combination of Artemisia annua extract, Centella asiatica extract and Gentiana yunnanensis extract has a synergistic effect, and Gentiana produced by the same fermentation process cannot achieve the inhibitory effect of Gentiana yunnanensis on TNF-α inflammatory factors as defined in the present invention.

[0120] 5. Effect of the composition on moisturizing zebrafish

[0121] The zebrafish epidermis has a certain osmotic pressure tolerance range, and water loss will occur if the tolerance range is exceeded. Using a high osmotic pressure solution to treat zebrafish will cause its tail to shrink and become smaller due to water loss. Therefore, an induced model of zebrafish hydration and moisturizing can be established to evaluate the effect of moisturizing products.

[0122] (1) Breeding: Zebrafish were paired at a male-female ratio of 2:1, and the eggs were collected and incubated overnight (28°C incubator).

[0123] (2) Experimental groups: The experiment set up a blank control group and an experimental group. The experimental groups included a model group, a positive group, and a sample group, with 8 2 dpf fry in each group.

[0124] (3) Solution preparation: Accurately weigh 1 mg of samples of Examples 5, 7-9 and Comparative Examples 1-16 and prepare the sample solution to be tested, and use urea (0.05%) as a positive control for the experiment, adding 1 mg to each well. After the administration is completed, place the 6-well plate in a 28°C incubator in the dark for 24 hours.

[0125] Formula (4)

[0126] Test results: as shown in Table 7 below.

[0127] Table 7 Zebrafish moisturizing effect test

[0128] Experimental Group Zebrafish tail area reduction inhibition rate Positive group 1 blank 0 Example 5 0.55 Example 7 0.43 Example 8 0.41 Example 9 0.37 Comparative Example 1 0.06 Comparative Example 2 0.21 Comparative Example 3 0.12 Comparative Example 4 0.05 Comparative Example 5 0.16 Comparative Example 6 0.25 Comparative Example 7 0.22 Comparative Example 8 0.11 Comparative Example 9 0.02 Comparative Example 10 0.01 Comparative Example 11 0.02 Comparative Example 12 0.07 Comparative Example 13 0.21 Comparative Example 14 0.42 Comparative Example 15 0.32 Comparative Example 16 0.11

[0129] Analysis of the results in Table 7: From the results of Examples 5 and 7 to 9, the composition of Artemisia annua extract, Centella asiatica extract and Gentiana yunnanensis extract has a significant effect of inhibiting the reduction of the zebrafish tail area. The effect of Example 5, i.e., when the mass fraction ratio of the composition is 1:0.1:1, the effect of inhibiting the reduction of the zebrafish tail area is the best.

[0130] Comparing Example 5 with Comparative Examples 1 to 12, Example 5 has the best effect, indicating that the composition of different components can have a certain degree of influence on the effect of inhibiting the reduction of the zebrafish tail area. Comparing Example 5 with Comparative Examples 1 to 4, Comparative Examples 5 to 8, and Comparative Examples 9 to 12 respectively show that the Artemisia annua extract, Centella asiatica extract, and Gentiana yunnanensis extract in the composition have a certain synergistic effect.

[0131] By comparing the results of Example 5 with Comparative Examples 13 to 16, it can be seen that the combination of Artemisia annua extract, Centella asiatica extract and Gentiana yunnanensis extract has a synergistic effect, and Gentiana yunnanensis produced by the same fermentation process cannot achieve the inhibitory effect of Gentiana yunnanensis on the reduction of zebrafish tail area as defined in the present invention.

[0132] VI. Effects of the composition on the scratch migration of Hacat cells

[0133] The degree of cell migration has an important impact on wound healing and skin repair. In the process of wound healing, cell migration is an important step for epidermal cells to cover the wound, so the higher the cell migration rate, the faster the wound heals. At the same time, cell migration is also involved in the repair process after skin damage. If cell migration is impaired, it will lead to skin repair disorders, which in turn cause various skin diseases.

[0134] Inoculate cells into 12-well plates at the corresponding inoculation density and incubate overnight at 37°C in a CO2 incubator. When the cell plating rate in the 12-well plate reaches about 80%, scratch the sample group and the solvent control group. After the scratching, wash the cells with PBS to remove dead cells, and then add fresh EGF-free keratinocyte culture fluid, add 50μL of 10mg / L according to Examples 5, 7-9, and Comparative Examples 1-16, and take photos and record using an inverted microscope. Incubate in a 37°C, 5% CO2 incubator for 24h, take photos and record again, and use ImageJ software for statistical analysis.

[0135] Table 8 Hacat cell migration rate

[0136]

[0137]

[0138] From the analysis of the results in Table 8: From the results of Examples 5 and 7 to 9, the composition of Artemisia annua extract, Centella asiatica extract and Gentiana yunnanensis extract has a significant effect of improving cell migration rate. The effect of Example 5, i.e., when the mass fraction ratio of the composition is 1:0.1:1, the effect of improving cell migration rate is the best.

[0139] Comparing Example 5 with Comparative Examples 1 to 12, Example 5 has the best effect, indicating that the combination of different components can have a certain degree of influence on the efficacy of improving cell migration rate. Comparing Example 5 with Comparative Examples 1 to 4, Comparative Examples 5 to 8, and Comparative Examples 9 to 12 respectively, it is shown that the Artemisia annua extract, Centella asiatica extract, and Gentiana yunnanensis extract in the composition have a certain synergistic effect.

[0140] By comparing the results of Example 5 with Comparative Examples 13 to 16, it can be seen that the combination of Artemisia annua extract, Centella asiatica extract and Gentiana yunnanensis extract has a synergistic effect, and Gentiana yunnanensis produced by the same fermentation process cannot achieve the effect of Gentiana yunnanensis on improving cell migration rate as defined in the present invention.

[0141] 7. Preparation and stability testing of skin care gel

[0142] Examples 10-14 were prepared and tested according to the above formulation design.

[0143] 1. Heat resistance test: The constant temperature incubator was adjusted to 40°C, and three samples of each of Examples 10 to 14 were placed in transparent glass bottles with a sample volume of 10 ml / bottle. After sealing, the bottles were placed in a constant temperature incubator. After three months, the bottles were taken out and returned to room temperature to observe the changes in appearance.

[0144] 2. Cold resistance test: The constant temperature incubator was adjusted to -10°C, and three samples of each of Examples 10 to 14 were placed in transparent glass bottles with a sample volume of 10 ml / bottle. After sealing, the bottles were placed in a constant temperature incubator. After three months, the bottles were taken out and returned to room temperature to observe the changes in appearance.

[0145] 3. Room temperature test: Three samples of each of Examples 10 to 14 were placed in transparent glass bottles with a sample volume of 10 ml / bottle. After sealing, they were placed at room temperature for 6 months and the changes in appearance were observed.

[0146] 4. Light aging test: Three samples of each of the above-mentioned Examples 10 to 14 were placed in a transparent glass bottle with a sample volume of 10 ml / bottle. After sealing, the bottles were placed under incandescent light for three months, and the appearance changes were observed.

[0147] No precipitation or settling was observed in the heat resistance test, cold resistance test, room temperature test, and light aging test, and the original physical properties were maintained.

[0148] 7. Test on the immediate repair effect of skin care gel on human body

[0149] Transepidermal water loss (TEWL) is an important parameter for evaluating skin barrier function. It indicates the loss of water from the deep dermis through evaporation of the epidermis. The higher the TEWL, the more water is lost through the skin and the weaker the skin's moisturizing ability.

[0150] The transepidermal water loss rate is closely related to the hydration of the stratum corneum and is a commonly used indicator reflecting the barrier function of the stratum corneum. In the development of cosmetics, the efficacy of moisturizing cosmetics can be evaluated by testing the TEWL value of skin water loss. In addition, the transepidermal water loss rate can also be used in allergic patch tests, contact dermatitis, physical therapy, burns and new tissue monitoring to promptly detect whether the protective function of the skin has been damaged.

[0151] Therefore, transepidermal water loss rate is an important parameter for assessing skin health and is of great significance to the research, development and evaluation of moisturizing products.

[0152] 60 healthy subjects aged between 18 and 60 years old with no injuries on the inner side of their arms were selected and divided into groups, with 10 subjects in each group, including 5 males and 5 females. A sample group and a blank group (i.e., negative control group: no test sample) were set up, and the test area was randomly distributed 1 cm inside the volunteers' arms. 2 Area. The forearms (two areas) of the above-mentioned subjects were torn off with medical tape 20 times to induce damage to the skin barrier function, and then 5 mg / mL histamine was added to model for 10 minutes (obvious erythema appeared). One area was treated with 1g of the test product, and the other area was blank as a negative control area. The transepidermal water loss rate (TEWL) of the forearm skin was collected before tape tearing off (T0), before using the product (i.e., immediately after histamine) (T1), and 2 hours after using the product (T2). The difference in skin index changes between the product-treated area and the negative control area before and after using the product was used to evaluate the short-term repair effect of the product on skin barrier damage caused by tape tearing and histamine-induced skin irritation, and the average value was taken. The results are shown in Table 3 below.

[0153] Formula (5)

[0154] Table 9 Relative water loss rate of epidermis

[0155] Experimental Group T0 T1 T2 Relative transepidermal water loss Example 10 8.14％ 21.82％ 7.98％ -8.97％ Embodiment 11 8.28％ 22.31％ 7.44％ -10.14％ Example 12 8.13％ 22.33％ 7.95％ -4.21％ Example 13 8.12％ 23.13％ 7.92％ -2.46％ Embodiment 14 8.26％ 22.16％ 8.36％ 1.21％ Comparative Example 17 8.17％ 22.29％ 8.39％ 0.69％

[0156] Result analysis: From the results of Examples 10 to 14, the gel prepared from the combination of Artemisia annua extract, Centella asiatica extract and Gentiana yunnanensis extract has significant moisturizing and repairing effects. Example 11, i.e., when the mass fraction of the composition is 5wt%, has the best moisturizing and repairing effects.

[0157] Comparing Example 11 with Comparative Example 17, Example 11 has the best effect, which shows that the composition can effectively improve the moisturizing and repairing effects of the gel matrix.

[0158] The embodiments described above are part of the embodiments of the present application, rather than all of the embodiments. The detailed description of the embodiments of the present application is not intended to limit the scope of the present application for protection, but merely represents the selected embodiments of the present application. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present application.

Claims

1. A soothing, moisturizing and repairing composition, characterized in that: Contains the following components in parts by weight: 0.1-2 parts of Artemisia annua extract; Centella asiatica extract 0.05-1 part; 0.1-2 parts of Gentiana yunnanensis extract; The Yunnan gentian extract is obtained by the following method: The whole herb of Gentiana yunnanensis is crushed and fermented by bacteria to obtain Gentiana yunnanensis extract; The preparation method of the gentiana yunnanensis extract specifically comprises the following steps: S1. Dry and crush the whole herb of Gentiana yunnanensis, add it into a specific culture medium for sterilization, and the material-liquid ratio is 1:1-5; S2, adding fermentation bacteria to the product of S1, and fermenting; S3, collecting the filtrate of the S2 product; S4, drying the product of S3 to prepare a gentiana yunnanensis extract; The specific culture medium in S1 is yeast extract peptone glucose medium; The fermentation strain in S2 is at least one of Saccharomyces cerevisiae and Lactobacillus plantarum. When Saccharomyces cerevisiae and Lactobacillus plantarum are used together, the mass ratio of Saccharomyces cerevisiae to Lactobacillus plantarum is 1:0.1-2. The oxygen content of the fermentation in S2 is 8% to 12%, the fermentation temperature is 28 to 32° C., and the fermentation time is 24 to 72 hours.

2. The soothing, moisturizing and repairing composition according to claim 1, characterized in that: The mass ratio of the Artemisia annua extract, Centella asiatica extract and Gentiana yunnanensis extract is 1:0.05-8:0.1-10.

3. The soothing, moisturizing and repairing composition according to claim 1, characterized in that: The composition contains the following parts by mass: 1 part of Artemisia annua extract, 1 part of Centella asiatica extract, and 0.1 part of Gentiana yunnanensis extract.

4. A soothing, moisturizing and repairing skin care product, characterized in that: It contains 2 to 10 wt % of the soothing, moisturizing and repairing composition according to any one of claims 1 to 3.

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