A soothing and repairing composition, preparation method and application

Abstract

The present invention belongs to the technical field of skin care, and discloses a soothing and repairing composition, a preparation method and an application thereof. The soothing and repairing composition comprises, by weight, 2-10 parts of Centella asiatica fermentation product, 5-20 parts of Emblica emblica fermentation product, 5-20 parts of Panax notoginseng root fermentation product, and 2-10 parts of Portulaca oleracea fermentation product. The present invention produces a synergistic effect of enhancing the soothing and repairing effects of the skin by compounding the Centella asiatica fermentation product, the Emblica emblica fermentation product, the Panax notoginseng root fermentation product, and the Portulaca oleracea fermentation product, thereby effectively improving the soothing and repairing effects of the composition.

Description

Technical Field

[0001] The present invention relates to the technical field of skin care, and in particular to a soothing and repairing composition, a preparation method and an application thereof. Background Art

[0002] Skin allergy refers to an allergic skin disease caused by allergens, which may be caused by exogenous allergens entering the body, allergic constitution, psychological stress, living environment, etc.; therefore, to relieve skin allergies, it is mainly necessary to remove allergens, care for the skin, and adjust lifestyle.

[0003] One way to care for the skin is to use mild skin care products to relieve itching, redness, swelling and other discomforts caused by skin allergies; further, you can use skin care products that soothe the skin to effectively regulate the skin's homeostasis, soothe the skin and keep it in a healthy state.

[0004] Based on this, many prior arts have proposed skin care formulas for soothing the skin, such as prior art 1: Chinese patent application 202110834663.3 discloses a skin soothing moisturizing cream, including a skin soothing conditioner, wherein the skin soothing conditioner includes the following components: component A, component B, component C, component D and component E, wherein component A comprises hydrogenated starch hydrolysate and water; and / or component B comprises butylene glycol, water, Astragalus membranaceus root extract, Saposhnikovia root extract, Calendula officinalis extract, Albizzia julibrissin flower extract, Gastrodia elata root extract; and / or component C comprises water, glycerin, β-glucan; and / or component D comprises water, glycerin, Dendrobium nobile stem extract, Aloe vera leaf extract, Sophora flavescens root extract, Lycium barbarum fruit extract, Echinacea purpurogenum extract; and / or component E comprises water, phospholipids, and (neuro)sphingolipids.

[0005] Existing technology 1 combines multiple plant extract components to form a multi-dimensional soothing effect system, which has the effects of instantly relieving stinging, instantly relieving itching, repairing skin barrier, improving sensitive skin conditions, relieving skin redness, and replenishing water and moisturizing. It is a very good technical solution.

[0006] However, as the development of plant extraction technology has entered a bottleneck period, this field has begun to study how to synergistically improve the skin care effects of plant ingredients through plant fermentation processes. Compared with plant extraction technology, plant fermentation processes can not only fully exert the effects of plant active ingredients, but also promote the growth and reproduction of microorganisms through certain ingredients in some plants, especially Chinese herbal medicines, so that natural plants and microorganisms can produce synergistic effects during the fermentation process, thereby further improving the skin care effects of plants.

[0007] As in prior art 2: Chinese patent application 202210313150.2 discloses a method for preparing a fermented product of Gegen Qinlian Decoction, comprising: providing a traditional Chinese medicine matrix, wherein the traditional Chinese medicine matrix is ​​composed of Gegen Qinlian Decoction and water, and the Gegen Qinlian Decoction is composed of Pueraria lobata, Scutellaria baicalensis, Coptis chinensis and Glycyrrhiza uralensis; using Ganoderma lucidum (Ganoderma lucidum) to perform a first fermentation step on the traditional Chinese medicine matrix at 20°C to 35°C for 24 to 144 hours to obtain a first culture; and using yeast to perform a second fermentation step on the first culture at 20°C to 35°C for 12 to 96 hours to obtain the fermented product of Gegen Qinlian Decoction, wherein the fermented product of Gegen Qinlian Decoction has whitening and soothing effects on the skin.

[0008] Existing technology 2 uses fermentation technology to ferment the traditional Chinese medicine matrix, which effectively improves the whitening and soothing effects of the fermented product of Gegenqinlian Decoction, and is a technology worthy of reference; but based on the diversity of natural plants, it is not enough to have only one skin-soothing fermentation product composition formula. In order to develop more natural plant fermentation products to improve the soothing effect of skin care products, it is necessary to develop the fermentation of multiple natural plants to fill the gap in the market for compound soothing natural plant fermentation products. Summary of the invention

[0009] One of the purposes of the present invention is to provide a soothing and repairing composition, which is compounded with fermentation products of various natural plants to provide a new skin soothing formula, which fills the gap in the market and synergistically improves the skin soothing effect through the compounding of various natural plant fermentation products.

[0010] Another object of the present invention is to provide a method for preparing a soothing and repairing composition, which effectively enhances the synergistic effect between microorganisms and natural plants through a secondary fermentation process, thereby greatly improving the skin soothing and repairing effects of the soothing and repairing composition.

[0011] At the same time, the present invention also provides an application of a soothing and repairing composition, which can be used to prepare skin care products and effectively improve the skin soothing and repairing effects of the skin care products.

[0012] To achieve the above objectives, the present invention provides a soothing and repairing composition, comprising, by weight, 2-10 parts of Centella asiatica fermentation product, 5-20 parts of Emblica emblica fermentation product, 2-10 parts of Panax notoginseng root fermentation product, and 5-20 parts of Portulaca oleracea fermentation product.

[0013] Furthermore, the Centella asiatica fermentation product, the Emblica officinalis fermentation product, the Notoginseng root fermentation product, and the Portulaca oleracea fermentation product are obtained by using Centella asiatica, the Emblica officinalis, the Notoginseng root, and the Portulaca oleracea as fermentation substrates and fermenting yeast or lactic acid bacteria in the fermentation substrates.

[0014] Preferably, the yeast is Saccharomyces cerevisiae, and the lactic acid bacteria is Lactobacillus.

[0015] The present invention also provides a method for preparing the soothing and repairing composition, comprising the following steps:

[0016] Step 1: inoculating yeast seed liquid or lactic acid bacteria seed liquid into a fermentation medium containing Centella asiatica and Emblica officinalis to perform a primary fermentation to obtain a primary fermentation product;

[0017] Step 2: adding Bifidobacterium longum fermentation agent, Panax notoginseng root and Portulaca oleracea to the primary fermentation product for secondary fermentation to obtain the soothing and repairing composition;

[0018] The mass ratio of the yeast seed liquid or lactic acid bacteria seed liquid to the fermentation medium is 0.1-1:1000; the addition amount of the Bifidobacterium longum fermentation agent is 0.1-2wt% of the primary fermentation product.

[0019] Furthermore, the yeast seed liquid or lactic acid bacteria seed liquid is obtained by pre-activating the corresponding yeast liquid or lactic acid bacteria liquid, and the activation treatment process is as follows:

[0020] Yeast liquid or lactic acid bacteria liquid is added to the sterilized activation culture medium, and a yeast activation liquid or a lactic acid bacteria activation liquid is obtained by primary culture.

[0021] Adding yeast activation liquid or lactic acid bacteria activation liquid to the sterilized seed culture medium, performing secondary culture to obtain yeast seed liquid or lactic acid bacteria seed liquid;

[0022] The activation culture medium and the seed culture medium both contain the following ingredients: 0.1-10wt% sugar, 0.1-5wt% malt extract, and the balance is water.

[0023] Preferably, the sugars in the activation medium and the seed medium are independently one or more combinations of glucose, lactose and galactose.

[0024] The live bacteria content per milliliter of yeast or lactic acid bacteria solution is 1×10 7 -1×10 8 CFU / mL.

[0025] Preferably, the specific operation of step 1 is: prepare a fermentation medium by mixing 20g of glucose, 10g of malt extract, 2-10g of Centella asiatica powder, 5-20g of Emblica officinalis powder, and the balance of water per 1kg, inoculate yeast seed liquid or lactic acid bacteria seed liquid into the fermentation medium, and ferment at 28°C for 24h.

[0026] Preferably, the specific process of a single fermentation is: when fermenting for 0-4 hours, set the speed to 50 rpm; when fermenting for 4-8 hours, set the speed to 80 rpm; when fermenting for 8-24 hours, set the dissolved oxygen content value of the fermentation tank to 30%, the stirring speed is associated with the dissolved oxygen content, and the speed is automatically compensated and adjusted; after 24 hours, the stirring is stopped when the single fermentation ends.

[0027] Preferably, the specific operation of step 2 is: adding Bifidobacterium longum fermentation agent, 5-20g Panax notoginseng root powder, and 2-10g Portulaca oleracea powder to the primary fermentation product, setting the constant culture temperature of the fermentation tank to 37°C, closing the air inlet channel, anaerobic fermentation, setting the speed to a constant 80 rpm, and fermenting for 24 hours to obtain a fermentation culture liquid of secondary fermentation; sterilizing the fermentation culture liquid at a high temperature and high pressure of 121°C for 20 minutes, and after sterilization, using a centrifuge at a speed of 14000 rpm to treat the fermentation culture liquid, collecting the culture supernatant after centrifugation, and clarifying the culture supernatant to obtain the soothing and repairing composition.

[0028] The present invention also discloses an application of the soothing and repairing composition in preparing skin care products. The soothing and repairing composition is applied in the preparation of skin care products.

[0029] Preferably, the skin care product is an aqueous solution, essence, facial mask, lotion, or cream.

[0030] Beneficial Effects

[0031] Compared with the prior art, the present invention has at least the following advantages:

[0032] (1) The present invention produces a synergistic effect of enhancing the skin soothing and repairing effects by compounding the fermented products of Centella asiatica, the fermented products of Phyllanthus emblica, the fermented products of the root of Panax notoginseng, and the fermented products of Portulaca oleracea, thereby effectively improving the skin soothing and repairing effects of the composition;

[0033] (2) The present invention discloses a soothing and repairing composition, which uses Centella asiatica, Phyllanthus emblica, Radix notoginseng and Portulaca oleracea as fermentation substrates and is obtained by fermentation with specific strains of bacteria, and can greatly improve the skin soothing and repairing effects of the soothing and repairing composition;

[0034] (3) The present invention discloses a method for preparing a soothing and repairing composition, wherein Centella asiatica and Emblica officinalis are used as fermentation substrates for the first fermentation to promote the growth and reproduction of microorganisms, and then Bifidobacterium longum fermentation agent, Panax notoginseng root, and Portulaca oleracea are added for the second fermentation to maximize the synergistic effect of Centella asiatica, Emblica officinalis, Panax notoginseng root, Portulaca oleracea and microorganisms, thereby further improving the synergistic skin soothing and repairing effects of the soothing and repairing composition. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] The present invention is further described below in conjunction with the accompanying drawings and embodiments;

[0036] Figure 1 This is a chicken embryo observation picture before using the soothing and repairing composition of Example 1 of the present invention;

[0037] Figure 2 This is a picture of chicken embryo observation after using the soothing and repairing composition of Example 1 of the present invention. DETAILED DESCRIPTION

[0038] The present invention will be further described below in conjunction with the embodiments, but it does not constitute any limitation to the present invention. Any limited number of modifications made within the scope of the claims of the present invention are still within the scope of the claims of the present invention.

[0039] In order to explain the technical content of the present invention in detail, further description is given below in conjunction with the implementation modes.

[0040] In the following examples and comparative examples, the brewer's yeast liquid was purchased from Guangdong Microbiological Culture Collection Center, and the strain number is CGMCC 2.1364.

[0041] In the following examples and comparative examples, the saccharomyces cerevisiae seed solution is obtained by pre-activating the saccharomyces cerevisiae bacterial solution, and the activation process is as follows:

[0042] The saccharomyces cerevisiae liquid is added to the sterilized activation medium, and the saccharomyces cerevisiae activation liquid is obtained by primary cultivation.

[0043] Adding the activating solution of brewer's yeast to the sterilized seed culture medium, and performing secondary culture to obtain the brewer's yeast seed solution;

[0044] The activation medium comprises the following ingredients: 1wt% glucose, 0.5wt% malt extract, and the balance water;

[0045] The seed culture medium comprises the following ingredients: 2 wt% glucose, 1 wt% malt extract, and the balance water;

[0046] The concentration of the brewer's yeast solution is 1×10 7 CFU / g, and the added amount was 0.2wt% of the activation medium.

[0047] In the following examples and comparative examples, the Centella asiatica powder, Emblica officinalis powder, Panax notoginseng root powder, Portulaca oleracea powder, Chlorella powder, oat powder, Ganoderma lucidum powder, and Licorice powder are all powders obtained by crushing and sieving to 100 mesh.

[0048] Example 1

[0049] A soothing and repairing composition is prepared by the following steps:

[0050] Step 1: prepare a fermentation medium containing 20g glucose, 10g malt extract, 5g Centella asiatica powder, 10g Phyllanthus emblica powder, 10g Panax notoginseng root powder, 5g Portulaca oleracea powder and 940g water, inoculate 1g of Saccharomyces cerevisiae seed liquid into the fermentation medium and ferment at 28°C for 24h, wherein the rotation speed is set to 50 rpm during fermentation for 0-4h; the rotation speed is set to 80 rpm during fermentation for 4-8h; the dissolved oxygen content of the fermentation tank is set to 30% during fermentation for 8-24h, the stirring speed is linked to the dissolved oxygen content, and the speed is automatically compensated and adjusted; after 24h, the stirring is stopped when the fermentation ends.

[0051] Example 2

[0052] A soothing and repairing composition is prepared by the following steps:

[0053] Step 1: prepare a fermentation medium according to a mixture containing 20g of glucose, 10g of malt extract, 5g of Centella asiatica powder, 10g of Phyllanthus emblica powder and 955g of water, inoculate 1g of saccharomyces cerevisiae seed liquid into the fermentation medium, and ferment for 24 hours at 28°C, wherein the rotation speed is set to 50 rpm during the fermentation for 0-4h; the rotation speed is set to 80 rpm during the fermentation for 4-8h; the dissolved oxygen content value of the fermentation tank is set to 30% during the fermentation for 8-24h, the stirring speed is associated with the dissolved oxygen content, and the speed is automatically compensated and adjusted; after 24h, the fermentation is stopped and stirring is stopped to obtain a fermentation product;

[0054] Step 2: Add Bifidobacterium longum fermentation agent, 10g Panax notoginseng root powder, and 5g Portulaca oleracea powder to the primary fermentation product, set the constant temperature of the fermentation tank to 37°C, close the air inlet channel, perform anaerobic fermentation, set the speed to a constant 80 rpm, and ferment for 24 hours to obtain a fermentation culture liquid of secondary fermentation; sterilize the fermentation culture liquid at a high temperature and high pressure of 121°C for 20 minutes, and after sterilization, use a centrifuge at a speed of 14000 rpm to treat the fermentation culture liquid, collect the culture supernatant after centrifugation, and clarify the culture supernatant to obtain the soothing repair composition;

[0055] The addition amount of the Bifidobacterium longum fermentation agent is 0.5wt% of the primary fermentation product.

[0056] Example 3

[0057] A soothing and repairing composition is prepared by the following steps:

[0058] Step 1: prepare a fermentation medium according to a mixture containing 20g of glucose, 10g of malt extract, 5g of Centella asiatica powder, 10g of Phyllanthus emblica powder and 955g of water, inoculate 1g of saccharomyces cerevisiae seed liquid into the fermentation medium, and ferment for 24 hours at 28°C, wherein the rotation speed is set to 50 rpm during the fermentation for 0-4h; the rotation speed is set to 80 rpm during the fermentation for 4-8h; the dissolved oxygen content value of the fermentation tank is set to 30% during the fermentation for 8-24h, the stirring speed is associated with the dissolved oxygen content, and the speed is automatically compensated and adjusted; after 24h, the fermentation is stopped and stirring is stopped to obtain a fermentation product;

[0059] Step 2: Add 5g of Panax notoginseng root powder and 10g of Portulaca oleracea powder to the primary fermentation product, set the fermentation tank to a constant temperature of 37°C, close the air inlet channel, perform anaerobic fermentation, set the rotation speed to a constant 80 rpm, and ferment for 24 hours to obtain a fermentation culture medium for secondary fermentation; sterilize the fermentation culture medium at a high temperature and high pressure of 121°C for 20 minutes. After sterilization, use a centrifuge at a speed of 14000 rpm to treat the fermentation culture medium, collect the culture supernatant after centrifugation, and clarify the culture supernatant to obtain the soothing and repairing composition.

[0060] Example 4

[0061] It is generally the same as Example 2, except that in step 1, the amount of Centella asiatica powder is 4 g, and the amount of Emblica officinalis powder is 11 g; in step 2, the amount of Panax notoginseng root powder is 10 g, and the amount of Portulaca oleracea powder is 5 g.

[0062] Example 5

[0063] It is generally the same as Example 2, except that in step 1, the amount of Centella asiatica powder is 8 g, and the amount of Emblica officinalis powder is 7 g; in step 2, the amount of Panax notoginseng root powder is 2 g, and the amount of Portulaca oleracea powder is 13 g.

[0064] Comparative Example 1

[0065] The method is generally the same as Example 2, except that in step 1, no emblica powder is added, and only 15 g of Centella asiatica powder is added.

[0066] Comparative Example 2

[0067] The method is generally the same as Example 2, except that in step 1, no Centella asiatica powder is added, and only 15 g of Emblica officinalis powder is added.

[0068] Comparative Example 3

[0069] It is generally the same as Example 2, except that in step 2, no notoginseng root powder is added, and the amount of purslane powder is changed to 15 g.

[0070] Comparative Example 4

[0071] It is generally the same as Example 2, except that in step 2, no purslane powder is added, and the amount of Panax notoginseng root powder is changed to 15 g.

[0072] Comparative Example 5

[0073] The method is generally the same as Example 1, except that the Centella asiatica powder in step 1 is replaced by Chlorella vulgaris powder.

[0074] Comparative Example 6

[0075] The method is generally the same as Example 1, except that the emblica powder in step 1 is replaced by oat powder.

[0076] Comparative Example 7

[0077] It is generally the same as Example 1, except that the notoginseng root powder in step 1 is replaced by Ganoderma lucidum powder.

[0078] Comparative Example 8

[0079] The method is substantially the same as Example 1, except that the purslane powder in step 1 is replaced by licorice powder.

[0080] Performance Testing

[0081] 1. Hyaluronidase inhibition rate

[0082] 1. Purpose and principle of the experiment

[0083] Hyaluronidase is a specific cleavage enzyme of hyaluronic acid, a participant in allergic reactions, and has a strong correlation with the release of histamine by mast cells. Whether the test sample has a soothing effect can be judged by the hyaluronidase inhibition rate. The higher the hyaluronidase inhibition rate, the stronger the soothing effect of the substance, and vice versa.

[0084] This test refers to the laboratory method (HMC-WI-029 hyaluronidase inhibition rate) and compares the hyaluronidase inhibition rate test results of the test sample with that of the negative control. If the inhibition 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 a certain soothing effect.

[0085] This test method is an in vitro method suitable for evaluating cosmetic products that claim to achieve soothing effects by inhibiting hyaluronidase.

[0086] 2 Test indicators

[0087] Hyaluronidase inhibition rate determination criteria: If the hyaluronidase inhibition rate of the 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 a certain soothing effect.

[0088] 3 Experimental materials and methods

[0089] 3.1 Instruments and Equipment

[0090] BSA224S Analytical Balance

[0091] L6s UV Spectrophotometer

[0092] 3.2 Reagents

[0093] Hyaluronidase, BR

[0094] Sodium Hyaluronate, BR

[0095] 3.3 Test methods

[0096] (1) Treatment of control and test samples

[0097] Sample group: 100% of Examples 1-5 and Comparative Examples 1-8;

[0098] Positive control (dipotassium glycyrrhizinate, purity ≥98%): diluted with water to a positive control concentration of 3%;

[0099] Negative control: pure water.

[0100] (2) Test operation steps

[0101] Set up a sample group, a sample background group, a solvent group and a solvent background group. Each group needs to have 3 parallels. Add different reagent solutions to the four groups respectively, shake well, and leave at room temperature for 30 minutes for color development. Use a UV spectrophotometer to measure the absorbance value at a wavelength of 528nm.

[0102] (3) Calculation formula

[0103]

[0104] Where: A—is the absorbance of the reaction solution without sample;

[0105] B—is the absorbance of the reaction solution without sample and enzyme;

[0106] C—is the absorbance of the reaction solution containing sample and enzyme;

[0107] D—is the absorbance of the reaction solution containing sample and without enzyme.

[0108] (4) Data analysis

[0109] SPSS was used as the statistical analysis software, and the independent sample t-test was used to compare the hyaluronidase inhibition rates among the test samples, positive controls, and negative controls.

[0110] The above statistical analyses were all two-tailed tests, with a significance level of α = 0.05. P>0.05 means there is no significant difference between the two groups; P<0.05 means there is a significant difference between the two groups.

[0111] The soothing and repairing compositions obtained in Examples 1-5 and Comparative Examples 1-8 were tested according to the above test method, and the results are shown in Table 1;

[0112] Table 1 Test results of hyaluronidase inhibition rate of the soothing repairing composition obtained in Examples 1-5 and Comparative Examples 1-8

[0113] Hyaluronidase inhibition rate % P-value Example 1 55.24 ＜0.050 Example 2 59.38 ＜0.050 Example 3 57.49 ＜0.050 Example 4 58.98 ＜0.050 Example 5 58.49 ＜0.050 Comparative Example 1 37.90 ＜0.050 Comparative Example 2 38.28 ＜0.050 Comparative Example 3 48.91 ＜0.050 Comparative Example 4 47.87 ＜0.050 Comparative Example 5 38.26 ＜0.050 Comparative Example 6 44.58 ＜0.050 Comparative Example 7 49.46 ＜0.050 Comparative Example 8 45.73 ＜0.050 Negative control -2.81 / Positive Control 62.92 ＜0.050

[0114] According to the results in Table 1, we can see that:

[0115] According to the data comparison between Example 2 and Example 3 and Example 1, it can be seen that the technical scheme of using a secondary fermentation process in Example 2 and Example 3 of the present invention, and using Centella asiatica and Emblica officinalis as the fermentation substrates for the first fermentation to carry out a fermentation to promote the growth and reproduction of microorganisms can improve the extraction effect of hyaluronidase inhibitory active ingredients in Centella asiatica, Emblica officinalis, Panax notoginseng root and Portulaca oleracea, thereby further improving the soothing effect of the composition; and according to the data comparison between Example 2 and Example 3, it can be seen that the technical scheme of adding Bifidobacterium longum fermentation agent and brewer's yeast for mixed fermentation in Example 2 during the second fermentation process can also further improve the fermentation effect.

[0116] According to the data of Example 2 and Comparative Examples 1 and 2, the lack of the fermentation of Phyllanthus emblica in Comparative Example 1 and the lack of the fermentation of Centella asiatica in Comparative Example 2 will significantly reduce the hyaluronidase inhibition rate of the composition. The reason is that:

[0117] (1) The lack of emblica buds or Centella asiatica will cause the composition to lose the synergistic effect of the Centella asiatica fermentation product, emblica buds fermentation product, Panax notoginseng root fermentation product, and Portulaca oleracea fermentation product in enhancing the soothing effect of the composition. The compositions of Comparative Example 1 and Comparative Example 2 can only produce a simple superposition of effects, and the hyaluronidase inhibition effect is significantly reduced compared with Example 2.

[0118] (2) The lack of the combination of Phyllanthus emblica and Centella asiatica during the first fermentation process will lead to a decrease in the growth and reproduction ability of the microorganisms, thereby making the fermentation effect of the final composition worse, affecting the hyaluronidase inhibition effect of the composition.

[0119] Similarly, the technical solution of using Chlorella powder instead of Centella asiatica powder in Comparative Example 5 and the technical solution of using oat powder instead of Emblica emblica powder in Comparative Example 6 also resulted in a significant decrease in the hyaluronidase inhibition rate of the composition. However, compared with Comparative Example 1 and Comparative Example 2, since the combination of oat and Centella asiatica should have a certain promoting effect on the growth and reproduction ability of microorganisms, Comparative Example 6 has better hyaluronidase inhibition performance than Comparative Example 1, Comparative Example 2 and Comparative Example 5.

[0120] According to the data comparison of Example 2 and Comparative Example 3 and Comparative Example 4, it can be seen that in the absence of Panax notoginseng root fermentation product or Portulaca oleracea fermentation product, the hyaluronidase inhibition effect of the composition is significantly reduced, indicating that the present invention adopts the Centella asiatica fermentation product, the Emblica fructus fermentation product, the Panax notoginseng root fermentation product, and the Portulaca oleracea fermentation product to produce a synergistic effect, which synergistically improves the hyaluronidase inhibition efficacy of the composition.

[0121] Similarly, the technical solutions of Comparative Example 7 using Ganoderma lucidum powder to replace Panax notoginseng root powder for fermentation and Comparative Example 8 using Licorice powder to replace Purslane powder for fermentation are different from those of Comparative Examples 3 and 4 in hyaluronidase inhibition rate, but their hyaluronidase inhibition efficacy performance is almost the same, indicating that other plant fermentation products cannot replace Panax notoginseng root fermentation products, Purslane fermentation products and Centella asiatica fermentation products, Emblica emblica fermentation products, etc. to produce synergistic effects.

[0122] 2. Elastase inhibition rate

[0123] 1. Purpose and principle of the experiment

[0124] The anti-wrinkle effect is mainly characterized by evaluating the inhibition rate of test samples on elastase. Elastase is mainly synthesized and secreted by fibroblasts, which can degrade elastin in the skin and cause skin aging. The experimental principle of inhibiting elastase is that porcine pancreatic elastase catalyzes the reaction with the enzyme substrate. After adding the active substance, the absorbance changes. The size of the absorbance change reflects the inhibition rate of the elastase inhibitor.

[0125] This test refers to the laboratory method (HMC-WI-028 elastase inhibition rate) and compares the elastase inhibition rate test results of the test sample with that of the negative control. If the inhibition 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 a certain anti-wrinkle effect.

[0126] This test method is an in vitro method suitable for evaluating cosmetic products that claim to achieve anti-wrinkle efficacy by inhibiting elastase.

[0127] 2 Test indicators

[0128] Elastase inhibition rate determination standard: If the elastase inhibition rate of the 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 a certain anti-wrinkle effect.

[0129] 3 Experimental materials and methods

[0130] 3.1 Instruments and Equipment

[0131] BSA224S Analytical Balance

[0132] RT-6100 ELISA Analyzer

[0133] 3.2 Reagents

[0134] Elastase (Porcine Pancreas), BR

[0135] N-Succinyl-L-Alanyl-L-Alanyl-L-Alanine, 98%

[0136] Epigallocatechin gallate (EGCG), 98%

[0137] 3.3 Test methods

[0138] (1) Treatment of control and test samples

[0139] Sample group: 100% of Examples 1-5 and Comparative Examples 1-8;

[0140] Positive control (EGCG): diluted with water to a positive control concentration of 0.1%;

[0141] Negative control: pure water.

[0142] (2) Test operation steps

[0143] Set up sample group, sample background group, solvent group and solvent background group. Each group needs to set up 3 parallels. Add different reagent solutions into the 96-well plate, shake gently, incubate at 25℃ for 15 minutes, place it in the microplate reader, and measure the absorbance at 410nm.

[0144] (3) Calculation formula

[0145]

[0146] Where: A—is the absorbance of the reaction solution without sample;

[0147] B—is the absorbance of the reaction solution without sample and enzyme;

[0148] C—is the absorbance of the reaction solution containing sample and enzyme;

[0149] D—is the absorbance of the reaction solution containing sample and without enzyme.

[0150] (4) Data analysis

[0151] The statistical analysis software was SPSS, and the comparison of the elastase inhibition rate between the test sample, positive control and negative control was performed by independent sample t test. The above statistical analysis was two-tailed test, and the significance level was α = 0.05. P>0.05 indicated that there was no significant difference between the two groups; P<0.05 indicated that there was a significant difference between the two groups.

[0152] The soothing and repairing compositions obtained in Examples 1-5 and Comparative Examples 1-8 were tested according to the above test method, and the results are shown in Table 2;

[0153] Table 2 Test results of elastase inhibition rate of the soothing repair composition obtained in Examples 1-5 and Comparative Examples 1-8

[0154]

[0155]

[0156] According to the results in Table 2:

[0157] According to the data comparison between Example 2 and Example 3 and Example 1, it can be seen that the technical solution of using a secondary fermentation process in Example 2 and Example 3 of the present invention, and using Centella asiatica and Emblica emblica as the fermentation substrates of the first fermentation to carry out a fermentation to promote the growth and reproduction of microorganisms can improve the extraction effect of the active ingredients inhibiting elastase in Centella asiatica, Emblica emblica, Notoginseng root and Portulaca oleracea, thereby further improving the anti-wrinkle effect of the composition; and according to the data comparison between Example 2 and Example 3, it can be seen that the technical solution of adding Bifidobacterium longum fermentation agent and saccharomyces cerevisiae fermentation in Example 2 during the second fermentation process can also further improve the fermentation effect.

[0158] According to the data of Example 2 and Comparative Examples 1 and 2, the lack of the fermentation of Phyllanthus emblica in Comparative Example 1 and the lack of the fermentation of Centella asiatica in Comparative Example 2 will significantly reduce the hyaluronidase inhibition rate of the composition. The reason is that:

[0159] (1) The lack of Phyllanthus emblica or Centella asiatica will cause the composition to lose the synergistic effect of the Centella asiatica fermentation product, Phyllanthus emblica fermentation product, Panax notoginseng root fermentation product, and Portulaca oleracea fermentation product in improving the anti-wrinkle effect of the composition. The compositions of Comparative Example 1 and Comparative Example 2 can only produce a simple superposition of effects, and the elastase inhibition effect is significantly reduced compared with Example 2.

[0160] (2) The lack of the combination of Phyllanthus emblica and Centella asiatica in the first fermentation process will lead to a decrease in the growth and reproduction capacity of the microorganisms, thereby making the fermentation effect of the final composition worse, affecting the elastase inhibition effect of the composition.

[0161] Similarly, the technical solution of using Chlorella powder instead of Centella asiatica powder in Comparative Example 5 and the technical solution of using oat powder instead of Emblica emblica powder in Comparative Example 6 also resulted in a significant decrease in the elastase inhibition rate of the composition. However, compared with Comparative Example 1 and Comparative Example 2, since the combination of oat and Centella asiatica should have a certain promoting effect on the growth and reproduction ability of microorganisms, Comparative Example 6 has better elastase inhibition performance than Comparative Example 1, Comparative Example 2 and Comparative Example 5.

[0162] According to the data comparison of Example 2 and Comparative Example 3 and Comparative Example 4, it can be seen that in the absence of Panax notoginseng root fermentation product or Portulaca oleracea fermentation product, the elastase inhibition effect of the composition is significantly reduced, indicating that the present invention adopts the Centella asiatica fermentation product, the Emblica fructus fermentation product, the Panax notoginseng root fermentation product, and the Portulaca oleracea fermentation product to produce a synergistic effect, which synergistically improves the elastase inhibition effect of the composition.

[0163] Similarly, the technical solutions of Comparative Example 7 using Ganoderma lucidum powder to replace Panax notoginseng root powder for fermentation and Comparative Example 8 using Licorice powder to replace Purslane powder for fermentation are different from those of Comparative Examples 3 and 4 in terms of elastase inhibition rate, but their elastase inhibition efficacy performance is almost the same, indicating that other plant fermentation products cannot replace Panax notoginseng root fermentation products, Purslane fermentation products, Centella asiatica fermentation products, Emblica emblica fermentation products, etc. to produce synergistic effects.

[0164] 3. DPPH free radical scavenging rate

[0165] 1. Purpose and principle of the experiment

[0166] Excessive production of free radicals can lead to natural aging and photoaging of the skin, thus causing wrinkles. Therefore, whether it has the ability to scavenge free radicals is one of the important indicators for evaluating anti-aging and anti-oxidant cosmetics (raw materials).

[0167] 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 a certain antioxidant effect.

[0168] This test method is an in vitro method and is suitable for evaluating cosmetic products that claim to achieve antioxidant effects by scavenging free radicals.

[0169] 2 Test indicators

[0170] DPPH free radical scavenging rate determination standard: If the DPPH scavenging rate of the 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 a certain antioxidant effect.

[0171] 3 Experimental materials and methods

[0172] 3.1 Instruments and Equipment

[0173] BSA224S Analytical Balance

[0174] L6s UV Spectrophotometer

[0175] 3.2 Reagents

[0176] DPPH (1,1-diphenyl-2-picrylhydrazyl), 98%

[0177] 3.3 Test methods

[0178] (1) Treatment of control and test samples

[0179] Sample group: 100% of Examples 1-5 and Comparative Examples 1-8;

[0180] Positive control (vitamin E, purity ≥96%): dilute with 95% ethanol to a positive control concentration of 0.1%;

[0181] Negative control: pure water.

[0182] (2) Test operation steps

[0183] Set up sample tubes, sample background tubes, DPPH tubes, and solvent background tubes. Set up 3 parallel tubes for each group. Add different reagent solutions to the four groups, shake gently, and let stand at room temperature for 5 minutes. Transfer the reaction solutions of each group into a 1 cm cuvette and measure the absorbance at 517 nm.

[0184] (3) Calculation formula

[0185]

[0186] Where: T—absorbance of sample tube, i.e. absorbance of solution after sample reacts with DPPH;

[0187] T0—sample background absorbance;

[0188] 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;

[0189] C0—Solvent background absorbance.

[0190] The soothing and repairing compositions obtained in Examples 1-5 and Comparative Examples 1-8 were tested according to the above test method, and the results are shown in Table 3;

[0191] Table 3 DPPH free radical scavenging rate test results of the soothing repairing compositions obtained in Examples 1-5 and Comparative Examples 1-8

[0192]

[0193]

[0194] According to the results in Table 3:

[0195] According to the data comparison of Example 2 and Example 3 with Example 1, it can be seen that Examples 2 and 3 of the present invention adopt a secondary fermentation process, and the technical solution of using Centella asiatica and Emblica officinalis as the fermentation substrate of the first fermentation to carry out a fermentation to promote the growth and reproduction of microorganisms can improve the extraction effect of the active ingredients for scavenging DPPH free radicals in Centella asiatica, Emblica officinalis, Panax notoginseng root, and Portulaca oleracea, thereby further improving the antioxidant efficacy of the composition; and according to the data comparison of Example 2 and Example 3, it can be seen that the technical solution of adding a long bifidobacterium fermentation agent and mixed fermentation of saccharomyces cerevisiae in Example 2 during the second fermentation process can also further improve the fermentation effect, but compared with the data in Tables 1 and 2, the improvement range is significantly lower than the difference between Examples 1 and 3, indicating that the addition of a long bifidobacterium fermentation agent and mixed fermentation of saccharomyces cerevisiae in Example 2 can only improve the extraction effect of the active ingredients inhibiting hyaluronidase and elastase, and the improvement of the extraction effect of the antioxidant active ingredients is relatively weak.

[0196] According to the data of Example 2 and Comparative Examples 1 and 2, the lack of fermentation of Phyllanthus emblica in Comparative Example 1 and the lack of fermentation of Centella asiatica in Comparative Example 2 will significantly reduce the DPPH radical scavenging rate of the composition. The reason is that:

[0197] (1) The lack of emblica buds or Centella asiatica will cause the composition to lose the synergistic effect of the four fermentation products of Centella asiatica, emblica buds, Panax notoginseng root fermentation products, and Portulaca oleracea to enhance the antioxidant efficacy of the composition. The compositions of Comparative Example 1 and Comparative Example 2 can only produce a simple efficacy superposition, and the effect of scavenging DPPH free radicals is significantly reduced compared with Example 2.

[0198] (2) The lack of the combination of Phyllanthus emblica and Centella asiatica in the first fermentation process will lead to a decrease in the growth and reproduction ability of the microorganisms, thereby making the fermentation effect of the final composition worse, affecting the DPPH free radical scavenging effect of the composition.

[0199] Similarly, the technical solution of using Chlorella powder instead of Centella asiatica powder in Comparative Example 5 and the technical solution of using oat powder instead of Emblica emblica powder in Comparative Example 6 also resulted in a significant decrease in the DPPH radical scavenging rate of the composition. However, compared with Comparative Examples 1 and 2, since the combination of oats and Centella asiatica should have a certain promoting effect on the growth and reproduction ability of microorganisms, Comparative Example 6 has better DPPH radical scavenging performance than Comparative Examples 1, 2 and 5.

[0200] According to the data comparison of Example 2 and Comparative Example 3 and Comparative Example 4, it can be seen that in the absence of Panax notoginseng root fermentation product or Portulaca oleracea fermentation product, the elastase inhibition effect of the composition is significantly reduced, indicating that the present invention adopts the Centella asiatica fermentation product, the Emblica fructus fermentation product, the Panax notoginseng root fermentation product, and the Portulaca oleracea fermentation product to produce a synergistic effect, which synergistically enhances the antioxidant efficacy of the composition.

[0201] Similarly, the technical solutions of Example 7 using Ganoderma lucidum powder to replace Panax notoginseng root powder for fermentation and Example 8 using licorice powder to replace Purslane powder for fermentation are different from those of Examples 3 and 4 in DPPH radical scavenging rate, but their antioxidant efficacy performance is almost the same, indicating that other plant fermentation products cannot replace Panax notoginseng root fermentation products, Purslane fermentation products and Centella asiatica fermentation products, Emblica emblica fermentation products, etc. to produce synergistic effects.

[0202] 4. Irritation test

[0203] Safety Testing

[0204] Test method: Chicken embryo chorioallantoic membrane test

[0205] 1. Test purpose and principle

[0206] The chicken embryo chorioallantoic membrane test is an early in vitro eye irritation assessment method. The chorioallantoic membrane (CAM) is a respiratory membrane that surrounds the chicken embryo. This test uses the characteristics of the intact, clear and transparent vascular system of the chorioallantoic membrane of the mid-stage incubated chicken embryo. A certain amount of the test substance is directly exposed to the chicken embryo chorioallantoic membrane. After a period of action, the changes in the toxic effect indicators of the chorioallantoic membrane (such as bleeding, coagulation and vascular melting) are observed. These indicators reflect the changes in the morphological structure, color and permeability of the blood vessels and vascular networks, as well as the phenomena such as the denaturation of the chorioallantoic membrane proteins and the degree of damage. Then, a score is obtained by combining them to evaluate the eye irritation of the test substance.

[0207] The purpose of this test is to test the ability of the test substance to cause toxic changes in the chicken embryo chorioallantoic membrane and to evaluate the elements and processes of the potential eye irritation of the evaluated substance.

[0208] 2. Experimental Group Treatment and Reagents

[0209] Sample group: the soothing and repairing compositions prepared in Examples 1-5 and Comparative Examples 1-8.

[0210] Negative control: 0.9% saline.

[0211] Positive control: 1% sodium dodecyl sulfate (SDS) solution.

[0212] 3. Experimental steps

[0213] In this test, 6 chicken embryos were selected for each group, and the conditions of the chorioallantoic membrane were recorded with a camera. A polytetrafluoroethylene resin ring was placed on the chorioallantoic membrane of the chicken embryo and photographed. The sample to be tested was added to the polytetrafluoroethylene resin ring, the time of adding the sample was recorded, and the air chamber was covered with a moistened plastic wrap. The chicken embryo was moved to a constant temperature and humidity chamber for culture, and the degree of change of each toxic effect was observed.

[0214] 4. Calculation of results

[0215] The endpoint evaluation method was used to perform the test and the endpoint evaluation (ES) was calculated with two decimal places. Each chicken embryo was scored as the sum of the bleeding, coagulation and vascular melting observed in each chicken embryo. The average value of the mathematical sum obtained for ES-6 chicken embryos was taken.

[0216] The average ES score was calculated according to the following formula.

[0217]

[0218] 5. Result judgment criteria

[0219] ES≤4, non-irritating;

[0220] 4<ES≤12, mild irritation;

[0221] 12<ES<16, moderate irritation;

[0222] ES ≥ 16, strong irritant / corrosive.

[0223] 2. Test Results

[0224] According to the above test method, samples of Examples 1-5 and Comparative Examples 1-8 were tested, and the ES were all less than or equal to 4, and the results were all judged to be non-irritating.

[0225] Among them, the results of chicken embryos before and after use in Example 1 are as follows Figure 1-Figure 2 As shown, the sample test result of Example 1 is ES=4.0;

[0226] This proves that the technical solution of the present invention is mild and non-irritating.

[0227] Application Examples

[0228] A soothing and repairing composition essence, the formula ratio of which is shown in Table 4.

[0229] Table 4

[0230]

[0231]

[0232] The embodiments presented herein are only embodiments selected according to the combination of all possible embodiments. The appended claims should not be limited by the embodiments describing the present invention. Some numerical ranges used in the claims include sub-ranges therein, and the changes in these ranges should also be covered by the appended claims.

Claims

1. A soothing and repairing composition, characterized in that: The invention comprises, by weight, 2-10 parts of Centella asiatica fermentation product, 5-20 parts of Emblica officinalis fermentation product, 5-20 parts of Panax notoginseng root fermentation product, and 2-10 parts of Portulaca oleracea fermentation product, wherein the Centella asiatica fermentation product, the Emblica officinalis fermentation product, the Panax notoginseng root fermentation product, and the Portulaca oleracea fermentation product are obtained by fermenting the Centella asiatica, the Emblica officinalis, the Panax notoginseng root, and the Portulaca oleracea with the seed liquid of Saccharomyces cerevisiae in the fermentation substrate; The brewer's yeast is a brewer's yeast liquid with a strain number of CGMCC 2.1364, wherein the brewer's yeast seed liquid is activated by the following process: Adding the saccharomyces cerevisiae liquid to the sterilized activation medium, and performing primary cultivation to obtain the saccharomyces cerevisiae activation liquid; Adding the activating solution of brewer's yeast to the sterilized seed culture medium, and performing secondary culture to obtain the brewer's yeast seed solution; The activation medium comprises the following ingredients: 1wt% glucose, 0.5wt% malt extract, and the balance water; The seed culture medium comprises the following ingredients: 2wt% glucose, 1wt% malt extract, and the balance is water; The concentration of the brewer's yeast solution is 1×10 7 CFU / g, the amount added was 0.2wt% of the activated medium; The soothing and repairing composition is prepared by the following steps: Step 1: inoculating a fermentation medium containing Centella asiatica and Emblica officinalis with a seed solution of Saccharomyces cerevisiae for primary fermentation to obtain a primary fermentation product; Step 2: adding Bifidobacterium longum fermentation agent, Panax notoginseng root and Portulaca oleracea to the primary fermentation product for secondary fermentation to obtain the soothing and repairing composition; The mass ratio of the saccharomyces cerevisiae to the fermentation medium is 0.1-1:1000; and the addition amount of the Bifidobacterium longum fermentation agent is 0.1-2wt% of the primary fermentation product.

2. The soothing and repairing composition according to claim 1, characterized in that: The specific operation of step 1 is: prepare a fermentation medium by mixing 20g of glucose, 10g of malt extract, 2-10g of Centella asiatica powder, 5-20g of Emblica officinalis powder, and the balance of water per 1kg, inoculate brewer's yeast seed liquid into the fermentation medium, and ferment at 28°C for 24h.

3. The soothing and repairing composition according to claim 2, characterized in that: The specific process of primary fermentation is: when fermenting for 0-4h, set the speed to 50 rpm; when fermenting for 4-8h, set the speed to 80 rpm; when fermenting for 8-24h, set the dissolved oxygen content in the fermentation tank to 30%, the stirring speed is linked to the dissolved oxygen content, and the speed is automatically compensated and adjusted; after 24h, the stirring is stopped when the primary fermentation ends.

4. The soothing and repairing composition according to claim 1, characterized in that: The specific operation of step 2 is: adding Bifidobacterium longum fermentation agent, 5-20g Panax notoginseng root powder, and 2-10g Portulaca oleracea powder to the primary fermentation product, setting the constant temperature of the fermentation tank to 37°C, closing the air inlet channel, anaerobic fermentation, setting the rotation speed to a constant 80 rpm, and fermenting for 24 hours to obtain the fermentation culture liquid of the secondary fermentation; sterilizing the fermentation culture liquid at a high temperature and high pressure of 121°C for 20 minutes, and after sterilization, using a centrifuge at a speed of 14000 rpm to treat the fermentation culture liquid, collecting the culture supernatant after centrifugation, and clarifying the culture supernatant to obtain the soothing and repairing composition.

5. An application of the soothing and repairing composition according to claim 1, characterized in that: The soothing and repairing composition is used in the preparation of skin care products.

6. The use of the soothing and repairing composition according to claim 5, characterized in that: The skin care products are water solution, essence, facial mask, lotion and cream.

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