Anti-wrinkle firming composition and application thereof

Abstract

The invention discloses an anti-wrinkle firming composition and application thereof, and particularly belongs to the technical field of skin care products. The composition is prepared from the following components in parts by mass: 0.1 to 10 parts of rhodiola rosea extract, 5 to 20 parts of hydroxypropyl tetrahydropyrantriol, 0.02 to 2 parts of polyrock extract and 0.5 to 2 parts of adenosine, wherein the polyrock extract is obtained by carrying out compound fermentation on lactobacillus acidophilus and bacillus licheniformis; according to the anti-wrinkle and firming composition provided by the invention, when the rhodiola rosea extract, the hydroxypropyl tetrahydropyrantriol, the polyrock extract and the adenosine are compounded for use, excellent effects of resisting oxidation and promoting collagen synthesis are shown; the anti-wrinkle and firming emulsion provided by the invention can effectively reduce wrinkles and improve skin elasticity, shows excellent performance, overcomes the problems of efficacy limitation and single action mechanism of a single component, and realizes efficient anti-wrinkle and firming effects under a multi-channel synergistic effect.

Description

Technical Field

[0001] The present invention belongs to the technical field of skin care products, and particularly relates to an anti-wrinkle and firming composition and its application. Background Art

[0002] During the skin aging process, the formation of wrinkles is closely related to multiple factors such as the decline of skin barrier function, collagen loss, degradation of the extracellular matrix, and oxidative stress damage. Existing anti-wrinkle and firming products mostly rely on a single active ingredient, and their action pathways are single, making it difficult to comprehensively cope with the complex skin aging mechanism. For example, although Rhodiola rosea extract has antioxidant and anti-inflammatory activities, its promoting effect on collagen synthesis is limited when used alone; although propyltetrahydropyranyl triol can activate fibroblasts to stimulate the synthesis of glycosaminoglycans (GAGs) and collagen, its permeability and long-term effect are insufficient; adenosine can improve skin microcirculation by promoting cell metabolism, but its effect on skin barrier repair and deep wrinkle intervention is weak.

[0003] Particularly noteworthy is that when traditional plant extracts (such as Polygalaceae extract) are not biotransformed, the molecular weight and polarity of their active ingredients (such as polyphenols and flavonoids) limit their transdermal absorption efficiency, resulting in low bioavailability. In recent years, research has shown that microbial fermentation can significantly enhance the efficacy of plant extracts, can directionally degrade macromolecules, produce small molecule active metabolites (such as short-chain fatty acids and enzymatic hydrolysis products), and increase the release amount of antioxidants (such as hydroxytyrosol).

[0004] However, the prior art has not systematically synergistically combined plant fermented extracts with other anti-wrinkle ingredients to achieve a multi-target and multi-path anti-wrinkle mechanism. Summary of the Invention

[0005] Aiming at the problems of the limited efficacy of a single ingredient and the single action mechanism, the purpose of the present invention is to provide an anti-wrinkle and firming composition that exerts its effects through multi-path synergy. By combining the antioxidant damage ability of Rhodiola rosea, the extracellular matrix strengthening effect of propyltetrahydropyranyl triol, the multi-effect repair ability (including barrier repair and anti-inflammatory) of Polygalaceae fermentation product extract, and the metabolism promoting effect of adenosine, a three-dimensional anti-wrinkle network of "antioxidation - promoting collagen synthesis" is formed.

[0006] In order to achieve the above purpose, the present invention discloses the following technical solutions:

[0007] In the first aspect, the present invention provides an anti-wrinkle and firming composition, and by mass fraction, the composition contains the following components:

[0008] Rhodiola rosea extract 0.1 - 10 parts;

[0009] Propyltetrahydropyranyl triol 5 - 20 parts;

[0010] Rhodiola crenulata extract 0.02 - 2 parts;

[0011] Adenosine 0.5 - 2 parts.

[0012] Preferably, the composition contains the following components:

[0013] Rhodiola rosea extract 6 - 10 parts;

[0014] Hydroxypropyltetrahydropyrantriol 10 - 20 parts;

[0015] Rhodiola crenulata extract 1 - 2 parts;

[0016] Adenosine 1 - 2 parts.

[0017] More preferably, the Rhodiola crenulata extract described in the present invention is a product obtained by subjecting Rhodiola crenulata to a fermentation step, and the preparation method of the Rhodiola crenulata extract includes the following steps:

[0018] Step 1. Take a certain amount of dried whole plant of Rhodiola crenulata and place it in a universal pulverizer for pulverization and sieving to obtain Rhodiola crenulata powder;

[0019] Step 2. Mix the activated liquid of Lactobacillus acidophilus and the activated liquid of Bacillus licheniformis according to a volume ratio of 2:1 to obtain a compound bacterial liquid;

[0020] Step 3. Preparation of the fermentation medium: Rhodiola crenulata powder 200 g / L, soybean meal powder 40 g / L, glucose 20 g / L, ammonium sulfate 5 g / L, MgSO4·7H2O 0.5 g / L, MnSO4·H2O 0.05 g / L, Tween 80 0.1 v / v%, the balance being water. The prepared fermentation medium is sterilized at 121°C under high-pressure steam for 20 min;

[0021] Step 4. Inoculate the compound bacterial liquid into the fermentation medium at an inoculation amount of 5 v / v%. First, ferment and culture at 37°C, 130 r / min, and an aeration rate of 3 vvm for 20 h, then reduce the temperature and aeration rate to 30°C and 1.5 vvm, and maintain the rotation speed at 130 r / min for fermentation and culture for 28 h to obtain a fermentation product;

[0022] Step 5. Centrifuge the fermentation product to separate the supernatant, filter and sterilize the supernatant successively with 0.45 μm and 0.22 μm filter membranes, concentrate the filtered supernatant under reduced pressure, and then freeze-dry it to a water content of ≤5 wt% to obtain Rhodiola crenulata extract.

[0023] Even more preferably, the preparation method of the activated liquid of Lactobacillus acidophilus in Step 2 includes the following steps:

[0024] Inoculate the cryopreserved strain into MRS liquid medium, and culture it in a shaking flask at 37 °C and 150 r / min for 24 h. The obtained bacterial liquid is inoculated into a new MRS liquid medium at an inoculation amount of 2 v / v%, and cultured in a shaking flask at 37 °C and 150 r / min for 18 h to obtain an activated bacterial liquid of Lactobacillus acidophilus.

[0025] More preferably, the preparation method of the activated bacterial liquid of Bacillus licheniformis in step 2 includes the following steps:

[0026] Inoculate by streaking on LB solid medium, culture at 30 °C for 24 h, pick a well-grown single colony, inoculate it into LB liquid medium containing 0.1% glucose, and culture in a shaking flask at 35 °C and 180 r / min for 48 h to obtain an activated bacterial liquid of Bacillus licheniformis.

[0027] In a second aspect, the present invention provides the use of the anti-wrinkle and firming composition described in the first aspect in the preparation of a skin care product having firming and anti-wrinkle effects.

[0028] In a third aspect, the present invention provides an anti-wrinkle and firming lotion, and the lotion contains the anti-wrinkle and firming composition described in the first aspect;

[0029] The addition amount of the composition in the lotion is 1-5 wt%.

[0030] Preferably, the lotion further contains a humectant, an emulsifier, squalane, an antioxidant, a pH regulator, and a solvent.

[0031] In a fourth aspect, the present invention provides a preparation method of the anti-wrinkle and firming lotion described in the third aspect, and the preparation method includes the following steps:

[0032] Step 2-1. Take a part of the solvent and mix it evenly with the humectant, emulsifier, squalane, antioxidant, and pH regulator by stirring at 60-80 °C to obtain a mixture;

[0033] Step 2-2. After cooling the mixture to 45 °C, add the anti-wrinkle and firming composition and the remaining solvent thereto, and stir evenly to obtain the anti-wrinkle and firming lotion.

[0034] In the present invention:

[0035] The rhodiola extract contains rich salidroside, flavonoid compounds (such as rutin), polysaccharides, phenolic acids (such as gallic acid), and various amino acids and trace elements. Its efficacy on the skin is as follows: reducing melanin production and brightening the skin tone by strong antioxidant (scavenging free radicals) and inhibiting the activity of tyrosinase; anti-inflammatory and soothing sensitive skin, and enhancing the skin barrier function; the polysaccharide component provides deep moisturization, promotes collagen synthesis, delays photoaging and wrinkle formation, and overall improves skin elasticity and glossiness.

[0036] By stimulating the synthesis of glycosaminoglycans and collagen, hydroxypropyltetrahydropyrantriol can enhance skin firmness and elasticity, and reduce fine lines; it promotes the synthesis of hyaluronic acid in the skin, can deeply moisturize and improve dryness, and at the same time enhances the barrier function by strengthening the epidermal-dermal junction layer, gently repairs photoaging and oxidative damage, and is especially suitable for the anti-aging needs of sensitive skin. It is a multi-functional active ingredient with anti-wrinkle, moisturizing and barrier repair effects.

[0037] After fermenting saxifraga aizoon l. with bacillus licheniformis and lactobacillus acidophilus, saxifraga aizoon l. extract is obtained. Its original components (such as polyphenols, tannins, polysaccharides) are decomposed into small molecule phenolic acids (such as gallic acid), free flavonoid aglycones (such as quercetin), and microbial metabolites (such as antibacterial peptides, lactic acid, exopolysaccharides, etc.) are generated, significantly enhancing antioxidant, antibacterial, moisturizing and anti-inflammatory activities. At the same time, it was found during the experiment of the present invention that the composite fermentation of bacillus licheniformis and lactobacillus acidophilus enhanced the performance of saxifraga aizoon l. extract in promoting skin collagen synthesis.

[0038] In skin care, adenosine regulates signal pathways by activating cell surface receptors (such as A2A receptors). Its main effects include: promoting the synthesis of collagen and elastic fibers, reducing fine lines and enhancing skin firmness; inhibiting the release of inflammatory factors (such as TNF-α, IL-6), soothing sensitive flushing and improving the barrier function; enhancing the lipid metabolism of the stratum corneum and strengthening the ability to lock in moisture; at the same time, as an intermediate of energy metabolism, it accelerates cell repair and renewal, and helps to improve photoaging and oxidative damage. It is a multi-functional active ingredient with anti-aging, anti-inflammatory and repair effects.

[0039] Advantages of the present invention:

[0040] 1. When the rhodiola rosea extract, hydroxypropyltetrahydropyrantriol, saxifraga aizoon l. extract, and adenosine in the anti-wrinkle and firming composition provided by the present invention are used in combination, they exhibit excellent antioxidant and collagen synthesis promoting effects;

[0041] 2. The saxifraga aizoon l. extract obtained by the composite fermentation of lactobacillus acidophilus and bacillus licheniformis has a certain enhancing effect on antioxidant and collagen synthesis;

[0042] 3. The anti-wrinkle and firming lotion provided by the present invention can effectively reduce wrinkles and improve skin elasticity, showing excellent performance. Specific embodiments

[0043] To better illustrate the purpose, technical solution and advantages of the present invention, the present invention will be further described below in conjunction with specific embodiments. Those skilled in the art should understand that the specific embodiments described here are only used to explain the present invention and are not used to limit the present invention.

[0044] Unless otherwise specified, the test methods used in the examples and comparative examples are conventional methods; the materials, reagents, etc. used can be obtained from commercial sources unless otherwise specified; the percentages mentioned in the examples and comparative examples are mass percentages unless otherwise specified.

[0045] In the present invention:

[0046] Rhodiola rosea extract: Extract of Rhodiola rosea, purchased from Beijing Orient Miao Sen Biotechnology Co., Ltd.;

[0047] Hydroxypropyltetrahydropyrantriol: HYDROXYPROPYL TETRAHYDROPYRANTRIOL, purchased from Tianjin Taipu Pharmaceutical Co., Ltd.;

[0048] Cistus monspeliensis L.: Scientific name Cistus monspeliensis L., purchased from BASF (China) Co., Ltd.;

[0049] Adenosine: ADENOSINE, purchased from Guangdong Dingchun Biopharmaceutical Technology Co., Ltd.;

[0050] Lactobacillus acidophilus: Purchased from Guangdong Provincial Microbial Culture Collection Center, with the preservation number GDMCC NO.1.1807;

[0051] Bacillus licheniformis: Purchased from China General Microbiological Culture Collection Center, with the preservation number CGMCCNO.1.807;

[0052] MRS liquid medium: Trypticase peptone 10.0 g / L, beef extract powder 10.0 g / L, yeast extract powder 4.0 g / L, ammonium citrate 2.0 g / L, sodium acetate 5.0 g / L, MgSO4·7H2O 0.2 g / L, MnSO4·4H2O

[0053] 0.05 g / L, dipotassium hydrogen phosphate 2.0 g / L, glucose 20.0 g / L, Tween-80 1.0 g / L;

[0054] LB solid medium: Tryptone 10 g / L, yeast extract powder 5 g / L, sodium chloride 10 g / L, agar 15 g / L;

[0055] LB liquid medium: Glucose 10 g / L, tryptone 10 g / L, yeast extract powder 5 g / L, sodium chloride 10 g / L.

[0056] Preparation of Cistus monspeliensis L. extract

[0057] Step 1-1. Take a certain amount of the dried whole plant of Cistus monspeliensis L. and place it in a universal pulverizer for pulverization, pass through a 40-mesh sieve to obtain Cistus monspeliensis L. powder, and irradiate and sterilize it for later use;

[0058] Step 1-2. Activation of Lactobacillus acidophilus: Take the cryopreserved strain and inoculate it into MRS liquid medium. Shake flask culture at 37°C and 150 r / min for 24 h. Inoculate the obtained bacterial liquid into a new MRS liquid medium at an inoculation amount of 2 v / v%, and shake flask culture at 37°C and 150 r / min for 18 h to obtain the activated bacterial liquid of Lactobacillus acidophilus;

[0059] Step 1-3. Activation of Bacillus licheniformis: Streak inoculate on LB solid medium and culture at 30°C for 24 h. Pick a well-grown single colony and inoculate it into LB liquid medium containing 0.1% glucose. Shake flask culture at 35°C and 180 r / min for 48 h to obtain the activated bacterial liquid of Bacillus licheniformis;

[0060] Step 1-4. Mix the activated bacterial liquid of Lactobacillus acidophilus obtained in Step 1-2 and the activated bacterial liquid of Bacillus licheniformis obtained in Step 1-3 according to a volume ratio of 2:1 to obtain a compound bacterial liquid. Among them, the viable count of the activated bacterial liquid of Lactobacillus acidophilus is 1×10 8 CFU / mL, and the viable count of the activated bacterial liquid of Bacillus licheniformis is 1×10 8 CFU / mL;

[0061] Step 1-5. Preparation of fermentation medium: 200 g / L of Polygonum bistorta powder, 40 g / L of soybean meal powder, 20 g / L of glucose, 5 g / L of ammonium sulfate, 0.5 g / L of MgSO4·7H2O, 0.05 g / L of MnSO4·H2O, 0.1 v / v% of Tween 80, and the balance is water. The prepared fermentation medium is sterilized by high-pressure steam at 121°C for 20 min;

[0062] Step 1-6. Inoculate the compound bacterial liquid obtained in Step 1-4 into the fermentation medium at an inoculation amount of 5 v / v%. First, ferment and culture at 37°C, 130 r / min, and an aeration rate of 3 vvm for 20 h, then lower the temperature to 30°C, reduce the aeration rate to 1.5 vvm, and maintain the rotation speed at 130 r / min for fermentation and culture for 28 h to obtain a fermentation product;

[0063] Step 1-7. Centrifuge the fermentation product to separate the supernatant. Filter and sterilize the supernatant successively with 0.45 μm and 0.22 μm filter membranes. Concentrate the filtered supernatant under reduced pressure, and then freeze-dry it until the water content ≤ 5 wt% to obtain Polygonum bistorta extract. Recorded as Polygonum bistorta extract ①.

[0064] In order to verify the difference between the Polygonum bistorta extract prepared by the above preparation method and the Polygonum bistorta extract obtained by other methods, the following operations are carried out:

[0065] Saxifraga umbellata extract ②: Replace the complex bacterial solution in steps 1-6 of the preparation method of the above Saxifraga umbellata extract ① with a single activated Lactobacillus acidophilus bacterial solution, that is, only ferment and extract Lactobacillus acidophilus, without fermenting the complex bacterial solution, and ensure that the number of viable bacteria inoculated during fermentation is the same. The remaining steps are also the same as those of the preparation method of Saxifraga umbellata extract ①.

[0066] Saxifraga umbellata extract ③: Replace the complex bacterial solution in steps 1-6 of the preparation method of the above Saxifraga umbellata extract ① with a single activated Bacillus licheniformis bacterial solution, that is, only ferment and extract Bacillus licheniformis, without fermenting the complex bacterial solution, and ensure that the number of viable bacteria inoculated during fermentation is the same. The remaining steps are also the same as those of the preparation method of Saxifraga umbellata extract ①.

[0067] Saxifraga umbellata extract ④: Mix the Saxifraga umbellata powder obtained in step 1-1 with deionized water at a material-liquid ratio of 1 g: 20 mL, soak and extract at 90 °C and 200 r / min for 6 h, and filter to obtain a filtrate and filter residue; the filtrate is successively filtered and sterilized using 0.45 μm and 0.22 μm filter membranes, and then concentrated under reduced pressure and freeze-dried until the water content ≤ 5 wt% to obtain a water extract for standby; the filter residue is mixed with an ethanol solution with a concentration of 70 v / v% at a material-liquid ratio of 1 g: 15 mL, refluxed and extracted 3 times at 70 °C and 150 r / min for 1 h each time. The extracted solution obtained by reflux is centrifuged at 10000 r / min for 10 min to take the supernatant, and the supernatant is concentrated under reduced pressure and then freeze-dried until the water content ≤ 5 wt% to obtain an ethanol extract. The water extract and the ethanol extract are combined to obtain Saxifraga umbellata extract ④.

[0068] Preparation of anti-wrinkle and firming composition

[0069] Accurately weigh according to the raw material mass ratio in Table 2, and place it in a high-speed mixing and stirring machine to mix evenly to obtain an anti-wrinkle and firming composition.

[0070] Table 1 Mass ratio of anti-wrinkle and firming composition

[0071]

[0072] Note: "-" in the table indicates no addition.

[0073] Composition performance test

[0074] 1 DPPH free radical scavenging rate test

[0075] Test principle: 1,1-Diphenyl-2-picrylhydrazyl (abbreviated as DPPH) is a stable long-lived free radical. Its ethanol solution is dark purple and has a strong absorption near 517 nm. When there is a free radical scavenger, the light absorption of the DPPH ethanol solution weakens due to the pairing of its single electron. The degree of fading of the DPPH ethanol solution is linearly related to the number of electrons it accepts. Thus, the ability of the test sample to scavenge free radicals, that is, the antioxidant activity, can be evaluated.

[0076] Test sample solution: Dilute compositions 1-11 with deionized water to obtain test sample solutions 1-11 with a concentration of 2 mg / mL.

[0077] DPPH ethanol solution: Take a certain amount of DPPH and prepare a 0.1 mmol / L DPPH ethanol solution with 95 v / v% ethanol aqueous solution.

[0078] Mix 2.0 mL of each test sample solution with 2.0 mL of 0.1 mmol / L DPPH ethanol solution, react in the dark at room temperature for 30 min, and measure the absorbance (OD) value at 517 nm and record it as A2. Mix 2.0 mL of each test sample solution with 2.0 mL of 95 v / v% ethanol solution, react in the dark at room temperature for 30 min, and measure the absorbance (OD) value at 517 nm and record it as A1. Mix 2.0 mL of 0.1 mmol / L DPPH ethanol solution with 2.0 mL of deionized water, stand still in the dark at room temperature for 30 min, and measure the absorbance (OD) value at 517 nm and record it as A0. Repeat each sample 3 times. Through the formula "R D / % = [1 - (A2 - A1) / A0] × 100%", calculate the DPPH free radical scavenging rate (R D ). The results are shown in Table 2.

[0079] 2ABTS + ABTS Free Radical Scavenging Rate Test

[0080] Test principle: ABTS + free radicals have a maximum absorption peak in the visible light region at 734 nm. When an antioxidant substance is present, the electrons of ABTS+ are captured and reduced to unactivated ABTS molecules, resulting in a decrease in absorbance, thereby reflecting the free radical scavenging ability of the antioxidant substance.

[0081] Mix 5 mL of ABTS (7 mmol·L -1 ) solution with 88 μL of potassium persulfate aqueous solution (140 mmol·L -1 ), react in the dark for 12 - 14 h to obtain ABTS free radicals (ABTS +) Reaction solution (to be prepared and used immediately). Take 1 mL of the reaction solution and dilute it with absolute ethanol to an OD value of 0.701 (at 734 nm) to obtain the ABTS working solution. Dilute compositions 1 - 11 to the test sample solutions with a concentration of 2 mg / mL using deionized water. Pipette 0.1 mL of each test sample solution or deionized water, mix well with 3.9 mL of the ABTS working solution, react in the dark at room temperature for 6 min, measure the OD value at 734 nm, and repeat each group of samples 3 times. Calculate the ABTS radical scavenging rate (S A ), and the results are shown in Table 2:

[0082] S A / % = [(T0 - T1) / T0] × 100%.

[0083] In the formula:

[0084] T0: OD value of 0.1 mL of deionized water + 3.9 mL of ABTS working solution;

[0085] T1: OD value of 0.1 mL of each test sample solution + 3.9 mL of ABTS working solution.

[0086] Table 2 Test results of antioxidant performance

[0087] Group <![CDATA[R D / %]]> <![CDATA[S A / %]]> Composition 1 76.34 84.15 Composition 2 88.17 91.03 Composition 3 88.93 92.18 Composition 4 87.56 90.57 Composition 5 57.49 62.82 Composition 6 63.77 69.58 Composition 7 51.23 58.39 Composition 8 60.08 66.23 Composition 9 73.96 78.42 Composition 10 71.15 75.89 Composition 11 69.43 72.35

[0088] 3 Promotion of type I collagen synthesis test

[0089] Test principle: Using fibroblasts as the research object, first, based on fibroblasts, conduct cytotoxicity detection (MTT) to determine the safe dosing concentration of the sample on fibroblasts; on this basis, using fibroblasts as the research object, establish a cell photoaging damage model through UVA irradiation, and detect the change in the content of type I collagen (Collagen I) after the action of the sample to evaluate the firming and anti-wrinkle efficacy of the test sample. The test refers to Xie Yuanfang's "Firming and Anti-wrinkle Efficacy of Black Truffle Extract" and T / SHRH 031 - 2020 "Test Methods for Firming and Anti-wrinkle Efficacy of Cosmetics - Determination of Type I Collagen Content in Vitro Fibroblasts".

[0090] 3.1 Cytotoxicity detection

[0091] Before the test, conduct cytotoxicity detection (MTT method) on mouse embryonic fibroblasts (3T3, purchased from Haixing Biology), and select a cell viability ≥ 90% as the safe range of the test substance. The test results determined that when the concentration of compositions 1 - 11 was ≤ 3 wt%, the cell viability was ≥ 90%, and the following tests were carried out at a working concentration of 1 wt%.

[0092] 3.2 Influence on type I collagen synthesis

[0093] (1) Inoculation: Inoculate cells into a 96-well plate at an inoculation density of 2.2×10 4 cells / well, and place it in an incubator to incubate overnight (37°C, 5 v / v% CO2).

[0094] (2) UVA irradiation: After washing the cells with PBS, according to the experimental grouping in Table 3, perform UVA irradiation of 30 J / cm 2 on the groups with UVA irradiation.

[0095] (2) Solution preparation and sample addition: Dilute Compositions 1-11 to a concentration of 1 wt% with DMEM cell culture medium respectively to obtain the test sample solutions, and use TGF-β1 (100 ng / mL) as the positive control for the experiment. Add 100 μL of sample to each well, and set 3 replicate wells for each group. After drug administration, place the 96-well plate in an incubator at 37°C in the dark for 24 h.

[0096] Table 3 Experimental grouping

[0097]

[0098] (4) Collection of cell supernatant: Take out the 96-well plate 24 h after drug administration, collect the corresponding cell supernatant of each sample group into a centrifuge tube, centrifuge at 1000 rpm for 10 min, collect the supernatant and place it in a 1.5 mL centrifuge tube, and store it at -20°C for later use.

[0099] (5) Determination of type I collagen concentration in cell supernatant by ELISA kit: Take out the kit and the test samples 30 min before the test, place them at room temperature and then use them. The test operation is carried out strictly according to the kit instructions, and the average value of each group of test results is taken.

[0100] The growth rate of type I collagen is calculated according to the following formula:

[0101] Growth rate of type I collagen (%) = (A 待测样品 / 阳性 - A 模型对照 ) / A 模型对照 × 100%

[0102] In the formula:

[0103] A 待测样品 / 阳性 : The average value of type I collagen content in the test sample / positive group;

[0104] A 模型对照 : The average value of type I collagen content in the model control.

[0105] The results are shown in Table 4.

[0106] Table 4 Detection results of type I collagen content

[0107]

[0108] 4 Result Analysis

[0109] According to the data in Table 2, Compositions 1-4 showed certain DPPH radical and ABTS + radical scavenging properties. Among them, the radical scavenging rates of Compositions 2-4 were better than that of Composition 1, and Composition 3 showed the best antioxidant effect; Rhodiola rosea extract, propyltetrahydropyranyl triol, Polygonum bistorta extract, and adenosine were missing in Compositions 5-8 respectively. The results showed that the radical scavenging rates of Compositions 5-8 decreased significantly compared with Composition 3, indicating that these four components, namely Rhodiola rosea extract, propyltetrahydropyranyl triol, Polygonum bistorta extract, and adenosine, play an essential role in antioxidant activity, and these four components have a synergistic effect of promoting each other. The Polygonum bistorta extract in Compositions 9-10 was replaced by single-strain fermentation or ordinary solvent extraction respectively. The results showed that the antioxidant performance also decreased compared with Composition 3, indicating that the antioxidant performance of the Polygonum bistorta extract after compound fermentation with Lactobacillus acidophilus and Bacillus licheniformis was significantly improved.

[0110] According to the data in Table 4, the positive control group showed a certain effect of promoting the synthesis of type I collagen, indicating that the test method was effective; Compositions 1-4 showed certain properties of promoting the synthesis of type I collagen. Among them, Compositions 2-4 showed better firming and anti-wrinkle properties; Rhodiola rosea extract, propyltetrahydropyranyl triol, Polygonum bistorta extract, and adenosine were missing in Compositions 5-8 respectively. The results showed that the growth rate of type I collagen in Compositions 5-8 decreased significantly compared with Composition 3, indicating that these four components, namely Rhodiola rosea extract, propyltetrahydropyranyl triol, Polygonum bistorta extract, and adenosine, play an essential role in promoting collagen synthesis, and these four components have a synergistic effect of promoting each other. The Polygonum bistorta extract in Compositions 9-10 was replaced by single-strain fermentation or ordinary solvent extraction respectively. The results showed that the collagen synthesis performance also decreased compared with Composition 3, indicating that the anti-wrinkle and firming performance of the Polygonum bistorta extract after compound fermentation with Lactobacillus acidophilus and Bacillus licheniformis was significantly improved.

[0111] Preparation of Anti-Wrinkle and Firming Lotion

[0112] Step 2-1. Stir and dissolve Component A completely at 85 °C and 200 r / min, and keep it warm for later use;

[0113] Step 2-2. Stir and dissolve Component B completely at 80 °C and 200 r / min, and keep it warm for later use;

[0114] Step 2-3. Add the mixed Component A and Component B to the emulsifying pot, and homogenize and disperse at 80 °C and 10000 r / min for 20 min to obtain the emulsified component AB;

[0115] Step 2-4. Cool the emulsifying components A and B to 65°C, add the pre-mixed component C and the pre-dissolved component D, and stir for 30 min at a rotation speed of 200 r / min to obtain component ABCD.

[0116] Step 2-5. Cool the component ABCD obtained in Step 2-4 to 45°C, add component E thereto, stir and mix at a rotation speed of 200 r / min for 30 min, cool to 38°C, pass through a 200-mesh sieve, take the semi-finished product for inspection, and discharge after passing the inspection to obtain the anti-wrinkle and firming emulsion.

[0117] Table 5 Raw materials of the anti-wrinkle and firming emulsion

[0118]

[0119]

[0120] Note: The unit in the table is mass percentage.

[0121] Emulsion performance test

[0122] 1 Patch test

[0123] Use the anti-wrinkle and firming emulsion test substances prepared in Examples 1-3 and the matrix group to conduct a human skin patch test according to the test method in the "Cosmetics Safety and Technology Specifications" 2015 edition.

[0124] Observe the skin reactions at 30 min (after the indentation disappears), 24 h, and 48 h after removing the test substance patch tester respectively according to the criteria in Table 6, and record the observation results. The results are shown in Table 7.

[0125] Table 6 Grading criteria for skin reactions in the skin closed patch test

[0126]

[0127] Table 7 Results of human safety test

[0128] Number 30min 24h 48h Example 1 Grade 0, 30 people Grade 0, 30 people Grade 0, 30 people Example 2 Grade 0, 30 people Grade 0, 30 people Grade 0, 30 people Example 3 Grade 0, 30 people Grade 0, 30 people Grade 0, 30 people Example 4 Grade 0, 30 people Grade 0, 30 people Grade 0, 30 people Example 5 Grade 0, 30 people Grade 0, 30 people Grade 0, 30 people Example 6 Grade 0, 30 people Grade 0, 30 people Grade 0, 30 people

[0129] 2 Evaluation of anti-wrinkle and firming effect

[0130] Test substances: Examples 1-3 and the matrix group

[0131] Test subjects: A total of 80 people, 20 people in each group, numbered 1-20 respectively. The test subjects use the test substance on their faces 2 times a day, 1 mL each time, evenly apply it on the face, and gently massage until completely absorbed, and use it continuously for 4 weeks.

[0132] Subject selection: Healthy women aged 25 to 55 years old; Skin photobiological types are type II to type IV skin; They can cooperate well with the experimenters, maintain a regular life during the research period and avoid sunlight and ultraviolet radiation; There are problems such as fine wrinkles, poor elasticity and dullness on the facial skin.

[0133] Evaluation method: Use a Visia skin detector to detect the improvement of skin wrinkles and elasticity in the subjects before and 28 days after using the test substance. The experimental results are shown in Table 8.

[0134] Table 8 Improvement effects of wrinkles and firmness

[0135] Group Wrinkle reduction / % Elasticity / % Example 1 77.19 74.35 Example 2 74.54 71.92 Example 3 65.73 60.78 Matrix group 8.62 10.38

[0136] Result analysis:

[0137] The anti-wrinkle and firming lotion provided by the present invention can effectively reduce wrinkles and improve the elasticity of the skin, showing excellent performance. The firming and anti-wrinkle performance of the lotion is positively correlated with the concentration of composition 3 and has a certain concentration dependence, indicating that the composition plays a key role in the overall lotion formulation.

[0138] Finally, it should be noted that the above are only the preferred embodiments of the present invention and are not used to limit the present invention. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments, or perform equivalent replacements on some of the technical features. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. An anti-wrinkle and firming composition, characterized in that, By mass parts, the composition contains the following components:

2. The anti-wrinkle and firming composition according to claim 1, characterized in that, The composition contains the following components:

3. The anti-wrinkle and firming composition according to claim 1 or 2, characterized in that The preparation method of the extract of *Rosa cymosa Tratt* contains the following steps: Step 1. Take a certain amount of the dried whole plant of *Rosa cymosa Tratt* and place it in a universal pulverizer to crush and sieve, obtaining *Rosa cymosa Tratt* powder; Step 2. Compound and mix the activated bacterial liquid of *Lactobacillus acidophilus* and the activated bacterial liquid of *Bacillus licheniformis* according to a volume ratio of 2:1 to obtain a compound bacterial liquid; Step 3. Preparation of the fermentation medium: 200 g / L of *Rosa cymosa Tratt* powder, 40 g / L of soybean meal powder, 20 g / L of glucose, 5 g / L of ammonium sulfate, 0.5 g / L of MgSO₄·7H₂O, 0.05 g / L of MnSO₄·H₂O, 0.1 v / v% of Tween 80, and the balance of water. The prepared fermentation medium is sterilized at 121 °C under high-pressure steam for 20 min; Step 4. Inoculate the compound bacterial liquid into the fermentation medium at an inoculation amount of 5 v / v%. First, ferment and culture at 37 °C, 130 r / min, and an aeration rate of 3 vvm for 20 h, then reduce the temperature and aeration rate to 30 °C and 1.5 vvm, and maintain the rotation speed at 130 r / min for fermentation and culture for 28 h to obtain a fermentation product; Step 5. Centrifuge the fermentation product to separate and take the supernatant. The supernatant is successively filtered and sterilized using 0.45 μm and 0.22 μm filter membranes. The filtered supernatant is concentrated under reduced pressure and then freeze-dried until the water content ≤ 5 wt% to obtain the extract of *Rosa cymosa Tratt*.

4. The anti-wrinkle and firming composition according to claim 3, wherein The preparation method of the activated bacterial liquid of *Lactobacillus acidophilus* in Step 2 includes the following steps: Inoculate the cryopreserved strain into MRS liquid medium and shake-flask culture at 37 °C and 150 r / min for 24 h. The obtained bacterial liquid is inoculated into a new MRS liquid medium at an inoculation amount of 2 v / v% and shake-flask cultured at 37 °C and 150 r / min for 18 h to obtain the activated bacterial liquid of *Lactobacillus acidophilus*.

5. The anti-wrinkle and firming composition according to claim 3, characterized in that, The preparation method of the activated bacterial liquid of *Bacillus licheniformis* in Step 2 includes the following steps: Inoculate by streaking on LB solid medium and culture at 30 °C for 24 h. Pick well-grown single colonies and inoculate them into LB liquid medium containing 0.1% glucose, and shake-flask culture at 35 °C and 180 r / min for 48 h to obtain the activated bacterial liquid of *Bacillus licheniformis*.

6. Use of the anti-wrinkle and firming composition according to any one of claims 1-5 in the preparation of a skin care product with firming and anti-wrinkle effects.

7. An anti-wrinkle and firming lotion, characterized in that, The emulsion contains the anti-wrinkle and firming composition according to any one of claims 1-5; The addition amount of the composition in the emulsion is 1-5 wt%.

8. The anti-wrinkle and firming emulsion according to claim 7, wherein The emulsion also contains a humectant, an emulsifier, squalane, an antioxidant, a pH regulator, and a solvent.

9. The preparation method of the anti-wrinkle and firming emulsion according to claim 8, characterized in that, The preparation method includes the following steps: Step 2-1. Take a part of the solvent and mix it with a humectant, an emulsifier, squalane, an antioxidant, and a pH regulator evenly by stirring at 60-80 °C to obtain a mixture; Step 2-2. After cooling the mixture to 45 °C, add the anti-wrinkle and firming composition and the remaining solvent thereto, and stir evenly to obtain the anti-wrinkle and firming emulsion.

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