Anti-wrinkle firming composition and use thereof

Abstract

The application discloses an anti-wrinkle firming composition and application thereof, and belongs to the technical field of skin care products. The composition comprises the following components in a mass ratio: 0.1-10 parts of rhodiola extract, 5-20 parts of hydroxypropyl tetrahydropyran triol, 0.02-2 parts of polygala tenuifolia extract and 0.5-2 parts of adenosine, wherein the polygala tenuifolia extract is obtained by compound fermentation of lactobacillus acidophilus and bacillus licheniformis. The rhodiola extract, hydroxypropyl tetrahydropyran triol, polygala tenuifolia extract and adenosine provided by the application have excellent antioxidant and collagen synthesis promoting effects when used in combination. The anti-wrinkle firming emulsion provided by the application can effectively reduce wrinkles and improve the elasticity of skin, has excellent performance, overcomes the problems of single component efficacy limitation and single action mechanism, and realizes efficient anti-wrinkle firming effect under the synergistic action of multiple pathways.

Description

Technical Field

[0001] This invention belongs to the field of skin care technology, specifically relating to an anti-wrinkle and firming composition and its application. Background Technology

[0002] During skin aging, wrinkle formation is closely related to multiple factors, including decreased skin barrier function, collagen loss, extracellular matrix degradation, and oxidative stress damage. Existing anti-wrinkle and firming products mostly rely on single active ingredients, with limited pathways of action, making it difficult to comprehensively address the complex mechanisms of skin aging. For example, while Rhodiola rosea extract possesses antioxidant and anti-inflammatory activities, its effect on promoting collagen synthesis is limited when used alone; hydroxypropyl tetrahydropyranotriol, although it can activate fibroblasts to stimulate glycosaminoglycans (GAGs) and collagen synthesis, suffers from insufficient permeability and long-lasting effects; adenosine can improve skin microcirculation by promoting cell metabolism, but its effects on skin barrier repair and deep wrinkle intervention are weak.

[0003] Of particular note is that, without biotransformation, the molecular weight and polarity of the active ingredients (such as polyphenols and flavonoids) in traditional plant extracts (e.g., polycarboxylic acid rose extract) limit their transdermal absorption efficiency, resulting in low bioavailability. Recent studies have shown that microbial fermentation can significantly enhance the efficacy of plant extracts, selectively degrading macromolecules, producing small-molecule active metabolites (such as short-chain fatty acids and enzymatic hydrolysates), and increasing the release of antioxidants (such as hydroxytyrosol).

[0004] However, existing technologies do not systematically and synergistically combine plant fermentation extracts with other anti-wrinkle ingredients to achieve a multi-target, multi-pathway anti-wrinkle mechanism. Summary of the Invention

[0005] To address the limitations of single-ingredient efficacy and singular mechanisms of action, this invention aims to provide an anti-wrinkle and firming composition based on the synergistic effects of multiple pathways. By combining the antioxidant damage-preventing capabilities of Rhodiola rosea, the extracellular matrix-strengthening effects of hydroxypropyl tetrahydropyranotriol, the multi-functional repairing power of polychaete rock rose ferment extract (including barrier repair and anti-inflammation), and the metabolic-promoting effects of adenosine, a three-dimensional anti-wrinkle network of "antioxidant-promoted collagen synthesis" is formed.

[0006] To achieve the above objectives, the present invention discloses the following technical solutions:

[0007] In a first aspect, the present invention provides an anti-wrinkle and firming composition, wherein the composition comprises, by parts by weight:

[0008] Rhodiola rosea extract 0.1-10 parts;

[0009] Hydroxypropyltetrahydropyranotriol 5-20 parts;

[0010] 0.02-2 parts of polychaete rose extract;

[0011] Adenosine 0.5-2 parts.

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

[0013] 6-10 parts of Rhodiola rosea extract;

[0014] 10-20 parts of hydroxypropyl tetrahydropyranotriol;

[0015] 1-2 parts of polystyrene rose extract;

[0016] Adenosine 1-2 parts.

[0017] More preferably, the polychaete rose extract of the present invention is a product obtained by fermentation of polychaete rose, and the preparation method of the polychaete rose extract includes the following steps:

[0018] Step 1. Take a certain amount of dried whole plant of Rosa polypore and place it in a universal pulverizer to pulverize and sieve it to obtain Rosa polypore powder;

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

[0020] Step 3. Preparation of fermentation medium: 200 g / L polycarbonate powder, 40 g / L soybean meal powder, 20 g / L glucose, 5 g / L ammonium sulfate, 0.5 g / L MgSO4·7H2O, 0.05 g / L MnSO4·H2O, 0.1 v / v% Tween 80, and the balance water. The prepared fermentation medium was autoclaved at 121℃ for 20 min.

[0021] Step 4. Take the compound bacterial solution and inoculate it into the fermentation medium at an inoculation rate of 5 v / v%. First, ferment and culture it at 37℃, 130 r / min and 3 vvm aeration for 20 h. Then, reduce the temperature and aeration to 30℃ and 1.5 vvm and maintain the rotation speed of 130 r / min for 28 h to obtain the fermentation product.

[0022] Step 5. Centrifuge the fermentation product to obtain the supernatant. Filter the supernatant through 0.45μm and 0.22μm filter membranes for sterilization. Concentrate the filtered supernatant under reduced pressure and then freeze-dry it until the water content is ≤5wt% to obtain polycarbonate rose extract.

[0023] More preferably, the method for preparing the activated Lactobacillus acidophilus bacterial solution in step 2 includes the following steps:

[0024] The frozen bacterial culture was inoculated into MRS liquid medium and cultured in a shake flask at 37℃ and 150 r / min for 24 h. The resulting bacterial culture was then inoculated into fresh MRS liquid medium at an inoculation rate of 2 v / v% and cultured in a shake flask at 37℃ and 150 r / min for 18 h to obtain activated Lactobacillus acidophilus bacterial culture.

[0025] More preferably, the method for preparing the activated Bacillus licheniformis bacterial solution in step 2 includes the following steps:

[0026] The bacteria were streaked onto LB solid medium and incubated at 30°C for 24 hours. Well-grown single colonies were picked and inoculated into LB liquid medium containing 0.1% glucose. The cultures were then shaken at 35°C and 180 rpm for 48 hours to obtain activated Bacillus licheniformis culture.

[0027] Secondly, the present invention provides the application of the anti-wrinkle and firming composition described in the first aspect in the preparation of skin care products with firming and anti-wrinkle effects.

[0028] Thirdly, the present invention provides an anti-wrinkle and firming emulsion, the emulsion comprising the anti-wrinkle and firming composition described in the first aspect;

[0029] The composition is added to the emulsion in an amount of 1-5 wt%.

[0030] Preferably, the emulsion further comprises a humectant, an emulsifier, squalane, an antioxidant, a pH adjuster, and a solvent.

[0031] Fourthly, the present invention provides a method for preparing the anti-wrinkle and firming emulsion described in the third aspect, the method comprising the following steps:

[0032] Step 2-1. Take a portion of the solvent and mix it with the humectant, emulsifier, squalane, antioxidant, and pH adjuster at 60-80℃ until homogeneous 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 to it, stir evenly to obtain the anti-wrinkle and firming emulsion.

[0034] In this invention:

[0035] Rhodiola rosea extract is rich in rhodioloside, flavonoids (such as rutin), polysaccharides, phenolic acids (such as gallic acid), and various amino acids and trace elements. Its effects on the skin include: reducing melanin production and brightening skin tone through powerful antioxidant effects (scavenging free radicals) and inhibition of tyrosinase activity; anti-inflammatory and soothing effects on sensitive skin, enhancing the skin barrier function; and polysaccharide components providing deep hydration, promoting collagen synthesis, delaying photoaging and wrinkle formation, and improving overall skin elasticity and radiance.

[0036] Hydroxypropyl tetrahydropyranotriol enhances skin firmness and elasticity and reduces fine lines by stimulating the synthesis of glycosaminoglycans and collagen. It promotes the synthesis of hyaluronic acid in the skin, which can deeply moisturize and improve dryness. At the same time, it strengthens the barrier function by reinforcing the epidermal-dermal junction and gently repairs photoaging and oxidative damage. It is especially suitable for the anti-aging needs of sensitive skin and is a multi-effect active ingredient that combines anti-wrinkle, moisturizing and barrier repair.

[0037] Polymeric rosehip extract is obtained by fermentation of Bacillus licheniformis and Lactobacillus acidophilus. Its original components (such as polyphenols, tannins, and polysaccharides) are decomposed into small molecule phenolic acids (such as gallic acid) and free flavonoid aglycones (such as quercetin), and microbial metabolites (such as antimicrobial peptides, lactic acid, and extracellular polysaccharides) are generated, which significantly enhances its antioxidant, antibacterial, moisturizing, and anti-inflammatory activities. At the same time, it was found in the experiment of this invention that the combined fermentation of Bacillus licheniformis and Lactobacillus acidophilus enhanced the performance of polymeric rosehip extract in promoting skin collagen synthesis.

[0038] In skin care, adenosine regulates signaling pathways by activating cell surface receptors (such as A2A receptors). Its main functions include: promoting collagen and elastin synthesis, reducing fine lines and improving skin firmness; inhibiting the release of inflammatory factors (such as TNF-α and IL-6), soothing sensitivity and redness and improving barrier function; enhancing lipid metabolism in the stratum corneum and strengthening moisturizing and water-locking capabilities; and as an energy metabolism intermediate, accelerating cell repair and renewal, and helping to improve photoaging and oxidative damage. It is a multifunctional active ingredient with anti-aging, anti-inflammatory and repairing properties.

[0039] The beneficial effects of this invention are:

[0040] 1. The anti-wrinkle and firming composition provided by the present invention contains Rhodiola rosea extract, hydroxypropyl tetrahydropyrantriol, polycarboxylic acid extract, and adenosine, which exhibit excellent antioxidant and collagen synthesis-promoting effects when used in combination.

[0041] 2. The polychaete extract obtained by co-fermentation of Lactobacillus acidophilus and Bacillus licheniformis has a certain enhancing effect on anti-oxidation and collagen synthesis;

[0042] 3. The anti-wrinkle and firming lotion provided by this invention can effectively reduce wrinkles and improve skin elasticity, exhibiting excellent performance. Detailed Implementation

[0043] To better illustrate the objectives, technical solutions, and advantages of this invention, the invention will be further described below with reference to specific embodiments. Those skilled in the art should understand that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

[0044] Unless otherwise specified, the test methods used in the examples and comparative examples are conventional methods; the materials and reagents used are commercially available unless otherwise specified; and the percentages mentioned in the examples and comparative examples are mass percentages unless otherwise specified.

[0045] In this invention:

[0046] Rhodiola rosea extract: Rhodiola rosea extract, purchased from Beijing Dongfang Miaosen Biotechnology Co., Ltd.;

[0047] Hydroxypropyltetrahydropyrantriol (HYDROXYPROPYL TETRAHYDROPYRANTRIOL) was purchased from Tianjin Taipu Pharmaceutical Co., Ltd.

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

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

[0050] Lactobacillus acidophilus: purchased from Guangdong Provincial Microbial Culture Collection Center, accession number GDMCC NO.1.1807;

[0051] Bacillus licheniformis: purchased from China General Microbiological Culture Collection Center, accession number CGMCCNO.1.807;

[0052] MRS liquid culture medium: casein digest 10.0 g / L, beef extract 10.0 g / L, yeast extract 4.0 g / L, triammonium 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 5 g / L, sodium chloride 10 g / L, agar 15 g / L;

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

[0056] Preparation of polychaete rose extract

[0057] Step 1-1. Take a certain amount of dried whole plant of Rosa polypore and put it into a universal pulverizer for pulverization. Pass it through a 40-mesh sieve to obtain Rosa polypore powder. Sterilize it by irradiation and set it aside for later use.

[0058] Step 1-2. Activation of Lactobacillus acidophilus: Take the frozen bacterial culture and inoculate it into MRS liquid medium. Incubate in a shake flask at 37℃ and 150r / min for 24h. Inoculate the obtained bacterial solution into a new MRS liquid medium at an inoculation rate of 2v / v% and incubate in a shake flask at 37℃ and 150r / min for 18h to obtain activated Lactobacillus acidophilus bacterial solution.

[0059] Steps 1-3. Activation of Bacillus licheniformis: Streak inoculated onto LB solid medium and incubated at 30°C for 24 hours. Select well-grown single colonies and inoculate them into LB liquid medium containing 0.1% glucose. Incubate in shake flasks at 35°C and 180 r / min for 48 hours to obtain activated Bacillus licheniformis bacterial solution.

[0060] Steps 1-4. Mix the activated Lactobacillus acidophilus solution obtained in Step 1-2 and the activated Bacillus licheniformis solution obtained in Step 1-3 at a volume ratio of 2:1 to obtain a compound bacterial solution, wherein the viable count of the activated Lactobacillus acidophilus solution is 1×10⁻⁶. 8 The viable count of the activated Bacillus licheniformis solution was 1×10⁻⁶ CFU / mL. 8 CFU / mL;

[0061] Steps 1-5. Preparation of fermentation medium: 200 g / L polycarbonate powder, 40 g / L soybean meal powder, 20 g / L glucose, 5 g / L ammonium sulfate, 0.5 g / L MgSO4·7H2O, 0.05 g / L MnSO4·H2O, 0.1 v / v% Tween 80, and the balance water. The prepared fermentation medium is autoclaved at 121℃ for 20 min.

[0062] Steps 1-6. Take the compound bacterial solution obtained in Steps 1-4 and inoculate it into the fermentation medium at an inoculation rate of 5 v / v%. First, ferment and culture it at 37℃, 130 r / min and 3 vvm aeration for 20 h. Then, lower the temperature to 30℃, reduce the aeration to 1.5 vvm, and maintain the rotation speed of 130 r / min for 28 h to obtain the fermentation product.

[0063] Steps 1-7. Centrifuge the fermentation product to separate the supernatant. Filter the supernatant through 0.45μm and 0.22μm filter membranes for sterilization. Concentrate the filtered supernatant under reduced pressure and then freeze-dry until the water content is ≤5wt% to obtain polychaete rose extract. This is designated as polychaete rose extract①.

[0064] To verify the difference between the polychaete rose extract prepared by the above method and the polychaete rose extract obtained by other methods, the following operations were performed:

[0065] Polymeric Rosa ferruginosa extract ②: Replace the compound bacterial solution in steps 1-6 of the above preparation method of polymeric Rosa ferruginosa extract ① with a separate activated Lactobacillus acidophilus bacterial solution, that is, only Lactobacillus acidophilus is fermented and extracted, and the fermentation of the compound bacterial solution is not carried out. The number of live bacteria inoculated during fermentation is the same, and the remaining steps are consistent with the preparation method of polymeric Rosa ferruginosa extract ①.

[0066] Polymeric Rosa licheniformis Extract ③: Replace the compound bacterial solution in steps 1-6 of the above preparation method of polymeric Rosa licheniformis Extract ① with a separate activated bacterial solution of Bacillus licheniformis. That is, only Bacillus licheniformis is fermented and extracted, and the fermentation of the compound bacterial solution is not carried out. The number of live bacteria inoculated during fermentation is the same. The remaining steps are also consistent with the preparation method of polymeric Rosa licheniformis Extract ①.

[0067] Polymeric Rosa polypore extract ④: The polymeric Rosa polypore powder obtained in step 1-1 was mixed with deionized water at a ratio of 1g:20mL. The mixture was soaked and extracted at 90℃ and 200r / min for 6h. The filtrate and residue were obtained by filtration. The filtrate was filtered through 0.45μm and 0.22μm filter membranes for sterilization, concentrated under reduced pressure, and then freeze-dried to a water content ≤5wt% to obtain the water extract for later use. The residue was mixed with a 70v / v% ethanol solution at a ratio of 1g:15mL. The mixture was refluxed and extracted 3 times at 70℃ and 150r / min for 1h each time. The refluxed extract was centrifuged at 10000r / min for 10min to obtain the supernatant. The supernatant was concentrated under reduced pressure and then freeze-dried to a water content ≤5wt% to obtain the alcohol extract. The water extract and alcohol extract were combined to obtain polymeric Rosa polypore extract ④.

[0068] Preparation of anti-wrinkle and firming composition

[0069] The raw materials were accurately weighed according to the mass ratio in Table 2, and then mixed evenly in a high-speed mixer 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 additions.

[0073] Composition performance testing

[0074] 1DPPH free radical scavenging rate test

[0075] Experimental Principle: 1,1-Diphenyl-2-trinitrophenylhydrazine (DPPH) is a stable, long-lived free radical. Its ethanol solution is deep purple and exhibits strong absorption around 517 nm. In the presence of a free radical scavenger, the light absorption of the DPPH ethanol solution decreases due to the pairing of unpaired electrons with the DPPH. The degree of fading of the DPPH ethanol solution is linearly related to the number of electrons it accepts, thus allowing evaluation of the test sample's ability to scavenge free radicals, i.e., its antioxidant activity.

[0076] Test sample solution: Test sample solution 1-11 is prepared by diluting composition 1-11 to a concentration of 2 mg / mL with deionized water;

[0077] DPPH ethanol solution: Prepare a 0.1 mmol / L DPPH ethanol solution by mixing a certain amount of DPPH with a 95 v / v ethanol aqueous solution;

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

[0079] 2ABTS + Free radical scavenging rate test

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

[0081] Add 5 mL of ABTS (7 mmol·L⁻¹) -1 ) solution and 88 μL potassium persulfate aqueous solution (140 mmol·L -1 Mix and react in the dark for 12–14 hours to obtain ABTS free radicals (ABTS). +) Reaction solution (must be prepared fresh for use). Take 1 mL of the reaction solution and dilute with anhydrous ethanol to an OD value of 0.701 (at 734 nm) to obtain the ABTS working solution. Dilute compositions 1-11 with deionized water to a concentration of 2 mg / mL for the test sample solution. Pipette 0.1 mL of each test sample solution or deionized water and mix thoroughly with 3.9 mL of ABTS working solution. React at room temperature in the dark for 6 min, and measure the OD value at 734 nm. Repeat each group of samples 3 times. Calculate the ABTS free radical scavenging rate (S) using the following formula. A 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 deionized water + 3.9 mL 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 Results of Antioxidant Performance Test

[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. Test to promote type I collagen synthesis

[0089] Experimental Principle: Using fibroblasts as the research object, the first step is to conduct a cytotoxicity test (MTT) on fibroblasts to determine the safe dosage concentration of the sample on fibroblasts. Based on this, a cell photoaging damage model is established using fibroblasts through UVA irradiation, and the changes in type I collagen content after sample treatment are detected to evaluate the firming and anti-wrinkle efficacy of the test sample. The experiment references Xie Yuanfang's "Firming and Anti-wrinkle Efficacy of Black Truffle Extract" and T / SHRH 031-2020 "Test of Firming and Anti-wrinkle Efficacy of Cosmetics - In Vitro Method for Determination of Type I Collagen Content in Fibroblasts".

[0090] 3.1 Cytotoxicity Detection

[0091] Before testing, mouse embryonic fibroblasts (3T3, purchased from Haixing Biotechnology) were subjected to cytotoxicity testing (MTT method). Cell viability ≥90% was selected as the safe range of the test substance. The test results determined that when the concentration of composition 1-11 was ≤3wt%, the cell viability was ≥90%. The following tests were conducted at a working concentration of 1wt%.

[0092] 3.2 Effects on Type I Collagen Synthesis

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

[0094] (2) UVA irradiation: After washing cells with PBS, according to the experimental groups in Table 3, the groups receiving UVA irradiation were subjected to 30 J / cm² irradiation. 2 UVA irradiation.

[0095] (2) Solution preparation and sample addition: The composition was diluted 1-11 to a concentration of 1 wt% with DMEM cell culture medium to obtain the test sample solution. TGF-β1 (100 ng / mL) was used as a positive control for the experiment. 100 μL of sample was added to each well, and 3 replicates were set up for each group. After the drug administration was completed, the 96-well plate was placed in an incubator at 37℃ and incubated in the dark for 24 h.

[0096] Table 3 Experimental Groups

[0097]

[0098] (4) Collection of cell supernatant: 24 h after drug administration, remove the 96-well plate and collect the 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. Store at -20℃ for later use.

[0099] (5) ELISA kit to determine the concentration of type I collagen in cell supernatant: Take out the kit and the sample to be tested 30 minutes before the test, place them at room temperature before use, and strictly follow the kit instructions for the test operation. Take the average value of each test result.

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

[0101] Type I collagen growth rate (%) = (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 Mean content of type I collagen in the tested samples / positive groups;

[0104] A 模型对照 Mean type I collagen content in the model control group.

[0105] The results are shown in Table 4.

[0106] Table 4. Results of Type I Collagen Content Detection

[0107]

[0108] 4 Results Analysis

[0109] According to the data in Table 2, compositions 1-4 exhibit certain scavenging effects on DPPH free radicals and ABTS. + The free radical scavenging performance of compositions 2-4 was superior to that of composition 1, while composition 3 exhibited the best antioxidant effect. Compositions 5-8, lacking Rhodiola rosea extract, hydroxypropyl tetrahydropyranotriol, polychaete rock rose extract, and adenosine, showed a significant decrease in free radical scavenging rate compared to composition 3. This indicates that these four components—Rhodiola rosea extract, hydroxypropyl tetrahydropyranotriol, polychaete rock rose extract, and adenosine—are indispensable for antioxidant activity, exhibiting a synergistic effect. Replacing the polychaete rock rose extract in compositions 9-10 with either single-strain fermentation or conventional solvent extraction also resulted in a decrease in antioxidant performance compared to composition 3, indicating a significant improvement in antioxidant performance after co-fermentation with Lactobacillus acidophilus and Bacillus licheniformis.

[0110] According to the data in Table 4, the positive control group showed a certain effect in promoting type I collagen synthesis, indicating that the experimental method was effective. Compositions 1-4 showed certain properties in promoting type I collagen synthesis, among which compositions 2-4 showed better firming and anti-wrinkle properties. Compositions 5-8 lacked Rhodiola rosea extract, hydroxypropyl tetrahydropyranotriol, polychaete rock extract, and adenosine, respectively. The results showed that the growth rate of type I collagen in compositions 5-8 was significantly lower than that in composition 3, indicating that Rhodiola rosea extract, hydroxypropyl tetrahydropyranotriol, polychaete rock extract, and adenosine are indispensable in promoting collagen synthesis, and these four components have a synergistic effect of mutual promotion. When the polychaete rose extract in compositions 9-10 was replaced with either single-strain fermentation or conventional solvent extraction, the results showed that the collagen synthesis performance also decreased compared to composition 3. This indicates that the anti-wrinkle and firming properties of the polychaete rose extract after co-fermentation with Lactobacillus acidophilus and Bacillus licheniformis were significantly improved.

[0111] Preparation of anti-wrinkle and firming emulsion

[0112] Step 2-1. Dissolve component A completely by stirring at 85℃ and 200r / min, and keep warm for later use;

[0113] Step 2-2. Dissolve component B completely by stirring at 80℃ and 200r / min, and keep warm for later use;

[0114] Steps 2-3. Add the mixed components A and B to the emulsification tank and homogenize them at 80℃ and 10000r / min for 20min to obtain emulsified components AB;

[0115] Steps 2-4. Cool the emulsified component AB to 65℃, add the pre-mixed component C and the pre-dissolved component D, and stir at 200 r / min for 30 min to obtain component ABCD;

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

[0117] Table 5 Raw Materials for Anti-wrinkle Firming Emulsion

[0118]

[0119]

[0120] Note: The units in the table are percentages by mass.

[0121] Emulsion performance testing

[0122] 1. Spot test

[0123] The anti-wrinkle and firming emulsions prepared in Examples 1-3 and the matrix group were subjected to human skin patch tests according to the test methods of the 2015 edition of the "Cosmetic Safety Technical Specifications".

[0124] Skin reactions were observed according to the standards in Table 6 at 30 min (after the indentation disappeared), 24 h and 48 h after the removal of the test sample applicator, and the results were recorded. The results are shown in Table 7.

[0125] Table 6. Grading Criteria for Skin Reactions in Closed Patch Tests

[0126]

[0127] Table 7 Results of Human Safety Tests

[0128] serial number 30min 24h 48h Example 1 Level 0, 30 people Level 0, 30 people Level 0, 30 people Example 2 Level 0, 30 people Level 0, 30 people Level 0, 30 people Example 3 Level 0, 30 people Level 0, 30 people Level 0, 30 people Example 4 Level 0, 30 people Level 0, 30 people Level 0, 30 people Example 5 Level 0, 30 people Level 0, 30 people Level 0, 30 people Example 6 Level 0, 30 people Level 0, 30 people Level 0, 30 people

[0129] 2. Evaluation of anti-wrinkle and firming effects

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

[0131] Subjects: A total of 80 people, 20 people in each group, numbered 1-20. Subjects applied the test substance to their face twice a day, 1 mL each time, evenly applied to the face, and gently massaged until fully absorbed, for 4 consecutive weeks.

[0132] Subject selection: Healthy women aged 25 to 55; skin photobiological type II to IV; able to cooperate well with the experimenters, maintain a regular lifestyle during the study period and avoid sun exposure; facial skin with fine wrinkles, poor elasticity and dullness.

[0133] Evaluation method: The Visia skin analyzer was used to assess the improvement in skin wrinkles and elasticity in subjects before and 28 days after using the test product. The results are shown in Table 8.

[0134] Table 8. Effects on wrinkle and firmness improvement

[0135] Group Light wrinkles / % elasticity / % Example 1 77.19 74.35 Example 2 74.54 71.92 Example 3 65.73 60.78 matrix 8.62 10.38

[0136] Results analysis:

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

[0138] Finally, it should be noted that the above descriptions are merely preferred embodiments of the present invention and are not intended 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 make equivalent substitutions for some of the technical features. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. An anti-wrinkle and firming composition, characterized in that, The composition comprises the following components in parts by weight: Rhodiola rosea extract 0.1-10 parts; Hydroxypropyltetrahydropyranotriol 5-20 parts; 0.02-2 parts of polychaete rose extract; Adenosine 0.5-2 parts; The preparation method of the polychaete rose extract includes the following steps: Step 1. Take a certain amount of dried whole plant of Rosa polypore and place it in a universal pulverizer to pulverize and sieve it to obtain Rosa polypore powder; Step 2. Mix the activated bacterial solutions of Lactobacillus acidophilus and Bacillus licheniformis at a volume ratio of 2:1 to obtain a compound bacterial solution; Step 3. Preparation of fermentation medium: 200 g / L polycarbonate powder, 40 g / L soybean meal powder, 20 g / L glucose, 5 g / L ammonium sulfate, 0.5 g / L MgSO4·7H2O, 0.05 g / L MnSO4·H2O, 0.1 v / v% Tween 80, and the remainder water. The prepared fermentation medium was autoclaved at 121℃ for 20 min. Step 4. Take the compound bacterial solution and inoculate it into the fermentation medium at an inoculation rate of 5v / v%. First, ferment and culture it at 37℃, 130r / min and 3vvm aeration for 20h. Then, reduce the temperature and aeration to 30℃ and 1.5vvm and maintain the rotation speed of 130r / min for 28h to obtain the fermentation product. Step 5. Centrifuge the fermentation product to obtain the supernatant. Filter the supernatant through 0.45μm and 0.22μm filter membranes for sterilization. Concentrate the filtered supernatant under reduced pressure and freeze-dry it until the water content is ≤5wt% to obtain polycarbonate rose extract. The Lactobacillus acidophilus was purchased from Guangdong Provincial Microbial Culture Collection Center, with accession number GDMCC NO.1.1807; The Bacillus licheniformis was purchased from the China General Microbiological Culture Collection Center, with accession number CGMCCNO.1.

807.

2. The anti-wrinkle and firming composition according to claim 1, characterized in that, The composition comprises the following components in parts by weight: 6-10 parts of Rhodiola rosea extract; 10-20 parts of hydroxypropyl tetrahydropyranotriol; 1-2 parts of polystyrene rose extract; Adenosine 1-2 parts.

3. The anti-wrinkle and firming composition according to claim 1, characterized in that, The preparation method of the activated Lactobacillus acidophilus bacterial solution in step 2 includes the following steps: The frozen bacterial culture was inoculated into MRS liquid medium and cultured in a shake flask at 37℃ and 150 r / min for 24 h. The resulting bacterial culture was then inoculated into new MRS liquid medium at an inoculation rate of 2 v / v% and cultured in a shake flask at 37℃ and 150 r / min for 18 h to obtain activated Lactobacillus acidophilus bacterial culture.

4. The anti-wrinkle and firming composition according to claim 1, characterized in that, The preparation method of the activated Bacillus licheniformis bacterial solution in step 2 includes the following steps: The bacteria were streaked onto LB solid medium and incubated at 30°C for 24 hours. Well-grown single colonies were picked and inoculated into LB liquid medium containing 0.1% glucose. The cultures were then shaken at 35°C and 180 rpm for 48 hours to obtain activated Bacillus licheniformis culture.

5. The use of the anti-wrinkle and firming composition according to any one of claims 1-4 in the preparation of skin care products with firming and anti-wrinkle effects.

6. An anti-wrinkle and firming emulsion, characterized in that, The emulsion contains the anti-wrinkle and firming composition according to any one of claims 1-4; The composition is added to the emulsion in an amount of 1-5 wt%.

7. The anti-wrinkle and firming emulsion according to claim 6, characterized in that, The emulsion also contains humectants, emulsifiers, squalane, antioxidants, pH adjusters, and solvents.

8. The method for preparing the anti-wrinkle and firming emulsion according to claim 7, characterized in that, The preparation method includes the following steps: Step 2-1. Take a portion of the solvent and mix it with the humectant, emulsifier, squalane, antioxidant, and pH adjuster at 60-80℃ until homogeneous 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 to it, stir evenly to obtain the anti-wrinkle and firming emulsion.

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