Fermentation product for enhancing anti-aging effect of hydroxypropyl tetrahydropyrantriol and combined application of fermentation product

Abstract

The invention provides application of bifidobacterium / lactobacillus / soybean seed extract fermentation product filtrate in preparation of a product for promoting transdermal penetration of hydroxypropyl tetrahydropyrantriol. The invention further provides a composition, application of the composition and a product. According to the application, the bifidobacterium / lactobacillus / soybean seed extract fermentation product filtrate is used as the penetration enhancer, so that the percutaneous absorption of the water-soluble hydroxypropyl tetrahydropyrantriol can be promoted at a lower concentration, and the penetration enhancer has a better promotion effect than high-concentration hydroxyacetic acid and other tartaric acid and a typical chemical penetration enhancer NeoSolue-Aqulio and the like at a lower concentration. And moreover, the filtrate of the fermentation product of the bifidobacterium / lactobacillus / soybean seed extract is relatively mild, so that irritation and discomfort are avoided during use.

Description

Technical Field

[0001] The present invention relates to the technical field of daily chemical industry, and particularly relates to a fermentation product for enhancing the anti-aging effect of hydroxypropyltetrahydropyrantriol and its combined application. Background Art

[0002] Hydroxypropyltetrahydropyrantriol, also known as pro-Xylane, has been recognized by the scientific community and the cosmetics industry for its important role and potential in the anti-aging field, and is known as "one of the three major anti-aging ingredients". The lipophilic-hydrophilic balance coefficient LogP of hydroxypropyltetrahydropyrantriol is -2.07 (from chemicalbook), and it has strong water solubility. The stratum corneum of the epidermis is the main part of the skin for absorbing external substances, accounting for more than 90% of the total absorption capacity of the skin. Since the interstices of the stratum corneum are mainly lipid-based, the stratum corneum mainly absorbs lipophilic substances. Therefore, water-soluble active substances are difficult to penetrate the lipophilic stratum corneum barrier on the outermost layer of the skin. The core action site of hydroxypropyltetrahydropyrantriol is the basement membrane and the extracellular matrix of the dermis, and the corresponding efficacy lies in promoting the production of type IV collagen and type VII collagen in the basement membrane, and promoting the production of mucopolysaccharides such as glycosaminoglycans in the extracellular matrix of the dermis (Eur J Dermatol 2011; 21(2): 191-6). Therefore, hydroxypropyltetrahydropyrantriol needs to penetrate deep into the epidermis and even reach the dermis to play its role well. However, studies have shown that the bioavailability of hydroxypropyltetrahydropyrantriol is only 3-7% (Eur J Dermatol 2011; 21(2): 191-6). At the same time, existing studies have shown that even if the dosage of hydroxypropyltetrahydropyrantriol is increased, the improvement in bioavailability is not significant, and as the concentration of hydroxypropyltetrahydropyrantriol increases, its viscosity will also increase, which will damage the skin sensation during use. Therefore, technologies for promoting the penetration of hydroxypropyltetrahydropyrantriol or ingredients for assisting the penetration of hydroxypropyltetrahydropyrantriol are the key means to improve the utilization rate of hydroxypropyltetrahydropyrantriol.

[0003] Currently, common transdermal penetration enhancement technologies for cosmetics include physical penetration enhancement methods such as iontophoresis, electroporation, ultrasound, microneedles, thermal ablation, magnetic induction, photoacoustic waves, and electron beam radiation. By creating tiny channels on the skin through physical methods, the transdermal delivery of drugs can be improved. However, physical penetration usually requires professional equipment and technology and may cause discomfort or side effects to varying degrees. It is also possible to use chemical penetration enhancers such as azone and alcohols to improve the transdermal absorption efficiency of drugs by disrupting the lipid structure or reducing the thickness of the stratum corneum. In addition, vesicle delivery systems and microemulsion / nanoemulsion delivery systems such as liposomes, transfersomes, microemulsions, and nanoemulsions can be used to improve the transdermal absorption efficiency of drugs. For example, Chinese Patent CN116528820A discloses a microemulsion or nanoemulsion technology that can effectively improve the transdermal penetration of water-soluble active substances such as propyltetrahydropyranyl triol. However, this technology generally requires precise regulation of the proportions of various oils, surfactants, etc., and sometimes even requires the assistance of some high-energy equipment, and the preparation process is relatively complex. Summary of the Invention

[0004] In view of this, the present invention provides a composition and its application. The composition provided by the present application is relatively mild, has little irritation to the skin, and can efficiently promote the transdermal absorption of water-soluble active substances such as propyltetrahydropyranyl triol, significantly improving the utilization rate of propyltetrahydropyranyl triol.

[0005] The present application provides the use of the fermentation product filtrate of Bifidobacterium / Lactobacillus / soybean seed extract in the preparation of a product for promoting the transdermal penetration of propyltetrahydropyranyl triol.

[0006] The fermentation product filtrate of Bifidobacterium / Lactobacillus / soybean seed extract is a raw material developed by the applicant. The national cosmetic raw material preparation number is 20230032. The Chinese name for the record is the fermentation product filtrate of Bifidobacterium / Lactobacillus / soybean seed extract. The INCI name is Bifidobacterium / Lactobacillus / Soybean Seed Extract Ferment Filtrate, and the trade name is "Huanyuan". Based on the fermentation process, it enhances the bioavailability of metabolites, making the effect of the product greater than the sum of its parts. Finally, a "highly active composite metabolite group" of more than 500 is obtained, such as small molecule fruit acids like "lactic acid, citric acid", and also includes small molecule amino acids such as "L-alanine, ornithine, glycine, lysine, serine". In some specific implementation manners, the preparation method of the fermentation product filtrate of Bifidobacterium / Lactobacillus / soybean seed extract mainly includes: soaking black beans and water in a certain proportion, obtaining soybean seed extract after heating and water extraction, sterilizing to obtain a fermentation substrate, then transferring it to a fermentation tank, adding a strain composition (containing Bifidobacterium and Lactobacillus) for fermentation, and obtaining a fermentation filtrate after sterilization and filtration.

[0007] The applicant has found that the fermentation product filtrate of Bifidobacterium / Lactobacillus / soybean seed extract is not only milder, but can also assist the transdermal absorption of water-soluble propylhydroxytetrahydropyrantriol by accelerating the metabolism of the stratum corneum through small-molecule fruit acids and influencing the lipid and protein structures by small-molecule amino acids, and has a better promoting effect at lower concentrations than fruit acids such as glycolic acid at high concentrations and typical chemical penetration enhancers such as Neosolue™-Aqulio.

[0008] In some specific implementation manners, the concentration of the fermentation product filtrate of Bifidobacterium / Lactobacillus / soybean seed extract in the product is 0.01 wt% to 5 wt%, preferably 0.032 wt% to 3.2 wt%, and more preferably 0.032 wt%, 0.32 wt% or 3.2 wt%.

[0009] In some specific implementation manners, the concentration of propylhydroxytetrahydropyrantriol in the product is 5 wt% to 15 wt%, preferably 8 wt% to 13 wt%, more preferably 10 wt% to 11 wt%, and most preferably 10.5 wt%.

[0010] In some specific implementation manners, the product uses water as a solvent.

[0011] The present application also provides a composition, comprising a fermentation product filtrate of Bifidobacterium / Lactobacillus / soybean seed extract and propylhydroxytetrahydropyrantriol;

[0012] The mass ratio of the fermentation product filtrate of Bifidobacterium / Lactobacillus / soybean seed extract to propylhydroxytetrahydropyrantriol is 0.01 to 5:5 to 15.

[0013] The mass ratio of the fermentation product filtrate of Bifidobacterium / Lactobacillus / soybean seed extract to propylhydroxytetrahydropyrantriol is preferably 0.01 to 5:10 to 11, more preferably 0.032 to 3.2:10 to 11, and most preferably 0.032:10.5, 0.32:10.5 or 3.2:10.5.

[0014] In some specific implementation manners, the composition further comprises a solvent, and the solvent is water.

[0015] The present application uses the fermentation product filtrate of Bifidobacterium / Lactobacillus / soybean seed extract as a penetration enhancer, which can promote the transdermal absorption of water-soluble propylhydroxytetrahydropyrantriol at a lower concentration, and has a better promoting effect at a lower concentration than fruit acids such as glycolic acid at high concentrations and typical chemical penetration enhancers such as Neosolue™-Aqulio. Further, the inventor has found that other lactic acid bacteria fermentation products containing small molecules of lactic acid, such as lactic acid bacteria fermentation lysate, etc., also have the effect of promoting the transdermal absorption of water-soluble propylhydroxytetrahydropyrantriol.

[0016] The present application also provides the use of the composition described in the above technical solution in the preparation of an anti-aging product. Among them, the filtrate of the bifidobacterium / lactobacillus / soybean seed extract fermentation product can promote the transdermal absorption of water-soluble propyltetrahydropyranyl glucoside at a lower concentration as a penetration enhancer, enabling propyltetrahydropyranyl glucoside to penetrate the stratum corneum barrier and enter the core action site of the skin, thereby exerting a better anti-aging effect. At the same time, the composition is relatively mild and will not cause a sense of irritation or discomfort during use.

[0017] The present application also provides a product, including the composition described in the above technical solution, and the product is preferably a cosmetic.

[0018] In the present invention, according to the classification principle of cosmetics, cosmetics can be classified into: cleansing cosmetics, caring cosmetics, and beauty / cosmetic cosmetics.

[0019] Among them, the cleansing cosmetics refer to cosmetics that are applied to the human body surface (such as the epidermis, hair, nails, lips, etc.) by means of smearing, spraying or other similar methods to play a role in cleaning and hygiene or eliminating unpleasant odors. The caring cosmetics refer to cosmetics that are applied to the human body surface (such as the epidermis, hair, nails, lips, etc.) by means of smearing, spraying or other similar methods to play a role in maintenance. The beauty / cosmetic cosmetics refer to cosmetics that are applied to the human body surface (such as the epidermis, hair, nails, lips, etc.) by means of smearing, spraying or other similar methods to play a role in beautifying, decorating and enhancing human charm.

[0020] The cleansing cosmetics suitable for the skin include but are not limited to facial cleanser, makeup remover (milk), cleansing cream (honey), facial mask, floral water, prickly heat powder, talcum powder or bath liquid; the caring cosmetics suitable for the skin include but are not limited to skin care creams, lotions or toners; the beauty / cosmetic cosmetics suitable for the skin include but are not limited to powder puffs, rouge, eye shadows, eyeliner pencils (liquids), eyebrow pencils, perfumes or colognes; the cleansing cosmetics suitable for the hair include but are not limited to shampoos, hair creams or shaving creams; the caring cosmetics suitable for the hair include but are not limited to hair conditioners, hair milks, hair oils / waxes or hair treatment creams; the beauty / cosmetic cosmetics suitable for the hair include but are not limited to styling mousses / gels, hair dyes, hair perming agents, mascara (cream), hair growth agents or hair removal agents; the cleansing cosmetics suitable for the nails include but are not limited to nail polish removers; the caring cosmetics suitable for the nails include but are not limited to nail care liquids (creams), nail hardeners; the beauty / cosmetic cosmetics suitable for the nails include but are not limited to nail polishes; the cleansing cosmetics suitable for the lips include but are not limited to lip makeup removers; the caring cosmetics suitable for the lips include but are not limited to lip balms; the beauty / cosmetic cosmetics suitable for the lips include but are not limited to lipsticks, lip glosses or lip liners.

[0021] In this application, the fermentation product filtrate of Bifidobacterium / Lactobacillus / soybean seed extract is used as a penetration enhancer, which can promote the transdermal absorption of water-soluble propylene glycol alginate at a lower concentration, and has a better promoting effect than fruit acids such as glycolic acid at a high concentration and typical chemical penetration enhancers such as Neosolue™-Aqulio at a lower concentration. Moreover, the fermentation product filtrate of Bifidobacterium / Lactobacillus / soybean seed extract is relatively mild and will not cause a sense of irritation or discomfort during use. Detailed implementation manners

[0022] It should be understood that the expression "one or more of..." individually includes each of the objects recited after the said expression and various different combinations of two or more of the recited objects, unless otherwise understood from the context and usage. The expression "and / or" in combination with three or more recited objects should be understood to have the same meaning, unless otherwise understood from the context.

[0023] The terms "comprising", "having" or "containing", including the use of their grammatical synonyms, should generally be understood as open and non-limiting, for example, not excluding other unrecited elements or steps, unless otherwise specifically stated or understood from the context.

[0024] It should be understood that as long as the present invention is still operable, the order of steps or the order of performing certain actions is not important. In addition, two or more steps or actions can be carried out simultaneously.

[0025] The use of any and all examples or exemplary language such as "for example" or "including" herein is only intended to better illustrate the present invention and does not limit the scope of the present invention unless a claim is made. Any language in this specification should not be construed as indicating that any unclaimed element is essential for the practice of the present invention.

[0026] In addition, the numerical ranges and parameters used to define the present invention are approximate values. The relevant values in the specific embodiments have been presented as precisely as possible herein. However, any value inherently inevitably contains standard deviations due to individual test methods. Therefore, unless otherwise clearly stated, it should be understood that all ranges, quantities, values and percentages used in this disclosure are modified by "about". Herein, "about" generally means that the actual value is within plus or minus 10%, 5%, 1% or 0.5% of a specific value or range.

[0027] This application provides the use of the fermentation product filtrate of Bifidobacterium / Lactobacillus / soybean seed extract in the preparation of a product for promoting the transdermal absorption of propylene glycol alginate.

[0028] The filtrate of the fermentation product of Bifidobacterium / Lactobacillus / soybean seed extract is a new raw material developed by the applicant, with the trade name "Huafuyuan". Based on the fermentation process, it enhances the bioavailability of metabolites, making the effect of the product greater than the sum of its parts. Finally, a "highly active composite metabolite group" of more than 500 is obtained, such as small molecule fruit acids like "lactic acid, citric acid", and also includes small molecule amino acids like "L-alanine, ornithine, glycine, lysine, serine". The applicant found that the filtrate of the fermentation product of Bifidobacterium / Lactobacillus / soybean seed extract is not only milder, but also can help accelerate the metabolism of the stratum corneum through small molecule fruit acids and affect the lipid and protein structures through small molecule amino acids, thereby assisting the transdermal absorption of water-soluble hydroxypropyltetrahydropyranyl triol, and has a better promoting effect at a lower concentration than high-concentration fruit acids such as glycolic acid and typical chemical penetration enhancers like Neosolue™-Aqulio.

[0029] The following further illustrates the compositions and their applications provided in this application in conjunction with examples.

[0030] In the following examples, the filtrate of the fermentation product of Bifidobacterium / Lactobacillus / soybean seed extract has the trade name "Huafuyuan" and is sourced from Guangzhou Yixian E-commerce Co., Ltd.;

[0031] Neosolue™-Aqulio is purchased from Nippon Fine Chemical.

[0032] Test Example 1 pH value experiment

[0033] This experiment explored the influence of the system pH when different penetration enhancers including the active substance of Huafuyuan were added to the hydroxypropyltetrahydropyranyl triol solution. At 25°C, using only single solvent water, solutions were prepared according to the formula shown in Table 1, and the pH values of the corresponding solutions were measured using a pH meter. See Table 1, where Table 1 shows the formula and pH value of different compositions.

[0034] Table 1 Influence of different penetration enhancers on the pH of the hydroxypropyltetrahydropyranyl triol solution system

[0035]

[0036] As can be seen from Table 1, when Huafuyuan was added to the hydroxypropyltetrahydropyranyl triol solution, although the pH value of the solution decreased, it could remain above 3. However, after adding 5% glycolic acid (hydroxyacetic acid), a high-concentration small molecule fruit acid, the pH value decreased more, to around 1.9. A pH value of around 1.9 is very close to strong acidity, which may more strongly stimulate the skin nerve endings, causing more obvious discomfort such as stinging and burning, and will also cause more serious skin dryness and desquamation, bringing greater side effects to the skin. Thus, it can be seen that Huafuyuan is milder than other high-concentration small molecule fruit acid penetration enhancers.

[0037] Experimental Example 2 Chicken Embryo Stimulation Test

[0038] This experiment further explored the evaluation of the irritation of the filtrate of the fermentation product of Bifidobacterium / Lactobacillus / soybean seed extract (Huanyuan) on the chorioallantoic membrane of chicken embryos, that is, to evaluate the safety of Huanyuan.

[0039] 1. Experimental materials:

[0040] 1.1 Chicken embryos: SPF White Leghorn chickens, purchased from Jiangsu Boehringer Ingelheim Vetong Biotechnology Co., Ltd., production license number: SCXK (Su) 2022-0002;

[0041] 1.2 Incubation conditions: Incubator temperature 37.5±0.5°C, relative humidity 55-70%.

[0042] 2. Test procedures:

[0043] 2.1 CAM preparation: Purchase 0-day-old chicken embryos. When they are hatched to 9 days old, check and discard defective chicken embryos. Mark the position of the air chamber on the surface of the eggshell of normal chicken embryos, peel off part of the eggshell to expose the white egg membrane; carefully remove the inner membrane with forceps to ensure that the vascular membrane is not damaged.

[0044] 2.2 Formal test

[0045] 2.2.1 Stimulation scoring method: For at least 6 chicken embryos in each group, take 0.3 mL of the test substance and directly drop it on the surface of the CAM. Observe the reaction of the CAM, and record the time when each toxic effect appears within 5 minutes, accurate to seconds, including three reactions: bleeding, coagulation, and vascular lysis, and record the degree of the reaction.

[0046] 2.2.2 Endpoint scoring method: For at least 6 chicken embryos in each group, take 0.3 mL of the test substance and directly drop it on the surface of the CAM. Observe the reaction of the CAM. After 3 minutes of action, gently rinse with physiological saline to remove the test substance, and observe the three reactions and their degrees of bleeding, coagulation, and vascular lysis within about 30 seconds after rinsing. If the observation shows that the score of at least one reaction in all 6 chicken embryos is above moderate (total score ≥ 12), the test should be repeated once.

[0047] 2.2.3 The positive control and negative control are operated by the stimulation scoring method.

[0048] 2.3 Data analysis

[0049] 2.3.1 Stimulation scoring method (IS): Record the time and degree of reaction at each detection endpoint, and calculate the stimulation score (IS) using formula (I), and the result is reserved to two decimal places:

[0050]

[0051] Formula (I);

[0052] In Formula (I), sec H, sec L, and sec C represent the average time (seconds) at which bleeding, vascular lysis, and blood coagulation start to occur as observed on the CAM membrane, respectively.

[0053] The results were evaluated according to the method shown in Table 2:

[0054] Table 2 Evaluation of Results by Stimulation Scoring Method

[0055]

[0056] 2.3.2 Endpoint Scoring Method (ES): Score for each chick embryo = sum of the degrees of bleeding, blood coagulation, and vascular lysis observed in each chick embryo; Endpoint score (ES) = sum of the scores of 6 chick embryos.

[0057] The results were evaluated according to the method shown in Table 3:

[0058] Table 3 Evaluation of Results by Endpoint Scoring Method

[0059]

[0060] 3. Data Results:

[0061] The stimulation scoring method was selected for the test samples. The results are shown in Table 4, which presents the stimulation scoring results of different active substances provided in this application.

[0062] Table 4 Scoring Results

[0063]

[0064] 4. Conclusion

[0065] According to the judgment criteria of SN / T 2329 - 2009, under the experimental conditions of this study, when the concentration of the test sample "Huanyuan" was 0.45% (w / w), the IS score was 0.00, indicating no irritation; when the concentrations of the test sample "Salicylic Acid" were 1% and 2% (w / w), the IS scores were above 12, indicating strong irritation / corrosion.

[0066] Based on Examples 1 and 2, it can be seen that although Huanyuan is a fermentation product filtrate of Bifidobacterium / Lactobacillus / soybean seed extract, it is milder and less sensitizing compared to other acid exfoliating components.

[0067] In the following examples and comparative examples, the focus is on exploring the permeation - enhancing effect of different permeation - enhancers on glucosamine hydrochloride.

[0068] Experimental Example 3 Safety Evaluation of Human Patch Test

[0069] This experiment further explores the safety evaluation study of the solution of propyltetrahydropyranyl triol containing a high concentration of "Huanyuan" Bifidobacterium / Lactobacillus / Soybean Seed Extract Fermentation Product Filtrate (Huanyuan) for consumers, that is, further evaluating the safety of Huanyuan on the human body.

[0070] Test method: Select 33 subjects and record the skin reactions of the subjects. Take 20 μl of the sample in Example 3 and place it in the patch device. The control hole is a blank control (without placing any substance). Apply the sample in Example 3 and the blank control to the flexor side of the forearm of the subject, and gently press with the palm to make it evenly adhere to the skin for 24 h. Observe the skin reactions at 30 min (after the indentation disappears), 24 h, and 48 h after removing the test patch device according to the following table standards. And record the observation results

[0071] Evaluation criteria: Cosmetics Safety Technical Specifications (2015 Edition)

[0072] Skin Reaction Grading Standard for Skin Occlusive Patch Test

[0073]

[0074] Test results: Among the 33 subjects, only 1 subject had a grade 1 adverse reaction 0.5 h after removing the patch device; based on the regulations in the latest "Technical Guidelines for Human Skin Patch Test", if within 5 cases there is a grade 1 reaction in the patch test and no grade 2 or higher-level reactions occur, it is generally considered that the test product does not cause obvious adverse reactions within the test range. In this patch test, in the test group of the solution of propyltetrahydropyranyl triol containing a high concentration of Bifidobacterium / Lactobacillus / Soybean Seed Extract Fermentation Product Filtrate (Example 3), only 1 case of grade 1 adverse reaction (i.e., only faint erythema) occurred at half an hour, and at 24 h and 48 h, it had completely subsided without further aggravation. Thus, it can be seen that this example is relatively mild;

[0075] Experimental Example 4

[0076] This experiment is based on the suckling pig skin-Franz cell system, and the concentration of propyltetrahydropyranyl triol in the receiving liquid at different time points is determined by high-performance liquid chromatography to evaluate the skin penetration behavior of different penetration enhancers containing Huanyuan on propyltetrahydropyranyl triol.

[0077] 4.1 Materials and Reagents

[0078] In this test, the back skin of 1-month-old suckling pigs was selected as the test model, and the suckling pig skin - Franz cell was selected as the test system. PBS buffer solution (Solarbio), Neosolue™-Aqulio (bis-diethoxydiglycol cyclohexane 1,4-dicarboxylate, from Nippon Fine Chemical), propylidene glycol-alpha-D-glucopyranoside (S / R racemic sample), high performance liquid chromatography (Thermo Fisher).

[0079] 4.2 Test device

[0080] TK-12D transdermal diffusion tester.

[0081] 4.3 Test samples

[0082] The test samples are shown in Table 5, and the solvent is water.

[0083] Table 5 Test sample information

[0084]

[0085] 4.4 Methods and procedures

[0086] 4.4.1 Preparation of ex vivo suckling pig skin

[0087] The back skin of 1-month-old suckling pigs stored at -20°C was thawed at room temperature with deionized water and repeatedly rinsed with PBS buffer solution.

[0088] 4.4.2 Permeation ability test

[0089] 1) Fix the suckling pig skin between the supply chamber and the receiving chamber of the Franz diffusion cell, with the stratum corneum of the skin facing the supply chamber and the dermal layer facing the receiving chamber;

[0090] 2) Add 7 mL of the receiving solution to the receiving chamber. After tightening and fixing the suckling pig skin, add 1 mL of the receiving solution (water) to the receiving chamber through a sampler, drain the air, and make the dermal layer of the skin in close contact with the receiving solution;

[0091] 3) Sample application: Add the sample to the skin surface in the supply chamber. The effective permeation area S is approximately 3.14 cm 2 . Add the sample to the surface of the suckling pig skin and spread the sample evenly from the central part of the skin radially towards the edge. There are 3 replicates and parallels for each sample;

[0092] 4) Permeation: Turn on the electromagnetic stirrer and stir at a speed of 300 rpm, maintain a constant water bath at (32 ± 1)°C, and ensure that there are no bubbles in the water bath sandwich;

[0093] 5) Samples were taken at 1h, 2h, 4h, 8h, and 24h respectively. Using a sampler, 2 mL of the receiving solution was extracted through the sampling tube, and then placed in a 2 mL centrifuge tube. In addition, an equal amount of the receiving solution was replenished after each sampling.

[0094] 4.4.3 Transdermal Content Test

[0095] After 24h of penetration, the contents of the target substance in the skin surface, in the skin, and under the skin of different experimental groups were detected by HPLC. The specific treatment methods are as follows:

[0096] 1) Test for the unpenetrated part on the skin surface: After 24h, use a 1 mL pipette to suck acetonitrile and repeatedly blow and wash the skin surface multiple times. Wash 5 times for a total of 5 mL, and place it in a 5 mL centrifuge tube for testing.

[0097] 2) Test for the part in the skin: Use a sterilized scalpel to cut the skin along the inner edge of the supply chamber. Cut the skin into pieces and add 5 mL of acetonitrile for ultrasonic extraction for 60 min. After ultrasonic extraction, suck out the supernatant for testing.

[0098] 3) Test for the part under the skin: At each time point, 2 mL of the receiving solution was taken from the receiving chamber and placed in a 2 mL centrifuge tube for testing.

[0099] 4.4.4 Statistics and Analysis

[0100] 1) Cumulative penetration amount Q = Cn×V + ∑Ci×V0 (i = 1···n - 1) Formula (II)

[0101] In the formula:

[0102] Q: Cumulative penetration amount;

[0103] V: Volume of the receiving solution in the receiving chamber. V0: Volume of each sampling;

[0104] Ci: Concentration of the sample in the receiving solution from the first sampling to the previous sampling;

[0105] Cn: Concentration of the sample measured at the nth sampling point.

[0106] 2) Penetration rate

[0107] Make a regression equation with the cumulative penetration amount (Y-axis) against the penetration time (X-axis). The slope of the equation is the penetration rate K. P .

[0108] 3) Recovery rate

[0109] Recovery rate = total content (skin surface + in the skin + under the skin) / sample loading amount × 100%.

[0110] 4) Analysis of statistical results

[0111] GraphPad Prism was used for plotting, and the results were expressed as Mean±SD.

[0112] 4.5 Test Results

[0113] 1) Cumulative Permeation Amount

[0114] According to formula (II), the cumulative permeation amount of the samples at different time points was obtained. The results are shown in Table 6, which is a summary table of the cumulative permeation amount results of the samples.

[0115] Table 6 Summary Table of Cumulative Permeation Amount Results

[0116]

[0117] Note: Q is the cumulative skin permeation amount (μg), and SD is the standard deviation.

[0118] 2) Permeation Rate

[0119] A regression equation was made with the cumulative permeation amount (Y-axis) against the permeation time (X-axis). The slope of the equation is the permeation rate Kp. The results are shown in Table 7 below, which is the permeation rate results of the samples.

[0120] Table 7 Permeation Rate Results of Samples

[0121]

[0122] 3) Recovery Rate:

[0123] The summary of the results of three parallel tests is shown in Table 8, which is the statistical table of the recovery rate of the samples.

[0124] Table 8 Recovery Rate of Samples

[0125]

[0126] 4.6 Conclusion

[0127] Referring to Table 6, it can be seen that at the test concentrations of 0.032% - 3.2%, Huanfuyuan is helpful for the permeation of propyltetrahydropyranyl triol. And this permeation effect can be significantly found after 1 hour of epidermal action, and it is significantly better than typical chemical penetration enhancers and also significantly better than high-concentration fruit acids. Moreover, the permeation amount gradually increases with time;

[0128] Referring to Table 7, it can be seen that after the addition of Huanfuyuan, the penetration rate of propyltetrahydropyranyl triol is increased by about 3 to 7 times, which is 1.3 to 3.3 times that of chemical penetration enhancers. Based on this, it can be known that the filtrate of the fermentation product of Bifidobacterium / Lactobacillus / Soybean Seed Extract, namely "Huanfuyuan", has a good effect on improving the penetration of water-soluble propyltetrahydropyranyl triol, thereby helping it penetrate into the basal layer and even the dermis of the skin, and better exerting the anti-aging effect.

[0129] The above are only the preferred embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. Use of the filtrate of the fermentation product of Bifidobacterium / Lactobacillus / soybean seed extract in the preparation of a product for promoting the transdermal penetration of propyltetrahydropyrantriol.

2. The application according to claim 1, wherein The concentration of the filtrate of the fermentation product of Bifidobacterium / Lactobacillus / soybean seed extract in the product is 0.01 wt% - 5 wt%.

3. The application according to claim 1, characterized in that, The concentration of propyltetrahydropyrantriol in the product is 5 wt% - 15 wt%.

4. A composition, characterized in that, It includes the filtrate of the fermentation product of Bifidobacterium / Lactobacillus / soybean seed extract and propyltetrahydropyrantriol; The mass ratio of the filtrate of the fermentation product of Bifidobacterium / Lactobacillus / soybean seed extract to propyltetrahydropyrantriol is 0.01 - 5:5 - 15.

5. The composition according to claim 4, wherein The mass ratio of the filtrate of the fermentation product of Bifidobacterium / Lactobacillus / soybean seed extract to propyltetrahydropyrantriol is 0.01 - 5:10 - 11.

6. The composition according to claim 5, characterized in that, The mass ratio of the filtrate of the fermentation product of Bifidobacterium / Lactobacillus / soybean seed extract to propyltetrahydropyrantriol is 0.032 - 3.2:10 - 11.

7. The composition according to claim 6, characterized in that, The mass ratio of the filtrate of the fermentation product of Bifidobacterium / Lactobacillus / soybean seed extract to propyltetrahydropyrantriol is 0.032:10.5, 0.32:10.5 or 3.2:10.

5.

8. The composition according to any one of claims 4 to 7, characterized in that, It also includes water.

9. Use of the composition according to any one of claims 4 - 8 in the preparation of an anti - aging product.

10. An anti-aging product, characterized in that, It includes the composition according to any one of claims 4 - 8.

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