A fermentation product for enhancing the anti-aging effect of hydroxypropyl tetrahydropyrantriol and its combined application

Abstract

The present application provides the application of bifidobacterium / lactobacillus / soybean seed extract fermentation product filtrate in the preparation of a product that promotes the transdermal absorption of hydroxypropyl tetrahydropyrantriol. The present application also provides a composition, its application and product. The present application uses bifidobacterium / lactobacillus / soybean seed extract fermentation product filtrate as a penetration enhancer, which can promote the transdermal absorption of water-soluble hydroxypropyl tetrahydropyrantriol at a lower concentration, and has a better promoting effect than high concentrations of glycolic acid and other fruit acids and typical chemical penetration enhancers such as Neosolue™-Aqulio at a lower concentration. Moreover, bifidobacterium / lactobacillus / soybean seed extract fermentation product filtrate is relatively mild and will not cause irritation or discomfort when used.

Description

Technical Field

[0001] The present invention relates to the technical field of daily chemical products, in particular to a fermentation product capable of enhancing the anti-aging effect of hydroxypropyl tetrahydropyrantriol and a combined application thereof. Background Art

[0002] Hydroxypropyl tetrahydropyrantriol, also known as hydroxypropyl tetrahydropyrantriol, has been recognized by the scientific community and the cosmetics industry for its significant role and potential in anti-aging, earning it the title "one of the three most powerful anti-aging ingredients." Hydroxypropyl tetrahydropyrantriol has a LogP of -2.07 (source: Chemicalbook), indicating strong water solubility. The stratum corneum is the primary site of absorption for external substances, accounting for over 90% of the skin's total absorption capacity. Because the interstices of the stratum corneum are primarily composed of lipids, the stratum corneum primarily absorbs fat-soluble substances. Consequently, water-soluble active ingredients have difficulty penetrating the lipid-soluble barrier of the outermost layer of the skin. The core action sites of hydroxypropyl tetrahydropyrantriol are the basement membrane and the extracellular matrix of the dermis. Its corresponding efficacy is to promote the production of type IV collagen and type VII collagen in the basement membrane and the production of mucopolysaccharides such as glycosaminoglycans in the extracellular matrix of the dermis (Eur J Dermatol 2011; 21(2): 191-6). Therefore, hydroxypropyl tetrahydropyrantriol needs to penetrate deep into the epidermis and even reach the dermis to work well. However, studies have shown that the bioavailability of hydroxypropyl tetrahydropyrantriol is only 3-7% (Eur J Dermatol 2011; 21(2): 191-6). At the same time, studies have shown that even if the dosage of hydroxypropyl tetrahydropyrantriol is increased, the improvement in bioavailability is not significant. Moreover, as the concentration of hydroxypropyl tetrahydropyrantriol increases, its viscosity will also increase, which will affect the skin feel when used. Therefore, the technology that promotes the penetration of hydroxypropyl tetrahydropyrantriol or the component that assists the penetration of hydroxypropyl tetrahydropyrantriol is the key means to improve the utilization rate of hydroxypropyl tetrahydropyrantriol.

[0003] Currently, common cosmetic transdermal penetration enhancement technologies include physical penetration enhancement methods, such as iontophoresis, electroporation, ultrasound, microneedles, thermal ablation, magnetic induction, photomechanical waves, and electron beam radiation. These methods physically create tiny channels in the skin, which can enhance drug transdermal delivery. However, physical penetration generally requires specialized equipment and techniques and may cause varying degrees of discomfort or side effects. Chemical penetration enhancers, such as azones and alcohols, can also be used to enhance drug transdermal absorption efficiency by disrupting lipid structures or reducing stratum corneum thickness. Vesicle delivery systems and microemulsion / nanoemulsion delivery systems, such as liposomes, transfersomes, microemulsions, and nanoemulsions, can also be used to enhance drug transdermal absorption efficiency. For example, Chinese patent CN116528820A discloses a microemulsion or nanoemulsion technology that effectively improves the transdermal delivery of water-soluble active ingredients, such as hydroxypropyl tetrahydropyrantriol. However, this technology generally requires precise control of the ratios of various oils, surfactants, and other ingredients, and sometimes even requires the use of high-energy equipment, resulting in a relatively complex preparation process. Summary of the Invention

[0004] In view of this, the present invention provides a composition and its application. The composition provided in the present application is relatively mild, has little irritation to the skin, and can effectively promote the transdermal absorption of water-soluble active ingredients such as hydroxypropyl tetrahydropyrantriol, thereby significantly improving the utilization rate of hydroxypropyl tetrahydropyrantriol.

[0005] The present application provides the use of a filtrate of a fermentation product of Bifidobacterium / Lactobacillus / soybean seed extract in the preparation of a product that promotes the transdermal penetration of hydroxypropyl tetrahydropyrantriol.

[0006] Bifidobacterium / Lactobacillus / Soybean Seed Extract Ferment Filtrate is a raw material developed by the applicant, with the national cosmetics registration number 20230032. The registered Chinese name is Bifidobacterium / Lactobacillus / Soybean Seed Extract Ferment Filtrate, the INCI name is Bifidobacterium / Lactobacillus / Soybean Seed Extract Ferment Filtrate, and the trade name is "Huanfuyuan". It is based on a fermentation process, which enhances the bioavailability of metabolites, making the product's effect greater than the sum of its parts, and ultimately obtaining 500+ "highly active complex metabolite groups", such as "lactic acid, citric acid, etc." small molecule fruit acids, as well as small molecule amino acids such as "L-alanine, ornithine, glycine, lysine, and serine." In some specific implementations, the preparation method of the bifidobacterium / lactobacillus / soybean seed extract fermentation product filtrate mainly includes: soaking black beans and water in a certain proportion, obtaining soybean seed extract after heating and water extraction, obtaining fermentation base after sterilization, and then transferring it into a fermentation tank, adding a strain composition (containing bifidobacteria and lactobacillus) for fermentation, and obtaining fermentation filtrate after sterilization and filtration.

[0007] The applicant discovered that the filtrate of the fermentation product of Bifidobacterium / Lactobacillus / soybean seed extract is not only milder, but also can help accelerate stratum corneum metabolism through small molecule fruit acids and small molecule amino acids to affect lipid and protein structure, thereby assisting the transdermal absorption of water-soluble hydroxypropyl tetrahydropyrantriol. At lower concentrations, it has a better promoting effect than high concentrations of fruit acids such as glycolic acid and typical chemical penetration enhancers such as Neosolue™-Aqulio.

[0008] In some specific implementations, the concentration of the bifidobacterium / lactobacillus / soybean seed extract fermentation product filtrate in the product is 0.01 wt% to 5 wt%, preferably 0.032 wt% to 3.2 wt%, more preferably 0.032 wt%, 0.32 wt% or 3.2 wt%.

[0009] In some specific implementations, the concentration of hydroxypropyl tetrahydropyrantriol 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 implementations, the product uses water as a solvent.

[0011] The present application also provides a composition comprising a bifidobacterium / lactobacillus / soybean seed extract fermentation product filtrate and hydroxypropyl tetrahydropyrantriol;

[0012] The mass ratio of the bifidobacterium / lactobacillus / soybean seed extract fermentation product filtrate to hydroxypropyl tetrahydropyrantriol is 0.01-5:5-15.

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

[0014] In some specific implementations, the composition further comprises a solvent, which is water.

[0015] This application uses the filtrate of the fermentation product of Bifidobacterium / Lactobacillus / soybean seed extract as a penetration enhancer, which can promote the transdermal absorption of water-soluble hydroxypropyl tetrahydropyrantriol at lower concentrations. At lower concentrations, it has a better promoting effect than high concentrations of glycolic acid and other fruit acids and typical chemical penetration enhancers such as Neosolue™-Aqulio. Furthermore, the inventors have discovered that other lactic acid bacteria fermentation products containing small molecule lactic acid, such as lactic acid bacteria fermentation lysate, also have the effect of promoting the transdermal absorption of water-soluble hydroxypropyl tetrahydropyrantriol.

[0016] The present application also provides the use of the composition described in the above technical solution in the preparation of anti-aging products. The filtrate of the fermentation product of Bifidobacterium / Lactobacillus / soybean seed extract acts as a penetration enhancer, which can promote the transdermal absorption of water-soluble hydroxypropyl tetrahydropyrantriol at relatively low concentrations, allowing hydroxypropyl tetrahydropyrantriol to penetrate the stratum corneum barrier and enter the core active areas of the skin, thereby exerting a good anti-aging effect. Furthermore, the composition is relatively mild and does not cause irritation or discomfort during use.

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

[0018] In the present invention, according to the cosmetic classification principle, cosmetics can be divided into: cleansing cosmetics, care cosmetics and beauty / decorative cosmetics.

[0019] Among them, the term "cleansing cosmetics" refers to cosmetics that are applied to the human body surface (such as the epidermis, hair, nails, lips, etc.) by smearing, spraying, or other similar methods to provide a cleansing and sanitation effect or eliminate bad odors. The term "care cosmetics" refers to cosmetics that are applied to the human body surface (such as the epidermis, hair, nails, lips, etc.) by smearing, spraying, or other similar methods to provide a maintenance effect. The term "beautifying / modifying cosmetics" refers to cosmetics that are applied to the human body surface (such as the epidermis, hair, nails, lips, etc.) by smearing, spraying, or other similar methods to provide a beautifying, modifying, or enhancing human attractiveness.

[0020] Skin-applicable cleansing cosmetics include but are not limited to facial cleansers, makeup removers (milk), cleansing creams (honey), facial masks, floral water, prickly heat powder, talcum powder or bath lotion; skin-care cosmetics include but are not limited to: skin care creams, lotions or toners; skin-beautifying / decorative cosmetics include but are not limited to pressed powder, rouge, eye shadow, eyeliner (liquid), eyebrow pencil, perfume or cologne; hair-applicable cleansing cosmetics include but are not limited to shampoo, shampoo cream or shaving cream; hair-care cosmetics include but are not limited to conditioner, hair cream, hair oil / wax or hair treatment cream; hair-applicable Beauty / decorative cosmetics include but are not limited to styling mousse / hair spray, hair dye, perm, mascara (cream), hair growth agent or depilatory agent; cleansing cosmetics for nails include but are not limited to nail polish remover; care cosmetics for nails include but are not limited to nail lotion (cream), nail hardener; beauty / decorative cosmetics for nails include but are not limited to nail polish; cleansing cosmetics for lips include but are not limited to lip makeup remover; care cosmetics for lips include but are not limited to lip balm; beauty / decorative cosmetics for lips include but are not limited to lipstick, lip gloss or lip liner.

[0021] This application uses a bifidobacterium / lactobacterium / soybean seed extract fermentation product filtrate as a penetration enhancer, capable of promoting transdermal absorption of water-soluble hydroxypropyl tetrahydropyrantriol at relatively low concentrations. This lower concentration demonstrates a superior effect compared to high-concentration glycolic acid and other fruit acids, as well as typical chemical penetration enhancers such as Neosolue™-Aqulio. Furthermore, the bifidobacterium / lactobacterium / soybean seed extract fermentation product filtrate is relatively mild, causing no irritation or discomfort upon use. DETAILED DESCRIPTION

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

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

[0024] It should be understood that the order of steps or the order in which certain actions are performed are not important as long as the present invention remains operable. Additionally, two or more steps or actions may be performed simultaneously.

[0025] The use of any and all examples or exemplary language, such as "such as" or "including," herein is intended merely to better illustrate the invention and does not limit the scope of the invention unless otherwise claimed. No language in this specification should be construed as indicating any non-claimed element as essential to the practice of the invention.

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

[0027] The present application provides the use of a filtrate of a fermentation product of Bifidobacterium / Lactobacillus / soybean seed extract in the preparation of a product that promotes the transdermal penetration of hydroxypropyl tetrahydropyrantriol.

[0028] The filtrate of the fermentation product of Bifidobacterium / Lactobacillus / soybean seed extract is a new raw material developed by the applicant, marketed as "Huanfuyuan." Based on a fermentation process, it enhances the bioavailability of metabolites, making the product more effective than the sum of its parts. The resulting product is a "highly active complex metabolite panel" of over 500, including small-molecule fruit acids such as lactic acid and citric acid, as well as small-molecule amino acids such as L-alanine, ornithine, glycine, lysine, and serine. The applicant discovered that the filtrate of the fermentation product of Bifidobacterium / Lactobacillus / soybean seed extract is not only milder, but also helps accelerate stratum corneum metabolism through small-molecule fruit acids and influences lipid and protein structure through small-molecule amino acids, thereby assisting in the transdermal absorption of water-soluble hydroxypropyl tetrahydropyrantriol. At lower concentrations, it exhibits a superior promoting effect compared to high-concentration fruit acids such as glycolic acid and typical chemical penetration enhancers such as Neosolue™-Aqulio.

[0029] The compositions and applications of the present invention are further described below with reference to the following examples.

[0030] In the following examples, the trade name of the filtrate of the fermentation product of Bifidobacterium / Lactobacillus / soybean seed extract is "Huanfuyuan", which is sourced from Guangzhou Yixian E-Commerce Co., Ltd.

[0031] Neosolue™-Aqulio was purchased from Nippon Seika.

[0032] Test Example 1 pH value experiment

[0033] This experiment investigated the effect of adding different penetration enhancers, including Huanfuyuan active ingredients, to a hydroxypropyl tetrahydropyrantriol solution on the pH of the system. Using only water as the solvent at 25°C, solutions were prepared according to the formula shown in Table 1. The pH values ​​of the corresponding solutions were measured using a pH meter. Table 1 shows the formulas and pH values ​​of the different compositions.

[0034] Table 1 Effects of different penetration enhancers on the pH of hydroxypropyl tetrahydropyrantriol solution

[0035]

[0036] As shown in Table 1, although the addition of Huanfuyuan to the hydroxypropyl tetrahydropyrantriol solution lowered the pH of the solution, the pH was maintained above 3. However, after the addition of 5% glycolic acid (hydroxyacetic acid), a high-concentration small-molecule fruit acid, the pH dropped even further, to around 1.9. A pH of around 1.9 is very close to strong acidity, which may more strongly stimulate the skin's nerve endings, causing more noticeable discomfort such as stinging and burning, and may also cause more severe skin dryness and desquamation, bringing greater side effects to the skin. This shows that Huanfuyuan is milder than other high-concentration small-molecule fruit acid penetration enhancers.

[0037] Test Example 2 Chicken embryo stimulation test

[0038] This study further explored the irritation of the "Huanfuyuan" Bifidobacterium / Lactobacillus / soybean seed extract fermentation product filtrate (Huanfuyuan) to the chicken embryo chorioallantoic membrane, that is, to evaluate the safety of Huanfuyuan.

[0039] 1. Experimental materials:

[0040] 1.1 Chicken embryos: SPF-grade Bailaihang chicken, purchased from Jiangsu Boehringer Ingelheim Weitong Biotechnology Co., Ltd., production license number: SCXK (Su) 2022-0002;

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

[0042] 2. Test steps:

[0043] 2.1 CAM Preparation: Purchase 0-day-old chick embryos. At 9 days of incubation, inspect and discard any defective embryos. Mark the location of the air cell on the surface of the eggshell of a normal chick embryo. Peel back the eggshell to expose the white membrane. Carefully remove the inner membrane with forceps, ensuring that the vascular membrane is not damaged.

[0044] 2.2 Formal test

[0045] 2.2.1 Irritation scoring method: For each group of at least 6 chicken embryos, 0.3 mL of the test substance was directly dripped onto the CAM surface. The CAM reaction was observed and the time of occurrence of each toxic effect within 5 minutes was recorded to the nearest second. The three reactions, including bleeding, coagulation, and vascular melting, were recorded, and the degree of the reaction was recorded.

[0046] 2.2.2 Endpoint Scoring Method: For at least six chick embryos per group, 0.3 mL of the test substance is dripped directly onto the CAM surface. The CAM reaction is observed. After a 3-minute exposure, the test substance is gently rinsed away with saline. Within approximately 30 seconds after rinsing, the three reactions (hemorrhage, coagulation, and vascular lysis) are observed for severity. If all six chick embryos demonstrate a score of moderate or above for at least one reaction (total score ≥ 12), the test should be repeated.

[0047] 2.2.3 Positive and negative controls were operated using the stimulation scoring method.

[0048] 2.3 Data Analysis

[0049] 2.3.1 Irritation Score (IS): Record the time of occurrence and degree of reaction of each test endpoint, and calculate the Irritation Score (IS) using formula (I). The result is rounded to two decimal places:

[0050]

[0051] Formula (I);

[0052] In formula (I), sec H, sec L, and sec C represent the average time (in seconds) to the onset of bleeding, vascular melting, and coagulation observed on the CAM membrane, respectively.

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

[0054] Table 2 Evaluation results of stimulation scoring method

[0055]

[0056] 2.3.2 Endpoint score (ES): The score for each chicken embryo = the sum of the degree of bleeding, coagulation and vascular lysis observed in each chicken embryo; the endpoint score (ES) = the sum of the scores of 6 chicken embryos.

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

[0058] Table 3 Endpoint scoring method evaluation results

[0059]

[0060] 3. Data results:

[0061] The test samples were selected using the stimulation scoring method. The results are shown in Table 4, which shows the stimulation scoring results of different active ingredients 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, the test sample "Huanfuyuan" has an IS score of 0.00 when the concentration is 0.45% (w / w), which is non-irritating; the test sample "salicylic acid" has an IS score of 12 or above when the concentration is 1% and 2% (w / w), which is strongly irritating / corrosive.

[0066] Based on Example 1 and Example 2, it can be seen that although Huanfuyuan is the filtrate of the fermentation product of Bifidobacterium / Lactobacillus / soybean seed extract, it is more gentle and hypoallergenic than other acid-based exfoliating ingredients.

[0067] The following examples and comparative examples focus on exploring the penetration enhancement effect of different penetration enhancers on hydroxypropyl tetrahydropyrantriol solution.

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

[0069] This study further explored the safety evaluation of hydroxypropyl tetrahydropyrantriol solution containing high concentration of "Huanfuyuan" Bifidobacterium / Lactobacillus / soybean seed extract fermentation product filtrate (Huanfuyuan) for consumers, that is, to further evaluate the safety of Huanfuyuan on humans.

[0070] Test method: 33 subjects were selected and their skin reactions were recorded. 20 μl of the sample from Example 3 was placed in the patch applicator. The control well served as a blank control (no substance was placed). Both the sample from Example 3 and the blank control were applied to the curved side of the subject's forearm. Gently press with the palm of your hand to evenly apply the sample to the skin for 24 hours. Skin reactions were observed according to the standards in the table below 30 minutes (after the indentation disappears), 24 hours, and 48 hours after removing the test substance patch applicator. The results were recorded.

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

[0072] Skin reaction grading standard for occlusive patch test

[0073]

[0074] Test results: Among 33 subjects, only one experienced a Grade 1 adverse reaction 0.5 hours after patch removal. Based on the latest "Technical Guidelines for Human Skin Patch Testing," if a patch test results in fewer than five Grade 1 reactions and no Grade 2 or higher reactions, it can generally be considered that the test product has not caused significant adverse reactions within the test range. In this patch test, the hydroxypropyl tetrahydropyrantriol solution containing a high-concentration Bifidobacterium / Lactobacillus / soybean seed extract fermentation product filtrate (Example 3) tested only one Grade 1 adverse reaction (i.e., only a mild erythema) at half an hour, which had completely subsided at 24 and 48 hours, with no further aggravation. This indicates that this example is relatively mild.

[0075] Test Example 4

[0076] This study was based on a suckling pig skin-Franz cell system. The concentration of hydroxypropyl tetrahydropyrantriol in the receiving fluid at different time points was determined by high performance liquid chromatography-mass spectrometry (HPLC-MS / MS). The skin penetration behavior of hydroxypropyl tetrahydropyrantriol by different penetration enhancers, including Huanfuyuan, was evaluated.

[0077] 4.1 Materials and reagents

[0078] This test used one-month-old piglet dorsal skin as the test model and a piglet skin-Franz cell system. PBS buffer (Solebol), Neosolue™-Aqulio (bis-diethoxydiglycol cyclohexane 1,4-dicarboxylate, sourced from Nippon Seika), hydroxypropyl tetrahydropyrantriol (S / R dual-configuration sample), and a high-performance liquid chromatograph (Thermo Fisher Scientific) were used.

[0079] 4.2 Test setup

[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 Steps

[0086] 4.4.1 Preparation of ex vivo suckling pig skin

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

[0088] 4.4.2 Penetration Test

[0089] 1) Fix the pig skin between the donor and receiver chambers of a Franz diffusion cell, with the stratum corneum facing the donor chamber and the dermis facing the receiver chamber.

[0090] 2) Add 7 mL of receiving solution to the receiving chamber. After tightening the pig skin, add 1 mL of receiving solution (water) to the receiving chamber through the sampler. Expel all air to ensure close contact between the dermis and the receiving solution.

[0091] 3) Sample loading: Add sample to the skin surface in the supply chamber. The effective penetration 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 center of the skin to the edge. Repeat 3 times for each sample in parallel.

[0092] 4) Infiltration: Turn on the electromagnetic stirrer and stir at 300 rpm. Maintain a constant temperature water bath at (32 ± 1)°C and ensure there are no bubbles in the water bath.

[0093] 5) Take sample liquid at 1h, 2h, 4h, 8h, and 24h respectively, use a sampler to extract 2mL of receiving liquid through the sampling tube, and then place it in a 2mL centrifuge tube. In addition, add an equal amount of receiving liquid after each sampling.

[0094] 4.4.3 Transdermal content test

[0095] After 24 hours of penetration, the target substance content in the skin, skin, and subcutaneous tissues of different experimental groups was tested by HPLC. The specific processing method is as follows:

[0096] 1) Testing the unpenetrated portion of the skin: After 24 hours, use a 1 mL pipette to repeatedly pipette acetonitrile to clean the skin surface. Wash five times with a total of 5 mL of acetonitrile, and place the solution in a 5 mL centrifuge tube for testing.

[0097] 2) Skin test: Use a sterile scalpel to cut the skin along the edge of the supply chamber, cut the skin into pieces, add 5mL acetonitrile and ultrasonically extract for 60 minutes. After the ultrasonication is completed, aspirate the supernatant and wait for the test.

[0098] 3) Subcutaneous test: At each time point, 2 mL of receiving solution was drawn from the receiving chamber and placed in a 2 mL centrifuge tube for testing.

[0099] 4.4.4 Statistics and Analysis

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

[0101] Where:

[0102] Q: cumulative penetration;

[0103] V: volume of receiving liquid in the receiving chamber V0: volume of each sampling;

[0104] Ci: sample concentration in the receiving solution from the first sampling to the last sampling;

[0105] Cn: Sample concentration measured at the nth sampling point.

[0106] 2) Permeation rate

[0107] The cumulative permeation amount (Y axis) is used to make a regression equation against the permeation time (X axis). The slope of the equation is the permeation rate K. P .

[0108] 3) Recovery rate

[0109] Recovery rate = total content (skin, skin, and subcutaneous tissue) / sample amount × 100%.

[0110] 4) Statistical analysis

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

[0112] 4.5 Test Results

[0113] 1) Cumulative penetration

[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 of cumulative penetration results

[0116]

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

[0118] 2) Permeation rate

[0119] A regression equation was made using the cumulative permeation amount (Y-axis) versus the permeation time (X-axis). The slope of the equation is the permeation rate Kp. The results are shown in Table 7 below. Table 7 shows the sample permeation rate results.

[0120] Table 7 Sample penetration rate results

[0121]

[0122] 3) Recovery rate:

[0123] The results of three parallel tests are summarized in Table 8, which is a statistical table of sample recovery rates.

[0124] Table 8 Sample recovery rate

[0125]

[0126] 4.6 Conclusion

[0127] Referring to Table 6, it can be seen that Huanfuyuan facilitates the penetration of hydroxypropyl tetrahydropyrantriol at test concentrations of 0.032% to 3.2%, and this penetration effect can be found to be significant within 1 hour of epidermal application. It is significantly better than typical chemical penetration enhancers and can also be significantly better than high-concentration fruit acid. The penetration amount gradually increases with time.

[0128] Referring to Table 7, we can see that the addition of Huanfuyuan increases the permeation rate of hydroxypropyl tetrahydropyrantriol by approximately 3-7 times, which is 1.3-3.3 times that of chemical penetration enhancers. Based on this, it can be seen that the filtrate of the fermentation product of Bifidobacterium / Lactobacillus / soybean seed extract, namely "Huanfuyuan", has a good effect on improving the permeation of water-soluble hydroxypropyl tetrahydropyrantriol, thereby helping it penetrate into the basal layer of the skin and even the dermis, thereby better exerting its anti-aging effects.

[0129] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

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

2. The use according to claim 1, characterized in that The concentration of the bifidobacterium / lactobacillus / soybean seed extract fermentation product filtrate in the product is 0.01 wt % to 5 wt %.

3. The use according to claim 1, characterized in that The concentration of the hydroxypropyl tetrahydropyrantriol in the product is 5 wt % to 15 wt %.

4. The use according to claim 1, characterized in that In the product, the mass ratio of the bifidobacterium / lactobacillus / soybean seed extract fermentation product filtrate to hydroxypropyl tetrahydropyrantriol is 0.01-5:5-15.

5. The use according to claim 1, characterized in that In the product, the mass ratio of the bifidobacterium / lactobacillus / soybean seed extract fermentation product filtrate to hydroxypropyl tetrahydropyrantriol is 0.01-5:10-11.

6. The use according to claim 1, characterized in that In the product, the mass ratio of the bifidobacterium / lactobacillus / soybean seed extract fermentation product filtrate to hydroxypropyl tetrahydropyrantriol is 0.032-3.2:10-11.

7. The use according to claim 1, characterized in that In the product, the mass ratio of the bifidobacterium / lactobacillus / soybean seed extract fermentation product filtrate to hydroxypropyl tetrahydropyrantriol is 0.032:10.5, 0.32:10.5 or 3.2:10.

5.

8. The use according to claim 1, characterized in that The product is a cosmetic.

9. The use according to claim 8, characterized in that The cosmetics are cleansing cosmetics.

10. The use according to claim 8, characterized in that The cosmetics are care cosmetics.

11. The use according to claim 8, characterized in that The cosmetics are beauty / decorative cosmetics.

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