Synergistic Compositions Based on Ginsenosides

Abstract

This invention discloses a synergistic composition based on ginsenosides, comprising ginsenosides and ergothioneine, wherein the weight ratio of ginsenosides to ergothioneine is 0.5-5:1-9, and the content of ginsenosides in the composition is 0.05-0.5 wt%. This invention also discloses the application of ginsenosides in inhibiting the photodegradation of ergothioneine, and the application of this composition in topical skin preparations.

Description

Technical Field

[0001] This invention relates to the field of cosmetic raw material technology, and to synergistic compositions based on ginsenosides, specifically to compositions based on synergistic inhibition of photodegradation by ginsenosides and ergothioneine. The compositions also have synergistic antioxidant effects and can be used as Collagen I promoters, Collagen III promoters, or Elastin promoters. In addition, this invention provides a technical method for stabilizing the decomposition of ergothioneine under light and high temperature conditions. Background Technology

[0002] Ginseng is a traditional Chinese medicine, and its uses and effects, such as regulating blood pressure, promoting metabolism, enhancing immunity, and promoting collagen production, have been extensively studied. The bioactive components of ginseng include saponins, polysaccharides, and phenols, among which ginsenosides have various effects such as anti-inflammatory, blood pressure-lowering, and anti-cancer properties, and are receiving increasing attention.

[0003] Current research on the structure, configuration, structure-activity relationship, pharmacodynamics, and kinetic properties of ginsenosides is gradually deepening. Structurally, ginsenosides consist of a steroidal skeleton and glycosyl groups linked at C3, C6, and C20 positions, forming different types such as Rb1, Rg1, Rg3, and Rd. However, rare ginsenosides exhibit stronger pharmacodynamics and superior pharmacokinetic properties compared to common ginsenosides. Common ginsenosides show significantly improved oral bioavailability and bioactivity after intestinal metabolism and deglycosylation. Although the human gut microbiota can convert ginsenosides, the conversion efficiency is low, with significant individual differences, and most conversion occurs in the large intestine, affecting the absorption of rare ginsenosides. Besides relying on the gut microbiota, ginsenoside conversion can also be achieved through hot water hydrolysis, acid hydrolysis, alkaline hydrolysis, microbial methods, and enzymatic methods. Heat treatment can alter the composition of ginsenosides, reducing the content of common ginsenosides Rb1, Rc, Rd, Re, and Rg1, while increasing the content of rarer ginsenosides such as Rg5, Rk1, Rk2, Rk3, and Rg3. These newly formed ginsenosides have lower polarity and stronger biological activity. Commercially available red and black ginseng are products of partial ginsenoside conversion after high-temperature treatment. Combining high temperature with acid treatment can promote oxidation, hydrolysis, and dehydration reactions, thereby improving the ginsenoside conversion efficiency.

[0004] Ginsenoside Rg3, CAS No. 14197-60-5, molecular weight 785.01. Based on the above-mentioned medicinal effects, this invention utilizes ginsenosides extracted from black ginseng, in powder form. It contains a high content of ginsenosides, particularly the rare ginsenoside Rg3. Applying ginsenosides to cosmetics can promote fibroblast proliferation and collagen synthesis, thereby delaying skin aging. Its main mechanism of action is through continuously activating the collagen synthesis process of fibroblasts.

[0005] Ginsenosides are large-molecule, hygroscopic white crystals with a distinctive odor. Due to their structural characteristics, they are unstable in the external environment, easily undergoing oxidation and degradation reactions, and have limited ability to penetrate the stratum corneum, resulting in low bioavailability. To address these limitations in the application of ginsenosides, this invention aims to combine appropriate component technologies to improve their chemical stability, facilitate the formulation of pharmaceutical preparations and cosmetics, and maximize their biological activity.

[0006] Ergothioneine (EGT) is a natural super antioxidant, CAS 497-30-3, molecular formula C9H 16 N3O2S + With a molecular weight of 230.10, its scientific name is 2-mercapto-histidine-trimethyl inner salt, also known as ergotamine or ergot sulfur base. It is a histidine derivative containing a thiol group. The pure product is a white crystal, easily soluble in water, with a melting point of 275-277℃.

[0007] Ergothionein was isolated from ergot in 1909, hence its name. It has been found to be synthesized in most fungi, some mushrooms, streptococci, mycobacteria, and other microorganisms, and can be absorbed and accumulated by plants and animals.

[0008] Ergothioneine is currently obtained through biosynthesis, exhibiting good safety, high fermentation efficiency, and high purity. Its antioxidant properties are demonstrated by targeting various oxidative reaction pathways within the skin, such as those involving singlet oxygen, superoxide ions, and oxygen free radicals, as well as metabolic pathways, showing a dose-response relationship. Literature reports that its antioxidant activity increases with increasing ergothioneine content. Furthermore, compared to several reported natural antioxidants, EGT-containing agents reduce cellular peroxides by 16%, while idebenone-containing agents reduce them by only 5%. Similarly, EGT exhibits excellent antioxidant properties even at extremely low concentrations, while antioxidants like coenzyme Q10 cannot achieve the same antioxidant effect at low concentrations. The primary energy source for human cells is adenosine triphosphate (ATP), generated in the mitochondria through fatty acid conversion, which inevitably leads to the production of free radicals. Ergothioneine, as a super antioxidant, is the only antioxidant with a clearly defined mechanism capable of repairing mitochondria. The human body cannot synthesize ergothioneine on its own and must obtain it from external sources.

[0009] Ergothioneine is a water-soluble amino acid molecule. A unique transport protein in the human body, OCTN1, can transport it to the desired site. In aqueous solution, ergothioneine exists as a tautomer of thiols and thioketones. This means that different pH conditions and concentrations can alter the conformation of ergothioneine in the application system, leading to problems such as odor and spoilage, inevitably posing challenges to its application.

[0010] Inhibiting the photodegradation (improving photostability), odor, and discoloration of ergothionein has always been a difficult problem and challenge for the industry in applying this ingredient.

[0011] In the prior art, Chinese patent application CN116370334A discloses an anti-aging composition and its application. The composition comprises components in the following mass ratio: bakuchiol: RAR agonist: ergothioneine: ginsenoside F11 = 1:0.1-10:0.1-5:0.5-15, with ergothioneine content at 1000-2000 ppm. By adding the active ingredient RAR agonist to bakuchiol, the anti-aging efficacy of bakuchiol is enhanced. The addition of ergothioneine increases the anti-aging effect of the composition. Simultaneously, ergothioneine increases the antioxidant rate of bakuchiol. It also includes ginsenoside F11, which has a synergistic effect with the other components. It has significant anti-aging effects, is gentle, and comfortable to use, making it widely applicable in cosmetics and other beauty and skincare products, and is of great significance in the research and development of anti-aging products. The application did not provide evidence that ginsenosides can help inhibit the photodegradation of ergothioneine, or that they can help ergothioneine synergistically enhance its antioxidant or collagen-promoting effects.

[0012] This invention unexpectedly discovered that under different pH conditions and concentrations, the content of ergothioneine in the system was significantly increased, and at the same time, it unexpectedly synergistically enhanced the antioxidant effect and the collagen-promoting effect of ginsenosides, which has broad application potential in the cosmetic field. Summary of the Invention

[0013] The purpose of this invention is to provide a combination of ergothioneine and ginsenosides, offering an extremely efficient and cost-effective method for inhibiting the photodecomposition and high-temperature instability of ergothioneine. Unexpectedly, under different pH conditions and concentrations, it significantly increases the content of ergothioneine in the system, while also unexpectedly enhancing its antioxidant effects and the collagen-promoting effects of ginsenosides. This invention has broad application potential in the cosmetics field.

[0014] This invention provides a synergistic composition based on ginsenosides, comprising: ginsenosides and ergothioneine, wherein the weight ratio of ginsenosides to ergothioneine is 0.5-5:1-9, and wherein the content of ginsenosides in the composition is 0.05-0.5 wt%.

[0015] In a preferred embodiment, the weight ratio of ginsenosides to ergothioneine is 0.5-5:1-2.

[0016] In a preferred embodiment, the composition further comprises ≤5 wt% of a C3-C6 diol.

[0017] Preferably, the C3-C6 diols are selected from: butanediol, propylene glycol, 1,3-propanediol, pentanediol, 1,2-hexanediol, glycerol, diglycerol, etc.

[0018] In a preferred embodiment, the composition further comprises ≤0.8 wt% of acrylamide dimethyl taurate ammonium / VP copolymer.

[0019] In a preferred embodiment, the composition further comprises ≤2 wt% bis-diethoxydiethylenecyclohexane 1,4-dicarboxylic acid ester.

[0020] In a preferred embodiment, the composition may further comprise hyaluronic acid.

[0021] Preferably, the hyaluronic acid has a molecular weight of 3K-10KDa.

[0022] Preferably, the weight ratio of hyaluronic acid to ginsenoside is 1:1.

[0023] In a preferred embodiment, the composition further comprises a carrier acceptable in the field of topical skin agents.

[0024] Preferably, water is an acceptable carrier in the field of topical skin agents.

[0025] Preferably, the water content in the composition is 91-95 wt%.

[0026] In a preferred embodiment, the pH value of the composition is 4-8.

[0027] Preferably, the pH value of the composition is 4-6.

[0028] This invention also provides the application of ginsenosides in inhibiting the photodegradation of ergothionein.

[0029] In a preferred embodiment, the weight ratio of ginsenosides to ergothioneine is 0.5-5:1-2.

[0030] In a preferred embodiment, the application is performed at a pH value of 4-8.

[0031] Preferably, the application is performed at a pH value of 4-6.

[0032] The present invention also provides the antioxidant use of ginsenoside-based synergistic compositions, wherein the compositions comprise ginsenosides and ergothioneine in a weight ratio of 0.5-5:1-2.

[0033] In a preferred embodiment, the pH value of the composition is 4-6.

[0034] The present invention also provides the use of ginsenoside-based synergistic compositions as Collagen I promoters, or Collagen III promoters, or Elastin promoters, wherein the compositions comprise ginsenosides and ergothioneine in a weight ratio of 0.5-5:1-2.

[0035] In a preferred embodiment, the pH value of the composition is 4-6.

[0036] This invention also provides the application of synergistic compositions based on ginsenosides in topical skin preparations.

[0037] In a preferred embodiment, the topical skin agent is selected from: face cream, lotion, gel, toner, serum, face mask, eye cream, aerosol (cleansing foam), spray, shower gel, facial cleanser, and essence water.

[0038] In a preferred embodiment, the composition is used in a topical skin preparation at an amount of 0.0001 wt%-90 wt%.

[0039] The preferred weight percentage is 0.001wt%-10wt%.

[0040] A more preferred weight percentage is 0.001wt%-5wt%.

[0041] The beneficial effects of this invention are:

[0042] 1. This invention is the first to protect the stability of ergothioneine using ginsenosides as a component. Through the special function of the chemical structure, the conformational stability of ergothioneine under light and high temperature can be protected.

[0043] 2. This invention unexpectedly discovered that ginsenosides and ergothioneine have a very good synergistic effect, which can improve the antioxidant efficacy by more than 78%.

[0044] 3. This invention unexpectedly discovered that ginsenosides and ergothioneine have a very good synergistic effect, which can improve the ability to promote skin collagen production by more than 48%.

[0045] 4. This invention unexpectedly discovered that ginsenosides and ergothioneine have a very good synergistic effect, which can improve the ability to promote penetration by more than 50%. Attached Figure Description

[0046] Figure 1 The results of ROS content detection in test example 4.

[0047] Figure 2 This is a schematic diagram of the diffusion cell operation. Detailed Implementation

[0048] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. While any methods and materials similar or equivalent to those described herein may be used to practice or test the invention, preferred methods and materials are described herein. For the purposes of this invention, the following terms are defined.

[0049] As used herein, the term "about" means a quantity, level, value, dimension, size, or amount that differs from that of a reference by as much as 30%, 20%, or 10%. Percentages used herein, unless otherwise stated, are by weight.

[0050] Throughout this specification and claims, unless otherwise required, the words “comprising” and its variations “containing” and “including” shall be understood to mean including the said whole or step, or a group of whole or steps, but not excluding any other whole or step, or other group of whole or steps.

[0051] C3-C6 diols

[0052] The C3-C6 diol mentioned in this invention is butanediol, and the C3-C6 diol can be selected from: butanediol, propylene glycol, 1,3-propanediol, pentanediol, 1,2-hexanediol, glycerol, diglycerol, etc.

[0053] The present invention is further illustrated below with reference to specific embodiments. However, it should be understood that these embodiments are for illustrative purposes only and do not constitute a limitation on the scope of the invention. Test methods in the following embodiments, unless otherwise specified, are generally performed under conventional conditions or as recommended by the manufacturer. Unless otherwise stated, all percentages and parts are by weight.

[0054] The main experimental materials and reagents used in the examples are as follows:

[0055] 1. Ginsenosides, purity ≥99.0%, Shanghai Kunqian Biotechnology Co., Ltd.

[0056] 2. Ergothioneine, purity ≥ 99.0%, Shanghai Ergothioneine Co., Ltd.

[0057] 3. Tromethamine, analytical grade, Sinopharm Group

[0058] 4. Citric acid, analytical grade, Sinopharm Group

[0059] 5. Acrylamide dimethyl taurate ammonium / VP copolymer (AVC), purity ≥99.0%, Clariant (China) Co., Ltd.

[0060] 6. Hydroxypropyl β-cyclodextrin, purity ≥99%, Shandong Binzhou Zhiyuan Biotechnology Co., Ltd.

[0061] 7. bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylic acid ester, purity >99.0%, Nippon Seika Co., Ltd.

[0062] 8. Hydrolyzed hyaluronic acid (molecular weight 3K-10KDa) (purity ≥99.0%, Bloomage Biotechnology Co., Ltd.)

[0063] 9. Sodium hyaluronate (molecular weight 10K-100KDa) (purity ≥99.0%, Bloomage Biotechnology Co., Ltd.)

[0064] 10. Hyaluronic acid (molecular weight 100K-1000KDa) (purity ≥99.0%, Bloomage Biotechnology Co., Ltd.)

[0065] 11. Formic acid, purity ≥ 98.0%, Sigma-Aldrich (Product No. 43804)

[0066] 12. Acetonitrile, purity ≥ 99.9%, Sigma-Aldrich (Product No. 34851)

[0067] 13. Methanol, purity ≥ 99.9%, Sigma-Aldrich (Product No. 34885)

[0068] 14. PBS (Beijing Solarbio Science & Technology Co., Ltd., batch number P1022)

[0069] 15. DCFH-DA culture medium, LMAI Bio (Shanghai Lianmai Biotechnology Co., Ltd.)

[0070] 16. Pancreatic enzyme, Beijing Solarbio Science & Technology Co., Ltd., product batch number T8151

[0071] 17. Methylglyoxal, 40% aqueous solution, Sigma-Aldrich, lot number M0252

[0072] Main experimental instruments

[0073] 1. Waters ARC high-performance liquid chromatograph with diode array detector (Waters)

[0074] 2. XS205 analytical balance (METTLER TOLEDO)

[0075] 3. KQ-800DE CNC Ultrasonic Cleaner (Kunshan Ultrasonic Instrument Co., Ltd.)

[0076] 4. CO2 incubator (Thermo, 150I)

[0077] 5. Clean bench (Suzhou Antai, SW-CJ-1F)

[0078] 6. Flow cytometer (Beckman, CytoFLEX)

[0079] 7. Inverted microscope (Olympus, CKX53)

[0080] 8. Incucell incubator (MMM GmbH, Germany)

[0081] 9. PERCIVAL CU41L5 Illuminated Incubator (PERCIVAL Corporation, USA)

[0082] 10. Microplate reader (BioTek, Epoch)

[0083] 11. Real-time PCR instrument (BioRad, CFX-96)

[0084] 12. Franz cell diffusion cell system (Shanghai Huanghai Pharmaceutical Testing Instrument Factory, TK6-500)

[0085] Example 1:

[0086] Accurately weigh 0.9g of ergothioneine and dissolve it in 5g of butanediol. Add 0.1g of ginsenosides and stir well. Then add 2.0g of bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylic acid ester and stir thoroughly. Add 0.8g of AVC and disperse thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 4.0 using citric acid and tromethamine. Pour into a volumetric flask and set aside for later use.

[0087] Example 2:

[0088] Accurately weigh 0.7g of ergothioneine and dissolve it in 5g of butanediol. Add 0.1g of ginsenosides and stir well. Then add 2.0g of bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylic acid ester and stir thoroughly. Add 0.8g of AVC and disperse thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 4.0 using citric acid and tromethamine. Pour into a volumetric flask and set aside for later use.

[0089] Example 3:

[0090] Accurately weigh 0.5g of ergothioneine and dissolve it in 5g of butanediol. Add 0.1g of ginsenosides and stir well. Then add 2.0g of bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylic acid ester and stir thoroughly. Add 0.8g of AVC and disperse thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 4.0 using citric acid and tromethamine. Pour into a volumetric flask and set aside for later use.

[0091] Example 4:

[0092] Accurately weigh 0.1g of ergothioneine and dissolve it in 5g of butanediol. Add 0.05g of ginsenosides and stir until homogeneous. Then add 2.0g of bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylic acid ester and stir thoroughly until homogeneous. Add 0.8g of AVC and disperse thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 4.0 using citric acid and tromethamine. Pour into a volumetric flask and set aside for later use.

[0093] Example 5:

[0094] Accurately weigh 0.1g of ergothioneine and dissolve it in 5g of butanediol. Add 0.1g of ginsenosides and stir until homogeneous. Then add 2.0g of bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylic acid ester and stir thoroughly until homogeneous. Add 0.8g of AVC and disperse thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 4.0 using citric acid and tromethamine. Pour into a volumetric flask and set aside for later use.

[0095] Example 6:

[0096] Accurately weigh 0.1g of ergothioneine and dissolve it in 5g of butanediol. Add 0.2g of ginsenosides and stir until homogeneous. Then add 2.0g of bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylic acid ester and stir thoroughly until homogeneous. Add 0.8g of AVC and disperse thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 4.0 using citric acid and tromethamine. Pour into a volumetric flask and set aside for later use.

[0097] Example 7:

[0098] Accurately weigh 0.1g of ergothioneine and dissolve it in 5g of butanediol. Add 0.5g of ginsenosides and stir well. Then add 2.0g of bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylic acid ester and stir thoroughly. Add 0.8g of AVC and disperse thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 4.0 using citric acid and tromethamine. Pour into a volumetric flask and set aside for later use.

[0099] Example 8:

[0100] Accurately weigh 0.2g of ergothioneine and dissolve it in 5g of butanediol. Add 0.3g of ginsenosides and stir well. Then add 2.0g of bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylic acid ester and stir thoroughly. Add 0.8g of AVC and disperse thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 4.0 using citric acid and tromethamine. Pour into a volumetric flask and set aside for later use.

[0101] Example 9:

[0102] Accurately weigh 0.1g of ergothioneine and dissolve it in 5g of butanediol. Add 0.2g of hydroxypropyl β-cyclodextrin and stir until homogeneous and clear. Add 0.8g of AVC and disperse it thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 4.0 using citric acid and tromethamine. Pour the solution into a volumetric flask and set aside for later use.

[0103] Example 10:

[0104] Accurately weigh 0.1g of ergothioneine and dissolve it in 5g of butanediol. Add 0.1g of hydrolyzed hyaluronic acid (molecular weight 3K-10KDa), stir well, add 0.8g of AVC and disperse thoroughly until uniform. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 4.0 using citric acid and tromethamine. Pour into a volumetric flask and set aside for later use.

[0105] Example 11:

[0106] Accurately weigh 0.1g of ergothioneine and dissolve it in 5g of butanediol. Add 0.1g of sodium hyaluronate (molecular weight 10K-100KDa) and stir until homogeneous and clear. Add 0.8g of AVC and disperse it thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 4.0 using citric acid and tromethamine. Pour into a volumetric flask and set aside for later use.

[0107] Example 12:

[0108] Accurately weigh 0.1g of ergothioneine and dissolve it in 5g of butanediol. Add 0.1g of hyaluronic acid (molecular weight 3K-10KDa) and 0.1g of ginsenosides, and stir until homogeneous and clear. Add 0.8g of AVC and disperse it thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 4.0 using citric acid and tromethamine. Pour into a volumetric flask and set aside for later use.

[0109] Example 13:

[0110] Accurately weigh 0.1g of ergothioneine and dissolve it in 5g of butanediol. Add 0.1g of hyaluronic acid (molecular weight 100K-1000KDa) and stir until homogeneous and clear. Add 0.8g of AVC and disperse it thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 4.0 using citric acid and tromethamine. Pour the solution into a volumetric flask and set aside for later use.

[0111] Example 14:

[0112] Accurately weigh 0.1g of ergothioneine and dissolve it in 5g of butanediol. Stir until homogeneous and clear. Add 0.8g of AVC and disperse thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 4.0 using citric acid and tromethamine. Transfer the solution to a volumetric flask for later use.

[0113] Example 15:

[0114] Accurately weigh 0.9g of ergothioneine and dissolve it in 5g of butanediol. Add 0.1g of ginsenosides and stir well. Then add 2.0g of bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylic acid ester and stir thoroughly. Add 0.8g of AVC and disperse thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 6.0 using citric acid and tromethamine. Pour into a volumetric flask and set aside for later use.

[0115] Example 16:

[0116] Accurately weigh 0.7g of ergothioneine and dissolve it in 5g of butanediol. Add 0.1g of ginsenosides and stir well. Then add 2.0g of bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylic acid ester and stir thoroughly. Add 0.8g of AVC and disperse thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 6.0 using citric acid and tromethamine. Pour into a volumetric flask and set aside for later use.

[0117] Example 17:

[0118] Accurately weigh 0.5g of ergothioneine and dissolve it in 5g of butanediol. Add 0.1g of ginsenosides and stir well. Then add 2.0g of bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylic acid ester and stir thoroughly. Add 0.8g of AVC and disperse thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 6.0 using citric acid and tromethamine. Pour into a volumetric flask and set aside for later use.

[0119] Example 18:

[0120] Accurately weigh 0.1g of ergothioneine and dissolve it in 5g of butanediol. Add 0.05g of ginsenosides and stir until homogeneous. Then add 2.0g of bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylic acid ester and stir thoroughly until homogeneous. Add 0.8g of AVC and disperse thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 6.0 using citric acid and tromethamine. Pour into a volumetric flask and set aside for later use.

[0121] Example 19:

[0122] Accurately weigh 0.1g of ergothioneine and dissolve it in 5g of butanediol. Add 0.1g of ginsenosides and stir until homogeneous. Then add 2.0g of bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylic acid ester and stir thoroughly until homogeneous. Add 0.8g of AVC and disperse thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 6.0 using citric acid and tromethamine. Pour into a volumetric flask and set aside for later use.

[0123] Example 20:

[0124] Accurately weigh 0.1g of ergothioneine and dissolve it in 5g of butanediol. Add 0.2g of ginsenosides and stir well. Then add 2.0g of bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylic acid ester and stir thoroughly. Add 0.8g of AVC and disperse thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 6.0 using citric acid and tromethamine. Pour into a volumetric flask and set aside for later use.

[0125] Example 21:

[0126] Accurately weigh 0.1g of ergothioneine and dissolve it in 5g of butanediol. Add 0.5g of ginsenosides and stir well. Then add 2.0g of bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylic acid ester and stir thoroughly. Add 0.8g of AVC and disperse thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 6.0 using citric acid and tromethamine. Pour into a volumetric flask and set aside for later use.

[0127] Example 22:

[0128] Accurately weigh 0.2g of ergothioneine and dissolve it in 5g of butanediol. Add 0.3g of ginsenosides and stir well. Then add 2.0g of bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylic acid ester and stir thoroughly. Add 0.8g of AVC and disperse thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 6.0 using citric acid and tromethamine. Pour into a volumetric flask and set aside for later use.

[0129] Example 23:

[0130] Accurately weigh 0.1g of ergothioneine and dissolve it in 5g of butanediol. Add 0.2g of hydroxypropyl β-cyclodextrin and stir until homogeneous and clear. Add 0.8g of AVC and disperse it thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 6.0 using citric acid and tromethamine. Pour the solution into a volumetric flask and set aside for later use.

[0131] Example 24:

[0132] Accurately weigh 0.1g of ergothioneine and dissolve it in 5g of butanediol. Add 0.1g of hydrolyzed hyaluronic acid (molecular weight 3K-10KDa), stir well, then add 0.8g of AVC and disperse thoroughly until uniform. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 6.0 using citric acid and tromethamine. Pour into a volumetric flask and set aside for later use.

[0133] Example 25:

[0134] Accurately weigh 0.1g of ergothioneine and dissolve it in 5g of butanediol. Add 0.1g of sodium hyaluronate (molecular weight 10K-100KDa) and stir until homogeneous and clear. Add 0.8g of AVC and disperse it thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 6.0 using citric acid and tromethamine. Pour the solution into a volumetric flask and set aside for later use.

[0135] Example 26:

[0136] Accurately weigh 0.1g of ergothioneine and dissolve it in 5g of butanediol. Add 0.1g of hyaluronic acid (molecular weight 3K-10KDa) and 0.1g of ginsenosides, and stir until homogeneous and clear. Add 0.8g of AVC and disperse it thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 6.0 using citric acid and tromethamine. Pour into a volumetric flask and set aside for later use.

[0137] Example 27:

[0138] Accurately weigh 0.1g of ergothioneine and dissolve it in 5g of butanediol. Add 0.1g of hyaluronic acid (molecular weight 100K-1000KDa) and stir until homogeneous and clear. Add 0.8g of AVC and disperse it thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 6.0 using citric acid and tromethamine. Pour the solution into a volumetric flask and set aside for later use.

[0139] Example 28:

[0140] Accurately weigh 0.1g of ergothioneine and dissolve it in 5g of butanediol. Stir until homogeneous and clear. Add 0.8g of AVC and disperse thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 6.0 using citric acid and tromethamine. Transfer the solution to a volumetric flask for later use.

[0141] Example 29:

[0142] Accurately weigh 0.9g of ergothioneine and dissolve it in 5g of butanediol. Add 0.1g of ginsenosides and stir well. Then add 2.0g of bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylic acid ester and stir thoroughly. Add 0.8g of AVC and disperse thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 8.0 using citric acid and tromethamine. Pour into a volumetric flask and set aside for later use.

[0143] Example 30:

[0144] Accurately weigh 0.7g of ergothioneine and dissolve it in 5g of butanediol. Add 0.1g of ginsenosides and stir well. Then add 2.0g of bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylic acid ester and stir thoroughly. Add 0.8g of AVC and disperse thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 8.0 using citric acid and tromethamine. Pour into a volumetric flask and set aside for later use.

[0145] Example 31:

[0146] Accurately weigh 0.5g of ergothioneine and dissolve it in 5g of butanediol. Add 0.1g of ginsenosides and stir well. Then add 2.0g of bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylic acid ester and stir thoroughly. Add 0.8g of AVC and disperse thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 8.0 using citric acid and tromethamine. Pour into a volumetric flask and set aside for later use.

[0147] Example 32:

[0148] Accurately weigh 0.1g of ergothioneine and dissolve it in 5g of butanediol. Add 0.05g of ginsenosides and stir until homogeneous. Then add 2.0g of bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylic acid ester and stir thoroughly until homogeneous. Add 0.8g of AVC and disperse thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 8.0 using citric acid and tromethamine. Pour into a volumetric flask and set aside for later use.

[0149] Example 33:

[0150] Accurately weigh 0.1g of ergothioneine and dissolve it in 5g of butanediol. Add 0.1g of ginsenosides and stir well. Then add 2.0g of bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylic acid ester and stir thoroughly. Add 0.8g of AVC and disperse thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 8.0 using citric acid and tromethamine. Pour into a volumetric flask and set aside for later use.

[0151] Example 34:

[0152] Accurately weigh 0.1g of ergothioneine and dissolve it in 5g of butanediol. Add 0.2g of ginsenosides and stir well. Then add 2.0g of bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylic acid ester and stir thoroughly. Add 0.8g of AVC and disperse thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 8.0 using citric acid and tromethamine. Pour into a volumetric flask and set aside for later use.

[0153] Example 35:

[0154] Accurately weigh 0.1g of ergothioneine and dissolve it in 5g of butanediol. Add 0.5g of ginsenosides and stir until homogeneous. Then add 2.0g of bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylic acid ester and stir thoroughly until homogeneous. Add 0.8g of AVC and disperse thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 8.0 using citric acid and tromethamine. Pour into a volumetric flask and set aside for later use.

[0155] Example 36:

[0156] Accurately weigh 0.2g of ergothioneine and dissolve it in 5g of butanediol. Add 0.3g of ginsenosides and stir well. Then add 2.0g of bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylic acid ester and stir thoroughly. Add 0.8g of AVC and disperse thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 8.0 using citric acid and tromethamine. Pour into a volumetric flask and set aside for later use.

[0157] Example 37:

[0158] Accurately weigh 0.1g of ergothioneine and dissolve it in 5g of butanediol. Add 0.2g of hydroxypropyl β-cyclodextrin and stir until homogeneous and clear. Add 0.8g of AVC and disperse it thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 8.0 using citric acid and tromethamine. Pour into a volumetric flask and set aside for later use.

[0159] Example 38:

[0160] Accurately weigh 0.1g of ergothioneine and dissolve it in 5g of butanediol. Add 0.1g of hydrolyzed hyaluronic acid (molecular weight 3K-10KDa), stir well, add 0.8g of AVC and disperse thoroughly until uniform. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 8.0 using citric acid and tromethamine. Pour into a volumetric flask and set aside for later use.

[0161] Example 39:

[0162] Accurately weigh 0.1g of ergothioneine and dissolve it in 5g of butanediol. Add 0.1g of sodium hyaluronate (molecular weight 10K-100KDa) and stir until homogeneous and clear. Add 0.8g of AVC and disperse it thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 8.0 using citric acid and tromethamine. Pour the solution into a volumetric flask and set aside for later use.

[0163] Example 40:

[0164] Accurately weigh 0.1g of ergothioneine and dissolve it in 5g of butanediol. Add 0.1g of hyaluronic acid (molecular weight 3K-10KDa) and 0.1g of ginsenosides, and stir until homogeneous and clear. Add 0.8g of AVC and disperse it thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 8.0 using citric acid and tromethamine. Pour into a volumetric flask and set aside for later use.

[0165] Example 41:

[0166] Accurately weigh 0.1g of ergothioneine and dissolve it in 5g of butanediol. Add 0.1g of hyaluronic acid (molecular weight 100K-1000KDa) and stir until homogeneous and clear. Add 0.8g of AVC and disperse it thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 8.0 using citric acid and tromethamine. Pour the solution into a volumetric flask and set aside for later use.

[0167] Example 42:

[0168] Accurately weigh 0.1g of ergothioneine and dissolve it in 5g of butanediol. Stir until homogeneous and clear. Add 0.8g of AVC and disperse thoroughly until homogeneous. Add water to adjust the total volume to 100g of aqueous solution. Adjust the pH of the solution to 8.0 using citric acid and tromethamine. Pour into a volumetric flask and set aside.

[0169] Test Example 1: Illumination Test Experiment

[0170] Test samples: Composition samples prepared in Examples 1-42

[0171] Product stability refers to the stability of a product in an inert container. In this experiment, a transparent glass bottle with a cylindrical diameter of about 5 cm was used. The composition samples prepared in Examples 1-42 were placed in the glass bottle and the cap was tightened. The bottle was continuously irradiated in a light test light box for 14 days. The test light box was equipped with a set of lamps, and the sample was about 30-40 cm away from the lamps. A suitable lamp for this purpose is POLARIT (daylight type), a 40-watt (Thorn-EMI) lamp tube with a length of 132 cm. A set of 12 lamps provides the light intensity and spectral distribution to simulate sunlight.

[0172] The aroma characteristics and intensity of the above samples were evaluated by professional fragrance tasters using direct olfaction. This evaluation method can be used to determine the stability and degree of change of the aroma. A three-level grading standard was adopted.

[0173] Classification Odor and strength standards Level 1 No discernible change in taste Level 2 Slight change in taste Level 3 Significant taste change

[0174] The results of the composition samples prepared in Examples 1-42 after 14 days of light exposure testing are shown in Table 1:

[0175] Table 1. Results of light irradiation tests on the composition samples prepared in Examples 1-42.

[0176]

[0177] After 14 days of simulated sunlight exposure testing, under acidic conditions (pH=4), the composition samples prepared in Examples 4-8, which contained ergothioneine, butylene glycol, AVC, ginsenosides, and water, showed no perceptible change in taste; the composition samples prepared in Examples 1-3 showed only slight changes in taste; and the composition sample prepared in Example 12, which contained ergothioneine, butylene glycol, AVC, ginsenosides, hyaluronic acid (molecular weight 3K-10KDa), and water, showed no perceptible change in taste.

[0178] At the same pH value (pH=4), when all or part of the ginsenosides in the composition were replaced with hydroxypropyl β-cyclodextrin, hydrolyzed hyaluronic acid (molecular weight 3K-10KDa), sodium hyaluronate (molecular weight 10K-100KDa), hyaluronic acid (molecular weight 3K-10KDa), or hyaluronic acid (molecular weight 100K-1000KDa), the composition samples showed different light irradiation test performance.

[0179] Specifically as follows:

[0180] Example 9 prepared a composition sample containing ergothioneine, butylene glycol, AVC, hydroxypropyl β-cyclodextrin, and water. Example 6 prepared a composition sample containing ergothioneine, butylene glycol, AVC, ginsenosides, bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylate, and water. Except that the former had the same hydroxypropyl β-cyclodextrin content as the latter, and the latter also contained an additional 2 wt% of bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylate, and with the remaining components in both samples having the same content, under light irradiation testing, the composition sample prepared in Example 6 performed significantly better than the composition sample prepared in Example 9. This indicates that the combination of ginsenosides and bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylate has a positive stabilizing effect on ergothioneine in the composition.

[0181] Example 10 prepared a composition sample containing ergothioneine, butylene glycol, AVC, hydrolyzed hyaluronic acid (molecular weight 3K-10KDa), and water. Example 5 prepared a composition sample containing ergothioneine, butylene glycol, AVC, ginsenosides, bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylate, and water. Except that the former had the same content of hydrolyzed hyaluronic acid (molecular weight 3K-10KDa) as the latter, the latter also contained an additional 2wt% of bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylate. With the remaining components in both examples having the same content, under light irradiation testing, the composition sample prepared in Example 5 performed significantly better than the composition sample prepared in Example 10. This indicates that the combination of ginsenosides and bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylate has a positive stabilizing effect on ergothioneine in the composition. Furthermore, it also shows that hydrolyzed hyaluronic acid (molecular weight 3K-10KDa) cannot inhibit the photodegradation of ergothioneine.

[0182] Example 11 prepared a composition sample containing ergothioneine, butylene glycol, AVC, sodium hyaluronate (molecular weight 10K-100KDa), and water. Example 5 prepared a composition sample containing ergothioneine, butylene glycol, AVC, ginsenosides, bis-diethoxydiethylene cyclohexane 1,4-dicarboxylate, and water. Except that the former had the same sodium hyaluronate (molecular weight 10K-100KDa) content and the latter had the same ginsenoside content, the latter also contained 2 wt% bis-diethoxydiethylene cyclohexane 1,4-dicarboxylate. With the remaining components in both examples having the same content, under light irradiation testing, the composition sample prepared in Example 5 performed significantly better than the composition sample prepared in Example 11. This indicates that the combination of ginsenosides and bis-diethoxydiethylene cyclohexane 1,4-dicarboxylate can help inhibit the photodegradation of ergothioneine in the composition. Furthermore, it was also stated that sodium hyaluronate (molecular weight 10K-100KDa) does not inhibit the photodegradation of ergothionein as disclosed in the prior art.

[0183] Example 12 prepared a composition sample containing ergothioneine, butylene glycol, AVC, hyaluronic acid (molecular weight 3K-10KDa), ginsenosides, and water. Under light irradiation, no noticeable change in taste was observed. This indicates that the combined use of ginsenosides and hyaluronic acid (molecular weight 3K-10KDa) can also inhibit the photodegradation of ergothioneine.

[0184] Example 13 prepared a composition sample containing ergothioneine, butylene glycol, AVC, hyaluronic acid (molecular weight 100K-1000KDa), and water. Example 5 prepared a composition sample containing ergothioneine, butylene glycol, AVC, ginsenosides, bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylate, and water. Except that the former contained the same amount of sodium hyaluronate (molecular weight 10K-100KDa) and the latter contained the same amount of ginsenosides, the latter also contained 2 wt% of bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylate. With the remaining components present in the same amounts, under light irradiation testing, the composition sample prepared in Example 5 performed significantly better than that prepared in Example 13. This indicates that the combination of ginsenosides and bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylate can help inhibit the photodegradation of ergothioneine in the composition. Furthermore, it was also stated that hyaluronic acid (molecular weight 100K-1000KDa) cannot help inhibit the photodegradation of ergothionein.

[0185] After 14 days of simulated sunlight exposure testing, under acidic conditions (pH=6), the composition samples containing ergothioneine, butylene glycol, AVC, ginsenosides, and water prepared in Examples 18-19 and 21-22 showed no perceptible change in taste; the composition samples prepared in Examples 16-17 and 20 showed only slight changes in taste; and the composition sample prepared in Example 15 showed a significant change in taste.

[0186] At the same pH value (pH=6), when all or part of the ginsenosides in the composition were replaced with hydroxypropyl β-cyclodextrin, hydrolyzed hyaluronic acid (molecular weight 3K-10KDa), sodium hyaluronate (molecular weight 10K-100KDa), or hyaluronic acid (molecular weight 100K-1000KDa), the composition samples showed different light irradiation test performance.

[0187] Specifically as follows:

[0188] Example 23 prepared a composition sample containing ergothioneine, butylene glycol, AVC, hydroxypropyl β-cyclodextrin, and water. Example 20 prepared a composition sample containing ergothioneine, butylene glycol, AVC, ginsenosides, bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylate, and water. Except that the former had the same hydroxypropyl β-cyclodextrin content as the latter, the latter also contained an additional 2 wt% of bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylate. With the remaining component contents identical, under light irradiation testing, the composition sample prepared in Example 20 performed better than that prepared in Example 23. This indicates that the combination of bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylate and ginsenosides can help inhibit the photodegradation of ergothioneine in the composition. Hydroxypropyl β-cyclodextrin, however, cannot help inhibit the photodegradation of ergothioneine.

[0189] Example 24 prepared a composition sample containing ergothioneine, butylene glycol, AVC, hydrolyzed hyaluronic acid (molecular weight 3K-10KDa), and water. Under light irradiation, the composition sample prepared in Example 24 showed a significant change in taste. This indicates that hydrolyzed hyaluronic acid (molecular weight 3K-10KDa) does not help inhibit the photodegradation of ergothioneine.

[0190] Example 25 prepared a composition sample containing ergothioneine, butylene glycol, AVC, sodium hyaluronate (molecular weight 10K-100KDa), and water. Under light irradiation, the composition sample prepared in Example 25 showed a significant change in taste. This indicates that sodium hyaluronate (molecular weight 10K-100KDa) does not help inhibit the photodegradation of ergothioneine.

[0191] Example 26 prepared a composition sample containing ergothioneine, butylene glycol, AVC, ginsenosides, hyaluronic acid (molecular weight 3K-10KDa), and water. Under light irradiation, the composition sample prepared in Example 26 showed a significant change in taste. This indicates that the combination of hyaluronic acid (molecular weight 3K-10KDa) and ginsenosides does not help inhibit the photodegradation of ergothioneine.

[0192] Example 27 prepared a composition sample containing ergothioneine, butylene glycol, AVC, hyaluronic acid (molecular weight 100K-1000KDa), and water. Under light irradiation, the composition sample prepared in Example 27 showed a significant change in taste. This indicates that hyaluronic acid (molecular weight 100K-1000KDa) does not help inhibit the photodegradation of ergothioneine.

[0193] Example 28 prepared a composition sample containing ergothioneine, butylene glycol, AVC, and water, while Example 20 prepared a composition sample containing ergothioneine, butylene glycol, AVC, ginsenosides, bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylate, and water. The difference between the two is that Example 20 contains 2.2 wt% of ginsenosides and bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylate. With the contents of the other components being the same, under light irradiation testing, the composition sample prepared in Example 20 performed better than the composition sample prepared in Example 28. This further clarifies that the combination of ginsenosides and bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylate can help inhibit the photodegradation of ergothioneine in the composition.

[0194] Under alkaline conditions (pH=8), the composition samples prepared in Examples 29-35 all showed significant taste changes, while the composition sample prepared in Example 36 showed no discernible taste change. This is mainly due to the decomposition of ergothionein in the samples under high-temperature stability testing; indicating that only compositions with appropriate amounts of ginsenosides or ginsenosides and bis-diethoxydiethylenecyclohexane 1,4-dicarboxylic acid ester can effectively inhibit the photodegradation of ergothionein. This information provides a very good solution and technique.

[0195] At the same pH value (pH=8), when all or part of the ginsenosides or the combination of ginsenosides and bis-diethoxydiethylenecyclohexane 1,4-dicarboxylic acid ester in the composition were replaced with hydroxypropyl β-cyclodextrin, hydrolyzed hyaluronic acid (molecular weight 3K-10KDa), sodium hyaluronate (molecular weight 10K-100KDa), hyaluronic acid (molecular weight 3K-10KDa), or hyaluronic acid (molecular weight 100K-1000KDa), the light irradiation test of the composition sample still showed a significant change in taste and could not effectively inhibit the photodegradation of ergothionein.

[0196] In conclusion, under acidic conditions, ginsenosides or combinations of ginsenosides and bis-diethoxydiethylenecyclohexane 1,4-dicarboxylic acid ester can significantly help inhibit the photodegradation of ergothionein.

[0197] Test Example 2: High Temperature Test Experiment

[0198] Test samples: Composition samples prepared in Examples 1-42

[0199] Generally speaking, the reaction rate roughly doubles for every 10°C increase in temperature. Many important properties of products, especially cosmetics, such as appearance, color, odor, and texture, are visually measured and not easily expressed numerically. If any changes are not described visually but recorded in five levels from slight to severe, it is easier to preserve records and evaluate results.

[0200] The samples from Examples 1-42 were poured into glass bottles, the caps were tightened, and the bottles were placed in an Incucell incubator at 48°C for 14 days to observe the changes in the products.

[0201] The aroma characteristics and intensity of the above samples were evaluated by professional fragrance tasters using direct olfaction. This evaluation method can be used to determine the stability and degree of change of the aroma. A three-level grading standard was adopted.

[0202] Classification Odor and strength standards Level 1 No discernible change in taste Level 2 Slight change in taste Level 3 Significant taste change

[0203] Table 2 High-temperature tests of the composition samples prepared in Examples 1-42

[0204]

[0205] After 14 days of high-temperature testing at 48°C, under acidic conditions (pH=4), the composition samples prepared in Examples 4, 6, and 8, which contained ergothioneine, butylene glycol, AVC, ginsenosides, and water, showed no perceptible change in taste; the composition samples prepared in Examples 1-3, 5, 7, and 12 showed only slight changes in taste.

[0206] At the same pH value (pH=4), when all or part of the ginsenosides in the composition were replaced with hydroxypropyl β-cyclodextrin, hydrolyzed hyaluronic acid (molecular weight 3K-10KDa), sodium hyaluronate (molecular weight 10K-100KDa), hyaluronic acid (molecular weight 3K-10KDa), or hyaluronic acid (molecular weight 100K-1000KDa), the composition samples showed different light irradiation test performance.

[0207] Specifically as follows:

[0208] Example 9 prepared a composition sample containing ergothioneine, butylene glycol, AVC, hydroxypropyl β-cyclodextrin, and water. Example 6 prepared a composition sample containing ergothioneine, butylene glycol, AVC, ginsenosides, bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylate, and water. Except that the former contained the same amount of hydroxypropyl β-cyclodextrin as the latter, and the latter contained an additional 2 wt% of bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylate, and with the remaining components in both samples having the same content, the composition sample prepared in Example 6 showed significantly better performance than the composition sample prepared in Example 9 under high-temperature stability testing. This indicates that the combination of ginsenosides and bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylate has a positive stabilizing effect on ergothioneine in the composition.

[0209] Example 10 prepared a composition sample containing ergothioneine, butylene glycol, AVC, hydrolyzed hyaluronic acid (molecular weight 3K-10KDa), and water. Example 5 prepared a composition sample containing ergothioneine, butylene glycol, AVC, ginsenosides, bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylate, and water. Except that the former had the same content of hydrolyzed hyaluronic acid (molecular weight 3K-10KDa) as the latter had the same content of ginsenosides, the latter also had an additional 2wt% of bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylate added. With the remaining components in both examples having the same content, the composition sample prepared in Example 5 performed better than the composition sample prepared in Example 10 under high-temperature stability testing. This indicates that the combination of ginsenosides and bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylate has a positive stabilizing effect on ergothioneine in the composition. Furthermore, it also shows that hydrolyzed hyaluronic acid (molecular weight 3K-10KDa) cannot inhibit the photodegradation of ergothioneine.

[0210] Example 11 prepared a composition sample containing ergothioneine, butylene glycol, AVC, sodium hyaluronate (molecular weight 10K-100KDa), and water. Example 5 prepared a composition sample containing ergothioneine, butylene glycol, AVC, ginsenosides, bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylate, and water. Except that the former had the same sodium hyaluronate (molecular weight 10K-100KDa) content and the latter had the same ginsenoside content, the latter also contained 2 wt% bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylate. With the remaining components in both examples having the same content, the composition sample prepared in Example 5 performed better than the composition sample prepared in Example 11 under high-temperature stability testing. This indicates that the combination of ginsenosides and bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylate can help inhibit the photodegradation of ergothioneine in the composition. Furthermore, it was also stated that sodium hyaluronate (molecular weight 10K-100KDa) does not inhibit the photodegradation of ergothionein as disclosed in the prior art.

[0211] Example 12 prepared a composition sample containing ergothioneine, butylene glycol, AVC, hyaluronic acid (molecular weight 3K-10KDa), ginsenosides, and water. Under high-temperature stability testing, a slight change in taste was observed. This indicates that the combined use of ginsenosides and hyaluronic acid (molecular weight 3K-10KDa) can also inhibit the photodegradation of ergothioneine.

[0212] Example 13 prepared a composition sample containing ergothioneine, butylene glycol, AVC, hyaluronic acid (molecular weight 100K-1000KDa), and water. Example 5 prepared a composition sample containing ergothioneine, butylene glycol, AVC, ginsenosides, bis-diethoxydiethylene cyclohexane 1,4-dicarboxylate, and water. Except that the former contained the same amount of sodium hyaluronate (molecular weight 10K-100KDa) and the latter contained the same amount of ginsenosides, the latter also contained 2 wt% of bis-diethoxydiethylene cyclohexane 1,4-dicarboxylate. With the remaining components in both examples having the same content, the composition sample prepared in Example 5 showed significantly better performance than the composition sample prepared in Example 13 under high-temperature stability testing. This indicates that the combination of ginsenosides and bis-diethoxydiethylene cyclohexane 1,4-dicarboxylate can help inhibit the photodegradation of ergothioneine in the composition. Furthermore, it was also stated that hyaluronic acid (molecular weight 100K-1000KDa) cannot help inhibit the photodegradation of ergothionein.

[0213] After 14 days of high-temperature stability testing, under acidic conditions (pH=6), the composition samples containing ergothioneine, butylene glycol, AVC, ginsenosides and water prepared in Examples 18-19 and 20-22 showed no perceptible change in taste; the composition sample prepared in Examples 16-17 showed only slight change in taste; and the composition sample prepared in Example 15 showed significant change in taste.

[0214] At the same pH value (pH=6), when all or part of the ginsenosides in the composition were replaced with hydroxypropyl β-cyclodextrin, hydrolyzed hyaluronic acid (molecular weight 3K-10KDa), sodium hyaluronate (molecular weight 10K-100KDa), or hyaluronic acid (molecular weight 100K-1000KDa), the composition samples showed different high-temperature stability test results.

[0215] Specifically as follows:

[0216] Example 23 prepared a composition sample containing ergothioneine, butylene glycol, AVC, hydroxypropyl β-cyclodextrin, and water. Example 20 prepared a composition sample containing ergothioneine, butylene glycol, AVC, ginsenosides, bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylate, and water. Except that the former had the same hydroxypropyl β-cyclodextrin content as the latter, and the latter also contained an additional 2 wt% of bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylate, with the remaining components present in identical amounts, the composition sample prepared in Example 20 performed better than the composition sample prepared in Example 23 under high-temperature stability testing. This indicates that the combination of bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylate and ginsenosides can help inhibit the photodegradation of ergothioneine in the composition. Hydroxypropyl β-cyclodextrin, however, cannot help inhibit the photodegradation of ergothioneine.

[0217] Example 24 prepared a composition sample containing ergothioneine, butylene glycol, AVC, hydrolyzed hyaluronic acid (molecular weight 3K-10KDa), and water. Under high-temperature stability testing, the composition sample prepared in Example 24 showed a significant change in taste. This indicates that hydrolyzed hyaluronic acid (molecular weight 3K-10KDa) does not help inhibit the photodegradation of ergothioneine.

[0218] Example 25 prepared a composition sample containing ergothioneine, butylene glycol, AVC, sodium hyaluronate (molecular weight 10K-100KDa), and water. Under high-temperature stability testing, the composition sample prepared in Example 25 showed a significant change in taste. This indicates that sodium hyaluronate (molecular weight 10K-100KDa) cannot help inhibit the photodegradation of ergothioneine.

[0219] Example 26 prepared a composition sample containing ergothioneine, butylene glycol, AVC, ginsenosides, hyaluronic acid (molecular weight 3K-10KDa), and water. Under high-temperature stability testing, the composition sample prepared in Example 26 showed a significant change in taste. This indicates that the combination of hyaluronic acid (molecular weight 3K-10KDa) and ginsenosides does not help inhibit the photodegradation of ergothioneine.

[0220] Example 27 prepared a composition sample containing ergothioneine, butylene glycol, AVC, hyaluronic acid (molecular weight 100K-1000KDa), and water. Under high-temperature stability testing, the composition sample prepared in Example 27 showed a significant change in taste. This indicates that hyaluronic acid (molecular weight 100K-1000KDa) does not help inhibit the photodegradation of ergothioneine.

[0221] Example 28 prepared a composition sample containing ergothioneine, butylene glycol, AVC, and water, while Example 20 prepared a composition sample containing ergothioneine, butylene glycol, AVC, ginsenosides, bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylate, and water. The difference between the two is that Example 20 contains 2.2 wt% of ginsenosides and bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylate. With the contents of the other components being the same, the composition sample prepared in Example 20 performed better than the composition sample prepared in Example 28 under high-temperature stability testing. This further clarifies that the combination of ginsenosides and bis-diethoxydiethylene glycol cyclohexane 1,4-dicarboxylate can help inhibit the photodegradation of ergothioneine in the composition.

[0222] Under alkaline conditions (pH=8), the compositions prepared in Examples 29-30 and 32-35 all showed significant changes in taste, while the composition prepared in Example 36 showed no noticeable change in taste. The composition prepared in Example 31 showed a slight change in taste; this was mainly due to the decomposition of ergothioneine in the sample under high-temperature stability testing. This indicates that only compositions with appropriate amounts of ginsenosides or a combination of ginsenosides and bis-diethoxydiethylenecyclohexane 1,4-dicarboxylic acid ester can effectively inhibit the photodegradation of ergothioneine. This information provides a very good solution and technique.

[0223] At the same pH value (pH=8), when all or part of the ginsenosides or the combination of ginsenosides and bis-diethoxydiethylenecyclohexane 1,4-dicarboxylic acid ester in the composition were replaced with hydroxypropyl β-cyclodextrin, hydrolyzed hyaluronic acid (molecular weight 3K-10KDa), sodium hyaluronate (molecular weight 10K-100KDa), hyaluronic acid (molecular weight 3K-10KDa), or hyaluronic acid (molecular weight 100K-1000KDa), the light irradiation test of the composition sample still showed a significant change in taste and could not effectively inhibit the photodegradation of ergothionein.

[0224] In conclusion, under acidic conditions, ginsenosides or combinations of ginsenosides and bis-diethoxydiethylenecyclohexane 1,4-dicarboxylic acid ester can significantly help inhibit the photodegradation of ergothionein.

[0225] Test Example 3: Detection of Ergothionein by Ultra-High Performance Liquid Chromatography

[0226] Test samples: Composition samples prepared in Examples 1-42 after the light irradiation test in Test Example 1; Composition samples prepared in Examples 1-42 after the high temperature stability test in Test Example 2.

[0227] Test standard: NY / T 3872-2021 (UPLC)

[0228] The instrument conditions were as follows: Agilent HILIC Plus 4.6×100 mm 3.5 μm, column temperature 30℃, mobile phase 0.1% formic acid acetonitrile solution-0.1% formic acid aqueous solution (80+20 V / V), flow rate 1.0 mL / min, detection wavelength 262 nm, and injection volume 10 μL.

[0229] Testing steps:

[0230] Preparation of ergothioneine standard stock solution (0.5 g / L): Weigh 50 mg of ergothioneine standard (accurate to 0.01 mg), dissolve in water, and dilute to 100 mL. Store at 4℃. Preparation of ergothioneine standard working solutions: Accurately transfer a certain amount of ergothioneine standard stock solution and serially dilute with 0.1% formic acid methanol solution to obtain a series of standard working solutions (mass concentrations of 5 μg / mL, 10 μg / L, 25 μg / mL, 50 μg / mL, and 100 μg / mL, respectively).

[0231] Standard curve plotting: The standard working solution was measured under the above chromatographic conditions, and the standard curve was plotted with the mass concentration of the standard working solution as the abscissa and the peak area as the ordinate.

[0232] Sample preparation and determination: Weigh 0.1–0.5 g (accurate to 0.1 mg) of sample into a 10 mL stoppered colorimetric tube, add 8 mL of 0.1% formic acid in methanol solution, and sonicate for 20 min. Make up to volume with 0.1% formic acid in methanol solution, shake well, and filter a portion of the liquid through a 0.22 μm filter membrane. Then, determine the sample under the chromatographic conditions described above. Quantify the sample using the standard curve. The analyte response value in the sample solution should be within the linear range of the standard curve.

[0233] Results Calculation: Quantitative analysis using the external standard method; the formula for calculating the ergothionein content in the sample is as follows:

[0234]

[0235] In the formula:

[0236] X — Mass fraction of the analyte in the sample, %

[0237] C—The concentration of ergothioneine in the sample solution obtained from the standard curve, in micrograms per milliliter (μg·mL). -1 );

[0238] V—The total volume of the sample after dilution, in milliliters (mL);

[0239] m — Sample mass, in grams (g);

[0240] The ergothioneine content in the composition samples after light exposure or high temperature stability testing is shown in the table below:

[0241] Table 3. Ergothionein content in the composition samples after light or high temperature stability tests.

[0242]

[0243] The results show that, after comprehensive investigation under various light conditions (as shown in Table 1), high temperature conditions (as shown in Table 2), different pH conditions, and different ergothioneine concentrations, very clear test results were obtained as shown in the table above.

[0244] First, under alkaline conditions, ergothioneine is more easily decomposed. Due to its unique structure, containing hydroxyl and carbonyl groups, ergothioneine reacts chemically with hydroxide ions under alkaline conditions, resulting in changes in its structure, properties, and odor. Regardless of concentration or under light and high temperature conditions, the structural integrity and existence of ergothioneine are significantly challenged, and its taste changes markedly, becoming noticeably foul. However, even under such stringent conditions, we unexpectedly discovered that the synergistic effect of ergothioneine and ginsenosides, as in Example 36, exhibited excellent stability, specifically at an ergothioneine:ginsenoside ratio of 2:3, i.e., a slight excess of ginsenosides. Ginsenosides are tetracyclic triterpenoid compounds, mainly in diol and triol configurations. The structural feature with better efficacy is the presence of hydroxyl substitutions at C-3, C-12, and C-20. Although there are no reports of ginsenosides using abundant hydroxyl groups on their surface to effectively protect ergothioneine through hydrogen bonding, based on their structural characteristics and experimental results, it is speculated that this condition and ratio provides a very good solution and technology for the application of ergothioneine in alkaline products.

[0245] Acidic pH is the recommended direction for ergothioneine application. A comparison of pH=4 and pH=6 shows that a lower pH under acidic conditions is more conducive to the stability of ergothioneine. Unexpectedly, by adjusting the proportions of different ergothioneine compositions at various concentrations, several different combinations and concentrations were found to have excellent protective functions. Examples 14, 28, and 42 demonstrate that under different pH conditions, the ergothioneine content degraded by more than 30-50% within two weeks.

[0246] In contrast to existing technologies, such as the Chinese patent application CN110327242B which discloses a method for inhibiting the photodegradation of ergothioneine using hyaluronic acid salts, the composition samples prepared in Examples 10-11, 13, 24-25, 27, 38-39, and 41 of this application were used to simultaneously investigate the inhibitory effect of hyaluronic acid and its derivatives on ergothioneine in the prior art. Compared with ergothioneine under the same concentration and pH conditions, no significant improvement in photodegradation and high-temperature degradation stability was observed. On the contrary, the content of ergothioneine under these conditions decreased to some extent. It is speculated that the possible reason is that different types of hyaluronic acid, including hyaluronic acid with a relatively small molecular weight but still having a molecular weight of several thousand Daltons, did not form a complete and tight encapsulation structure, resulting in ergothioneine concentration differences and escape.

[0247] Under pH=4 conditions, the composition samples prepared in Examples 4-8 unexpectedly achieved excellent protective effects against ergothioneine. After 14 days of light exposure or 14 days of high-temperature stability testing, the content in the samples remained at over 74%.

[0248] The composition sample prepared in Example 12 also showed an inhibitory effect on ergothioneine. After 14 days of light exposure or 14 days of high temperature stability test, the content in the sample was maintained at more than 78%.

[0249] Under pH=6 conditions, the compositions prepared in Examples 16-22 also unexpectedly exhibited excellent protective effects against ergothioneine. After 14 days of light exposure or 14 days of high-temperature stability testing, the ergothioneine content in the samples remained above 70%.

[0250] In summary, when the pH is acidic, the ginsenoside molecules in the composition sample have abundant hydroxyl groups. Within the cyclodextrin inclusions, there are numerous binding sites for both. This binding is a spontaneous reaction primarily driven by hydrophobic interactions. Reports on molecular docking suggest that the addition of ginsenosides may have altered the internal water transport environment of ergothioneine, leading to a more stable binding. This also essentially ensured that the ergothioneine content remained above 70%.

[0251] For example, a 2:1 mass ratio of ergothioneine to ginsenosides is considered an optimal concentration. However, as shown in Example 4, a higher concentration of ergothioneine resulted in accelerated degradation. Even when the concentration of ginsenosides was further reduced to a 5:1 or 4:1 mass ratio, a relatively good effect was maintained. However, a 10:1 ratio did not provide adequate protection. Therefore, given the relatively high cost of ginsenosides, the fact that they can achieve good protective effects at lower concentrations is an unexpected achievement.

[0252] Compared to Example 9, simply using cyclodextrin to encapsulate ergothioneine has proven to be an ineffective way to address its photodegradation and high-temperature stability.

[0253] In terms of different concentrations, the ratio of ergothioneine to ginsenosides is best when the ratio is between 1:2 and 0.5:5. In addition, Example 12 illustrates the synergistic effect and usefulness of hyaluronic acid when ginsenosides are combined with hyaluronic acid.

[0254] Test Example 4: Evaluation of Antioxidant Performance

[0255] Test samples: Composition samples prepared in Examples 4, 6, 8, 14, 18-19, 21-22, 28, 36, and 42.

[0256] Experimental methods:

[0257] 1) Cell seeding: at 2.2 × 10⁻⁶ 5 Seed keratinocytes at a seeding density of cells / well into 6-well plates and incubate overnight in an incubator (37°C, 5% CO2).

[0258] 2) Drug administration: According to the test group, when the cell deposition rate in the 6-well plate reaches 30%-50%, the drugs are administered to the groups (the compositions of different embodiments are added), 2 mL of sample is added to each well, and each group has 3 replicates. After drug administration, the 6-well plate is placed in an incubator (37℃, 5% CO2) for 24 h.

[0259] 3) UVB irradiation: According to the test groups, the groups exposed to UVB irradiation were subjected to 300mJ / cm² irradiation. 2 UVB irradiation.

[0260] 4) ROS content test: After irradiation, wash each well of cells three times with PBS, add 1 mL of 10 μM DCFH-DA probe to each well, incubate in an incubator (37℃, 5% CO2) for 30 min, discard the culture medium containing DCFH-DA, wash three times with PBS, digest the cells with trypsin (0.25%), wash the cells once with PBS, add a certain amount of fresh PBS, and perform flow cytometry detection.

[0261] 5) Statistical analysis of results: GrapHPad Prism plots were used, and results are expressed as Mean ± SD. t-tests were used for comparisons between groups. All statistical analyses were two-tailed. P < 0.05 was considered statistically significant, and P < 0.01 was considered highly statistically significant.

[0262] Table 4. Evaluation results of antioxidant performance of some examples

[0263]

[0264] Note: Mean fluorescence intensity (MFI) reflects the ROS content. When performing statistical analysis using the t-test method, significance compared to the BC group is indicated by # (P-value < 0.05 is indicated by #, P-value < 0.01 is indicated by ##); significance compared to the NC group is indicated by * (P-value < 0.05 is indicated by *, P-value < 0.01 is indicated by **).

[0265] Ergothioneine regulates intracellular redox reactions and participates in intracellular energy regulation, acting as a physiological protectant for cells. It is safe and stable in both biological and pharmacological properties. Simultaneously, it maintains the stability of other components and promotes the absorption of other active ingredients by cells, exhibiting excellent synergistic effects, achieving a synergistic effect greater than the sum of its parts (1+1>2). Not only does it exert a powerful antioxidant effect, but it also enhances the efficacy and activity of other substances.

[0266] For the preferred combination of embodiments (mainly in terms of photodegradation stability and high temperature stability), the antioxidant properties were tested. The antioxidant test adopted a more reasonable and scientific detection method, which is based on the content of reactive oxygen species (ROS) produced by keratinocytes under UVB stimulation. This method is a more accurate way to evaluate the skin's oxidation mechanism and antioxidant mechanism.

[0267] Comparing the antioxidant properties of ergothioneine under different pH conditions, it was found that commonly used vitamin E (as a control group) exhibited superior antioxidant performance. Vitamin E is a substance with strong antioxidant activity and is a crucial antioxidant in the human body, blocking free radical chain reactions. It plays an indispensable role in stabilizing and protecting cell membranes and in normal human metabolism. Vitamin E plays an important role in inhibiting inflammatory responses, fighting free radicals, anti-aging, and anti-cancer processes by influencing inflammatory factors and regulating cell signal transduction. Vitamin E has been widely used to treat various skin diseases, including photoaging, inflammatory skin diseases, connective tissue diseases, viral skin diseases, and herpes. Regarding ergothioneine itself, the compositions prepared in Examples 14, 28, and 42 also significantly improved antioxidant properties compared to vitamin E.

[0268] Comparing the combinations of ergothioneine and ginsenosides under different pH conditions, a significant increase in antioxidant capacity was unexpectedly found in all cases. In most examples, the antioxidant capacity increased by more than 60%, especially in Example 8, where the antioxidant capacity at pH=4 improved to 78.8%, a 75.23% increase compared to the 44.97% antioxidant capacity prepared in Example 14. Figure 1 As shown.

[0269] The composition samples prepared in Example 8 and Example 22 differed only in their pH values, and their antioxidant capacity was improved by 78.80% and 72.20%, respectively.

[0270] Test Example 5: Efficacy Evaluation Based on Fibroblasts

[0271] Test examples: Composition samples prepared in Examples 8, 22, and 36;

[0272] Test Plan

[0273] 1) Vaccination: 2 x 10 5 Seed cells (batch number: Fb19052002, provided by Guangdong Boxi Biotechnology Co., Ltd.) at a seeding density of cells / well into 6-well plates and incubate overnight in an incubator (37℃, 5% CO2).

[0274] 2) Solution preparation: TGF-β1 (Peprotech) was used as a positive control (PC) at a concentration of 100 ng / mL. The sample and control were prepared at 0.2% (v / v). The detection indicators were Collagen I, Collagen III, and Elastin. The detection method was qRT-PCR.

[0275] 3) Drug administration: According to the test protocol, when the cell deposition rate in the 6-well plate reached 40%~60%, drug administration was performed in groups (compositions added according to different embodiments), with 2 mL of drug administered to each well, and 3 replicates per group. After drug administration, the 6-well plate was placed in an incubator (37℃, 5% CO2). 2 Incubate for 24 hours.

[0276] 4) Cell collection: After culturing for 24 hours, wash twice with 1 mL / well PBS (Solepro), add 1 mL RNAisoPlus (Takara) to each well, lyse the cells by pipetting, and collect the samples.

[0277] 5) Gene expression detection: RNA was extracted, reverse transcribed into cDNA (using the Takara reverse transcription kit), and then detected by real-time PCR. The results were calculated using the 2-ΔΔCT method.

[0278] 6) Statistical analysis of results: GrapHPad Prism plots were used, and results are expressed as Mean ± SD. t-tests were used for comparisons between groups. All statistical analyses were two-tailed. P < 0.05 was considered statistically significant, and P < 0.01 was considered highly statistically significant.

[0279] Based on the testing method, fibroblasts were treated with RNAiso Plus and then collected. Following the kit instructions, RNA extraction, reverse transcription, and quantitative real-time PCR were performed. The detection results are shown in the table below: Table 5: Detection results of Collagen I gene in some examples.

[0280] Group Mean value SD P-value Upward adjustment rate (vsBC) BC 1.00 0.03 / / TGF-β1 (PC) 1.93 0.19 0.001* 93.00% Ginsenosides (comparison) 1.49 0.11 0.002** 49.00% Example 8 2.13 0.06 0.000** 113.00% Example 22 1.75 0.19 0.003** 75.00% Example 36 1.64 0.21 0.007** 64.00%

[0281] Note: When performing statistical analysis using the t-test method, significance compared to group BC is indicated by *, P-value < 0.05 is indicated by *, and P-value < 0.01 is indicated by **.

[0282] Table 6 shows the Collagen III gene detection results in some examples.

[0283] Group Mean value SD P-value Upward adjustment rate (vsPC) BC 1.00 0.06 / / TGF-β1 (PC) 1.78 0.04 0.000* 78.00% Ginsenosides (comparison) 1.55 0.04 0.000** 55.00% Example 8 2.03 0.12 0.000** 103.00% Example 22 1.72 0.05 0.001** 72.00% Example 36 1.61 0.03 0.001** 61.00%

[0284] Note: When performing statistical analysis using the t-test method, significance compared to group BC is indicated by *, P-value < 0.05 is indicated by *, and P-value < 0.01 is indicated by **.

[0285] Table 7. Results of Elastin gene detection in some examples.

[0286] Group Mean value SD P-value Upward adjustment rate (vsPC) BC 1.00 0.03 / / TGF-β1 (PC) 1.51 0.10 0.001* 51.00% Ginsenosides (comparison) 1.14 0.03 0.004** 14.00% Example 8 1.65 0.13 0.001** 65.00% Example 22 1.21 0.07 0.002** 21.00% Example 36 1.09 0.10 0.339 7.00%

[0287] Note: When performing statistical analysis using the t-test method, significance compared to group BC is indicated by *, P-value < 0.05 is indicated by *, and P-value < 0.01 is indicated by **.

[0288] Compared with the PC group, it was unexpectedly found that the combination of ergothioneine and ginsenosides significantly and synergistically enhanced the expression of collagen and elastin genes by ginsenosides. Among the examples under different pH conditions, the synergistic improvement effect was better under acidic conditions.

[0289] Changes in the skin's internal composition and structure are the primary cause of skin aging. These changes ultimately lead to visible signs of aging such as wrinkles and sagging. These changes include alterations in the content or density of hyaluronic acid and collagen, changes in the skin's internal structure including changes in the density of dermal papillae and dermal capillaries, and "dermal cavitation." Aging is a key research area in dermatology, and from a skin research perspective, how to combat skin aging more scientifically and effectively remains an important challenge.

[0290] Collagen is an important structural component of the skin. Types I, III, IV, V, VII, XVII, and XVIII collagen are known to exist in the skin. Types I and III collagen are found in the reticular fibrous layer of the dermis. Based on fibroblasts, the translation and expression product of the Collagen I gene is a triple helix structure formed by three identical or different peptide chains. With age, collagen degrades, becomes brittle, and undergoes glycation, resulting in signs of aging. Collagen degradation occurs through matrix metalloproteinases (MMPs). Types I, II, and III collagen can be cleaved into 1 / 4 and 3 / 4 size fragments by MMP1, MMP8, and MMP13, respectively, and are further cleaved by the membrane-anchoring protein MMP14. Simultaneously, MMP2 can also cleave type I collagen.

[0291] Experimental results showed that ginsenosides had a good effect on promoting Collagen I genes. However, ergothioneine was unexpectedly found to have an excellent synergistic effect, which could improve the gene improvement effect by 64%. In the same composition, the improvement of collagen type I was different under different pH conditions, such as pH 6 and pH 8, and the efficacy was not as good as under pH 4.

[0292] Collagen III fibers are relatively small, compared to the coarser type I collagen (which constitutes the main body of the skin), and are a major component of the skin's reticular structure. They form fine, sharply layered bands, providing support for the dermis. Experimental results showed that ginsenosides also have a good ability to promote collagen III gene expression; however, ergothioneine unexpectedly enhanced the effect of ginsenosides significantly, increasing collagen III expression by 48%. Similar to the trend in collagen I gene expression, a pH-dependent effect on the growth-promoting effect was also unexpectedly observed.

[0293] Elastin is the main component of elastic fibers, accounting for up to 90%. Its precursor is soluble tropoelastin (TE), which is mainly synthesized by fibroblasts in the skin. Intracellularly, TE binds to elastin-binding protein (EBP). EBP protects TE from unwanted intracellular degradation and aggregation and is also responsible for transporting TE to the cell membrane. Subsequently, TE is secreted onto the extracellular microfibril backbone, where it is converted into insoluble elastin polymers through cross-linking by lysyl oxidase (LOX). Experimental results show that ginsenosides also have a good ability to promote elastin gene expression. However, it was unexpectedly found that ergothioneine can also significantly enhance the effect of ginsenosides, increasing elastin expression by 51%.

[0294] Test Example 6: Evaluation of Penetration Effect

[0295] This test used piglet skin (purchased from Shanghai Institute of Biomedical Engineering) as the test model and the piglet skin-Franz cell diffusion cell system as the test system. Figure 2 A schematic diagram of a Franz diffusion cell for transdermal drug absorption:

[0296] (1) The test samples were selected from Examples 8, 22 and 36, and the ergothioneine standard sample was used as a control. The concentration was diluted to the same concentration as the above examples (i.e., the theoretical concentration).

[0297] (2) Test scheme: Ergothionein was used as the target substance. The cumulative permeability of the samples in the pig skin-Franz cell diffusion cell system after 24 hours was tested to describe its in vitro transdermal behavior.

[0298] (3) Test procedure: First, add a quantitative amount of receiving solution (PBS phosphate buffer pH 7.2-7.4) to the receiving chamber and place the matching magnetic stir bar into the receiving chamber; second, after activating the pig skin with PBS solution, fix it between the supply chamber and the receiving chamber of the Franz diffusion cell, with the stratum corneum facing the supply chamber and the dermis facing the receiving chamber, and tighten and fix the skin; third, use a sampler to inject the receiving solution (PBS phosphate buffer) into the receiving chamber through the sampling tube, remove the air, and make the dermis of the skin in close contact with the receiving solution; then, add different samples to the skin surface in the supply chamber (150mg is used as the appropriate amount of sample to be added in this test), and spread the sample evenly from the center of the skin to the edge. Each sample was repeated 3 times in parallel. Next, the Franz cell diffusion cell was fixed in the transdermal absorption diffusion apparatus, and the electromagnetic stirrer was turned on to stir at a speed of 440 rpm / min. The water bath was kept at a constant temperature of (32±1)℃, and it was ensured that there were no air bubbles in the water bath jacket. Finally, for sampling, the sample liquid at the 24-hour time point was taken, and the receiving liquid was extracted through the sampling tube using a sampler. The receiving cell and one side of the dermal layer of piglet skin were then rinsed with PBS buffer solution. After rinsing 3 times, the receiving liquids were combined, the mass was recorded, and the concentration of ergothioneine in the receiving liquid was tested.

[0299] Table 8 Penetration Effect

[0300]

[0301] Diethoxydiethylene cyclohexane 1,4-dicarboxylic acid ester is an amphiphilic permeation-enhancing component that can reconstruct the cells of the stratum corneum of the skin, thereby forming intercellular spaces that allow the ginsenoside-ergothioneine combination to pass through smoothly, thus promoting the permeation effect. In Examples 8, 22, and 36, it can be found that the permeation efficiency is significantly improved compared to the system with ergothioneine alone, with an improvement efficiency exceeding 50%. The significant effect of pH value on the permeation system was also observed.

[0302] Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Anyone skilled in the art can make various modifications and alterations without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be determined by the claims.

[0303] The composition samples prepared in Examples 4-8, 12, 16-22, and 36 were used for the preparation of a topical skin agent. The topical skin agent is preferably a cosmetic composition, such as a lotion, serum, or cream. The composition samples prepared in Examples 4-8, 12, 16-22, and 36 have a weight percentage of 0.0001 wt%-90 wt% in the topical skin agent. A preferred weight percentage is 0.001 wt%-10 wt%. A more preferred weight percentage is 0.001 wt%-5 wt%.

[0304] The following are examples of the specific applications of the compositions prepared in Examples 4-8, 12, 16-22, and 36 in topical skin preparations, along with the formulations and preparation methods of these dosage forms. In the following tables, "-" indicates no additives.

[0305] Application Example 1: Preparation of Face Cream

[0306]

[0307] Application Example 2: Emulsion Preparation

[0308]

[0309] Application Example 3: Preparation of Gel

[0310]

[0311] Application Example 4: Preparation of Toner

[0312]

[0313] Application Example 5: Preparation of Serum

[0314]

[0315] Application Example 6: Preparation of Facial Masks

[0316]

[0317] Application Example 7: Preparation of Eye Cream

[0318]

[0319] Application Example 8: Preparation of Spray

[0320]

[0321] Application Example 9: Preparation of Shower Gel

[0322]

[0323] Application Example 10: Preparation of Facial Cleanser

[0324]

[0325] Application Example 11: Preparation of Essence Water

[0326]

[0327] In the examples of specific applications of all the above topical skin agents, the composition samples of Examples 4-8, 12, 16-22 and 36 can be used directly in the formulation, either partially or completely, to replace deionized water, or can be used directly as a formulation.

Claims

1. A synergistic composition based on ginsenosides, comprising ginsenosides, ergothioneine, butanediol, bis-diethoxydiethylene cyclohexane 1,4-dicarboxylic acid ester, acrylamide dimethyl taurate ammonium / VP copolymer, and water, wherein, The weight ratio of ginsenosides to ergothioneine is 0.5-5:1-2. The content of ginsenosides is 0.05-0.5 wt%. The content of butanediol is 5 wt%. The content of the acrylamide dimethyl taurate ammonium / VP copolymer is 0.8 wt%. The content of the bis-diethoxydiethylenecyclohexane 1,4-dicarboxylic acid ester is 2 wt%. The ergothioneine content is 0.1-0.2 wt%. The pH value of the composition is 4-6.

2. The application of ginsenosides and bis-diethoxydiethylenecyclohexane 1,4-dicarboxylic acid ester in inhibiting the photodegradation of ergothionein, wherein the weight ratio of ginsenosides to ergothionein is 0.5-5:1-2, the content of bis-diethoxydiethylenecyclohexane 1,4-dicarboxylic acid ester is 2 wt%, the content of ginsenosides is 0.05-0.5 wt%, the content of ergothionein is 0.1-0.2 wt%, and the pH value of the application is 4-6.

3. Antioxidant use of the composition as claimed in claim 1 for non-therapeutic purposes.

4. Non-therapeutic use of the composition of claim 1 for anti-skin aging purposes.

5. The use of the composition of claim 1 in a non-therapeutic topical skin preparation.

6. The application as described in claim 5, characterized in that, The topical skin agents are selected from: face creams, lotions, gels, toners, face masks, eye creams, sprays, shower gels, and facial cleansers.

7. The application as described in claim 5, wherein the composition is used in a topical skin preparation in an amount of 0.0001 wt% to 90 wt%.

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