A synergistic whitening composition and method of making and using same

Abstract

The present application belongs to the technical field of cosmetics, and discloses a synergistic whitening composition, a preparation method and application thereof. The composition comprises components: glabridin, polydatin and resveratrol. Glabridin, polydatin and resveratrol in the composition of the present application exert a synergistic effect, regulate three whitening mechanism paths of inhibiting tyrosinase activity, inhibiting melanin production and resisting oxidative stress, thereby having a whitening effect. Further, the composition of the present application exhibits a whitening effect at a lower concentration, reduces side effects such as irritation caused by using a single high-concentration chemical component, and is safer.

Description

Technical Field

[0001] This invention belongs to the field of cosmetic technology, and specifically relates to a highly effective synergistic whitening composition, its preparation method, and its application. Background Technology

[0002] With the improvement of people's living standards and aesthetic standards, skin whitening has become a popular topic and pursuit among consumers. In recent years, the research and development of skin whitening skincare products has become increasingly active, and safety and efficacy have received more and more attention from consumers and researchers.

[0003] A person's skin color is fundamentally determined by the amount and distribution of melanin in the skin. Melanin is mainly produced by melanocytes in the basal layer of the skin. These melanocytes secrete melanin granules, which are transported to keratinocytes via cell dendrites. Keratinocytes absorb and take in the melanin granules, and as they proliferate and differentiate, they ascend to the stratum corneum, eventually being excreted with the shedding of the stratum corneum. Problems in any of these steps can lead to abnormal accumulation and deposition of melanin in the skin, affecting skin color and causing skin problems such as age spots, freckles, and pigmentation. Traditional whitening agents such as lead powder, hydroquinone, and rhododendron alcohol have strong cytotoxic side effects, easily causing local allergic reactions such as itching and stinging, and even potentially leading to hyperpigmentation or other skin problems, such as vitiligo and increased wrinkle formation. Hydroquinone, in particular, has been listed as a potential carcinogen and is prohibited in skincare products in both the EU and China. Currently used skin whitening agents include niacinamide, arbutin, kojic acid, licorice root extract, ascorbic acid, and phenylethyl resorcinol, mostly chemical ingredients. These ingredients have been shown to inhibit tyrosinase activity. The use of these substances, especially in appropriate amounts, requires careful consideration. Excessive amounts can easily cause skin irritation, allergies, and even severely interfere with normal melanin production, leading to white patches on the skin. Some still pose certain safety risks; for example, kojic acid may increase the risk of cancer.

[0004] Therefore, developing more efficient, gentle, and safe whitening agents is a technical challenge that urgently needs to be solved by those skilled in the art. Summary of the Invention

[0005] This invention aims to at least solve one of the technical problems existing in the prior art. To this end, this invention provides a composition in which glycyrrhizin, polygalactosin, and resveratrol exert a synergistic effect, regulating three major whitening mechanism pathways: inhibition of tyrosinase activity, inhibition of melanin production, and anti-oxidative stress, thereby achieving a highly effective whitening effect. Furthermore, this composition exhibits a highly effective whitening effect at lower concentrations, reducing the irritation and other side effects caused by using a single high-concentration chemical ingredient, thus improving safety.

[0006] A first aspect of the present invention provides a composition comprising the components: glycyrrhizin, polydipsia glycoside, and resveratrol.

[0007] According to some embodiments of the present invention, the amount of glycyrrhizin added to the composition is 10wt%-70wt%.

[0008] According to some embodiments of the present invention, the amount of polygalactoside added to the composition is 10wt%-50wt%.

[0009] According to some embodiments of the present invention, the amount of resveratrol added to the composition is 10wt%-80wt%.

[0010] A second aspect of the present invention provides a method for preparing the composition, comprising the following steps:

[0011] The composition is obtained by mixing the glycyrrhizin, polydipsia glycoside, and resveratrol.

[0012] A third aspect of the present invention provides a cosmetic product comprising the composition described herein.

[0013] According to some embodiments of the present invention, the dosage form of the cosmetic is selected from creams, lotions, serums, aqueous solutions, oils, gels, or masks.

[0014] According to some embodiments of the present invention, the cosmetic also includes cosmetic-acceptable excipients.

[0015] According to some embodiments of the present invention, the excipients include at least one of emollients, emulsifiers, thickeners, humectants, pH adjusters, and preservatives.

[0016] According to some embodiments of the present invention, the amount of the composition added to the cosmetic is ≤20wt%. Further, the amount of the composition added to the cosmetic is 0.01wt%-20wt%. Even further, the amount of the composition added to the cosmetic is 0.1wt%-20wt%. Still further, the amount of the composition added to the cosmetic is 0.3wt%-10wt%.

[0017] A fourth aspect of the present invention provides an emulsion comprising the composition described herein.

[0018] According to some embodiments of the present invention, the emulsion comprises the following components: glycerol, betaine, sodium acrylate copolymer, lecithin, cetearyl glucoside, cetearyl alcohol, polydimethylsiloxane, isohexadecane, dioctyl carbonate, tocopherol (vitamin E), 1,3-propanediol, p-hydroxyacetophenone, 1,2-hexanediol, glycyrrhizin, polydextrin, resveratrol, and water.

[0019] A fifth aspect of the present invention provides an essence comprising the composition described herein.

[0020] According to some embodiments of the present invention, the essence comprises the following components: disodium EDTA, betaine, ammonium acryloyl dimethyl taurate / behenyl ether-25 methacrylate crosspolymer, PEG / PPG / polybutylene glycol-8 / 5 / 3 glycerin, butylene glycol, bis-PEG-18 methyl ether dimethyl silane, tocopherol (vitamin E), sodium hyaluronate, 1,3-propanediol, p-hydroxyacetophenone, 1,2-hexanediol, glycyrrhizin, polyoxin, resveratrol, and water.

[0021] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0022] 1) The composition of the present invention, through the combination of glycyrrhizin, polygalactoside, and resveratrol, has a significant synergistic effect on the inhibition of melanin content and tyrosinase activity. The effect of the composition is significantly better than that of each individual component. Moreover, the composition still maintains its high efficiency in inhibiting melanin and tyrosinase activity at extremely low concentrations (after dilution of 10,000 times), which is a characteristic that individual components do not possess.

[0023] 2) The composition of the present invention, through the combination of glycyrrhizin, polydipsia glycoside and resveratrol, has a significant synergistic effect in antioxidant activity, including the ability to scavenge DPPH free radicals and hydroxyl free radicals. The effect of the composition is significantly better than that of each individual component, and also better than any combination of two components.

[0024] 3) Combating oxidative stress, inhibiting tyrosinase activity, and inhibiting melanin production are the three major pathways for whitening efficacy. The composition of this invention combines glycyrrhizin, polygalactosidase and resveratrol, which show significant synergy in all three pathways. Even at low concentrations, it can still show significant effects in inhibiting melanin and tyrosinase. While ensuring the whitening effect, this greatly reduces the probability of adverse reactions caused by using high concentrations of single chemical ingredients.

[0025] 4) The preparation method of the composition of the present invention is simple and easy to implement, the raw materials are readily available, and it can be produced on a large scale. Detailed Implementation

[0026] To enable those skilled in the art to more clearly understand the technical solutions described in this invention, the following embodiments are provided for illustration. It should be noted that the following embodiments do not constitute a limitation on the scope of protection claimed by this invention.

[0027] Unless otherwise specified, the raw materials, reagents or devices used in the following examples are available from conventional commercial sources or can be obtained by existing known methods.

[0028] Example 1

[0029] This embodiment provides a composition composed of powders of glycyrrhizin, polydipsia glycoside, and resveratrol mixed in a molar ratio of 1:1:1 (mass percentages: glycyrrhizin 34.4 wt%, polydipsia glycoside 41.4 wt%, resveratrol 24.2 wt%).

[0030] Example 2

[0031] This embodiment provides a composition composed of powders of glycyrrhizin, polydipsia glycoside, and resveratrol in a molar ratio of 4.9:1.2:1 (mass percentages: glycyrrhizin 70wt%, polydipsia glycoside 20wt%, resveratrol 10wt%), and the amount used is the same as that of the composition in Example 1.

[0032] Example 3

[0033] This embodiment provides a composition composed of powders of glycyrrhizin, polydipsia rubescens, and resveratrol in a molar ratio of 1:4.2:5.7 (mass percentages: glycyrrhizin 10 wt%, polydipsia rubescens 50 wt%, resveratrol 40 wt%), the amounts of which are the same as those in the composition of Example 1.

[0034] Example 4

[0035] This embodiment provides a composition composed of powders of glycyrrhizin, polydipsia glycoside, and resveratrol in a molar ratio of 1.2:1:13.7 (mass percentages: glycyrrhizin 10wt%, polydipsia glycoside 10wt%, resveratrol 80wt%), the amount of which is the same as that of the composition in Example 1.

[0036] Comparative Example 1

[0037] This comparative example provides a single component containing only glycyrrhizin powder, in the same amount as the composition of Example 1.

[0038] Comparative Example 2

[0039] This comparative example provides a single component containing only polydipsia glycoside powder, in the same amount as the composition of Example 1.

[0040] Comparative Example 3

[0041] This comparative example provides a single component containing only resveratrol powder, in the same amount as the composition of Example 1.

[0042] Comparative Example 4

[0043] This comparative example provides a single component containing only tranexamic acid powder, in the same amount as the composition of Example 1.

[0044] Comparative Example 5

[0045] This comparative example provides a single component containing only nicotinamide powder, in the same amount as the composition of Example 1.

[0046] Comparative Example 6

[0047] This comparative example provides a single component containing only phloretin powder, in the same amount as the composition of Example 1.

[0048] Comparative Example 7

[0049] This comparative example provides a composition composed of glycyrrhizin and polydipsia glycoside powders mixed in a molar ratio of 1:1 (glycyrrhizin 54.6 wt% and polydipsia glycoside 45.4 wt%), the amount of which is the same as that of the composition in Example 1.

[0050] Comparative Example 8

[0051] This comparative example provides a composition composed of glycyrrhizin and resveratrol powders mixed in a molar ratio of 1:1 (glycyrrhizin 41.3 wt% and resveratrol 58.7 wt%), the amounts of which are the same as those in the composition of Example 1.

[0052] Comparative Example 9

[0053] This comparative example provides a composition composed of powders of glycyrrhizin, nicotinamide, and resveratrol mixed in a molar ratio of 1:1:1 (mass percentages: glycyrrhizin 19.7 wt%, nicotinamide 52.3 wt%, resveratrol 28 wt%), in the same amounts as the composition of Example 1. The only difference from Example 1 is that nicotinamide is used instead of polygalactosidase.

[0054] Comparative Example 10

[0055] This comparative example provides a composition composed of powders of glycyrrhizin, tranexamic acid, and resveratrol in a molar ratio of 1:1:1 (mass percentages: glycyrrhizin 25.7 wt%, tranexamic acid 53 wt%, resveratrol 21.3 wt%), in the same amounts as the composition of Example 1. The only difference from Example 1 is that resveratrol is replaced with tranexamic acid.

[0056] Comparative Example 11

[0057] This comparative example provides a composition composed of phlorizin, polydipsia glycoside, and resveratrol powders mixed in a molar ratio of 1:1:1 (phlorizin 34.5 wt%, polydipsia glycoside 24.2 wt%, resveratrol 41.3 wt%), in the same amounts as the composition of Example 1. The only difference from Example 1 is that glycyrrhizin is replaced with phlorizin.

[0058] Experiment 1: Inhibition of Melanin Synthesis and Tyrosinase Activity

[0059] 1.1 Preparation of sample solution

[0060] Using dimethyl sulfoxide (DMSO) as a solvent, Comparative Example 1 (glycyrrhizin powder) was prepared into sample solutions with molar concentrations of 0.0001 μM, 0.1 μM, and 1 μM, respectively, for testing.

[0061] Using DMSO as a solvent, Comparative Example 2 (Polygonin powder) was prepared into sample solutions with molar concentrations of 0.0001 μM, 0.1 μM, and 1 μM, respectively, for testing.

[0062] Using DMSO as a solvent, Comparative Example 3 (resveratrol powder) was prepared into sample solutions with molar concentrations of 0.0001 μM, 0.1 μM, and 1 μM, respectively, for testing.

[0063] Using deionized water as a solvent, Comparative Example 4 (tranexamic acid powder) was prepared into sample solutions with molar concentrations of 0.0001 μM, 0.1 μM, and 1 μM, respectively, for testing.

[0064] Using deionized water as a solvent, Comparative Example 5 (nicotinamide powder) was prepared into sample solutions with molar concentrations of 0.0001 μM, 0.1 μM, and 1 μM, respectively, for testing.

[0065] Using DMSO as a solvent, Comparative Example 6 (phlorizin powder) was prepared into sample solutions with molar concentrations of 0.0001 μM, 0.1 μM, and 1 μM, respectively, for testing.

[0066] Using DMSO as a solvent, Comparative Example 7 (made by mixing glycyrrhizin and polydipsia glycoside powders in an equimolar ratio) was prepared into sample solution 1 (containing 0.0001 μM glycyrrhizin and 0.0001 μM polydipsia glycoside), sample solution 2 (containing 0.1 μM glycyrrhizin and 0.1 μM polydipsia glycoside), and sample solution 3 (containing 1 μM glycyrrhizin and 1 μM polydipsia glycoside), respectively, and then tested.

[0067] Using DMSO as a solvent, Comparative Example 8 (a mixture of glycyrrhizin and resveratrol powders in an equimolar ratio) was prepared into sample solution 1 (containing 0.0001 μM glycyrrhizin and 0.0001 μM resveratrol), sample solution 2 (containing 0.1 μM glycyrrhizin and 0.1 μM resveratrol), and sample solution 3 (containing 1 μM glycyrrhizin and 1 μM resveratrol), respectively, for testing.

[0068] Using DMSO and deionized water as solvents, Comparative Example 9 (a mixture of equimolar amounts of glycyrrhizin, nicotinamide, and resveratrol powders) was prepared into sample solutions 1 (containing 0.0001 μM glycyrrhizin, 0.0001 μM nicotinamide, and 0.0001 μM resveratrol), 2 (containing 0.1 μM glycyrrhizin, 0.1 μM nicotinamide, and 0.1 μM resveratrol), and 3 (containing 1 μM glycyrrhizin, 1 μM nicotinamide, and 1 μM resveratrol), respectively, for testing.

[0069] Using DMSO and deionized water as solvents, Comparative Example 10 (a mixture of equimolar amounts of glycyrrhizin, polydipsia glycoside, and tranexamic acid powders) was prepared into sample solutions 1 (containing 0.0001 μM glycyrrhizin, 0.0001 μM polydipsia glycoside, and 0.0001 μM tranexamic acid), 2 (containing 0.1 μM glycyrrhizin, 0.1 μM polydipsia glycoside, and 0.1 μM tranexamic acid), and 3 (containing 1 μM glycyrrhizin, 1 μM polydipsia glycoside, and 1 μM tranexamic acid), respectively, and then tested.

[0070] Using DMSO as a solvent, Comparative Example 11 (a mixture of phlorizin, polydipsia glycoside, and resveratrol powders in equimolar ratio) was prepared into sample solution 1 (containing 0.0001 μM phlorizin, 0.0001 μM polydipsia glycoside, and 0.0001 μM resveratrol), sample solution 2 (containing 0.1 μM phlorizin, 0.1 μM polydipsia glycoside, and 0.1 μM resveratrol), and sample solution 3 (containing 1 μM phlorizin, 1 μM polydipsia glycoside, and 1 μM resveratrol), respectively, for testing.

[0071] Using DMSO as a solvent, the composition of Example 1 (a mixture of equimolar amounts of glycyrrhizin, polydipsia glycoside, and resveratrol powders) was prepared into sample solution 1 (containing 0.0001 μM glycyrrhizin, 0.0001 μM polydipsia glycoside, and 0.0001 μM resveratrol), sample solution 2 (containing 0.1 μM glycyrrhizin, 0.1 μM polydipsia glycoside, and 0.1 μM resveratrol), and sample solution 3 (containing 1 μM glycyrrhizin, 1 μM polydipsia glycoside, and 1 μM resveratrol) for testing.

[0072] 1.2 Experimental Principle

[0073] Melanin is a biological pigment synthesized and secreted by melanocytes. Within melanocytes, tyrosine is catalyzed by tyrosinase and undergoes a series of reactions to ultimately produce melanin.

[0074] B16-F10 mouse melanoma cells are widely used as test cells for assessing the efficacy of skin-whitening chemicals. α-MSH, an α-melanocyte-stimulating agent, can promote melanin secretion from B16-F10 cells and increase intracellular tyrosinase activity. By inducing B16-F10 cells with α-MSH, the inhibitory effects of test substances on melanin synthesis and tyrosinase activity can be measured.

[0075] 1.3 Experimental Methods

[0076] (1) Cell Culture and Treatment

[0077] B16-F10 cells were cultured in 1640 complete medium (1640 medium + 10% FBS + antibiotics) in a carbon dioxide incubator at 37°C and 5% CO2, and passaged every 2-3 days.

[0078] (2) Measurement of intracellular melanin content

[0079] B16-F10 cells in logarithmic growth phase were selected, digested with 0.25% trypsin, and seeded into 6-well plates with 2 mL of culture medium per well. The plates were incubated at 37°C in a 5% CO2 incubator. The day after seeding, culture medium containing different concentrations of the test substance and 1 μM α-MSH was added, and a control group (containing only 1 μM α-MSH) was set up with 2 mL per well. Each concentration was repeated in triplicate. After incubation at 37°C in a 5% CO2 incubator for 48 h, the supernatant was discarded, and 0.5 mL of 0.25% trypsin digestion solution was added to each well. Digestion was carried out at room temperature for 1 min. Digestion was stopped by adding 1 mL of culture medium, and the cells were pipetted into a single-cell suspension. Take 20 μL for cell counting, centrifuge 0.7 mL of cell suspension at 2000 r / min for 5 min, discard the supernatant, add 1 mL of 1 mol / L NaOH solution (containing 10% DMSO by mass), and incubate in an 80℃ water bath for 30 min. Then measure the absorbance at 490 nm using an ELISA reader.

[0080] (3) Measurement of intracellular tyrosinase activity

[0081] After the cells were treated as described above, they were digested and counted with trypsin (specific steps are the same as in 1.3-(2)). 0.7 mL of the cell suspension was centrifuged at 2000 r / min for 5 min, the supernatant was discarded, and 1 mL of 0.5% sodium deoxycholate solution was added. The cells were incubated on ice for 15 min to lyse them and prepare an extract containing tyrosinase. After preheating at 37℃, 0.5 mL of 0.3% dopa solution was added, and the reaction was carried out at 37℃ for 10 min. The absorbance was then measured at 475 nm using an ELISA reader.

[0082] (4) Results Statistics

[0083] The OD value is calibrated by the number of cells.

[0084] Melanin synthesis inhibition rate (%) = [1 - (As / Ds) / (Ac / Dc)] × 100

[0085] Tyrosinase activity inhibition rate (%) = [1 - (As / Ds) / (Ac / Dc)] × 100

[0086] In the formula: As is the absorbance of the sample group, Ac is the absorbance of the control group, Ds is the cell concentration of the sample group, and Dc is the cell concentration of the control group.

[0087] 1.4 Experimental Results

[0088] The experimental results are shown in Tables 1-3.

[0089] Table 1. Results of intracellular melanin content and tyrosinase activity at 0.0001 μM

[0090]

[0091] Table 2. Results of intracellular melanin content and tyrosinase activity at 0.1 μM.

[0092]

[0093] Table 3. Results of intracellular melanin content and tyrosinase activity at 1 μM.

[0094]

[0095]

[0096] Melanin is primarily produced and secreted by melanocytes in the basal layer of the skin. The level of melanin directly affects our skin tone. Dullness, age spots, and pigmentation are all related to abnormal melanin production and metabolism. Tyrosinase, as the rate-limiting enzyme regulating melanin production, directly influences melanin synthesis. Therefore, cellular methods are commonly used to detect the effects of cosmetic ingredients on melanin content and tyrosinase activity, thereby assessing their whitening effects.

[0097] As shown in Table 1, when the concentration of glycyrrhizin is greater than or equal to 0.1 μM, it can inhibit tyrosinase activity and melanin production, and exhibits a certain dose-dependent relationship. Low concentration of glycyrrhizin (0.0001 μM) has no significant inhibitory effect on melanin production and tyrosinase activity. When the concentration of polygalactoside is greater than or equal to 0.0001 μM, it can inhibit melanin production. When its concentration is greater than or equal to 0.1 μM, it can inhibit tyrosinase activity, and the two show a certain dose-dependent relationship. Although low concentration of polygalactoside (0.0001 μM) can inhibit melanin content to a certain extent, it does not show an inhibitory effect on tyrosinase activity. When the concentration of resveratrol is greater than or equal to 0.0001 μM, it can inhibit tyrosinase activity and melanin production. The combination of glycyrrhizin, resveratrol, and polygalactoside (Example 1) showed superior inhibitory effects on melanin production and tyrosinase activity compared to either ingredient alone, and also superior to the combination of the two. Furthermore, Example 1 exhibited a significant inhibitory effect at a low concentration (0.0001 μM) (while glycyrrhizin and resveratrol alone showed little effect). Replacing any one of the ingredients (glycyrrhizin, resveratrol, or polygalactoside) with other whitening active ingredients did not result in a significant synergistic inhibitory effect on melanin production and tyrosinase activity.

[0098] In summary, the combination of glycyrrhizin, resveratrol, and polygalactoside showed a significant synergistic effect in inhibiting melanin synthesis and tyrosinase activity, which is an efficacy characteristic not found in other types of compositions. This makes the composition more effective in whitening and allows for a reduction in the amount of each individual ingredient added, thus reducing irritation and lowering costs.

[0099] Experimental Example 2: DPPH Free Radical Scavenging Experiment

[0100] 2.1 Preparation of sample solution

[0101] Using DMSO as a solvent, Comparative Example 1 (glycyrrhizin powder) was prepared into sample solutions with molar concentrations of 0.033 mM and 3.33 mM, respectively, for testing.

[0102] Using DMSO as a solvent, Comparative Example 2 (Polygonin powder) was prepared into sample solutions with molar concentrations of 0.033 mM and 3.33 mM, respectively, for testing.

[0103] Using DMSO as a solvent, Comparative Example 3 (resveratrol powder) was prepared into sample solutions with molar concentrations of 0.033 mM and 3.33 mM, respectively, for testing.

[0104] Using DMSO as a solvent, Comparative Example 7 (made by mixing equimolar amounts of glycyrrhizin and polydipsia glycoside powders) was prepared into sample solution 1 (containing 0.033 mM glycyrrhizin and 0.033 mM polydipsia glycoside) and sample solution 2 (containing 3.33 mM glycyrrhizin and 3.33 mM polydipsia glycoside) for testing.

[0105] Using DMSO as a solvent, Comparative Example 8 (a mixture of glycyrrhizin and resveratrol powders in an equimolar ratio) was prepared into sample solution 1 (containing 0.033 mM glycyrrhizin and 0.033 mM resveratrol) and sample solution 2 (containing 3.33 mM glycyrrhizin and 3.33 mM resveratrol), and then tested.

[0106] Using DMSO and deionized water as solvents, Comparative Example 9 (a mixture of equimolar amounts of glycyrrhizin, nicotinamide, and resveratrol powders) was prepared into sample solution 1 (containing 0.033 mM glycyrrhizin, 0.033 mM nicotinamide, and 0.033 mM resveratrol) and sample solution 2 (containing 3.33 mM glycyrrhizin, 3.33 mM nicotinamide, and 3.33 mM resveratrol) for testing.

[0107] Using DMSO and deionized water as solvents, Comparative Example 10 (a mixture of equimolar amounts of glycyrrhizin, polydipsia glycoside, and tranexamic acid powders) was prepared into sample solution 1 (containing 0.033 mM glycyrrhizin, 0.033 mM polydipsia glycoside, and 0.033 mM tranexamic acid) and sample solution 2 (containing 3.33 mM glycyrrhizin, 3.33 mM polydipsia glycoside, and 3.33 mM tranexamic acid) for testing.

[0108] Using DMSO as a solvent, Comparative Example 11 (a mixture of phlorizin, polydipsia glycoside, and resveratrol powders in equimolar ratio) was prepared into sample solution 1 (containing 0.033 mM phlorizin, 0.033 mM polydipsia glycoside, and 0.033 mM resveratrol) and sample solution 2 (containing 3.33 mM phlorizin, 3.33 mM polydipsia glycoside, and 3.33 mM resveratrol) for testing.

[0109] Using DMSO as a solvent, the composition of Example 1 (a mixture of equimolar amounts of glycyrrhizin, polydipsia glycoside, and resveratrol powders) was prepared into sample solution 1 (containing 0.033 mM glycyrrhizin, 0.033 mM polydipsia glycoside, and 0.033 mM resveratrol) and sample solution 2 (containing 3.33 mM glycyrrhizin, 3.33 mM polydipsia glycoside, and 3.33 mM resveratrol) for testing.

[0110] Using DMSO as a solvent, the composition of Example 2 (a mixture of powders of glycyrrhizin, polygalactosidase, and resveratrol in a molar ratio of 4.9:1.2:1) was prepared into sample solutions of the same concentration as that of Example 1, and then tested.

[0111] Using DMSO as a solvent, Example 3 (a composition consisting of powders of glycyrrhizin, polydipsia oleracea glycoside, and resveratrol mixed in a molar ratio of 1:4.2:5.7) was prepared into sample solutions of the same concentration as Example 1 and tested.

[0112] Using DMSO as a solvent, the composition of Example 4 (a mixture of powders of glycyrrhizin, polygalactosidase, and resveratrol in a molar ratio of 1.2:1:13.7) was prepared into sample solutions of the same concentration as that of Example 1, and then tested.

[0113] 2.2 Experimental Principle

[0114] DPPH is a stable nitrogen-centered free radical with maximum absorption at a wavelength of 519 nm. DPPH solutions in ethanol / methanol are deep purple. When a free radical scavenger is added to a DPPH solution, the unpaired electrons of DPPH are paired, causing the color to lighten and the absorbance at the maximum absorption wavelength to decrease. Therefore, the degree of decrease in absorbance can be used to determine the ability of a substance to scavenge DPPH free radicals.

[0115] 2.3 Experimental Methods

[0116] (1) Preparation of DPPH solution

[0117] Weigh 0.05 g of DPPH, dissolve it in an appropriate amount of anhydrous ethanol, sonicate for 30 min, and then bring the volume to 100 mL. Store the DPPH solution away from light, prepare it fresh, and use it within 3.5 hours.

[0118] (2) Formal Experiment

[0119] Measurement of A0 value: Add 50 μL of anhydrous ethanol to the microplate (one sample is made in 3 replicates), then add 100 μL of DPPH solution to each well, seal the plate with plastic wrap, let it stand at 37℃ for 30 min, and measure the absorbance A0 at 519 nm in the microplate reader.

[0120] Measurement of A value: Add 50 μL of sample solution of different concentrations to each well of the microplate (one sample is made into 3 replicates), then add 100 μL of DPPH solution to each well, seal the plate with plastic wrap, let it stand at 37℃ for 30 min, and measure the absorbance A value at 519 nm in the microplate reader.

[0121] (3) Results Statistics

[0122] DPPH clearance rate (%) = (A0 - A) / A0 × 100

[0123] In the formula: A0 is the absorbance value of the control group, and A is the absorbance value of the sample group.

[0124] 2.4 Experimental Results

[0125] The experimental results are shown in Table 4.

[0126] Table 4. Results of DPPH free radical scavenging rate detection

[0127]

[0128]

[0129] When cells receive external stimuli (such as UV-induced oxidation), they experience oxidative stress, generating a large number of free radicals. Furthermore, melanin production is an aerobic reaction, and the presence of excessive free radicals accelerates melanin production. Therefore, the antioxidant activity of raw materials can indirectly affect melanin production. The antioxidant capacity can be assessed using the DPPH free radical scavenging method.

[0130] As shown in Table 4, glycyrrhizin, polygalactoside, and resveratrol all possess a certain ability to scavenge DPPH free radicals. However, the combination of the three (glycyrrhizin + polygalactoside + resveratrol) is significantly more effective than the combination of two (Comparative Example 7: glycyrrhizin + polygalactoside; Comparative Example 8: glycyrrhizin + resveratrol). Furthermore, replacing any one of the components among glycyrrhizin, resveratrol, and polygalactoside with other whitening active ingredients did not show any significant synergistic effect in scavenging DPPH free radicals.

[0131] The above results indicate that the combination of glycyrrhizin, polygalactoside, and resveratrol can significantly enhance its antioxidant effect, and the combination of the three has a synergistic effect on DPPH free radical scavenging rate.

[0132] Experimental Example 3: Hydroxyl Radical Scavenging Experiment

[0133] 3.1 Preparation of sample solution

[0134] Using DMSO as a solvent, Comparative Example 1 (glycyrrhizin powder) was prepared into a sample solution with a molar concentration of 16.67 mM for testing.

[0135] Using DMSO as a solvent, Comparative Example 2 (Polygonin powder) was prepared into a sample solution with a molar concentration of 16.67 mM for testing.

[0136] Using DMSO as a solvent, Comparative Example 3 (resveratrol powder) was prepared into a sample solution with a molar concentration of 16.67 mM for testing.

[0137] Using DMSO as a solvent, Comparative Example 7 (a mixture of equimolar amounts of glycyrrhizin and polydipsia glycoside powders) was prepared into a sample solution (containing 16.67 mM glycyrrhizin and 16.67 mM polydipsia glycoside) for testing.

[0138] Using DMSO as a solvent, Comparative Example 8 (a mixture of glycyrrhizin and resveratrol powders in an equimolar ratio) was prepared into a sample solution (containing 16.67 mM glycyrrhizin and 16.67 mM resveratrol) for testing.

[0139] Using DMSO as a solvent, the sample solution (containing 16.67 mM glycyrrhizin, 16.67 mM polydextrin and 16.67 mM resveratrol) of Example 1 (a composition consisting of equimolar mixtures of powders of glycyrrhizin, polydextrin and resveratrol) was prepared and tested.

[0140] 3.2 Experimental Principle

[0141] Hydroxyl radicals (·OH) are classic reactive oxygen species (ROS). ·OH possesses strong oxidizing power and high toxicity, posing significant harm to the body and exhibiting good biological representativeness. This reaction utilizes the Fenton reaction (Fe... 2+ +H₂O₂→·OH+H₂O+Fe 3+ The process involves generating ·OH radicals, followed by the addition of salicylic acid. The salicylic acid reacts with the generated ·OH radicals to form a colored compound, 2,3-dihydroxybenzoic acid, which has a maximum absorption peak at 510 nm. The absorbance at 510 nm is used to determine the hydroxyl radical scavenging ability of the test compound, and the level of hydroxyl radical scavenging ability reflects the antioxidant activity of the test compound.

[0142] 3.3 Experimental Methods

[0143] (1) Reagent preparation

[0144] 9mM ethanol-salicylic acid solution: Weigh 1.243g of salicylic acid, dissolve it in ethanol, bring the volume to 100mL, and then dilute it 10 times.

[0145] 9mM ferrous sulfate solution: Weigh 2.502g FeSO4·7H2O, dissolve in deionized water, bring to a final volume of 100mL, and then dilute 10 times.

[0146] 8.8 mL H₂O₂ solution: Weigh 9.973 g of 30% H₂O₂, bring the volume to 100 mL with deionized water, and then dilute 100 times.

[0147] (2) Formal Experiment

[0148] The amounts of each reagent added are shown in Table 5. After adding the reagents to the ELISA plate, seal the plate with plastic wrap, incubate at 37°C for 15 minutes, then remove it and measure the absorbance A0, Ax, and Ax0 at 510 nm using an ELISA reader. Each group needs to be performed in triplicate.

[0149] Table 5 Sample dosage for hydroxyl radical scavenging ability test

[0150] Reagent (μL) <![CDATA[Blank control group (A0)]]> Sample group (Ax) <![CDATA[Sample control group (Ax0)]]> <![CDATA[9 mmol / L FeSO4 solution]]> 10 10 10 9 mmol / L ethanol-salicylic acid solution 10 10 10 <![CDATA[8.8 mmol / L H2O2 solution]]> 10 10 0 Deionized water 120 0 10 sample 0 120 120 total 150 150 150

[0151] (3) Results Statistics

[0152] Hydroxyl radical scavenging rate (%) = [A0 - (Ax - Ax0)] / A0 × 100;

[0153] In the formula: A0 is the absorbance value of the blank control group, which refers to the system without the addition of the sample.

[0154] Ax represents the absorbance of the sample group.

[0155] Ax0 is the absorbance value of the sample control group, which refers to the system without H2O2 and with the sample.

[0156] 3.4 Experimental Results

[0157] The experimental results are shown in Table 6.

[0158] Table 6. Results of Hydroxyl Radical Scavenging Rate Detection

[0159]

[0160]

[0161] Hydroxyl radicals are a type of reactive oxygen species (ROS) that can cause significant damage to the body. We can use in vitro ·OH scavenging to assess the antioxidant activity of test samples. The higher the ·OH scavenging rate, the stronger the antioxidant activity.

[0162] As shown in Table 6, all three of them, including glycyrrhizin, polydipsia glycoside, and resveratrol, have a certain ability to scavenge ·OH. In the composition, the effect of the combination of the three (Example 1: glycyrrhizin + polydipsia glycoside + resveratrol) is significantly better than that of the individual components or the combination of the two (Comparative Example 7: glycyrrhizin + polydipsia glycoside; Comparative Example 8: glycyrrhizin + resveratrol).

[0163] The above results indicate that the combination of glycyrrhizin, polygalactoside, and resveratrol significantly enhances its antioxidant effect, and the three components exhibit synergistic effects in ·OH scavenging. This enhanced antioxidant efficacy further contributes to its skin-whitening effect, helping to improve uneven skin tone and dullness.

[0164] Application Example 1

[0165] This application example provides an emulsion containing the composition of the present invention, the specific formulation of which is shown in Table 7, with the total mass fraction being 100%.

[0166] Table 7 Emulsion Formulations

[0167]

[0168]

[0169] The method for preparing the above emulsion includes the following steps:

[0170] 1. Dissolve the p-hydroxyacetophenone in 1,2-hexanediol by heating until clear and transparent, and set aside;

[0171] 2. Heat and stir the aqueous phase to 80°C to ensure uniform dispersion, then add the additive phase;

[0172] 3. Add the oil phase to the oil phase pot and heat and stir until 80°C;

[0173] 4. Transfer the material from the oil phase pot into the emulsification pot, stir evenly, then vacuum the emulsification pot, homogenize and emulsify for 10 minutes, keep warm for 15 minutes, then turn on the cooling water to cool to 40℃, then add the additive phase and water phase mixture, and stir evenly.

[0174] 5. Cool down to 35℃, and discharge the material after passing the inspection.

[0175] Application Example 2

[0176] This application example provides an essence containing the composition of the present invention. The specific formula of the essence is shown in Table 8, and the total mass fraction is 100%.

[0177] Table 8 Serum Formula

[0178]

[0179]

[0180] The preparation method of the above-mentioned serum includes the following steps:

[0181] 1. Dissolve the p-hydroxyacetophenone in 1,2-hexanediol by heating until clear and transparent, and set aside;

[0182] 2. Preheat to dissolve the oil phase until it becomes transparent;

[0183] 3. Add the aqueous phase to the deionized water, stir well, and heat and stir to 80℃;

[0184] 4. Keep warm for 10 minutes, then turn on the cooling water to cool down to 60°C. While stirring, slowly add the p-hydroxyacetophenone that has been dissolved in 1,2-hexanediol and the oil phase, and stir until it is evenly dissolved.

[0185] 5. Cool to 40°C and add the additive phase composition;

[0186] 6. Stir well, cool to 35℃, and discharge through a 300-mesh filter after the pH test is qualified.

[0187] It is not difficult to imagine that the lotion in Application Example 1 and the serum in Application Example 2, since both use the composition of the present invention, have a synergistic effect due to the synergistic effect of glycyrrhizin, polydipsia glycoside, and resveratrol in the composition. By regulating the three major whitening mechanism pathways of inhibiting tyrosinase activity, inhibiting melanin production, and combating oxidative stress, they have a highly effective whitening effect. Therefore, these cosmetics, because they contain the composition of the present invention, also have a highly effective whitening effect.

[0188] The preferred embodiments of the present invention have been described in detail above, but the present invention is not limited to the embodiments described. Those skilled in the art can make various equivalent modifications or substitutions without departing from the spirit of the present invention, and these equivalent modifications or substitutions are all included within the scope defined by the claims of this application.

Claims

1. A composition characterized in that, The composition consists of 10wt%-70wt% glabridin, 10wt%-50wt% polydatin and 10wt%-80wt% resveratrol.

2. Process for the preparation of the composition according to claim 1, characterized in that, The method comprises the following steps: Mixing the glabridin, polydatin and resveratrol to obtain the composition.

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