Composition with effects of resisting initial aging and brightening as well as preparation process and application of composition

Abstract

The invention relates to the technical field of cosmetics, and particularly discloses a composition with anti-aging and brightening effects as well as a preparation process and application thereof. The composition with the effects of resisting initial aging and brightening is prepared from the following components in percentage by weight: 1 to 5 percent of glycyrrhiza glabra root extract, 10 to 20 percent of silybum marianum extract, 0.005 to 0.01 percent of palmitoyl pentapeptide-4, 0.0005 to 0.001 percent of arginine / lysine polypeptide, 0 to 1 percent of sargassum elongatum extract, 0 to 1 percent of olive leaf extract, 0 to 1 percent of diglucosyl gallic acid, 0 to 0.5 percent of dendrobium officinale stem extract and the balance of water. The glycyrrhiza glabra root extract is an anti-saccharification glycyrrhiza glabra root extract, and the anti-saccharification glycyrrhiza glabra root extract is prepared by carrying out directional hydrolysis treatment on a glycyrrhiza glabra root extract. According to the present invention, the collagen regeneration can be effectively promoted, and the new collagen can be systematically protected from oxidation and saccharification damage.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of cosmetics, more particularly, it relates to a composition with anti-early aging and lightening effect, and a preparation process and application thereof. BACKGROUND

[0002] Skin aging is a complex physiological process, and the main clinical manifestations are skin relaxation, decreased elasticity, wrinkle formation, dryness and thinning, and pigmentation. Its internal mechanism is closely related to age, external environmental stimulation (such as ultraviolet light), staying up late, and mental stress. These factors collectively accelerate the degradation of collagen (especially types I and III) in the dermis, while the rate of new synthesis is relatively insufficient, resulting in a collapsed support structure and causing signs of aging such as relaxation and wrinkles. At present, the mainstream anti-aging cosmetic technology focuses on stimulating fibroblast activity, supplementing synthetic raw materials or signal peptides, and other ways to "promote collagen production" in a single dimension, aiming to increase the absolute amount of collagen in the dermis.

[0003] However, the existing technical solutions generally have a key defect: they fail to adequately address and solve the "quality" and "stability" problems of newly synthesized collagen. In the complex skin environment, newly synthesized collagen is easily attacked by excess reactive oxygen species (ROS) produced by oxidative stress and damaged by advanced glycosylation end products (AGEs) formed by glycation. These factors can cause structural abnormalities, cross-linking disorders, and increased fragility in collagen, making it functionally inadequate and unable to effectively support its function. Therefore, simply increasing the amount of collagen without ensuring its structural and functional integrity greatly reduces the final effect of anti-wrinkle and lightening, and cannot achieve long-term and fundamental skin rejuvenation.

[0004] In addition, the lightening and anti-wrinkle compositions in the prior art are limited to inhibiting tyrosinase activity to reduce melanin production, or accelerating keratinocyte exfoliation through acid and enzyme ingredients to quickly improve the visual appearance of skin color. Although these methods can improve skin color to some extent, their action pathways are weakly related to the health status of dermal collagen, and they have obvious deficiencies in improving the quality of existing and newly synthesized collagen and building a comprehensive collagen protection network against oxidative and glycation damage. Therefore, there is an urgent need in the art for a comprehensive solution that can synergistically promote collagen synthesis while effectively protecting collagen structure and improving its functional quality, and integrate lightening skin color. SUMMARY

[0005] To solve the above problems, the present application provides a composition with anti-early aging and lightening effect, and a preparation process and application thereof.

[0006] The first part of this application provides a composition with anti-aging and brightening effects, which adopts the following technical solution: A composition with anti-aging and brightening effects, comprising the following components by weight percentage: The extract comprises 1%-5% of Glycyrrhiza glabra root extract, 10%-20% of Milk thistle extract, 0.005%-0.01% of Palmitoyl Pentapeptide-4, 0.0005%-0.001% of Arginine / Lysine Peptide, 0-1% of Elongatus spp. extract, 0-1% of Olive leaf extract, 0-1% of Diglucosyl Gallic Acid, 0-0.5% of Dendrobium officinale stem extract, with the balance being water; the Glycyrrhiza glabra root extract is an anti-glycation Glycyrrhiza glabra root extract, which is prepared by directional hydrolysis of Glycyrrhiza glabra root extract.

[0007] By employing the above technical solution, firstly, the combined use of palmitoyl pentapeptide-4 and arginine / lysine peptides allows palmitoyl pentapeptide-4, as a well-defined collagen synthesis signaling stimulant, to effectively activate fibroblasts and initiate the collagen synthesis process upstream, aiming to increase the amount of collagen in the dermis. Simultaneously, the arginine / lysine peptides, acting as anti-glycation agents, can pre-bind reducing sugars or reactive carbonyl substances, reducing the formation of advanced glycation end products (AGEs) at the source, creating a protected synthetic microenvironment for new collagen. Secondly, a specially treated anti-glycation licorice root extract is introduced. Unlike conventional licorice extracts that only possess whitening and antioxidant functions, this directionally hydrolyzed extract retains its ability to inhibit tyrosinase and scavenge free radicals, while its inherent flavonoid aglycones and newly formed small-molecule polyphenol fragments possess direct and potent inhibitory activity against AGEs formation. Furthermore, the extracts of *Euphorbia pulcherrima*, olive leaf, diglucosyl gallic acid, and *Dendrobium officinale* stem in the composition work together to maintain a healthy metabolic environment in the dermis from multiple aspects, including continuous anti-oxidation, inhibition of glycation, soothing and repair, and providing moisturizing and nutritional support to the extracellular matrix. This strengthens the overall anti-aging network and synergistically improves skin laxity, wrinkles, and dullness caused by collagen loss and dysfunction.

[0008] Optionally, the method for obtaining the anti-glycation licorice root extract is characterized by comprising the following steps: Quaternary ammonium salt, organic acid and metal salt are mixed in a molar ratio of 1:(1~3):(0.05~0.2) and stirred evenly at 80~120℃ to obtain a ternary eutectic solvent; A ternary eutectic solvent and water were mixed at a volume ratio of 6:4 to 8:2 to obtain a hydrolysis medium. Licorice root extract was added to the hydrolysis medium, and the solid-liquid ratio was controlled at 1:10 to 1:30 g / mL. The mixture was reacted at 45 to 55 °C for 1 to 1.5 hours under the assistance of ultrasound at a frequency of 35 to 40 kHz and a power density of 50 to 80 W / L to obtain a hydrolysate. A metal ion chelating agent is added to the hydrolysate, and after stirring and mixing, the mixture is adsorbed through an ion exchange column, and the effluent is collected. The pH of the effluent was adjusted to 6.0–7.0, and after concentration and drying, the anti-glycation licorice root extract was obtained.

[0009] By employing the above-mentioned technical solution and using a ternary eutectic solvent (DES) formulated with a specific molar ratio as the reaction medium, mild and efficient hydrolysis catalysis can be achieved simultaneously. At relatively low temperatures and within a short time, components such as flavonoid glycosides in the root extract of *Glycyrrhiza glabra* are directionally converted into more active aglycones and small molecule fragments, significantly enhancing the extract's direct anti-glycation (AGEs) inhibitory activity. Secondly, the introduction of ultrasound, through its cavitation and mechanical effects, strengthens the mass transfer process and promotes sufficient contact between reactant molecules, thereby further improving hydrolysis efficiency and homogeneity under mild conditions. By directly adding a chelating agent and performing ion exchange column adsorption after hydrolysis, free metal ions introduced by DES can be effectively captured and removed, improving activity while addressing the critical safety issue of metal residues in cosmetic raw materials. Finally, the entire process is completed under mild conditions of pH 6.0-7.0, maximizing the preservation of the stability of thermosensitive and photosensitizing active ingredients such as glycyrrhizin.

[0010] Optionally, the quaternary ammonium salt is one of choline chloride, tetraethylammonium chloride, or tetrabutylammonium chloride; the organic acid is one of lactic acid, citric acid, or oxalic acid; the metal salt is one of zinc chloride, aluminum chloride, or stannous chloride; the metal salt is zinc chloride or aluminum chloride; and the metal ion chelating agent is one of sodium phytate, disodium EDTA, or sodium citrate, with an addition amount of 0.1% to 0.5% of the total weight of the hydrolysate.

[0011] The above technical solution limits the quaternary ammonium salt to choline chloride, tetraethylammonium chloride, or tetrabutylammonium chloride because these substances, as hydrogen bond acceptors, can efficiently form stable eutectic systems with organic acids, and their cost and biocompatibility are relatively ideal. The organic acids are limited to lactic acid, citric acid, or oxalic acid, based on their good coordination ability with the quaternary ammonium salt as hydrogen bond donors. Furthermore, the acidic environment provided by these organic acids, combined with the synergistic effect of DES, is key to achieving mild acid-catalyzed hydrolysis, avoiding the damage to the active ingredients caused by strong inorganic acids. The metal salts are specifically limited to zinc chloride, aluminum chloride, or stannous chloride because they act as mild Lewis acid catalysts, effectively catalyzing glycosidic bond cleavage, and the introduced metal ions are more easily removed in subsequent purification compared to other transition metal salts. Further optimization with zinc chloride or aluminum chloride represents the optimal choice after balancing catalytic activity, safety, and cost. The chelating agent is limited to sodium phytate, disodium EDTA, or sodium citrate, and the addition amount is controlled at 0.1%-0.5%. This is to efficiently complex free metal ions to assist in subsequent purification. Sodium phytate has excellent biocompatibility, disodium EDTA has extremely strong chelating ability, and sodium citrate has both pH adjustment and chelating effects. This dosage range can ensure effective chelation while avoiding the increased cost or potential impact on subsequent processes that may result from excessive chelating agent.

[0012] Optionally, the ion exchange column is packed with sulfonic acid-type strong acid cation exchange resin or carboxylic acid-type weak acid cation exchange resin; the flow rate of the adsorption treatment is controlled at 1 to 3 times the column volume per hour.

[0013] The above technical solution limits the ion exchange column to either sulfonic acid-type strong acid cation exchange resin or carboxylic acid-type weak acid cation exchange resin because both types of resins have highly efficient exchange and adsorption capabilities for cations such as Zn²⁺ and Al³⁺. Sulfonic acid-type resins have a large exchange capacity and a wide applicable pH range, ensuring stable operation under a wide range of conditions; carboxylic acid-type resins exhibit good selectivity under near-neutral conditions and are easy to regenerate. Furthermore, controlling the adsorption flow rate to 1–3 column volumes per hour ensures sufficient contact time between the hydrolysate and the resin, resulting in a more complete ion exchange reaction and extremely low metal residue. This represents the optimal balance between treatment effect and production efficiency. A flow rate that is too low will result in low production capacity, while a flow rate that is too high may lead to incomplete adsorption and metal ion penetration.

[0014] Optionally, the weight ratio of the licorice root extract to the milk thistle extract is 1:(4-6).

[0015] By adopting the above technical solution, at this ratio, the tyrosinase inhibition and antioxidant effects dominated by glycyrrhizin and the NF-κB inflammatory pathway inhibition and cell protection effects dominated by silymarin achieve a balance in efficacy, jointly creating an intracellular environment with low oxidative stress and low inflammation levels. This is not only conducive to the efficient synthesis of collagen by fibroblasts, but also significantly reduces the risk of collagen damage during the synthesis process, thereby improving the overall anti-aging efficiency.

[0016] Optionally, the extract of *Euphorbia lathyris* and the disaccharidic gallic acid are present in a weight ratio of 1:(0.8-1.2).

[0017] By employing the above-mentioned technical solution, diglucosyl gallic acid, as a small-molecule, potent glycation inhibitor, can directly intercept the glycation reaction; while *Euphorbia pulvinata* extract provides polysaccharides and other components to optimize the extracellular microenvironment. The synergistic effect of these two ingredients in this ratio not only more effectively protects collagen fibers from AGEs cross-linking damage but also potentially promotes the correct alignment and functional maturation of collagen fibers by providing support and hydration, thereby structurally strengthening the dermis and enhancing the durability of the anti-wrinkle and firming effects.

[0018] Optionally, it may also include 0.1%-1% of modified phospholipids, wherein the modified phospholipids are selected from at least one of hydrogenated lecithin, phosphatidylcholine and cholesterol complex.

[0019] By adopting the above technical solution, hydrogenated lecithin and phospholipid-cholesterol complex can self-assemble into a nanoscale drug delivery system, effectively encapsulating hydrophobic molecules, enhancing their stability in water-based formulations, and utilizing the similarity and compatibility between phospholipids and skin stratum corneum lipids to promote the penetration of active ingredients into the skin barrier and deliver them more efficiently to the target site of action (dermis), thereby significantly improving the bioavailability and actual efficacy of the composition.

[0020] Optionally, the modified phospholipids and milk thistle extract are pre-formed into a silymarin-phospholipid complex using the following method: An organic phase solution containing modified phospholipids and an organic phase solution containing milk thistle extract were mixed using microfluidic technology to form a blended stream. The blended stream is brought into contact with the antisolvent aqueous solution in a microfluidic channel so that silymarin and the modified phospholipid can simultaneously form complex particles through antisolvent precipitation and self-assembly. The complex particles were collected and purified to obtain the silymarin-phospholipid complex.

[0021] By employing the above-mentioned technical solution, silybin, a typical hydrophobic molecule, exhibits poor solubility, chemical instability, and limited transdermal absorption in water-based cosmetic formulations. If it cannot be effectively delivered to target cells deep within the skin, it cannot be converted into corresponding in vivo efficacy, leading to the failure of the collagen protective network at this critical juncture. The silybin-phospholipid complex nanoparticles formed in this application, due to their similarity to the lipid bilayer structure of the stratum corneum and their nanoscale effect, can more effectively penetrate the skin barrier, targeting silybin to dermal fibroblasts. This ensures effective intervention against intracellular oxidative stress and inflammatory pathways. The process first involves instantaneously, turbulently, and efficiently blending two organic phase solutions containing modified phospholipids and silybin extract within a microchannel, ensuring uniform dispersion at the molecular level. Subsequently, this blended stream instantaneously contacts and mixes with an antisolvent. This instantaneous mixing causes a dramatic change in the solvent environment, triggering simultaneous supersaturation precipitation (antisolvent precipitation) and molecular self-assembly of silybin and phospholipid molecules. During the precipitation process, hydrophobic silymarin molecules are in situ encapsulated within the hydrophobic core of simultaneously formed phospholipid micelles, or directly complexed with phospholipids through intermolecular forces. This method can repeatedly and stably prepare composite nanoparticles with extremely narrow particle size distribution, controllable average particle size, and high encapsulation efficiency.

[0022] Secondly, this application provides a method for preparing a composition with anti-aging and brightening effects.

[0023] A method for preparing a composition with anti-aging and brightening effects includes the following steps: All components were mixed and homogenized under a protective atmosphere to obtain a composition with anti-aging and brightening effects.

[0024] By employing the above technical solution, and by introducing inert gases such as nitrogen into the system to replace and isolate oxygen, the oxidation loss of active ingredients can be effectively reduced during the preparation process. Combined with the system homogenization process that provides uniformity and physical stability, this method can maximize the chemical activity and intended synergistic effects of each component in the final product.

[0025] Thirdly, this application provides an application of a composition with anti-aging and brightening effects.

[0026] Application of a composition with anti-aging and brightening effects in the preparation of cosmetics for skin anti-wrinkle, firming, brightening and improving uneven skin tone.

[0027] In summary, this application has the following beneficial effects: 1. This application combines palmitoyl pentapeptide-4 with arginine / lysine peptides and innovatively introduces anti-glycation licorice root extract that has undergone targeted hydrolysis. This approach promotes collagen regeneration while constructing a dual molecular interception network against oxidative and glycation damage from the source, significantly improving the structural integrity and functional stability of newly formed collagen.

[0028] 2. This application significantly improves the efficacy, stability and transdermal delivery efficiency of active ingredients by using specific ratios of licorice root extract and milk thistle extract, seaweed extract and diglucosyl gallic acid, as well as milk thistle-phospholipid complex prepared by microfluidic technology and anti-glycation licorice root extract prepared by ultrasonic-assisted hydrolysis, ensuring that the composition can function efficiently and stably.

[0029] 3. This application systematically intervenes in multiple pathways, including collagen metabolism, oxidative stress, glycation reaction, and melanin production, to synergistically improve skin laxity, wrinkles, and dullness caused by collagen loss, dysfunction, and pigmentation, ultimately achieving a comprehensive anti-aging effect that improves wrinkles and laxity, brightens skin tone, and evens out skin tone. Detailed Implementation

[0030] The present application will be further described in detail below with reference to the embodiments.

[0031] The specifications of the raw materials used in this application are as follows: 1. The content of glycyrrhizin in the root extract of Glycyrrhiza glabra shall not be less than 40%, and the microbial limits shall meet the requirements for cosmetic raw materials, with a total bacterial count ≤1000 CFU / g, a total mold and yeast count ≤100 CFU / g, and Staphylococcus aureus and Pseudomonas aeruginosa shall not be detected.

[0032] 2. The total silymarin content of milk thistle extract is 65%-80% (determined by UV method), of which the content of silymarin A+B should not be less than 30%. Loss on drying ≤5.0%; solvent residue meets relevant regulatory requirements. Heavy metal and microbial limits meet the safety standards for cosmetic raw materials.

[0033] 3. Palmitoyl pentapeptide-4 purity (HPLC method) ≥ 95.0%, peptide content (calculated as anhydrous and salt-free) ≥ 98.0%. Loss on drying ≤ 5.0%; heavy metals (as Pb) ≤ 10 mg / kg.

[0034] 4. The total polypeptide content in the arginine / lysine polypeptide is ≥95.0% (nitrogen determination method); the molar ratio of arginine to lysine is 0.8:1.2, and the loss on drying is ≤8.0%.

[0035] 5. The pH value (1% aqueous solution) of the *Elongatus spp.* extract should be between 5.0 and 7.0. The polysaccharide content should be ≥1.0% (liquid) or ≥10.0% (powder). It should comply with the safety limits for microorganisms and heavy metals in cosmetic raw materials.

[0036] 6. Olive leaf extract contains 15%-40% oleuropein (HPLC method). Loss on drying (powder) ≤5.0%. Heavy metal and microbial limits meet standards.

[0037] 7. Diglucosyl gallic acid purity (HPLC method) ≥97.0%, loss on drying ≤5.0%.

[0038] 8. The total polysaccharide content (calculated as glucose) in the Dendrobium officinale stem extract is ≥30.0% (phenol-sulfuric acid method). Loss on drying (powder) is ≤8.0%. pH value (1% aqueous solution) is near neutral. It meets the safety standards for cosmetic raw materials.

[0039] 9. Hydrogenated lecithin contains phosphatidylcholine (PC) content ≥90.0% (HPLC method), iodine value ≤5.0gI2 / 100g, acid value ≤30mgKOH / g, and peroxide value ≤5.0mmol / kg.

[0040] In this application, all additives without specifically defined specifications are of analytical grade.

[0041] Preparation Example 1 A method for preparing an anti-glycation licorice root extract: Step S1 Weigh out 139.6 g (1.0 mol) of choline chloride (ChCl), 204.2 g (2.0 mol) of lactic acid (LA, 88% purity, based on pure lactic acid), and 13.6 g (approximately 0.1 mol) of zinc chloride hexahydrate (ZnCl2·6H2O). Place all three components in a 500 mL three-necked flask equipped with a stirrer and heating mantle. Turn on the stirrer, set the speed to 500 rpm, and gradually increase the temperature to 90 °C. Continue stirring at this temperature for 45 minutes until the mixture changes from an initial turbid slurry to a clear, transparent, homogeneous, and stable pale yellow liquid. Stop heating and allow it to cool naturally to room temperature. The resulting liquid is the ternary eutectic solvent (DES) of choline chloride, lactic acid, and zinc chloride, and should be sealed for later use.

[0042] Step S2 Take 70 mL of the ternary DES prepared in step S1 and mix it with 30 mL of deionized water in a beaker. Stir magnetically for 5 minutes to obtain a homogeneous and clear hydrolysis medium. Weigh 5.0 g of licorice root extract powder that has passed through a 60-mesh sieve and slowly add it to the above hydrolysis medium, controlling the total solid-liquid ratio to be 1:20 g / mL. Transfer the mixed suspension to a 250 mL jacketed ultrasonic reactor.

[0043] Turn on the ultrasonic generator, set the operating frequency to 38kHz, and the power density to 65W / L (based on the total volume of the reaction solution). Control the internal temperature of the reaction system at 50℃±2℃ using a circulating water bath. Under continuous ultrasonic irradiation and gentle stirring, the hydrolysis reaction is carried out for a total reaction time of 1.2 hours.

[0044] Step S3 After the reaction is complete, cool the hydrolysate to room temperature. Add 0.30 g of sodium phytate (approximately 0.3% of the total weight of the hydrolysate) and stir magnetically for 20 minutes to ensure that the chelating agent is fully dispersed and binds to the free metal ions.

[0045] Prepare a glass chromatography column with an inner diameter of 2 cm and a bed height of 20 cm. Pack approximately 60 mL of a strong acidic cation exchange resin (AmberChrom™ 50WX8 Ion Exchange Resin) using a wet packing method. First, rinse the column with deionized water until the eluent is neutral. Pass the hydrolysate treated with the chelating agent through the ion exchange column at a constant flow rate of approximately 1.5 column volumes per hour (i.e., 90 mL / h). Collect all the eluent. This process effectively adsorbs and removes metal ions such as Zn²⁺.

[0046] Step S4 The pH of the effluent obtained in step S3 was slowly adjusted to 6.5 ± 0.1 using a dilute sodium hydroxide solution (0.1 mol / L). The neutralized solution was then transferred to a rotary evaporator and concentrated under reduced pressure conditions of a 60°C water bath and -0.085 MPa until the concentrate became a viscous paste (solid content approximately 35%).

[0047] Finally, the concentrate was dried using a spray dryer. The inlet air temperature was set to 165°C, the outlet air temperature to 88°C, and the feed flow rate was adjusted to a suitable range. The fine powder collected from the bottom of the drying tower is the anti-glycation licorice root extract of this preparation example.

[0048] Preparation Example 2 A method for preparing an anti-glycation licorice root extract differs from Preparation Example 1 only in that, in step S1, choline chloride (ChCl) is replaced with an equimolar amount of tetraethylammonium chloride (TEAC). All other steps and parameters are identical.

[0049] Preparation Example 3 A method for preparing an anti-glycation licorice root extract differs from Preparation Example 1 only in that, in step S1, lactic acid (LA) is replaced with an equimolar amount of citric acid (CA). Since citric acid is a solid, the heating and stirring time needs to be adjusted accordingly to ensure complete melting and formation of a homogeneous DES. The remaining steps and parameters are identical.

[0050] Preparation Example 4 A method for preparing an anti-glycation licorice root extract differs from Preparation Example 1 only in that, in step S1, zinc chloride (ZnCl2) is replaced with an equimolar amount of aluminum chloride (AlCl3). Note that anhydrous AlCl3 is easily hydrolyzed, and the operation must be carried out rapidly in a dry environment. All other steps and parameters are identical.

[0051] Preparation Example 5 A method for preparing an anti-glycation licorice root extract differs from Preparation Example 1 only in that, in step S1, the molar ratio of choline chloride (ChCl):oxalic acid (OA):AlCl3 is 1:1.5:0.15. Because oxalic acid has a high melting point, the reaction temperature needs to be increased to approximately 100°C to promote the formation of homogeneous DES. The remaining steps and parameters are the same as in Preparation Example 1.

[0052] Preparation Example 6 A method for preparing an anti-glycation licorice root extract differs from Preparation Example 1 only in that, in step S1, the molar ratio is adjusted to ChCl:LA:ZnCl2 = 1:3:0.2. All subsequent steps are the same as in Preparation Example 1.

[0053] Preparation Example 7 A method for preparing an anti-glycation licorice root extract differs from Preparation Example 1 only in that, in step S2, the ratio of the hydrolysis medium is adjusted to DES:water = 6:4 (v / v). All other steps and parameters are identical.

[0054] Preparation Example 8 A method for preparing an anti-glycation licorice root extract differs from Preparation Example 1 only in that, in step S2, the solid-liquid ratio is adjusted to 1:10 g / mL. Specifically, 5.0 g of licorice root extract powder is mixed with 50 mL of hydrolysis medium (pre-mixed at a 7:3 ratio). All other steps and parameters are identical.

[0055] Preparation Example 9 A method for preparing an anti-glycation licorice root extract differs from Preparation Example 1 only in that, in step S2, the ultrasonic conditions are adjusted to a frequency of 35 kHz and a power density of 50 W / L. All other steps and parameters are identical.

[0056] Preparation Example 10 A method for preparing an anti-glycation licorice root extract differs from Preparation Example 1 only in that, in step S2, the reaction conditions are adjusted to react at 45°C for 1.5 hours. All other steps and parameters are identical.

[0057] Preparation Example 11 A method for preparing an anti-glycation licorice root extract differs from Preparation Example 1 only in that, in step S3, sodium phytate is replaced with disodium EDTA, and the amount added is adjusted to 0.1% of the total weight of the hydrolysate. The remaining steps and parameters are exactly the same.

[0058] Preparation Example 12 A method for preparing an anti-glycation licorice root extract differs from Preparation Example 1 only in that, in step S3, the sulfonic acid-type strong acid cation exchange resin is replaced with a carboxylic acid-type weak acid cation exchange resin (Amberlite™ IRC86). All other steps and parameters are identical.

[0059] Preparation Example 13 A method for preparing an anti-glycation licorice root extract differs from Preparation Example 1 only in that, in step S3, the flow rate of the adsorption treatment is controlled at 3 column volumes per hour. This high flow rate is used to verify whether the metal ion removal efficiency can still meet the requirements at a faster processing speed. The remaining steps and parameters are exactly the same.

[0060] Preparation Example 14 A method for preparing an anti-glycation licorice root extract differs from preparation example 1 only in that: in step S2, the ratio of the hydrolysis medium is adjusted to DES:water = 8:2 (v / v).

[0061] Preparation Example 15 A method for preparing an anti-glycation licorice root extract differs from Preparation Example 1 only in that, in step S2, the solid-liquid ratio is adjusted to 1:30 g / mL. Specifically, 5.0 g of licorice root extract powder is mixed with 150 mL of hydrolysis medium (at a 7:3 ratio).

[0062] Preparation Example 16 A method for preparing an anti-glycation licorice root extract differs from preparation example 1 only in that: in step S1, the molar ratio of the DES components is adjusted to ChCl:LA:ZnCl2=1:2:0.05.

[0063] Preparation Example 17 A method for preparing an anti-glycation licorice root extract differs from preparation example 1 only in that the reaction time in step S2 is adjusted to 0.5 hours.

[0064] Preparation Example 18 A method for preparing an anti-glycation licorice root extract differs from preparation example 1 only in that: in step S3, the amount of sodium phytate added is adjusted to 0.5% of the total weight of the hydrolysate.

[0065] Preparation Example 19 A method for preparing an anti-glycation licorice root extract differs from Preparation Example 1 only in that, in step S1, the metal salt is replaced with an equimolar amount of stannous chloride (SnCl2). Since SnCl2 is easily oxidized, the operation must be carried out under an inert gas atmosphere.

[0066] Preparation Example 20 A method for preparing an anti-glycation licorice root extract differs from preparation example 1 only in that, in step S1, the quaternary ammonium salt is replaced with an equimolar amount of tetrabutylammonium chloride (TBAC).

[0067] Preparation Example 21 A method for preparing an anti-glycation licorice root extract differs from preparation example 1 only in that: in step S3, the chelating agent is replaced with sodium citrate, and the amount added is still 0.3% of the total weight of the hydrolysate.

[0068] Preparation Example 22 Example of preparation of a silymarin-phospholipid complex: Step S1: Phospholipid phase solution: Accurately weigh 10.0 g hydrogenated lecithin and 1.5 g cholesterol (molar ratio approximately 1:0.3), dissolve them in 200 mL anhydrous ethanol, and stir magnetically at room temperature until completely dissolved and clear to obtain the phospholipid phase stock solution.

[0069] Milk thistle extract phase solution: Accurately weigh 20.0g milk thistle extract (total silymarin content 65%), dissolve in 200mL anhydrous ethanol, and sonicate until completely clear to obtain the extract phase stock solution.

[0070] Antisolvent aqueous solution: Measure 2000 mL of deionized water, preheat to 55°C, and add 10.0 g of glycerol (0.5% w / v) to it, stirring until dissolved.

[0071] Step S2: A glass microfluidic chip with a "Y"-shaped injection and "T"-shaped precipitation node structure (main channel hydraulic diameter 200 μm) was used.

[0072] Two precision injection pumps were used to pump the phospholipid stock solution and the milk thistle extract stock solution into the "Y"-shaped inlet at a volumetric flow rate of 1:1 (2 mL / min each). The two liquid streams were instantly mixed in the microchannel to form a uniform blend.

[0073] A third precision injection pump is used to pump the preheated antisolvent aqueous solution (containing glycerol) into the "T" junction at a flow rate of 20 mL / min.

[0074] At the "T" junction, the generated 4 mL / min blended stream and 20 mL / min antisolvent aqueous solution came into instantaneous contact and mixed at a volumetric flow rate ratio of 1:5. In the instant of mixing, the solution rapidly changed from clear to opalescent, indicating that silymarin and phospholipids simultaneously formed nanoparticles through antisolvent precipitation and molecular self-assembly.

[0075] Step S3: Collect the opalescent liquid. Transfer the liquid to a rotary evaporator and slowly evaporate it under reduced pressure in a 40°C water bath to remove ethanol, yielding a concentrated aqueous dispersion of the silymarin-phospholipid complex.

[0076] Step S4: The above concentrated dispersion system was replenished to its original volume with deionized water, and then dialyzed and concentrated using a 100kDa molecular weight cutoff ultrafiltration membrane tangential flow filter to remove residual free small molecules, glycerol and salts, to obtain a silymarin-phospholipid complex nanodispersion.

[0077] Preparation Example 23 A method for preparing a silymarin-phospholipid complex differs from Preparation Example 14 in that the modified phospholipid is a complex of phosphatidylcholine and cholesterol, wherein the molar ratio of phosphatidylcholine to cholesterol is 1:0.2. In the microfluidic blending, the flow rate ratio of the phospholipid phase to the extract phase is 1:2 (i.e., 1 mL / min: 2 mL / min), and the volumetric flow rate ratio of the blended stream to the antisolvent aqueous solution is 1:8.

[0078] Preparation Example 24 A method for preparing a silymarin-phospholipid complex differs from Preparation Example 14 in that the modified phospholipid is a complex of phosphatidylcholine and cholesterol, wherein the molar ratio of phosphatidylcholine to cholesterol is 1:0.5. The organic solvent used is acetone. The temperature of the antisolvent aqueous solution is 40°C, and it does not contain glycerol.

[0079] Preparation Example 25 A method for preparing a silymarin-phospholipid complex differs from Preparation Example 14 in that, in the microfluidic blending, the flow rate ratio of the phospholipid phase to the extract phase is 1.5:1 (i.e., 3 mL / min: 2 mL / min). The temperature of the antisolvent aqueous solution is 60°C, and the content of glycerol is 1.0% (w / v). The volumetric flow rate ratio of the blend stream to the antisolvent aqueous solution is 1:15.

[0080] Example 1 A method for preparing a composition with anti-aging and brightening effects The formula for this embodiment, based on a total weight of 1000g, is as follows: The composition includes: 30.0 g of Glycyrrhiza glabra root extract, 150.0 g of Milk thistle extract, 0.075 g of Palmitoyl pentapeptide-4, 0.0075 g of Arginine / Lysine polypeptide, 5.0 g of Echinochloa crus-galli extract, 5.0 g of Olive leaf extract, 5.0 g of Diglucosyl Gallic acid, 2.5 g of Dendrobium officinale stem extract, and the balance being deionized water. The Glycyrrhiza glabra root extract is the anti-glycation Glycyrrhiza glabra root extract prepared by the method in Preparation Example 1.

[0081] Add approximately 80% of the formula amount of deionized water to a mixing pot, heat to 75-80℃, and while stirring, add the following extracts in sequence: Glycyrrhiza glabra root extract, Milk thistle extract, Elongatus spp. extract, Olive leaf extract, Diglucosyl gallic acid, and Dendrobium officinale stem extract, and continue stirring until evenly dispersed.

[0082] Lower the temperature of the aqueous phase to below 40°C, and add a solution of palmitoyl pentapeptide-4 and arginine / lysine peptides that has been dissolved in a small amount of deionized water beforehand while stirring.

[0083] Add deionized water to bring the total weight to 1000g, and adjust the stirring speed to make the system homogeneous. Seal the mixing pot, purge the air with nitrogen, and homogenize at 2000rpm for 5 minutes under a protective atmosphere.

[0084] The homogenized material is cooled to below 30°C and discharged to obtain the composition with anti-aging and brightening effects.

[0085] Example 2: The preparation method of a composition with anti-aging and brightening effects differs from that of Example 1 in that the raw materials used are: 10g of Glycyrrhiza glabra root extract, 100g of milk thistle extract, 0.05g of palmitoyl pentapeptide-4, 0.005g of arginine / lysine polypeptide, 10g of Elongatus spp. extract, 10g of olive leaf extract, 10g of diglucosyl gallic acid, 5g of Dendrobium officinale stem extract, and the remainder is water.

[0086] Example 3 The preparation method of a composition with anti-aging and brightening effects differs from that of Example 1 in that the raw materials used are 50g of Glycyrrhiza glabra root extract, 200g of milk thistle extract, 0.1g of palmitoyl pentapeptide-4, 0.01g of arginine / lysine polypeptide, and the remainder is water.

[0087] Example 4-23 The preparation method of a composition with anti-aging and brightening effects differs from that of Example 1 in that: the licorice root extract is the anti-glycation licorice root extract obtained by the method of Preparation Examples 2-21.

[0088] Example 24 A method for preparing a composition with anti-aging and brightening effects differs from Example 1 in that the weight ratio of licorice root extract to milk thistle extract is adjusted to 1:4. Specifically, the composition consists of 30.0g of licorice root extract and 120.0g of milk thistle extract.

[0089] Example 25 A method for preparing a composition with anti-aging and brightening effects differs from Example 1 in that the weight ratio of licorice root extract to milk thistle extract is adjusted to 1:6. Specifically, the composition consists of 30.0g of licorice root extract and 180.0g of milk thistle extract.

[0090] Example 26 The preparation method of a composition with anti-aging and brightening effects differs from that of Example 1 in that: the amount of licorice root extract 30.0g and milk thistle extract 200.0g is adjusted.

[0091] Example 27 The preparation method of a composition with anti-aging and brightening effects differs from that of Example 1 in that: the amount of licorice root extract 30.0g and milk thistle extract 100.0g is adjusted.

[0092] Example 28 The preparation method of a composition with anti-aging and brightening effects differs from that of Example 1 in that: the amount of *Euphorbia pulcherrima* extract is adjusted to 5.0g and the amount of diglucosyl gallic acid to 4.0g.

[0093] Example 29 The preparation method of a composition with anti-aging and brightening effects differs from that of Example 1 in that: the amount of *Euphorbia pulcherrima* extract is adjusted to 5.0g and the amount of diglucosyl gallic acid to 6.0g.

[0094] Example 30 The preparation method of a composition with anti-aging and brightening effects differs from that of Example 1 in that: the amount of *Euphorbia pulcherrima* extract is adjusted to 5.0g and the amount of diglucosyl gallic acid to 3.0g.

[0095] Example 31 The preparation method of a composition with anti-aging and brightening effects differs from that of Example 1 in that: the amount of *Euphorbia pulcherrima* extract is adjusted to 5.0g and the amount of diglucosyl gallic acid to 7.0g.

[0096] Example 32 A method for preparing a composition with anti-aging and brightening effects differs from Example 1 in that: 1.0 g of modified phospholipid is added, and the modified phospholipid is hydrogenated lecithin. During preparation, the hydrogenated lecithin is added together with the aqueous phase raw materials in step 1.

[0097] Example 33 A method for preparing a composition with anti-aging and brightening effects differs from Example 1 in that 5.0 g of hydrogenated lecithin is added. During preparation, the hydrogenated lecithin is added together with the aqueous phase raw materials in step 1.

[0098] Example 34 A method for preparing a composition with anti-aging and brightening effects differs from Example 1 in that 10.0 g of hydrogenated lecithin is added. During preparation, the hydrogenated lecithin is added together with the aqueous phase raw materials in step 1.

[0099] Example 35 A method for preparing a composition with anti-aging and brightening effects differs from Example 1 in that: an additional 5.0 g of modified phospholipid is added. The modified phospholipid is a complex of phosphatidylcholine and cholesterol, wherein the molar ratio of phosphatidylcholine to cholesterol is 1:0.3. During preparation, this complex is added together with the aqueous phase raw materials in step 1.

[0100] Example 36 A method for preparing a composition with anti-aging and brightening effects differs from Example 1 in that: an additional 5.0g of modified phospholipid is added. The modified phospholipid is a complex of hydrogenated lecithin and cholesterol, wherein the molar ratio of hydrogenated lecithin to cholesterol is 1:0.2. During preparation, this complex is added together with the aqueous phase raw materials in step 1.

[0101] Example 37 A method for preparing a composition with anti-aging and brightening effects differs from Example 1 in that an additional 5.0 g of modified phospholipids is added. During preparation, the modified phospholipids and milk thistle extract are pre-prepared using the method described in Example 22 to form a milk thistle-phospholipid complex before being added to the aqueous phase raw materials.

[0102] Example 38 A method for preparing a composition with anti-aging and brightening effects differs from Example 1 in that an additional 5.0 g of modified phospholipids is added. During preparation, the modified phospholipids and milk thistle extract are pre-prepared using the method described in Example 23 to form a milk thistle-phospholipid complex before being added to the aqueous phase raw materials.

[0103] Example 39 A method for preparing a composition with anti-aging and brightening effects differs from Example 1 in that an additional 5.0 g of modified phospholipids is added. During preparation, the modified phospholipids and milk thistle extract are pre-prepared using the method described in Example 24 to form a milk thistle-phospholipid complex before being added to the aqueous phase raw materials.

[0104] Example 40 A method for preparing a composition with anti-aging and brightening effects differs from Example 1 in that an additional 5.0 g of modified phospholipids is added. During preparation, the modified phospholipids and milk thistle extract are pre-prepared using the method described in Example 25 to form a milk thistle-phospholipid complex before being added to the aqueous phase raw materials.

[0105] Comparative Example 1 The only difference from Example 1 is that the "anti-glycation licorice root extract" is replaced with an equal amount of licorice root extract that has not undergone any special directional hydrolysis treatment. All other components and preparation methods are exactly the same.

[0106] Comparative Example 2 The difference from Example 1 is that arginine / lysine peptides are not added, and this portion is replaced with deionized water. Other components and preparation methods are the same.

[0107] Comparative Example 3 The difference from Example 1 is that palmitoyl pentapeptide-4 is not added, and this portion is replaced with deionized water. All other components and preparation methods are the same.

[0108] Comparative Example 4 This comparative example represents a conventional method for promoting collagen production in a single dimension. The difference between this example and Example 1 is the composition of the raw materials: 0.1g palmitoyl pentapeptide-4, 50.0g glycerol, 1.0g sodium hyaluronate, and deionized water to a final volume of 1000g. The preparation method is the same as that in this application.

[0109] Comparative Example 5 This comparative example is a conventional solution from the prior art for inhibiting melanin production and anti-oxidation. The difference from Example 1 is that the raw material composition is 30.0g of licorice root extract, 30.0g of nicotinamide, 20.0g of vitamin C ethyl ether, and deionized water to a final volume of 1000g. The preparation method is the same as in this application.

[0110] Performance testing Experiment 1: Evaluation of the synergistic effect of promoting collagen synthesis and inhibiting glycation damage in vitro To verify the synergistic effect of the composition in promoting collagen regeneration and protecting its quality, the following in vitro experiments were conducted. First, collagen synthesis was detected: 5th-8th generation normal human dermal fibroblasts were used at a concentration of 1×10⁻⁸. 4Cells / well were seeded at a density of 1 / well in 96-well plates and cultured at 37°C and 5% CO2 for 24 hours. After adhesion, the culture medium was replaced with serum-free medium containing 1% (v / v) sterile filtered test sample, and cultured for another 48 hours. The cell culture supernatant was collected, and the absorbance was measured at 450 nm using a human type I collagen propeptide ELISA kit, strictly following the instructions. The PIP content in the supernatant was calculated based on the standard curve to characterize the rate of type I collagen synthesis. The results were expressed as a percentage relative to the blank control group. Next, in vitro anti-glycation activity was evaluated: a bovine serum albumin-fructose glycation reaction model was established. 10 mg / mL bovine serum albumin (BSA) and 0.5 M fructose were dissolved together in phosphate-buffered saline (PBS) at pH 7.4. The test sample was added to the reaction system to a final concentration of 1% (w / v). The BSA-fructose system without the sample served as the model control, and the system without BSA and fructose served as the blank control. After incubating each reaction system at 37°C in the dark for 7 days, the fluorescence intensity was measured using a fluorescence microplate reader at an excitation wavelength of 370 nm and an emission wavelength of 440 nm. The AGEs generation inhibition rate was calculated using the formula: Inhibition rate (%) = [1 - (FI sample - FI blank) / (FI model - FI blank)] × 100%, where FI represents fluorescence intensity.

[0111] Experiment 2: Evaluation of scavenging intracellular reactive oxygen species and protecting cells from damage caused by advanced glycation end products (AGEs). To evaluate the composition's ability to resist oxidative stress and counteract the direct cytotoxicity of AGEs, the following cell experiments were performed. Intracellular reactive oxygen species (ROS) scavenging assay: Dermal fibroblasts were seeded in black 96-well plates and cultured overnight. Cells were then stimulated with medium containing 200 μM H2O2 for 1 hour to induce oxidative stress. The H2O2-containing medium was removed and replaced with medium containing 1% of the test sample, and the cells were cultured for another 2 hours. Subsequently, a final concentration of 10 μM DCFH-DA fluorescent probe was added, and the cells were incubated at 37°C in the dark for 30 minutes. After washing with PBS, the fluorescence intensity was detected using a fluorescence microplate reader at an excitation wavelength of 485 nm and an emission wavelength of 535 nm. Wells stimulated only with H2O2 without sample treatment served as a model control group, and the ROS scavenging rate of each sample group was calculated. AGEs-induced cell damage protection assay: Cells were seeded in 96-well plates. After adhesion, the culture medium was changed. The experimental group received medium containing 100 μg / mL commercial AGE-BSA and 1% of the test sample. The positive control group received medium containing only an equal amount of AGE-BSA. The normal control group contained neither AGE-BSA nor the test sample. After 72 hours of culture, CCK-8 solution was added to each well, and incubation was continued for 2 hours. The absorbance was measured at 450 nm. The relative cell viability was calculated as: (OD experimental group - OD blank) / (OD normal control group - OD blank) × 100%.

[0112] Experiment 3: Evaluation of an in vitro cell model inhibiting melanin production To verify the skin-brightening effect of the composition, a melanin production inhibition experiment was conducted using a B16 melanoma cell model. B16 cells in logarithmic growth phase were seeded at a density of 5 × 10³ cells / well in 96-well plates. After 24 hours of culture and adherence, the culture medium was replaced with fresh medium. The experimental group received 100 nM α-melanocyte-stimulating hormone (α-MSH) and 1% of the test sample in the culture medium. The model control group received only an equal amount of α-MSH, and the normal control group received neither the inducing agent nor the test sample. After 72 hours of further culture in the dark, the culture medium was discarded, cells were washed with PBS, and lysed in a 1 M NaOH solution at 80°C for 1 hour. After centrifugation of the lysate, the supernatant was measured at 405 nm to reflect the relative melanin content within the cells. The melanin production inhibition rate was calculated using the formula: Inhibition rate (%) = [1 - (OD experimental group - OD normal control group) / (OD model control group - OD normal control group)] × 100%.

[0113] Experiment 4: Safety Evaluation of Human Skin – Closed Patch Test The safety of the finished product composition was assessed according to the occlusive patch test method for human skin in the "Cosmetic Safety Technical Specifications" (2015 edition). Thirty volunteers with healthy skin and no history of allergies were recruited, and the test was conducted on normal skin areas on the backs of the subjects. Approximately 0.02 mL of the test substance (the finished product from the example), negative control (physiological saline), and positive control (0.5% sodium lauryl sulfate aqueous solution) were placed in separate patch applicators and applied to the skin. The patch applicators were removed after 24 hours, and skin reactions were observed at 30 minutes, 24 hours, and 48 hours after removal. Results were recorded using a 0-4 rating scale (0 for no reaction, 4 for severe erythema, edema, or blisters). The primary observation indicators were the incidence and intensity of skin irritation.

[0114] Experiment 5: Evaluation of an in vitro three-dimensional skin model for improving skin elasticity The effect of the composition on improving skin elasticity was evaluated using an in vitro reconstructed full-thickness skin model (EpidermFT™ model). The skin model was placed in a 6-well plate culture insert and cultured at the gas-liquid interface until complete differentiation and maturity. During the experiment, 50 μL of the test sample solution (1%) was uniformly applied to the surface of the skin model once daily, while the control group was treated with an equal volume of PBS buffer for 7 consecutive days. After treatment, three points were randomly selected on the surface of the skin model using a micromechanical testing instrument equipped with a spherical indenter (500 μm diameter). A normal pressure of 10 mN was applied to the surface of the skin model using microindentation mode, and the force-displacement curves during loading-unloading were recorded. The tangential modulus within the 0-10% strain range was calculated as the elastic modulus value. The relative increase in elastic modulus was calculated using the formula: Increase (%) = (Average elastic modulus of the treated group - Average elastic modulus of the control group) / Average elastic modulus of the control group × 100%.

[0115] Experiment 6: In vitro biochemical evaluation of inhibition of tyrosinase activity To elucidate the brightening mechanism of the composition at the enzymatic level, an in vitro tyrosinase activity inhibition experiment was conducted. Mushroom tyrosinase or recombinant human tyrosinase was used. The total reaction volume was 200 μL, containing 50 U / mL tyrosinase, 0.5 mL L-DOPA (dissolved in pH 6.8 phosphate buffer), and 1% of the test sample solution. The control group sample solution was replaced with an equal volume of PBS. The reaction system was incubated at 37°C for 30 minutes, and the absorbance was immediately measured at 475 nm using a microplate reader. This absorbance value was directly proportional to the amount of dopaquinone produced by the enzyme. The tyrosinase activity inhibition rate was calculated using the following formula: Inhibition rate (%) = (1 - Sample A / Control A) × 100%, where Sample A and Control A are the average absorbance values ​​of the experimental and control groups, respectively.

[0116] Table 1: Effects on type I collagen synthesis (PIP content) and in vitro anti-glycation (AGEs inhibition) activity in dermal fibroblasts Group Collagen type I propeptide (PIP) content (relative to blank control) AGEs production inhibition rate (%) Blank control 100.0 ± 5.2 - Comparative Example 1 135.5 ± 6.8 38.2 ± 3.1 Comparative Example 2 152.7 ± 7.1 15.4 ± 2.5 Comparative Example 3 105.3 ± 4.9 65.8 ± 4.0 Comparative Example 4 142.1 ± 6.5 8.1 ± 1.8 Comparative Example 5 103.8 ± 5.1 41.5 ± 3.5 Example 1 168.9 ± 8.2 72.5 ± 4.8 Example 2 161.2 ± 7.5 69.8 ± 4.3 Example 3 165.5 ± 7.9 70.1 ± 4.5 Examples 4-23 163.1 - 167.5 68.9 - 73.8 Example 24 166.3 ± 7.8 71.0 ± 4.6 Example 25 169.8 ± 8.1 72.8 ± 4.7 Example 26 162.5 ± 7.6 69.5 ± 4.2 Example 27 158.7 ± 7.3 67.2 ± 4.0 Example 28 167.2 ± 7.9 73.1 ± 4.9 Example 29 166.8 ± 7.8 72.9 ± 4.8 Example 30 164.1 ± 7.6 69.8 ± 4.3 Example 31 163.9 ± 7.5 70.1 ± 4.4 Example 32 172.5 ± 8.5 73.5 ± 4.9 Example 33 175.8 ± 8.7 74.2 ± 5.0 Example 34 176.1 ± 8.8 74.0 ± 5.1 Example 35 174.2 ± 8.6 73.8 ± 4.9 Example 36 173.9 ± 8.5 73.6 ± 4.8 Example 37 181.5 ± 9.2 75.8 ± 5.3 Example 38 179.2 ± 9.0 75.2 ± 5.1 Example 39 178.6 ± 8.9 74.9 ± 5.0 Example 40 180.1 ± 9.1 75.5 ± 5.2 Table 2: Intracellular ROS clearance rate and protective effect against AGEs-induced cell damage Group ROS clearance rate (%) Cell relative survival rate (vs. AGEs damage model, %) H202 / AGEs model 0.0 ± 2.1 58.3 ± 3.8 Comparative Example 1 45.2 ± 3.8 72.1 ± 4.2 Example 1 58.8 ± 4.7 85.6 ± 5.1 Example 24 59.0 ± 4.7 85.9 ± 5.1 Example 25 59.5 ± 4.8 86.2 ± 5.2 Example 27 57.0 ± 4.5 83.1 ± 4.9 Example 28 59.2 ± 4.7 86.0 ± 5.1 Example 33 61.0 ± 4.9 87.5 ± 5.3 Example 34 62.0 ± 5.0 88.2 ± 5.4 Example 37 63.5 ± 5.2 89.4 ± 5.5 Table 3: Inhibitory effect of α-MSH on melanin production in B16 cells Group Melanin production inhibition rate (%) Comparative Example 1 40.5 ± 3.5 Comparative Example 5 65.8 ± 5.0 Example 1 55.2 ± 4.5 Example 25 56.8 ± 4.6 Example 37 59.1 ± 4.8 Table 4: Safety Evaluation on Human Skin Test group Number of subjects Number of positive cases of skin irritation Skin irritation incidence rate Maximum irritation grade Negative control (physiological saline) 30 0 0% 0 Positive control (0.5% SLS) 30 30 100% 2 Example 1-40 finished products 30 (each group) 0 0% 0 Table 5: Effect on improving the elastic modulus of the three-dimensional full-thickness skin model Group Relative modulus increase rate (%) Comparative Example 4 15.2 ± 2.8 Example 1 28.7 ± 3.5 Example 24 29.0 ± 3.5 Example 25 29.5 ± 3.6 Example 28 29.2 ± 3.6 Example 34 31.5 ± 3.7 Example 37 33.8 ± 3.9 Table 6: In vitro inhibition rate of tyrosinase activity Group Tyrosinase inhibition rate (%) Comparative Example 1 35.8 ± 3.0 Example 1 48.5 ± 3.8 Example 37 50.1 ± 3.9 As can be seen from Examples 1 and Comparative Examples 2 and 3, and in conjunction with Table 1, the combined use of palmitoyl pentapeptide-4 and arginine / lysine peptides in the composition of this application produces a significant synergistic effect. Example 1 demonstrates excellent performance in both promoting collagen synthesis and inhibiting glycation. While Comparative Example 2 shows a reasonable collagen-promoting effect, its anti-glycation ability is severely lacking; Comparative Example 3, on the other hand, hardly promotes collagen regeneration. This proves that both are indispensable, together constituting a dual guarantee mechanism of stimulating synthesis and protecting quality.

[0117] Combining Example 1 and Comparative Example 1 with Tables 1 and 6, it can be seen that the anti-glycation licorice root extract of Example 1, prepared using a targeted hydrolysis and purification process, exhibited significantly higher tyrosinase inhibition rates and AGEs inhibition rates than Comparative Example 1, which used a conventional extract. This indicates that the special processing effectively enhances the bioactivity of glycyrrhizin, resulting in a simultaneous synergistic effect in both brightening and anti-glycation dimensions, rather than merely providing traditional whitening and antioxidant benefits.

[0118] Combining Examples 1, 24, 25, and 27 with Tables 1 and 2, it can be seen that there is a clear optimal range for the weight ratio of Glycyrrhiza glabra root extract to Milk thistle extract. When the ratio is between 1:4 and 1:6, the indicators such as collagen promotion, anti-glycation, and cell protection all remain at peak levels. Once the ratio deviates from this range, as in Example 27, all indicators show a downward trend. This provides direct data support for the ratio range described in the claims, demonstrating the optimizing effect of this specific ratio on the synergistic effect.

[0119] Combining Examples 1, 33, 34, and 37 with Tables 1, 2, and 5, it can be seen that introducing a delivery system and optimizing its preparation process can significantly improve the final efficacy of the composition. Simply adding phospholipids already provides some improvement, but the silymarin-phospholipid complex prepared using microfluidic technology achieves a significant leap forward. Example 37 achieved optimal results in key indicators such as collagen promotion, skin elasticity improvement, ROS scavenging, and cell protection. This demonstrates that the microfluidic co-precipitation self-assembly process is a key technology for improving the delivery efficiency and bioavailability of hydrophobic active ingredients.

[0120] Combining Example 1 and Comparative Example 4 with Tables 1 and 5, it can be seen that the comprehensive solution of this application far surpasses single-dimensional collagen-promoting strategies in achieving the final anti-wrinkle and firming effect. Although Comparative Example 4 can promote collagen synthesis to a certain extent, its lack of a protective network for collagen quality ultimately results in a very limited improvement in the elasticity of the skin model. In contrast, Example 1, while effectively promoting collagen production, achieves nearly a twofold increase in elasticity thanks to its strong anti-glycation and antioxidant protection.

[0121] As can be seen from the data in Table 4, none of the finished products of Examples 1-40 caused any irritation in the human skin closed patch test, indicating that the composition has good skin safety and tolerability within the range of formulation, process and content provided in this application.

[0122] This specific embodiment is merely an explanation of this application and is not intended to limit it. After reading this specification, those skilled in the art can make modifications to this embodiment without contributing any inventive step, but such modifications are protected by patent law as long as they fall within the scope of the claims of this application.

Claims

1. A composition with anti-aging and brightening effects, characterized in that, By weight percentage, it includes the following components: The extract comprises 1%-5% of Glycyrrhiza glabra root extract, 10%-20% of Milk thistle extract, 0.005%-0.01% of Palmitoyl Pentapeptide-4, 0.0005%-0.001% of Arginine / Lysine Peptide, 0-1% of Elongatus spp. extract, 0-1% of Olive leaf extract, 0-1% of Diglucosyl Gallic Acid, 0-0.5% of Dendrobium officinale stem extract, with the balance being water; the Glycyrrhiza glabra root extract is an anti-glycation Glycyrrhiza glabra root extract, which is prepared by directional hydrolysis of Glycyrrhiza glabra root extract.

2. The composition with anti-aging and brightening effects according to claim 1, characterized in that, The method for extracting the anti-glycation licorice root extract is characterized by comprising the following steps: Quaternary ammonium salt, organic acid and metal salt are mixed in a molar ratio of 1:(1~3):(0.05~0.2) and stirred evenly at 80~120℃ to obtain a ternary eutectic solvent; A ternary eutectic solvent and water were mixed at a volume ratio of 6:4 to 8:2 to obtain a hydrolysis medium. Licorice root extract was added to the hydrolysis medium, and the solid-liquid ratio was controlled at 1:10 to 1:30 g / mL. The mixture was reacted at 45 to 55 °C for 1 to 1.5 hours under the assistance of ultrasound at a frequency of 35 to 40 kHz and a power density of 50 to 80 W / L to obtain a hydrolysate. A metal ion chelating agent is added to the hydrolysate, and after stirring and mixing, the mixture is adsorbed through an ion exchange column, and the effluent is collected. The pH of the effluent was adjusted to 6.0–7.0, and after concentration and drying, the anti-glycation licorice root extract was obtained.

3. The composition with anti-aging and brightening effects according to claim 2, characterized in that: The quaternary ammonium salt is one of choline chloride, tetraethylammonium chloride, or tetrabutylammonium chloride; the organic acid is one of lactic acid, citric acid, or oxalic acid; the metal salt is one of zinc chloride, aluminum chloride, or stannous chloride; the metal salt is zinc chloride or aluminum chloride; the metal ion chelating agent is one of sodium phytate, disodium EDTA, or sodium citrate, and its addition amount is 0.1% to 0.5% of the total weight of the hydrolysate.

4. The composition with anti-aging and brightening effects according to claim 2, characterized in that: The ion exchange column is packed with sulfonic acid-type strong acid cation exchange resin or carboxylic acid-type weak acid cation exchange resin; the flow rate of the adsorption treatment is controlled at 1 to 3 times the column volume per hour.

5. The composition with anti-aging and brightening effects according to claim 1, characterized in that, The extract of *Euphorbia lathyris* and the diglucosyl gallic acid are present in a weight ratio of 1:(0.8-1.2).

6. The composition with anti-aging and brightening effects according to claim 1, characterized in that, The weight ratio of the licorice root extract to the milk thistle extract is 1:(4-6).

7. The composition with anti-aging and brightening effects according to claim 1, characterized in that, It also includes 0.1%-1% of modified phospholipids, wherein the modified phospholipids are selected from at least one of hydrogenated lecithin, phosphatidylcholine and cholesterol complex.

8. The composition with anti-aging and brightening effects according to claim 7, characterized in that, The modified phospholipids and milk thistle extract are pre-formed into a silymarin-phospholipid complex using the following method: An organic phase solution containing modified phospholipids and an organic phase solution containing milk thistle extract were mixed using microfluidic technology to form a blended stream. The blended stream is brought into contact with the antisolvent aqueous solution in a microfluidic channel so that silymarin and the modified phospholipid can simultaneously form complex particles through antisolvent precipitation and self-assembly. The complex particles were collected and purified to obtain the silymarin-phospholipid complex.

9. A method for preparing a composition with anti-aging and brightening effects as described in any one of claims 1-8, characterized in that: Includes the following steps: All components were mixed and homogenized under a protective atmosphere to obtain a composition with anti-aging and brightening effects.

10. The composition having anti-aging and brightening effects as described in any one of claims 1-8 is used in the preparation of cosmetics for skin anti-wrinkle, firming, brightening and improving uneven skin tone.