A composition with anti-glycation and yellow-removing effects, and a preparation method and application thereof

Abstract

The application belongs to the field of cosmetics, and specifically discloses a composition with anti-glycation and yellow-removing effects, a preparation method and application thereof. The composition with anti-glycation and yellow-removing effects comprises the following components in parts by weight: tripterospermum sterol-flavonoid compound 0.5-5 parts, pterostilbene 0.5-5 parts, ganoderma lucidum extract 1-10 parts, and 80-98 parts by weight of a solvent. In the composition, the tripterospermum sterol-flavonoid compound, pterostilbene and ganoderma lucidum extract synergize with each other, inhibit the generation of AGEs, block the AGEs-RAGE signal transduction and reduce the content of CML in multiple links, so that the composition and the cosmetic containing the composition have remarkable anti-glycation and yellow-removing effects.

Description

Technical Field

[0001] This invention belongs to the field of cosmetics, specifically relating to a composition with anti-glycation and anti-yellowing effects, its preparation method, and its application. Background Technology

[0002] Skin glycation is a key endogenous factor inducing skin aging, and its non-enzymatic reaction has been widely confirmed to damage skin structure and function. Studies have shown that reducing sugars undergo irreversible covalent cross-linking reactions with structural proteins such as collagen and elastin in the skin, generating advanced glycation end products (AGEs) after complex multi-stage reactions. The accumulation of AGEs not only directly reduces skin permeability through their own brownish-yellow pigment deposition, leading to dull and yellowish skin tone, but also significantly weakens the mechanical strength and elasticity of collagen fibers through irreversible cross-linking, accelerating skin laxity and wrinkle formation. Furthermore, the binding of AGEs to the receptor for advanced glycation end products (RAGE) on the cell surface can activate downstream inflammatory signaling pathways, inducing oxidative stress and chronic inflammatory responses, further exacerbating the skin aging process and pigmentation problems. Currently, commercially available anti-glycation cosmetics still have many technical shortcomings: most products use a single active ingredient, which can only target a single stage of the glycation reaction and cannot comprehensively intervene in the skin glycation process from multiple stages, resulting in poor effects.

[0003] *Tetrastigma hemsleyanum* Diels et Gilg, a commonly used medicinal and edible plant, possesses extracts and active ingredients with diverse biological activities, exhibiting high medicinal, cosmetic, economic, and industrial value. However, its application in cosmetics is largely limited to conventional uses such as anti-inflammation and acne treatment, with limited development of other active effects, particularly its anti-glycation value, which requires further research.

[0004] Therefore, developing a multi-component, multi-target, safe and effective natural plant-derived anti-glycation and anti-yellowing composition is of great scientific value and market potential for breaking through existing technological bottlenecks and developing highly efficient and safe anti-glycation and anti-yellowing skin care products. Summary of the Invention

[0005] In view of the problems mentioned above in the prior art, the present invention will provide a composition with anti-glycation and anti-yellowing effects, its preparation method and application.

[0006] To achieve the above objectives, the following technical solutions are specifically included: In a first aspect, the present invention provides a composition having anti-glycation and anti-yellowing effects, comprising the following components in parts by weight: 0.5-5 parts of trifolin sterol-flavonoid complex, 0.5-5 parts of pterostilbene, 1-10 parts of Ganoderma lucidum extract, and 80-98 parts by weight of solvent; wherein the trifolin sterol-flavonoid complex comprises trifolin sterol component and trifolin flavonoid component, wherein the trifolin sterol component comprises sterol compounds, and the trifolin flavonoid component comprises flavonoid compounds, wherein the total mass percentage of sterol compounds in the trifolin sterol component is greater than or equal to 15%, and the total mass percentage of flavonoid compounds in the trifolin flavonoid component is greater than or equal to 60%.

[0007] It should be noted that the sterol component of *Trifolium repens* refers to substances containing sterol compounds extracted from *Trifolium repens* plants, and the total sterol content is ≥15%; the flavonoid component of *Trifolium repens* refers to substances containing flavonoid compounds extracted from *Trifolium repens* plants, and the total flavonoid content is ≥60%.

[0008] The inventors of this invention have discovered that the trifolin sterol-flavonoid complex contains high levels of sterols and flavonoids, exhibiting excellent activity in resisting skin glycation (anti-glycation), inhibiting skin aging, and reducing pigmentation. Furthermore, when used in combination with extracts of pterostilbene and Ganoderma lucidum, it achieves anti-glycation, improves skin brightness, and reduces skin yellowing (anti-yellowing) effects through multiple mechanisms, including inhibiting AGEs formation, blocking AGEs-RAGE signal transduction, and reducing carboxymethyl lysine (CML) content. The inventors have also found that the aforementioned functions and effects cannot be obtained by adding only trifolin sterol components or trifolin flavonoid components to the composition of this invention, except in the form of the trifolin sterol-flavonoid complex.

[0009] Preferably, in the composition, the weight parts of the trifolin sterol-flavonoid complex can be 0.5 parts, 0.8 parts, 1.0 parts, 1.2 parts, 1.5 parts, 1.8 parts, 2.0 parts, 2.2 parts, 2.5 parts, 2.8 parts, 3.0 parts, 3.2 parts, 3.5 parts, 3.8 parts, 4.0 parts, 4.2 parts, 4.5 parts, 4.8 parts, 5.0 parts, or a range between the above two points.

[0010] Preferably, in the composition, the weight parts of pterostilbene can be 0.5 parts, 0.8 parts, 1.0 parts, 1.2 parts, 1.5 parts, 1.8 parts, 2.0 parts, 2.2 parts, 2.5 parts, 2.8 parts, 3.0 parts, 3.2 parts, 3.5 parts, 3.8 parts, 4.0 parts, 4.2 parts, 4.5 parts, 4.8 parts, 5.0 parts, or a range between the above two points.

[0011] Preferably, in the composition, the weight parts of the Ganoderma lucidum extract can be 1.0 parts, 1.5 parts, 2.0 parts, 2.5 parts, 3.0 parts, 3.5 parts, 4.0 parts, 4.5 parts, 5.0 parts, 5.5 parts, 6.0 parts, 6.5 parts, 7.0 parts, 7.5 parts, 8.0 parts, 8.5 parts, 9.0 parts, 9.5 parts, 10.0 parts, or a range between the above two points.

[0012] Preferably, the mass ratio of the trifolinol-flavonoid complex to pterostilbene is (0.25-4):1, more preferably (1.5-3):1; more specifically, it can be 0.25:1, 0.30:1, 0.35:1, 0.40:1, 0.45:1, 0.50:1, 0.55:1, 0.60:1, 0.65:1, 0.70:1, 0.75:1, 0.80:1, 0.85:1, 0.90:1, 0.95:1, 1.00:1, 1.05:1, 1.10:1, 1.15:1, 1.20:1, 1.25:1, 1.30:1, 1.35:1, 1.40:1, 1.45:1, 1.50:1, 1.55:1, 1.60:1, 1.65:1, 1.70:1, 1.75:1, 1.80:1, 1.85:1, 1.90:1, 1.95: 1, 2.00:1, 2.05:1, 2.10:1, 2.15:1, 2.20:1, 2.25:1, 2.30:1, 2.35:1, 2.40:1, 2.45:1, 2.50:1, 2.55:1, 2.60:1, 2.65:1, 2.70:1, 2.75:1, 2.80:1, 2.85:1, 2.90:1, 2.95:1, 3.00:1, 3. The mass ratios of trifolinol-flavonoid complex and pterostilbene within the above ranges are 0.5:1, 3.10:1, 3.15:1, 3.20:1, 3.25:1, 3.30:1, 3.35:1, 3.40:1, 3.45:1, 3.50:1, 3.55:1, 3.60:1, 3.65:1, 3.70:1, 3.75:1, 3.80:1, 3.85:1, 3.90:1, 3.95:1, 4.00:1, or any range between the two. The inventors of this invention have discovered that when the mass ratio of trifolinol-flavonoid complex to pterostilbene is within the above ranges, the composition exhibits superior anti-glycation and anti-yellowing effects.

[0013] Preferably, the mass ratio of Ganoderma lucidum extract to pterostilbene is (0.75-8):1, more preferably (2-5):1; more specifically, it can be 0.75:1, 1.00:1, 1.25:1, 1.50:1, 1.75:1, 2.00:1, 2.25:1, 2.50:1, 2.75:1, 3.00:1, 3.25:1, 3.50:1, 3. The mass ratios of trifolinol-flavonoid complex and pterostilbene within the above ranges are 75:1, 4.00:1, 4.25:1, 4.50:1, 4.75:1, 5.00:1, 5.25:1, 5.50:1, 5.75:1, 6.00:1, 6.25:1, 6.50:1, 6.75:1, 7.00:1, 7.25:1, 7.50:1, 7.75:1, 8.00:1, or any range between the two. The inventors of this invention have discovered that when the mass ratio of trifolinol-flavonoid complex to pterostilbene is within the above ranges, the composition exhibits superior anti-glycation and anti-yellowing effects.

[0014] Preferably, in the trifolin sterol component, the total mass percentage of sterol compounds is greater than or equal to 20%, and the total mass percentage of sterol compounds in the trifolin sterol component is 20.5%-30%, more preferably 23%-26%. The inventors of this invention have found that when the total content of sterol compounds in the trifolin sterol component is within the above range, the composition exhibits superior anti-glycation and anti-yellowing effects.

[0015] Preferably, the total mass percentage of flavonoids in the *Trifolium repens* flavonoid component is greater than or equal to 70%, more preferably 71%-79%, and even more preferably 73%-77%.

[0016] Preferably, based on the total mass of the trifolin sterol-flavonoid complex, the total mass percentage of sterol compounds in the trifolin sterol-flavonoid complex is greater than 1.5%, more preferably 2%-23%, more preferably 2.6%-21.5%, or even more preferably 4%-16.1%. The inventors of this invention have found that when the total sterol content in the trifolin sterol-flavonoid complex is within the above range, the composition exhibits superior anti-glycation and anti-yellowing effects.

[0017] Preferably, based on the total mass of the trifolinose sterol-flavonoid complex, the total mass percentage of flavonoids in the trifolinose sterol-flavonoid complex is greater than 5%, more preferably 7%-70%, more preferably 8.2%-66.1%, or even more preferably 24.7%-62%. The inventors of this invention have found that when the total flavonoid content in the trifolinose sterol-flavonoid complex is within the above range, the composition exhibits superior anti-glycation and anti-yellowing effects.

[0018] Preferably, in the trifolin sterol-flavonoid complex, the mass ratio of the trifolin sterol component to the trifolin flavonoid component is (0.125-8):1, more preferably (0.2-6):1, even more preferably (0.5-5):1, and still more preferably (0.8-3):1. The inventors of this invention have found that when the mass ratio of the trifolin sterol component to the trifolin flavonoid component in the trifolin sterol-flavonoid complex is within the above range, the composition exhibits superior anti-glycation and anti-yellowing effects.

[0019] Preferably, the trifolinol component includes at least one of β-sitosterol and stigmasterol; more preferably, the trifolinol component includes β-sitosterol and stigmasterol, and the total mass percentage of β-sitosterol and stigmasterol is 5%-13% based on the dry weight of the trifolinol component, and more preferably 8%-10%.

[0020] Preferably, the flavonoid component of *Trifolium repens* comprises flavonoid C glycosides. Specifically, the flavonoid component of *Trifolium repens* comprises at least one of purslane, isopurslane, vitexin, and isovitexin. More preferably, the flavonoid component of *Trifolium repens* comprises purslane, isopurslane, vitexin, and isovitexin, and the total mass percentage of flavonoid C glycosides of purslane, isopurslane, vitexin, and isovitexin, based on the weight of the flavonoid component of *Trifolium repens*, is 30%-35%.

[0021] Preferably, the solvent includes at least one of alcohols, betaine, and water.

[0022] More preferably, the solvent comprises the following components in parts by weight: 30-50 parts of 1,3-propanediol, 20-40 parts of betaine, and 10-40 parts of water.

[0023] Secondly, the present invention provides a method for preparing the composition having anti-glycation and anti-yellowing effects, comprising the following steps: S1. Obtain the trifoliol component and the trifoliol flavonoid component separately, and mix the trifoliol component and the trifoliol flavonoid component evenly to obtain the trifoliol-flavonoid complex. S2. Dissolve the trifolinol-flavonoid complex and pterostilbene in a solvent, then add Ganoderma lucidum extract and mix well to obtain the composition.

[0024] More specifically, in step S1, the trichomesterol component and the trichome flavonoid component are obtained using both commercially available and self-made methods. For example, they can be obtained using the following self-made method: (1-1) The powder of the stems and leaves of *Trifolium repens* was mixed with a mixed solution of ethanol and water, and after reflux extraction, filtration and concentration, the first type of extract was obtained. (1-2) Disperse the first type of extract in water, then add petroleum ether for extraction, and collect the petroleum ether phase and the aqueous alcohol phase; (1-3) The petroleum ether phase is concentrated to obtain a second type of extract. The second type of extract is mixed with an alkaline solution and subjected to a saponification reaction to obtain a saponified product. The saponified product is then subjected to dilution, extraction, collection of extract, washing and concentration to obtain a third type of extract. The third type of extract is dissolved in ethyl acetate and then filtered, crystallized and dried to obtain the trifolin sterol component. (1-4) The aqueous alcohol phase is dried to remove petroleum ether, ethanol is added and mixed evenly, and then allowed to stand, solid-liquid separation is performed, the supernatant is collected and concentrated to obtain the fourth type of extract; the fourth type of extract, a mixed solvent of ethyl acetate and n-butanol and sodium chloride are mixed, extracted, the organic phase is collected and concentrated to obtain the fifth type of extract; the fifth type of extract is dispersed in water, then eluted, the eluent is collected and concentrated and freeze-dried to obtain the trifolin flavonoid component.

[0025] In the above-mentioned self-made process, the above-ground parts of *Trifolium repens* are first extracted with an aqueous ethanol solution and concentrated to obtain a crude extract. The aqueous dispersion of the crude extract is then extracted with petroleum ether to obtain a petroleum ether phase (source of sterol components) and a hydro-alcohol phase (source of flavonoid components). The petroleum ether phase is subjected to saponification, extraction, and crystallization to obtain the sterol components. The hydro-alcohol phase is subjected to alcohol precipitation, extraction, and other processes to obtain the flavonoid components.

[0026] More specifically, in step (1-1), the powder of *Trifolium repens* stems and leaves has a mesh size of 20-100 mesh; based on 1000g of *Trifolium repens* stems and leaves powder, the volume of the mixed solution of ethanol and water is 5-50L; in the mixed solution of ethanol and water, the volume ratio of ethanol to water is 1:(0.5-1.2); the temperature of the reflux extraction is 60-80℃, the reflux extraction is repeated multiple times, the number of reflux extractions is 1-5 times, the time of each reflux extraction is 1-3h, and the extracts from multiple reflux extractions are combined and filtered.

[0027] More specifically, in steps (1-1)-(1-4), each concentration is independently selected from vacuum concentration at 40-60°C.

[0028] More specifically, in steps (1-2), the volume of water used in the extraction is 1-3 times that of the first type of extract; the volume of petroleum ether used in the extraction is 0.8-2 times the total volume of the first type of extract and water; the extraction is repeated multiple times, with 2-3 extractions and each extraction lasting 10-30 minutes; after extraction, the petroleum ether phase and the aqueous alcohol phase are collected by allowing the mixture to stand and separate; then the petroleum ether phase and the aqueous alcohol phase from the multiple extractions are combined separately.

[0029] More specifically, in steps (1-3), the alkaline solution comprises an ethanol solution of potassium hydroxide or sodium hydroxide, and the alkaline content in the alkaline solution is 5%-20% (w / v); the volume of the alkaline solution is 8-15 times the volume of the second type of extract; the temperature of the saponification reaction is 50-70℃, and the time of the saponification reaction is 0.5-3 hours; in the dilution process, the solvent used is water, and the volume of water is 0.8-1.2 times the volume of the saponified product; the solvent used in the extraction process is petroleum ether, and the extraction is repeated multiple times, with 2-3 extractions per extraction. The extraction time is 10-30 min. After extraction, the extracts from multiple extractions are combined. The washing solution is water, and the mixture is washed until neutral. The dissolution temperature is 40-60℃, and the filtration temperature is 30-60℃. The volume of ethyl acetate is 5-15 times the volume of the third type of extract. The crystallization temperature is 1-6℃, and the crystallization is performed multiple times, 2-5 times, with each crystallization lasting 12-24 hours. After crystallization, the crystals from multiple crystallizations are combined. The drying is performed under reduced pressure at a temperature of 40-60℃.

[0030] More specifically, in steps (1-4), the drying is performed by vacuum rotary evaporation at a temperature of 40-60°C for 5-20 minutes; the final concentration of ethanol in the system after uniform mixing is 50%-80% (v / v); the settling time is 12-24 hours; the solid-liquid separation is performed by centrifugation at a speed of 3000-5000 rpm for 1-10 minutes; in collecting and concentrating the supernatant, the supernatant is concentrated to 0.3-0.8 times its original volume; the volume of the mixed solvent of ethyl acetate and n-butanol is 1-3 times the volume of the fourth type of extract.

[0031] More specifically, in steps (1-4), the volume ratio of ethyl acetate to n-butanol in the mixed solvent of ethyl acetate and n-butanol is (2-10):1.

[0032] More specifically, in steps (1-4), the sodium chloride content is 1%-5% (w / v) based on the total mass of the fourth type of extract, the mixed solvent of ethyl acetate and n-butanol, and sodium chloride.

[0033] More specifically, in steps (1-4), the extraction is repeated multiple times, the number of extractions is 2-3 times, the extraction time for each extraction is 10-30 minutes, after the extraction, the organic phases after multiple extractions are combined, each extraction is carried out under shaking conditions, and after each extraction, the organic phase is collected by standing to achieve layering.

[0034] More specifically, in steps (1-4), in the water dispersion of the fifth type of extract, the mass-to-volume ratio of the fifth type of extract to water is 1:(3-8), in g / mL.

[0035] More specifically, in steps (1-4), during the elution, the stationary phase is a resin column, the mass of which is 5-10 times the mass of the fifth type of extract, and the mobile phase is an aqueous solution of ethanol. The elution process includes a first elution and a second elution. In the first elution, the mobile phase is an aqueous solution of ethanol with a volume fraction of 20%-30%, and in the second elution, the mobile phase is an aqueous solution of ethanol with a volume fraction of 60%-80%.

[0036] Preferably, in step S1, the mixing method of the trifolin sterol component and the trifolin flavonoid component includes grinding, stirring, shaking, etc.

[0037] Preferably, in step S2, the dissolution temperature is 50-60°C.

[0038] Thirdly, the present invention provides the application of the composition having anti-glycation and anti-yellowing effects in at least one of the following I-III for non-disease treatment purposes, or in the preparation of products having at least one of the following I-VII functions. I. Inhibit the formation of advanced glycation end products (AGEs); II. Inhibit the expression of receptor for advanced glycation end products (RAGE); III. Inhibits the formation of carboxymethyl lysine (CML); IV. Improves skin glycation reaction; V. Improves skin pigmentation; VI. Improve skin radiance; VII. Reduce skin yellowing.

[0039] The present invention also provides a cosmetic product comprising the aforementioned composition having anti-glycation and anti-yellowing effects.

[0040] Preferably, based on the total mass of the cosmetic, the mass percentage of the composition having anti-glycation and anti-yellowing effects is 0.1%-10%, more specifically, it includes 0.1%, 0.5%, 1%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, 5.5%, 6.0%, 6.5%, 7.0%, 7.5%, 8.0%, 8.5%, 9.0%, 9.5%, 10%, or a range between any two of the above points.

[0041] Preferably, the cosmetic dosage form includes one of serum, lotion, cream, mask, or toner. The composition of the present invention can be combined with cosmetic-acceptable excipients to form cosmetics of various dosage forms, enabling these cosmetics to provide customized skincare effects based on different skin types, needs, and scenarios, while also offering anti-glycation, brightening, and anti-skin-darkening benefits.

[0042] For example, when the cosmetic product is in the form of an essence lotion, the essence lotion comprises the following components by weight percentage: 1%-20% moisturizer, 1%-10% emollient, 0.1%-2% emulsifier, 0.01%-0.5% thickener, 0.01%-2% preservative, 0.01%-2% pH adjuster, 0-1% chelating agent, and 0.1%-10% of the anti-glycation and anti-yellowing composition. More specifically, in the essence lotion, the moisturizer includes, but is not limited to, glycerin, butylene glycol, pentylene glycol, hexanediol, and sodium hyaluronate; the emollient includes, but is not limited to, shea butter, polydimethylsiloxane, and caprylic / capric triglyceride; the emulsifier includes, but is not limited to, glyceryl stearate and PEG-100 stearate; the thickener includes, but is not limited to, xanthan gum and carbomer; the preservative includes, but is not limited to, p-hydroxyacetophenone; the pH adjuster includes, but is not limited to, tromethamine; and the chelating agent includes, but is not limited to, EDTA-2Na.

[0043] For example, the preparation method of the essence lotion includes the following steps: S1. Mix water, chelating agent, humectant, thickener and preservative at 70-90℃ until homogeneous to obtain phase A; S2. Mix the emulsifier and emollient evenly at 70-90℃ to obtain phase B; S3. Add phase B to phase A at 80-90℃, homogenize, then cool to 40-50℃, then add pH adjuster and the anti-glycation and anti-yellowing composition in sequence, mix evenly, cool to room temperature, filter, and obtain essence emulsion.

[0044] For example, when the cosmetic product is a face cream, the face cream comprises the following components in weight percentage: 1%-20% moisturizer, 1%-10% emulsifier, 0.1%-2% thickener, 0.01%-0.5% preservative, 0.01%-2% pH adjuster, 0-1% chelating agent, and 0.1%-10% of the anti-glycation and anti-yellowing composition. More specifically, in face creams, moisturizers include, but are not limited to, glycerin, 1,3-butanediol, 1,2-hexanediol, and sodium hyaluronate; emollients include, but are not limited to, shea butter, jojoba seed oil, polydimethylsiloxane, and caprylic / capric triglyceride; emulsifiers include, but are not limited to, glyceryl stearate, PEG-100 stearate, and hydrogenated lecithin; thickeners include, but are not limited to, ammonium acryloyl dimethyl taurate / VP copolymer, acrylate / C10-30 alkanol acrylate crosspolymer, and cetearyl alcohol; preservatives include, but are not limited to, phenoxyethanol; pH adjusters include, but are not limited to, tromethamine; and chelating agents include, but are not limited to, EDTA-2Na.

[0045] More specifically, the preparation method of the face cream includes the following steps: mixing moisturizer, a portion of thickener, chelate and water at 60-85℃ to obtain phase A; mixing emulsifier, remaining thickener and moisturizer at 60-85℃ to obtain phase B; adding phase B to phase A at 80-85℃ for homogenization, then cooling to 40-50℃, then sequentially adding pH adjuster, preservative and the anti-glycation and anti-yellowing composition, mixing evenly, and then cooling, inspecting and filtering to obtain the face cream.

[0046] Compared with the prior art, the present invention has the following beneficial effects: In the composition of the present invention, the trifolin sterol-flavonoid complex, pterostilbene, and Ganoderma lucidum extract work synergistically to enhance the effect of the composition in improving skin brightness and reducing yellowing by inhibiting AGEs generation, blocking AGEs-RAGE signal transduction, and reducing CML content, thereby making it and cosmetics containing it have significant anti-glycation and yellowing-reducing effects. Attached Figure Description

[0047] Figure 1 The bar chart shows the comparison of RAGE inhibition rates between the positive control (group) and the corresponding sample groups of Examples 1-6 and Comparative Examples 1-6.

[0048] Figure 2 The bar chart shows the comparison of CML inhibition rates between the positive control (group) and the corresponding sample groups of Examples 1-6 and Comparative Examples 1-6.

[0049] Figure 3 Comparison of facial skin tone improvement before (left) and after (right) 28 days of using serum 1 (containing 2wt% of the composition of Example 1) on a typical subject. Detailed Implementation

[0050] To better illustrate the purpose, technical solution, and advantages of this invention, specific embodiments will be used to further explain the invention below. Unless otherwise specified, the test methods used in the embodiments and / or comparative examples are conventional methods; the materials and reagents used, unless otherwise specified, are commercially available.

[0051] In this article, "v / v" represents volume ratio, "wt%" represents mass percentage, and "w / v" represents weight / volume percentage.

[0052] The trifolin sterol and trifolin flavonoid components used in the following examples and comparative examples were prepared in-house by the following methods: (1) Take the stems and leaves of the plant *Trifolium repens*, wash them, dry them naturally in the sun, crush them, and pass them through a 40-mesh sieve to obtain *Trifolium repens* powder. (2) Weigh 1000g of the *Trifolium repens* powder obtained in step S1, add 20L of 50% ethanol aqueous solution (v / v), and reflux extract twice in a constant temperature water bath at 70℃ for 2 hours each time. The reflux extraction conditions are the same each time. Combine the extracts, filter through a 200-mesh filter, and concentrate under reduced pressure at 55℃ until there is no alcohol taste to obtain the first type of extract. (3) The first type of extract was suspended in 2 times the volume of deionized water to obtain a mixed system. An equal volume of petroleum ether was added to the mixed system and the mixture was shaken and extracted for 20 minutes. The mixture was allowed to stand and separate into layers. The upper petroleum ether phase was collected and the lower water-alcohol phase was kept for later use. The petroleum ether phase was extracted twice more under the same conditions each time. The petroleum ether phase or water-alcohol phase extracted twice were combined.

[0053] (4) The petroleum ether phase was concentrated under reduced pressure at 40°C to a paste-like state to obtain a second type of paste; 10 times the volume of 12wt% potassium hydroxide ethanol solution was added, and the reaction was carried out at 65°C for 1.5 hours. After cooling to room temperature, a mixed system was obtained, and an equal volume of deionized water was added to dilute the mixed system; an equal volume of petroleum ether was added to the mixed system and the mixture was shaken and extracted for 20 minutes. The mixture was allowed to stand and separate into layers, and the upper petroleum ether phase was collected; the extraction was repeated once with petroleum ether under the same conditions each time. The petroleum ether phases from the repeated extractions were combined, washed with water until neutral, and concentrated under reduced pressure at 45°C to dryness to obtain a third type of paste. The obtained product was dissolved in 10 times the volume of ethyl acetate at 55°C and filtered while hot. The filtrate was allowed to stand at 4°C to crystallize for 18 hours. The crystals were collected by filtration and repeated twice under the same conditions each time. The crystals were combined and dried under reduced pressure at 45°C to obtain 10.94 g of trifolin sterol fraction. Using β-sitosterol as a reference, the trifolin sterol component was quantitatively analyzed by GC-MS using the external standard method. The results showed that the main detectable sterol components were β-sitosterol and stigmasterol, with a combined content of 9.71 wt%. The remaining components were mainly fat-soluble components coexisting with sterols, such as fatty acids and triterpenoids. Simultaneously, using β-sitosterol as a reference, the total sterol content of the trifolin sterol component was determined to be 24.12 wt% (all on a dry weight basis) using the vanillin-perchloric acid colorimetric method.

[0054] (5) Remove residual petroleum ether by rotary evaporation under reduced pressure at 45°C for 10 minutes, add anhydrous ethanol, and adjust the concentration of ethanol in the mixture to 75% (v / v). Stir well and let stand at room temperature for 18 hours. Centrifuge at 4000 rpm for 10 minutes to remove the precipitate and collect the supernatant.

[0055] (6) Concentrate the supernatant under reduced pressure to 1 / 2 of its original volume to obtain the fourth type of extract (aqueous phase); add ethyl acetate-n-butanol mixed solvent (volume ratio 4:1), the amount of mixed solvent is 1.5 times the volume of the fourth type of extract, and add 3wt% sodium chloride to the fourth type of extract, shake thoroughly for 15 minutes, let stand to separate the layers, collect the upper organic phase, repeat the extraction 3 times, and combine the organic phases.

[0056] (7) The organic phase was concentrated under reduced pressure at 45°C to a paste-like consistency, yielding the fifth type of extract. Deionized water was added to the fifth type of extract at a mass-to-volume ratio of 1:5 (g / mL) to disperse and dissolve the extract. The solution was then passed through a D101 macroporous adsorption resin column (resin volume was 8 times the mass of the extract) as the stationary phase. Impurities were first eluted with 4 times the column volume of 25 vol% ethanol aqueous solution (mobile phase), followed by 4 times the column volume of 70% ethanol aqueous solution (v / v) to elute flavonoids. The eluent was collected, concentrated under reduced pressure at 45°C, and freeze-dried to obtain 29.83 g of the *Trifolium repens* flavonoid fraction. Using rutin as a control, the total content of flavonoids in the obtained *Trifolium repens* flavonoid fraction was determined by ultraviolet-visible spectrophotometry (UV-Vis). The test results showed that the total content of flavonoids was 74.35 wt%. Meanwhile, high performance liquid chromatography (HPLC) was used to analyze the composition of flavonoids in the flavonoid fraction. The flavonoid fraction contained arbutin, isoarbutin, vitexin and isovitexin, with a total content of 33.47 wt% for the four flavonoid C glycosides; the remaining components were mainly phenolic acids (such as chlorogenic acid) and polysaccharides that coexisted with the flavonoids.

[0057] The pterostilbene used in the following examples and comparative examples was purchased from Sabinsa and is Pterowhite™, an extract of Pterocarpus marsupium bark, with a purity of ≥90% for the pterostilbene monomer in the extract.

[0058] The Ganoderma lucidum extracts used in the following examples and comparative examples were purchased from the Nanjing Institute of Wild Plants. They were obtained by water extraction and alcohol precipitation of Ganoderma lucidum fruiting bodies and are rich in water-soluble Ganoderma lucidum polysaccharides and fat-soluble Ganoderma lucidum triterpenes and other active ingredients.

[0059] Unless otherwise specified, the same raw materials are used in the following examples and comparative examples.

[0060] Examples 1-6 and Comparative Examples 1-6 A composition with anti-glycation and anti-yellowing effects is prepared by the following steps: S1. Mix the trifolin sterol component and the trifolin flavonoid component at a mass ratio of 0.5:1, place them in a mortar and grind until they are evenly mixed to obtain the trifolin sterol-flavonoid complex.

[0061] S2. According to the weight parts shown in Table 1, add betaine to a mixed solvent of 1,3-propanediol and deionized water, and stir in a water bath at 55°C until completely dissolved; then add trifolinol-flavonoid complex and pterostilbene, and continue stirring at 55°C until completely dissolved to form a homogeneous solution.

[0062] S3. Finally, add Ganoderma lucidum extract, stir thoroughly, and cool to room temperature to obtain a composition with anti-glycation and anti-yellowing effects.

[0063] Table 1 The compositions of the above examples and comparative examples were subjected to in vitro anti-glycation experiments, specifically including inhibition of AGEs, RAGE gene expression, and carboxymethyl lysine (CML) assays. Parallel blank assays were performed separately on the 1,3-propanediol, betaine, and water mixed solvent system used in this invention. The results confirmed that these solvents themselves had no significant effect on AGEs formation, RAGE gene expression, or CML accumulation, thus eliminating the interference of solvents on the activity assay results.

[0064] (1) The effect of the composition on AGEs was evaluated using the bovine serum albumin-glucose in vitro glycosylation assay model. Phosphate-buffered saline (PBS, pH 7.4) was used to prepare glucose solutions with a concentration of 200 mmol / L and bovine serum albumin (BSA, Solarbio, A8015) solutions with a mass concentration of 40 mg / mL. The solutions were then mixed to obtain BSA-glucose reaction solutions.

[0065] The experiment included a blank group, a background group, a control group, and a sample group. The active ingredients were trifolinol-flavonoid complex, pterostilbene, and Ganoderma lucidum extract. Based on the total active ingredient content of the stock solutions of the examples or comparative examples, the dilution factors were adjusted, and the solutions were diluted with PBS to a total active ingredient content of 2 mg / mL (i.e., 0.2%, w / v) to prepare the test sample solution. 0.5 mL of this diluted solution was added to 4.5 mL of BSA-glucose reaction solution to prepare the sample group (final active ingredient concentration in the reaction system was 0.2 mg / mL). The blank group received only PBS, the background group received the test sample solution and PBS (without BSA-glucose), and the control group received PBS and BSA-glucose reaction solution (without active ingredient). In addition, the blank group, background group, and control group received the same amount of propylene glycol and betaine (diluted by the same factor) as the sample group to eliminate solvent interference; the final volume for each group was 5 mL.

[0066] All groups were incubated at 37℃ in the dark for 30 days. Fluorescence intensity (F) was measured using a fluorescence spectrophotometer at an excitation wavelength of 370 nm and an emission wavelength of 440 nm. Each group had three replicates, and the experiment was independently repeated three times. Results are expressed as mean ± standard deviation. AGEs inhibition rate was calculated using the following formula. Independent samples t-test was used for comparisons between groups; p < 0.05 was considered statistically significant; p < 0.01 was considered highly significant. The test results are shown in Table 2.

[0067] AGEs inhibition rate (%) = 1 - [(F 样品组 -F背景 ) / (F 对照 -F 空白 (×100%)

[0068] Table 2 Results of AGEs inhibition rate Table 2 shows that the combinations of any one or two of the following in Comparative Examples 1-6—the trifolinol-flavonoid complex, pterostilbene, and Ganoderma lucidum extract—exhibited lower AGEs inhibition rates. Among the pairwise combinations, Comparative Example 2, containing both pterostilbene and Ganoderma lucidum extract, exhibited the highest inhibition rate (45.77%), significantly higher than Comparative Example 1 (40.45%) and Comparative Example 3 (43.20%), indicating that the combination of pterostilbene and Ganoderma lucidum extract is the optimal synergistic solution in this binary combination. Furthermore, when all three were present simultaneously, the AGEs inhibition rate was significantly higher, and significantly higher than that of Comparative Example 2 (p<0.01). These results indicate that the trifolinol-flavonoid complex can significantly enhance the anti-glycation efficacy of pterostilbene and Ganoderma lucidum extract, demonstrating a synergistic effect when used in combination.

[0069] (2) RAGE inhibition experiment Human skin fibroblasts in the logarithmic growth phase (Synthetic Biotechnology, SNP-H023) were digested with 0.25 mg / mL trypsin (Solarbio, T1300), centrifuged, and then cultured in complete medium (Synthetic Biotechnology, SNPM-H023) to a density of 1.0 × 10⁻⁶ cells / mL. 5 Cell suspension of 10 cells / mL was seeded into 12-well culture plates, 1 mL per well, and cultured at 37°C in a 5% CO2 incubator for 24 h.

[0070] The active ingredients were trifolinol-flavonoid complex, pterostilbene, and Ganoderma lucidum extract. The total active ingredient content was calculated by summing the corresponding mass percentages of the three. Based on the total active ingredient content of the stock solutions of Examples 1-6 or Comparative Examples 1-6, the solutions were diluted with serum-free DMEM medium (Shangen Biotechnology, SNM-002D) to a total active ingredient concentration of 0.1 mg / mL to prepare the test sample solution (containing serum-free medium and the sample of the examples or comparative examples). Simultaneously, a propylene glycol-betaine mixed solution of the same concentration and proportion as the sample group was prepared using serum-free DMEM medium as a solvent control (containing serum-free medium and solvent). Discard the culture medium in the cell culture plate and add 1 mL of treatment solution to each well according to the following groups: blank control group: add the solvent control mentioned above; model group: add 0.5 mM methylglyoxal (MGO) and the solvent control mentioned above; positive control group: add 0.5 mM MGO and 0.5 mM aminoguanidine hydrochloride and the solvent control mentioned above; sample group: add 0.5 mM MGO and the above test sample solution (final total concentration of active ingredient is 0.1 mg / mL). Each group has 3 replicates and is incubated at 37°C in a 5% CO2 incubator for 4 h.

[0071] After incubation, discard the culture medium and wash each well twice with 1 mL of PBS. Collect cells and extract total RNA according to the RNA extraction kit (Linke Biotech, MK0203). Reverse transcribe the total RNA into cDNA according to the cDNA first-strand synthesis kit (Linke Biotech, MK0601), using GAPDH (synthesized by Sangon Biotech) as an internal control, and employ 2... ⁻ΔΔCt The relative expression level of the RAGE gene was calculated using the following method (with the blank control group set at 1.0). The RAGE inhibition rate was calculated according to the following formula. Examples 1-6 were compared with Comparative Example 2 using an independent samples t-test to evaluate the statistical significance of the differences.

[0072] RAGE inhibition rate (%) = 1 - [(model group - sample group) / (model group - blank group) × 100%].

[0073] The results of the RAGE inhibition experiment are as follows: Figure 1 As shown, the inhibition rate of the trifolinol-flavonoid complex alone (Comparative Example 5) was 15.45%, indicating that it has a certain RAGE inhibitory ability. Among the pairwise combinations, Comparative Example 2, containing pterostilbene and Ganoderma lucidum extracts, had the highest inhibition rate (27.38%), which was better than the other binary combinations, Comparative Example 1 and Comparative Example 3. Based on this, after introducing the trifolinol-flavonoid complex, the RAGE inhibition rates of Examples 1-6 were all significantly higher than those of Comparative Example 2 (p<0.01). This indicates that the trifolinol-flavonoid complex can significantly enhance the inhibitory effect of pterostilbene and Ganoderma lucidum extracts on RAGE expression, and the combination of the three produces an overall effect superior to the pairwise combinations.

[0074] (3) Carboxymethyl lysine (CML) inhibition experiment Human skin fibroblasts in the logarithmic growth phase (Synthetic Biotechnology, SNP-H023) were digested with 0.25 mg / mL trypsin (Solarbio, T1300), centrifuged, and then cultured in complete medium (Synthetic Biotechnology, SNPM-H023) to a density of 1.0 × 10⁻⁶ cells / mL. 5 Cell suspension of 10 cells / mL was seeded into 12-well culture plates, 1 mL per well, and cultured at 37°C in a 5% CO2 incubator for 24 h.

[0075] The active ingredients were trifolinol-flavonoid complex, pterostilbene, and Ganoderma lucidum extract. The total active ingredient content was calculated by summing the corresponding mass percentages of the three. Based on the total active ingredient content of the stock solutions of Examples 1-6 or Comparative Examples 1-6, the solutions were diluted with serum-free DMEM medium (Shangen Biotechnology, SNM-002D) to a total active ingredient concentration of 0.1 mg / mL to prepare the test sample solution (containing serum-free medium and the sample of the examples or comparative examples). Simultaneously, a propylene glycol-betaine mixed solution of the same concentration and proportion as the sample group was prepared using serum-free DMEM medium as a solvent control (containing serum-free medium and solvent). Discard the culture medium from the cell culture plate and add 1 mL of treatment solution to each well according to the following groups: Blank control group: add the solvent control described above; Model group: add 0.5 mM methylglyoxal (MGO) and the solvent control described above; Positive control group: add 0.5 mM MGO and 0.5 mM aminoguanidine hydrochloride and the solvent control described above; Sample group: add 0.5 mM MGO and the above-mentioned test sample solution (final total concentration of active ingredient is 0.1 mg / mL). Each group has 3 replicates. Incubate at 37℃ in a 5% CO2 incubator for 48 h.

[0076] After incubation, discard the culture medium and wash each well once with 1 mL of PBS. Add 150 μL of radioimmunoprecipitation assay (RIPA) lysis buffer (Lianke Biotechnology, WB020) containing 10 μL / mL benzyl sulfonyl fluoride (PMSF) to each well, lyse on ice for 30 min, collect the cell lysate, centrifuge at 12000 rpm for 10 min at 4 °C, and collect the supernatant. Use a portion of the supernatant to determine the total protein concentration using a quinolinic acid (BCA) protein quantification kit (Lianke Biotechnology, PQ0012). For the remaining supernatant, follow the instructions for the human CML enzyme-linked immunosorbent assay (ELISA) kit (Cell Biolabs, STA-816), measuring the absorbance of each well at 450 nm. Calculate the CML content according to the standard curve and correct it with the total protein concentration. The result is expressed as "ng CML / mg total protein". The CML inhibition rate is calculated using the following formula. The independent samples t-test between groups was used to compare Examples 1-6 with Comparative Example 2 to evaluate the statistical significance of the differences.

[0077] CML inhibition rate (%) = 1 - [(model group - sample group) / (model group - blank group) × 100%].

[0078] CML is a characteristic advanced product of glycation, directly reflecting the degree of intracellular glycation damage accumulation. It is directly related to skin sallowness and collagen cross-linking, thus enhancing its efficacy. CML content detection results are as follows... Figure 2 As shown, the CML inhibition rate of Comparative Example 5 was 11.98%, indicating that the trifolinol-flavonoid complex has a certain CML inhibitory ability. Among the pairwise combinations, Comparative Example 2, containing pterostilbene and Ganoderma lucidum extracts, had the highest inhibition rate (19.43%), which was better than the other binary combinations in Comparative Examples 1 and 3. Moreover, the CML inhibition rate of the trifolinol-flavonoid complex, pterostilbene and Ganoderma lucidum extracts in Examples 1-6 was significantly higher than that of Comparative Example 2 (p<0.01). This indicates that trifolinol-flavonoids can significantly enhance the CML inhibitory ability of pterostilbene and Ganoderma lucidum extracts, and the combination of the three produces an overall effect superior to the pairwise combinations.

[0079] Application Example 1 An essence lotion was prepared according to the formulations in Table 3, comprising the compositions of Example 1, Example 2, Comparative Example 2, and a blank matrix, and labeled as Essence Lotion 1, Essence Lotion 2, Essence Lotion 3, and Essence Lotion 4, respectively. Essence Lotion 4 contained no active ingredients (trifolinol-flavonoid complex, pterostilbene and Ganoderma lucidum extracts); the amount of active ingredients added to the other essence lotions was adjusted according to the dilution factor based on the total active ingredient content of their original solutions, so that the total active ingredient content in the final products was consistent (0.2 wt%). The specific preparation process includes the following steps: (1) According to Table 3, the raw materials of phase A were added to the reactor in sequence, stirred and heated to 85°C, and kept at the temperature to mix and stir evenly to obtain phase A; (2) According to Table 3, mix the raw materials of phase B, stir and heat to 85°C until completely melted and uniform to obtain phase B; (3) Keep at 85°C, slowly add phase B to phase A, homogenize at 8000 rpm for 5 minutes to form a uniform emulsion; stir and cool to 45°C, then add phase C and phase D raw materials in sequence, and mix and stir evenly. (4) Cool to room temperature and filter through a 200-mesh filter to obtain the essence lotion.

[0080] Table 3. Essence Lotion Formula Note: The total active ingredient content of the composition stock solution in Example 1 is 10%, the total active ingredient content of the composition stock solution in Example 2 is 13%, and the total active ingredient content of the composition stock solution in Comparative Example 2 is 7%. The addition amounts of each group were converted to ensure that the total active ingredient content in the final product is consistent (all are 0.2wt%).

[0081] Human efficacy evaluation of essence lotion (1) Test method Eighty healthy female volunteers aged 30-50 years with glycation symptoms such as dullness and uneven skin tone were recruited. They were randomly divided into four groups of 20 each. Each group applied serum 1, serum 2, serum 3, and serum 4 to the entire face twice daily, morning and evening, for 28 consecutive days. Facial L* (brightness) and b* (yellowing) values ​​were measured using a skin colorimeter before product use (day 0), after 14 days, and after 28 days. Paired t-tests were used to compare differences before and after use within the same group; independent samples t-tests were used between groups at the same time point, with changes in serum 1, serum 2, and serum 3 compared to serum 4, and serum 1 and serum 2 compared to serum 3. p < 0.05 was considered statistically significant, and p < 0.01 was considered highly statistically significant. After 28 days of use, participants completed a self-assessment questionnaire, evaluating indicators such as skin brightening, improvement in dullness, skin radiance, and skin texture. The colorimetric test results are shown in Table 4, and the subjects' self-evaluation results are shown in Table 5.

[0082] (2) Experimental results Table 4. Skin brightness (L* value) and yellowness (b* value) of subjects after using the serum containing the composition. Note: Data in Table 4 are expressed as mean ± standard deviation (n=20 / group). Change = Measured value at each time point − Measured value before use (day 0); Rate of change = Change / Measured value before use × 100%. Compared with the same group before use,* p<0.05, ** p<0.01; compared with serum 4 (blank group) at the same time point, △ p<0.05, △△ p<0.01; compared with serum 3 (comparative example 2) at the same time, # p<0.05, ## p<0.01. No * was marked at any time point in the control group, indicating that there was no statistically significant improvement in skin color compared to before and after treatment within the same group (p>0.05).

[0083] Table 5. Self-assessment results of subjects after 28 days of using the serum containing the composition. Note: The data in Table 5 represent the proportion of subjects in each experimental group who rated their condition as "significantly improved / enhanced" or "satisfied" (n=20 / group). After 28 days of use, subjects self-evaluated using a 4-point scale (significantly improved, somewhat improved, no change, worsened). The table shows the combined proportion of those who rated their condition as "significantly improved" and those who rated it as "somewhat improved."

[0084] As shown in Table 4, after 28 days of use, the L* value of the blank matrix group (Essence 4) increased by only 0.27 (+0.48%) and the b* value decreased by only 0.17 (-1.16%), and neither was statistically significant (p>0.05), indicating that the matrix itself had no significant effect on skin color.

[0085] After 28 days of use, the serum lotion 3 containing the composition of Comparative Example 2 (a combination of pterostilbene and Ganoderma lucidum extracts, without trifolinol-flavonoids) showed an increase in L* value of 1.78 (+3.27%, p<0.01) and a decrease in b* value of 0.99 (-6.51%, p<0.01), and its improvement effect was significantly better than that of the blank matrix (p<0.01), indicating that the combined use of pterostilbene and Ganoderma lucidum extracts has a certain effect on improving skin tone.

[0086] After 28 days of use, the L* value of serum 1 containing the composition of Example 1 increased by 2.86 (+5.07%, p<0.01), and the b* value decreased by 1.46 (-10.05%, p<0.01). After 28 days of use, the L* value of serum 2 containing the composition of Example 2 increased by 1.97 (+3.38%, p<0.01), and the b* value decreased by 1.09 (-7.89%, p<0.01). Both groups of examples containing trifolinol-flavonoids showed significantly better increases in L* value and decreases in b* value than the blank matrix (p<0.01) and Comparative Example 2 (p<0.05 or p<0.01), indicating that the combined use of trifolinol-flavonoids, pterostilbene, and Ganoderma lucidum extract has anti-glycation and brightening effects, and is significantly superior to the binary combination containing only pterostilbene and Ganoderma lucidum extract.

[0087] Table 5 shows that in Essence Lotion 1 and Essence Lotion 2 groups, 90%-100% of the subjects reported improvements in skin tone brightening, dullness reduction, radiance enhancement, and skin texture refinement, with overall satisfaction reaching 100%. In contrast, the satisfaction rate for Essence Lotion 3 (Comparative Example 2) was only 70%-90%, and the satisfaction rate for the blank matrix group was only 15%-30%. These subjective user feedbacks demonstrate that the trifolin sterol-flavonoids in the composition of this invention have a positive synergistic effect on enhancing anti-glycation and brightening / skin-brightening efficacy. Furthermore, a comparison of typical subject facial images is provided (…). Figure 3 The results showed that after 128 days of continuous use of the serum, the subjects' dull complexion improved significantly and their skin brightness increased markedly.

[0088] Application Example 2 A face cream, the preparation process of which is shown below: (1) According to Table 6, the raw materials of phase A were added to the reactor in sequence, stirred and heated to 85°C, and kept at the temperature to mix and stir evenly to obtain phase A; (2) According to Table 6, mix the raw materials of phase B, stir and heat to 85°C until completely melted and homogeneous to obtain phase B; (3) Keep at 85°C, slowly add phase B to phase A, homogenize at 8000 rpm for 5 minutes to form a uniform emulsion; stir and cool to 45°C, then add phase C, phase D and phase E raw materials in sequence, and mix and stir evenly. (4) After cooling to room temperature and passing the inspection, filter the material through a 200-mesh filter to obtain the face cream.

[0089] Table 6 Face Cream Recipes According to the formulation and preparation method of Application Example 2 above, a stable and uniform face cream product can also be obtained, and this face cream product also has anti-glycation, yellowing removal and brightening effects on the skin comparable to the essence lotion 1 in Application Example 1 above; it can be shown that the composition of the present invention can be adapted in both lotion and face cream cosmetic bases, and also exhibits obvious anti-glycation, yellowing removal and brightening effects on the skin.

[0090] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. A composition having anti-glycation and anti-yellowing effects, characterized in that, The product comprises the following components in parts by weight: 0.5-5 parts of trifolin sterol-flavonoid complex, 0.5-5 parts of pterostilbene, 1-10 parts of Ganoderma lucidum extract, and 80-98 parts by weight of solvent; the trifolin sterol-flavonoid complex comprises trifolin sterol component and trifolin flavonoid component, the trifolin sterol component comprises sterol compounds, the trifolin flavonoid component comprises flavonoid compounds, the total mass percentage of sterol compounds in the trifolin sterol component is greater than or equal to 15%, and the total mass percentage of flavonoid compounds in the trifolin flavonoid component is greater than or equal to 60%.

2. The composition with anti-glycation and anti-yellowing effects as described in claim 1, characterized in that, The mass ratio of the trifolin sterol-flavonoid complex to pterostilbene was (0.25-4):

1.

3. The composition with anti-glycation and anti-yellowing effects as described in claim 1, characterized in that, The mass ratio of Ganoderma lucidum extract to pterostilbene is (0.75-8):

1.

4. The composition with anti-glycation and anti-yellowing effects as described in claim 1, characterized in that, Based on the total mass of the *Trifoliol-Flavone Complex*, the total mass percentage of sterols in the *Trifoliol-Flavone Complex* is greater than 1.5%, and the total mass percentage of flavonoids is greater than 5%.

5. The composition with anti-glycation and anti-yellowing effects as described in claim 1, characterized in that, In the trifoliol-flavonoid complex, the mass ratio of the trifoliol component to the trifoliol flavonoid component is (0.125-8):

1.

6. The composition with anti-glycation and anti-yellowing effects as described in claim 1, characterized in that, The solvent includes at least one of alcohols, betaine, and water.

7. A method for preparing a composition with anti-glycation and anti-yellowing effects as described in any one of claims 1-6, characterized in that, Includes the following steps: S1. Obtain the trifoliol component and the trifoliol flavonoid component separately, and mix the trifoliol component and the trifoliol flavonoid component evenly to obtain the trifoliol-flavonoid complex. S2. Dissolve the trifolinol-flavonoid complex and pterostilbene in a solvent, then add Ganoderma lucidum extract and mix well to obtain the composition.

8. The use of the composition having anti-glycation and anti-yellowing effects according to any one of claims 1-6 in at least one of the following I-III for non-disease treatment purposes, or in the preparation of a product having at least one of the following I-VII functions, characterized in that, I. Inhibits the formation of advanced glycation end products; II. Inhibit the expression of receptors for advanced glycation end products; III. Inhibits the formation of carboxymethyl lysine; IV. Improves skin glycation reaction; V. Improves skin pigmentation; VI. Improve skin radiance; VII. Reduce skin yellowing.

9. A cosmetic product, characterized in that, The composition comprising any one of claims 1-6 having anti-glycation and anti-yellowing effects.

10. The cosmetic product as described in claim 9, characterized in that, Based on the total mass of the cosmetics, the composition having anti-glycation and anti-yellowing effects comprises 0.1%-10% by mass. And / or, the cosmetic dosage form includes one of serum, lotion, cream, mask or toner.

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