A composition for microecological balance and circadian rhythm repair and application

Abstract

The application relates to the cosmetic technical field, and particularly discloses a composition for micro-ecological balance and circadian rhythm repair and lightening and application. The composition for micro-ecological balance and circadian rhythm repair and lightening comprises the following components in mass percentage: glycyrrhiza glabra root extract 0.001%-1%, olea europaea leaf extract 0.1%-5%, hydraena elongata extract 0.1%-2%, gentiana rigescens 0.5%-3%, inulin 0.1%-5%, tripeptide-3 0.001%-0.01%, jasminum nudiflorum extract 0.5%-2%, and the balance is water. The composition has the advantages of systematically solving multiple problems such as skin redness, dullness and rhythm disorder.

Description

Technical Field

[0001] This application relates to the field of cosmetic technology, and more specifically, to a composition and its application for microecological balance and circadian rhythm repair and brightening. Background Technology

[0002] With increasing environmental pollution, life stress, and increasingly complex skincare ingredients, the proportion of people with sensitive skin continues to rise. This manifests as impaired skin barrier function, imbalanced microecology, and a tendency to experience redness, stinging, and other problems. Furthermore, these individuals often have difficulty tolerating traditional, highly effective whitening ingredients (such as high-concentration acids and retinol), indicating a significant unmet need in the field of whitening and fading dark spots.

[0003] Most brightening products on the market today have a single mechanism, focusing primarily on inhibiting tyrosinase activity or blocking melanin transfer. They generally neglect the root cause impact of skin microecological health on inflammatory factors and redness, and fail to consider the disruption of the skin's circadian rhythm (biological clock) caused by modern lifestyle factors such as staying up late, blue light, and ultraviolet light damage. Disruption of this rhythm directly leads to decreased skin repair ability and dull, sallow skin tone.

[0004] Current technologies typically involve simply layering soothing, whitening, and antioxidant ingredients, lacking core compositions that can simultaneously achieve synergistic effects across multiple dimensions, including the skin's microecological barrier, inflammatory factors, melanin pathways, and cellular rhythms. For example, inulin, as a prebiotic, is known to regulate the skin's microecology, but its synergistic effect with rhythm-regulating ingredients in brightening compositions has not been reported.

[0005] Therefore, there is an urgent need in this field for an innovative composition that can systematically solve multiple problems such as skin redness, dullness, and rhythm disorders. Summary of the Invention

[0006] To address the aforementioned issues, this application provides a composition and its application for restoring and brightening the microecological balance and circadian rhythm.

[0007] Part One, this application provides a composition for microecological balance and circadian rhythm repair and brightening: A composition for balancing the gut microbiome and restoring and brightening the circadian rhythm, comprising the following components by weight percentage: Licorice root extract 0.001%-1%, olive leaf extract 0.1%-5%, seaweed extract 0.1%-2%, gentian root extract 0.5%-3%, inulin 0.1%-5%, tripeptide-32 0.001%-0.01%, magnolia flower extract 0.5%-2%, balance being water.

[0008] By employing the above-mentioned technical solutions, licorice root extract, as the core whitening ingredient, can achieve multi-pathway whitening by inhibiting tyrosinase activity and scavenging free radicals within an addition range of 0.001%-1%, while its anti-inflammatory properties can alleviate inflammatory reactions in sensitive skin. Olive leaf extract, within a range of 0.1%-5%, is rich in oleuropein and hydroxytyrosol, effectively inhibiting the release of inflammatory factors and reducing skin redness. Elongated seaweed extract, within a range of 0.1%-2%, provides abundant polysaccharides and trace elements, enhancing skin barrier function and promoting cell metabolism. Gentiana scabra, within a range of 0.5%-3%, possesses both anti-inflammatory and antioxidant effects, relieving immediate discomfort in sensitive skin. Inulin, as a prebiotic, within a range of 0.1%-5%, can selectively proliferate beneficial skin flora, fundamentally improving inflammation caused by microecological imbalance. Tripeptide-32, as a rhythm-regulating peptide, can activate clock genes and enhance nighttime cell repair capabilities at extremely low concentrations of 0.001%-0.01%. Magnolia officinalis extract, at concentrations ranging from 0.5% to 2%, can repair photodamage and inhibit photoaging. Through the synergistic effect of the above components, this composition achieves comprehensive effects of balancing the microecology, repairing rhythm, and brightening through multiple pathways at low concentrations tolerable for sensitive skin.

[0009] Optional, by weight, it may also include 0.1%-2% arnica flower extract, 0.1%-1% bisabolol, and 0.05%-0.5% dipotassium glycyrrhizate.

[0010] By employing the above-mentioned technical solutions, arnica flower extract, within the range of 0.1%-2%, exhibits significant anti-inflammatory and microcirculation-promoting effects, accelerating the reduction of redness. Bisabolol, within the range of 0.1%-1%, serves as a classic soothing ingredient, inhibiting the release of inflammatory mediators and relieving stinging and burning sensations. Dipotassium glycyrrhizate, within the range of 0.05%-0.5%, possesses anti-inflammatory effects similar to glucocorticoids but without side effects, effectively suppressing allergic reactions. These three ingredients, synergistically combined with gentian extract and olive leaf extract in the base components, significantly enhance the immediate redness-reducing and soothing effects of the composition, making the product more suitable for use on sensitive skin during the acute redness phase.

[0011] Optionally, the olive leaf extract is subjected to metal ion chelation treatment, wherein the metal ion chelation treatment method includes: Olive leaf extract was mixed with an aqueous solution of sodium phytate or disodium ethylenediaminetetraacetate at a molar concentration of 0.01%-0.1% at a mass ratio of 1:8-15, stirred at 40-50℃ for 90-110 minutes, purified by membrane separation technology and then dried.

[0012] By employing the above-mentioned technical solution, the active polyphenolic components (such as oleuropein) in olive leaf extract are easily complexed with metal ions, thus reducing their activity. Treating the extract with a 0.01%-0.1% sodium phytate or disodium EDTA solution at 40-50°C for 90-110 minutes effectively removes or complexes the metal ions introduced during extraction, preventing the oxidative degradation of polyphenolic components. An optimized mass ratio of 1:8-15 ensures that the chelating agent can fully contact the extract without excessively diluting the active ingredients. The application of membrane separation technology avoids the loss of active ingredients caused by traditional precipitation methods. The stability of the olive leaf extract treated in this way is significantly improved, and its antioxidant and anti-inflammatory effects in the composition are maintained for a long time.

[0013] Optionally, the *Elongatus spp.* extract undergoes the following antioxidant stabilization pretreatment: Under nitrogen or argon protection, the extract of *Elongata spp.* is mixed with 3%-8% of natural tocopherol and ascorbate tetraisopalmitate at a mass ratio of 1:1-3, and stirred at 20-30°C for 20-40 minutes.

[0014] By employing the above-mentioned technical solution, the extract of *Euphorbia pulcherrima*, rich in unsaturated fatty acids and polysaccharides, is prone to oxidation and deterioration. Under inert gas protection, it is mixed with 3%-8% of natural tocopherol and ascorbate tetraisopalmitate at a mass ratio of 1:1-3, achieving a dual protection mechanism: tocopherol, as a fat-soluble antioxidant, scavenge lipid peroxidation free radicals, while ascorbate tetraisopalmitate, as an oil-soluble vitamin C derivative, regenerates tocopherol and provides its own antioxidant capacity. Mild conditions of 20-30℃ prevent the destruction of heat-sensitive components. The stability of the *Euphorbia pulcherrima* extract in the composition is significantly improved after this treatment, allowing its barrier repair and cell metabolism-promoting effects to be fully realized.

[0015] Optionally, the inulin is enzymatically modified to obtain fructooligosaccharides with a degree of polymerization of 2-10, wherein the enzymatic modification method includes: Inulin was prepared into an aqueous solution with a mass fraction of 10%-15%, and inulinase was added at a mass fraction of 0.2%-0.4% of the inulin. The mixture was enzymatically hydrolyzed at 53-58℃ for 3-5 hours. After enzyme inactivation, the mixture was spray-dried to obtain modified inulin with an oligofructose content ≥85%.

[0016] By employing the above technical solution, ordinary inulin (degree of polymerization > 20) exhibits limited water solubility and prebiotic activity. By preparing inulin into a 10%-15% aqueous solution, adding 0.2%-0.4% inulinase, and enzymatically hydrolyzing at 53-58℃ for 3-5 hours, it can be degraded into fructooligosaccharides with a degree of polymerization of 2-10. The spray-drying process ensures good flowability and solubility of the product. The resulting modified inulin has an fructooligosaccharide content ≥ 85%, and its prebiotic activity is 2-3 times higher than that of ordinary inulin, enabling more efficient proliferation of beneficial bacteria such as Bifidobacteria and inhibition of harmful bacteria growth. When used in compositions, it can achieve superior microecological regulation effects with lower addition amounts, while simultaneously improving product texture and skin feel.

[0017] Optionally, the Magnolia officinalis extract is prepared by a low-temperature dynamic countercurrent extraction process, with the extraction solvent being a 60%-75% (v / v) butanediol aqueous solution, the extraction temperature being 42-48℃, the extraction time being 2.5-3.5 hours, and the material-to-liquid ratio being 1:(8-12).

[0018] By employing the above technical solution, the active ingredients in Magnolia officinalis (such as flavonoids and phenylethanoids) are heat-sensitive, and traditional thermal extraction easily causes degradation. Using a 60%-75% (v / v) butylene glycol aqueous solution as the extraction solvent provides both good solubility and cosmetic compatibility. The extraction temperature is controlled within a mild range of 42-48℃, combined with 2.5-3.5 hours of dynamic countercurrent extraction, ensuring both mass transfer efficiency and avoiding damage to heat-sensitive components. An optimized solid-liquid ratio of 1:8-12 ensures a balance between extraction completeness and cost-effectiveness.

[0019] Optionally, the composition further comprises 0.001%-0.1% ergothioneine and 0.01%-0.5% evening primrose extract by weight. By employing the above-mentioned technical solution, ergothioneine, at a concentration of 0.001%-0.1%, acts as a super antioxidant, capable of penetrating cell membranes and entering mitochondria to scavenge free radicals. Its antioxidant capacity is more than five times that of glutathione, and it also possesses anti-blue light damage and anti-aging effects. Evening primrose extract, at a concentration of 0.01%-0.5%, is rich in gamma-linolenic acid and flavonoids, which can regulate the release of inflammatory mediators, improve skin microcirculation, and alleviate dullness caused by staying up late. The mitochondrial-targeted antioxidant effect of ergothioneine and the anti-inflammatory regulation of evening primrose extract create a synergistic effect, improving skin rhythm disorders through both cellular energy metabolism and inflammatory responses. These two components, along with tripeptide-32 and gentian extract in the base composition, further enhance the repair effect on skin damaged by lack of sleep and photoreactive substances.

[0020] Secondly, this application provides an application of a composition for microecological balance and circadian rhythm repair and brightening.

[0021] A composition for balancing the skin's microbiome and restoring its circadian rhythm and brightening its complexion is used in cosmetics to protect the skin from blue light damage.

[0022] In summary, this application has the following beneficial effects: 1. This application uses a scientifically formulated blend of licorice root extract, olive leaf extract, inulin, tripeptide-32, and magnolia flower extract. Through the synergistic effects of inhibiting inflammatory factors, regulating the skin microecology, activating circadian rhythm genes, inhibiting melanin through multiple pathways, and repairing photodamage, it achieves for the first time a systematic solution to the problems of redness, dullness, and rhythm disorder in sensitive skin from four dimensions: "microecological barrier - inflammation relief - rhythm repair - pigment metabolism". It can exert significant effects even at low concentrations, ensuring the product's extreme gentleness.

[0023] 2. This application features targeted process optimization for key components: olive leaf extract undergoes metal ion chelation treatment, significantly improving the stability of polyphenolic components; seaweed extract undergoes antioxidant stabilization pretreatment, effectively preventing the oxidation of unsaturated fatty acids; inulin is enzymatically modified into fructooligosaccharides, increasing prebiotic activity by 2-3 times. These optimized processes ensure the long-lasting efficacy of the active ingredients during long-term storage and use.

[0024] 3. Based on the basic formula, this application further adds ergothioneine and evening primrose extract. The two work synergistically to exert the dual effects of mitochondrial-targeted antioxidant and anti-inflammatory regulation. Combined with the rhythm activation effect of tripeptide-32, it synergistically improves the skin rhythm disorder and dullness caused by modern life factors such as staying up late and blue light from three levels: cellular energy metabolism, inflammatory response and biological clock regulation. It significantly enhances the repair effect of the composition on photo-damaged skin and skin damaged by staying up late. Detailed Implementation

[0025] The following detailed description of this application is provided in conjunction with the embodiments. It should be noted that: unless otherwise specified, the conditions in the following embodiments are performed under conventional conditions or conditions recommended by the manufacturer. Unless otherwise specified, the raw materials used in the following embodiments are all from commercially available sources.

[0026] Example 1 This embodiment provides a composition for microecological balance and circadian rhythm repair and brightening. The preparation method is as follows, based on a total composition volume of 10 kg: 1. Formula Composition Licorice root extract: 50.05g (40% purity, Shanxi Yuning Biotechnology Co., Ltd.); Olive leaf extract: Dosage 255g (model SC-SL053HEALTH, Xi'an Shangcheng Biotechnology Co., Ltd.); Himanthalia esculenta extract: 105g (trade name: AQUALGAE Himanthalia, Lessonia, France). Gentiana scabra: Dosage 175g (Manufacturer: Shanghai Jiayu Biotechnology Co., Ltd., Trade name: Gentiana scabra vesicles); Inulin: Dosage 255g (Manufacturer: NIPPON FINE CHEMICAL CO., LTD. Trade name: Inulin-SC); Tripeptide-32: Dosage 0.55g (Manufacturer: Shandong Jitai Biotechnology Co., Ltd., Trade Name: Tripeptide-32); Magnolia officinalis extract: Dosage 125g (Manufacturer: Shanghai Jiakai Biotechnology Co., Ltd., Trade name: Solarepare MB); The remainder is water.

[0027] 2. Preparation steps Step 1: Weigh out olive leaf extract, inulin, magnolia flower extract, gentian root extract, and licorice root extract, and set aside. Add 5 kg of purified water to the reaction vessel and start stirring at 30-40 rpm. Slowly add the weighed water-soluble raw materials to the reaction vessel while simultaneously starting the heating system to gradually raise the system temperature to 70-75℃. Once the temperature reaches 70-75℃, turn on the homogenizer at 2500-3000 rpm and homogenize for 5-8 minutes to ensure all water-soluble components are fully dissolved and evenly dispersed, with no visible particles. Turn off the homogenizer, but continue stirring (30-40 rpm) at 70-75℃ for 30 minutes to ensure full hydration and blending of the components.

[0028] Step 2: Turn off the heating system and turn on the circulating cooling water to slowly lower the system temperature to 45℃. Control the cooling rate at 1-2℃ / minute to avoid excessive temperature difference affecting the stability of the material. When the temperature stabilizes at 45℃, add the elongated seaweed extract. Maintain a stirring speed of 30-40 rpm and continue stirring for 15-20 minutes to ensure that the elongated seaweed extract is completely and evenly dispersed and the system is homogeneous.

[0029] Step 3: Further reduce the system temperature to below 40℃. Add tripeptide-32. Since the amount of tripeptide-32 used is extremely small, to ensure uniform dispersion, pre-dissolve tripeptide-32 in 50-100g of purified water at below 40℃ before slowly adding it to the reactor. Reduce the stirring speed to 20-25 rpm and stir slowly for 10-15 minutes until tripeptide-32 is completely dissolved and the system is uniformly mixed. Avoid high-speed stirring during this stage to prevent introducing excessive air bubbles or causing shear damage to heat-sensitive components.

[0030] Step 4: Add the remaining purified water to the reactor to bring the total composition volume to 10 kg. Continue stirring (20-25 rpm) during the water addition process to ensure system homogeneity. Continue stirring and cooling until the system temperature drops below room temperature (below 25°C). Turn off the stirring and let the composition stand for 30-60 minutes to allow any air bubbles introduced during stirring to rise and dissipate naturally. Filter the mixture through a 200-mesh filter to obtain the composition used for microecological balance and circadian rhythm repair and brightening.

[0031] Example 2 A composition for restoring and brightening the microecological balance and circadian rhythm, differing from Example 1 in that: based on a preparation of 10 kg, its formulation is as follows: The ingredients included 0.1g of Glycyrrhiza glabra root extract, 450g of olive leaf extract, 150g of Echinochloa chinensis extract, 50g of Gentiana scabra, 450g of inulin, 0.1g of tripeptide-32, 120g of Magnolia officinalis extract, and the remainder being water. The preparation method was the same as in Example 1.

[0032] Example 3 A composition for restoring and brightening the microecological balance and circadian rhythm, differing from Example 1 in that: based on a preparation of 10 kg, its formulation is as follows: The ingredients included 80g of Glycyrrhiza glabra root extract, 50g of olive leaf extract, 30g of Echinochloa chinensis extract, 280g of Gentiana scabra, 50g of inulin, 0.9g of tripeptide-32, 180g of Magnolia officinalis extract, and the remainder being water. The preparation method was the same as in Example 1.

[0033] Example 4 A composition for microecological balance and circadian rhythm repair and brightening differs from Example 1 in that it also contains 105g of arnica flower extract (purity specification 10:1, i.e., 10 kg of raw material to extract 1 kg of powder), 55g of bisabolol (98% α-bisabolol, Shaanxi Haochen Biotechnology), and 27.5g of dipotassium glycyrrhizate (98% purity (HPLC high performance liquid chromatography)).

[0034] Furthermore, arnica flower extract is added in preparation step two, bisabolol is added in preparation step three, and dipotassium glycyrrhizate is added in preparation step four.

[0035] Example 5 A composition for microecological balance and circadian rhythm repair and brightening, which differs from Example 4 in that it also contains 10g of arnica flower extract, 10g of bisabolol, and 5g of dipotassium glycyrrhizate.

[0036] Example 6 A composition for microecological balance and circadian rhythm repair and brightening, which differs from Example 4 in that it also contains 200g of arnica flower extract, 100g of bisabolol, and 50g of dipotassium glycyrrhizate.

[0037] Example 7 A composition for microecological balance and circadian rhythm repair and brightening differs from Example 1 in that: in the second preparation step, 5.05g of ergothioneine (purity 98%) and 25.5g of evening primrose extract (extraction ratio 10:1, Shaanxi Snowt Biotechnology) are added.

[0038] Example 8 A composition for microecological balance and circadian rhythm repair and brightening differs from Example 7 in that 0.1g of ergothioneine and 1g of evening primrose extract are added in preparation step two.

[0039] Example 9 A composition for microecological balance and circadian rhythm repair and brightening differs from Example 7 in that 10g of ergothioneine and 50g of evening primrose extract are added in preparation step two.

[0040] Example 10 A composition for microecological balance and circadian rhythm repair and brightening differs from Example 1 in that the olive leaf extract in this example undergoes metal ion chelation treatment, the specific steps of which are as follows: Take 1000g of olive leaf extract; prepare an aqueous solution of sodium phytate with a molar concentration of 0.055% by weighing 5.5g of sodium phytate and dissolving it in 10kg of purified water, stirring until homogeneous.

[0041] Add 1000g of olive leaf extract and 10kg of sodium phytate aqueous solution (mass ratio 1:10) to a reaction vessel. Start stirring at 40-50 rpm and heat to 45℃. Maintain this temperature and stir for 100 minutes to allow the metal ions to fully combine with the chelating agent.

[0042] After the reaction, the solution was purified using a membrane separation system. A nanofiltration membrane with a molecular weight cutoff of 2000 Daltons was used to remove unreacted small molecule chelating agents and free metal ions. The retentate, i.e., the purified olive leaf extract solution, was collected.

[0043] The purified extract solution was spray-dried. The inlet air temperature was 160-180℃, the outlet air temperature was 80-90℃, and the atomizer speed was 15000-20000 rpm. The dried powder was collected to obtain the olive leaf extract treated with metal ion chelation.

[0044] Example 11 A composition for microecological balance and circadian rhythm repair and brightening differs from Example 10 in that: olive leaf extract is mixed with a sodium phytate aqueous solution with a molar concentration of 0.01% at a mass ratio of 1:8.

[0045] Example 12 A composition for microecological balance and circadian rhythm repair and brightening differs from Example 10 in that: olive leaf extract is mixed with a 0.1% aqueous solution of disodium ethylenediaminetetraacetate at a mass ratio of 1:15.

[0046] Example 13 A composition for restoring and brightening the microecological balance and circadian rhythm, differing from Example 1 in that: the extract of *Euphorbia pulcherrima* in this example has undergone an antioxidant stabilization pretreatment, the specific steps of which are as follows: Prepare 1000g of *Euphorbia lathyris* extract; prepare a mixture of natural tocopherol and ascorbate tetraisopalmitate, the total amount of which is 5.5% of the mass of *Euphorbia lathyris* extract, i.e., 55g. The natural tocopherol and ascorbate tetraisopalmitate are mixed in a mass ratio of 1:2, i.e., 18.3g of natural tocopherol and 36.7g of ascorbate tetraisopalmitate.

[0047] The extract of *Elongatus spp.* was placed in a reaction vessel equipped with a stirrer. High-purity nitrogen (≥99.99%) was continuously introduced into the vessel to purge the air inside and maintain a positive nitrogen pressure environment.

[0048] Under nitrogen protection, slowly add 55g of the antioxidant mixture to the *Elongatus spp.* extract. Start stirring at 30-40 rpm, maintaining the system temperature at 25°C. Keep stirring at this temperature for 30 minutes to ensure the antioxidants and extract are thoroughly mixed.

[0049] After stirring, the mixture was cooled to room temperature under nitrogen protection to obtain the pretreated Elongatus extract.

[0050] Example 14 A composition for microecological balance and circadian rhythm repair and brightening differs from Example 13 in that: under nitrogen protection, extract of Elongatus spp. is mixed with 3% of natural tocopherol and ascorbate tetraisopalmitate at a mass ratio of 1:1.

[0051] Example 15 A composition for microecological balance and circadian rhythm repair and brightening differs from Example 13 in that: under argon protection, extract of Elongatus spp. is mixed with 8% of natural tocopherol and ascorbate tetraisopalmitate at a mass ratio of 1:3.

[0052] Example 16 A composition for restoring and brightening the microecological balance and circadian rhythm differs from Example 1 in that: in this example, the inulin undergoes enzymatic hydrolysis modification to prepare oligofructose with a degree of polymerization of 2-10. The specific steps are as follows: Prepare 1000g of inulin; prepare 3g of inulinase, which is 0.3% of the mass of inulin.

[0053] Add inulin to the reaction vessel, then add purified water to prepare a 12.5% ​​aqueous solution (i.e., add 7000g of purified water). Start stirring at 50-60 rpm and heat to 55-60℃ to completely dissolve the inulin.

[0054] Adjust the solution temperature to 55.5℃. Add inulinase and stir well. Incubate at 55.5℃ for 4 hours for enzymatic hydrolysis, and take samples periodically to detect the fructooligosaccharide content.

[0055] After enzymatic hydrolysis, rapidly raise the solution temperature to 85-90℃ and maintain this temperature for 10-15 minutes to inactivate the inulinase. Take samples for testing to ensure the fructooligosaccharide content is ≥85%.

[0056] The enzyme-inactivated liquid is spray-dried. The inlet air temperature is 170-180℃, the outlet air temperature is 85-90℃, and the atomizer speed is 18000-20000 rpm. The dried powder is collected to obtain modified inulin, with an oligofructose content ≥85% and a degree of polymerization of 2-10.

[0057] Example 17 A composition for microecological balance and circadian rhythm repair and brightening differs from Example 16 in that: inulin is prepared into a 10% aqueous solution and inulinase is added at 0.2% of the inulin mass.

[0058] Example 18 A composition for microecological balance and circadian rhythm repair and brightening differs from Example 16 in that: inulin is prepared into an aqueous solution with a mass fraction of 15%, and inulinase is added at a mass fraction of 0.4% of the inulin.

[0059] Example 19 A composition for microecological balance and circadian rhythm repair and brightening differs from Example 1 in that: the Magnolia officinalis extract in this example is prepared using a low-temperature dynamic countercurrent extraction process, the specific steps of which are as follows: Prepare 1000g of dried Magnolia officinalis raw material; the extraction solvent is a 67.5% (v / v) butanediol aqueous solution.

[0060] The raw material of Magnolia officinalis was fed into a dynamic countercurrent extraction device. 10 kg of the above-mentioned butanediol aqueous solution was added at a material-to-liquid ratio of 1:10. The extraction temperature was set to 45℃. The dynamic countercurrent extraction system was started, and the extraction time was 3 hours. During the extraction process, the solvent and material flowed continuously in opposite directions, maintaining dynamic contact.

[0061] After extraction, the extract is separated into solid and liquid components by pressure filtration to remove residue and collect the clear extract.

[0062] The extract was concentrated under reduced pressure at 55°C to recover the butanediol solvent, and concentrated to a relative density of 1.10 (measured at 60°C). The concentrate was then freeze-dried at -10°C to obtain a dry powder of Magnolia officinalis extract.

[0063] Example 20 A composition for microecological balance and circadian rhythm repair and brightening, which differs from Example 19 in that it is extracted by dynamic countercurrent extraction at a material-to-liquid ratio of 1:8.

[0064] Example 21 A composition for microecological balance and circadian rhythm repair and brightening, which differs from Example 19 in that it is extracted by dynamic countercurrent extraction at a material-to-liquid ratio of 1:12.

[0065] Comparative Example 1 A method for preparing a composition for microecological balance and circadian rhythm repair and brightening differs from Example 1 in that: licorice root extract is not used, but an equal amount of deionized water is substituted; the remaining components and preparation methods are the same as in Example 1.

[0066] Comparative Example 2 A method for preparing a composition for microecological balance and circadian rhythm repair and brightening differs from Example 1 in that: olive leaf extract is not used, but an equal amount of deionized water is used instead; the remaining components and preparation methods are the same as in Example 1.

[0067] Comparative Example 3 A method for preparing a composition for microecological balance and circadian rhythm repair and brightening differs from Example 1 in that: the extract of *Euphorbia pulcherrima* is not used, but an equal amount of deionized water is used instead; the remaining components and preparation methods are the same as in Example 1.

[0068] Comparative Example 4 A method for preparing a composition for microecological balance and circadian rhythm repair and brightening differs from Example 1 in that: inulin is not used, but is replaced by an equal amount of deionized water; the remaining components and preparation methods are the same as in Example 1.

[0069] Comparative Example 5 A method for preparing a composition for microecological balance and circadian rhythm repair and brightening differs from Example 1 in that: tripeptide-32 is not used, but replaced with an equal amount of deionized water; the remaining components and preparation methods are the same as in Example 1.

[0070] Comparative Example 6 A method for preparing a composition for microecological balance and circadian rhythm repair and brightening differs from Example 1 in that: Magnolia officinalis extract is not used; instead, an equal amount of deionized water is substituted. All other components and preparation methods are the same as in Example 1.

[0071] Performance testing 1. Detection of the efficacy of microecological regulation Detection indicator: Changes in the ratio of Staphylococcus epidermidis (beneficial bacteria) to Staphylococcus aureus (harmful bacteria).

[0072] Detection method: Ten subjects (aged 25-45 years) with sensitive facial skin and microecological imbalance were randomly selected for each example / comparative case. The subjects used the test sample for 28 consecutive days, once in the morning and once in the evening.

[0073] Facial stratum corneum samples were collected using sterile tape before use (D0) and 28 days after use (D28).

[0074] 16S rDNA high-throughput sequencing technology was used to detect bacterial abundance and calculate the ratio of Staphylococcus epidermidis to Staphylococcus aureus.

[0075] The results are expressed as the ratio of D28 to D0. A ratio >1 indicates the proliferation of beneficial bacteria, while a ratio <1 indicates the inhibition of harmful bacteria.

[0076] 2. Detection of the efficacy of circadian rhythm repair Detection indicators: relative mRNA expression levels of clock genes CLOCK, BMAL1, and PER1.

[0077] Detection method: An in vitro circadian rhythm disorder model was established using HaCaT immortalized human keratinocytes. Cells were treated with 100 μM luzindole, a melatonin antagonist, for 12 hours to induce circadian rhythm disorder. Subsequently, cells were treated with 0.5% of the test sample for 24 hours, and total RNA was extracted. The expression levels of CLOCK, BMAL1, and PER1 genes were detected by qRT-PCR. GAPDH was used as an internal control. Results are expressed as relative expression levels (treated / model control), with >1 indicating activation of the circadian rhythm genes.

[0078] 3. Brightening effect test Detection indicators: melanin content inhibition rate, tyrosinase activity inhibition rate.

[0079] Detection method: In vitro tyrosinase activity inhibition rate: The mushroom tyrosinase and L-DOPA reaction system was used. The sample solution was treated with 0.5% of the sample solution, and the change in absorbance at 405 nm was measured to calculate the tyrosinase activity inhibition rate.

[0080] In vitro melanin production inhibition rate: B16-F10 mouse melanoma cells were used to induce melanin production with 10 ng / mL α-MSH. After 72 hours of treatment with 0.5% sample, the absorbance of the cell lysate at 405 nm was measured, and the melanin production inhibition rate was calculated.

[0081] 4. Anti-inflammatory and soothing efficacy testing Detection indicators: redness improvement rate (change in a* value), and inhibition rate of inflammatory factor IL-6.

[0082] Detection method: Improvement of facial redness: 10 subjects in each group used the product continuously for 28 days. The facial a value (redness value) was measured using a skin colorimeter. The difference Δa between D28 and D0 was calculated. A negative value indicates a reduction in redness.

[0083] Inhibition of inflammatory factors: The RAW264.7 macrophage inflammation model stimulated by LPS was used. The cells were treated with 0.5% sample for 24 hours. The level of IL-6 in the cell supernatant was detected by ELISA and the inhibition rate was calculated.

[0084] 5. Mildness test Testing indicator: Lactic acid stinging test score.

[0085] Detection method: Thirty subjects with positive lactic acid stinging test results (sensitive skin) were selected for each group. A 5% lactic acid solution was applied to the nasolabial folds, and the stinging sensation was rated at 30 minutes (0-5 points, 0 for no sensation, 3 for obvious stinging, and 5 for severe stinging). After the subjects used the test sample for 7 consecutive days, the same lactic acid stinging test was performed again.

[0086] Calculate the rate of decrease in stinging score before and after use and take the average of each group (round to the nearest integer). The higher the rate of decrease, the more significant the relief and improvement.

[0087] sample Microbial community ratio Relative expression of rhythm genes Tyrosinase inhibition rate (%) Melanin production inhibition rate (%) Δa* (Redness Improvement) IL-6 inhibition rate (%) The rate of decrease in lactic acid stinging score (%) Example 1 1.68 2.15 62.5 58.3 -1.52 54.2 62 Example 2 1.52 1.93 55.2 51.5 -1.48 52.5 57 Example 3 1.75 2.28 65.8 62.2 -1.75 59.3 65 Example 4 1.65 2.08 60.8 56.5 -2.18 68.5 78 Example 5 1.52 1.92 52.3 48.2 -1.85 58.2 65 Example 6 1.70 2.12 63.2 59.1 -2.45 75.6 85 Example 7 1.72 2.58 68.2 64.5 -1.65 56.8 64 Example 8 1.60 2.25 58.5 55.2 -1.42 48.5 55 Example 9 1.75 2.85 70.5 67.8 -1.72 59.2 66 Example 10 1.75 2.22 68.5 64.2 -1.68 61.5 69 Example 11 1.70 2.18 65.2 61.0 -1.60 58.0 66 Example 12 1.78 2.25 70.2 66.0 -1.72 63.5 71 Example 13 1.80 2.28 66.5 62.8 -1.70 58.5 67 Example 14 1.73 2.20 64.2 60.5 -1.62 56.2 64 Example 15 1.85 2.32 68.0 64.5 -1.75 60.0 69 Example 16 2.35 2.20 64.0 60.0 -1.60 56.0 64 Example 17 2.05 2.18 63.2 59.0 -1.55 55.0 63 Example 18 2.58 2.22 64.5 60.5 -1.65 56.5 64 Example 19 1.70 2.35 70.2 65.5 -1.68 58.5 66 Example 20 1.69 2.28 66.8 62.0 -1.60 56.5 64 Example 21 1.71 2.42 72.5 68.0 -1.72 60.0 67 Comparative Example 1 1.55 1.95 18.5 22.3 -1.25 48.5 55 Comparative Example 2 1.52 1.92 55.2 52.5 -0.85 32.5 42 Comparative Example 3 1.25 1.85 58.5 55.2 -1.35 49.5 58 Comparative Example 4 0.95 2.05 60.2 56.5 -1.45 52.5 60 Comparative Example 5 1.62 1.15 60.8 57.2 -1.48 53.5 61 Comparative Example 6 1.60 1.45 48.5 45.2 -1.42 51.2 59 Combining Example 1 and Comparative Examples 1-6 with Table 1, it can be seen that Example 1 is significantly superior to the comparative examples in all indicators, including bacterial community ratio, relative expression of rhythm genes, tyrosinase inhibition rate, melanin production inhibition rate, Δa* redness improvement, IL-6 inhibition rate, and lactic acid stinging score reduction rate. Specifically, in Comparative Example 1, the absence of *Glycyrrhiza glabra* root extract resulted in a sharp drop in tyrosinase inhibition rate to 18.5% and melanin production inhibition rate to 22.3%, confirming that *Glycyrrhiza glabra* root extract is the core brightening component. In Comparative Example 2, the absence of *Olive oleifera* leaf extract resulted in a decrease in IL-6 inhibition rate to 32.5%, a weakening of Δa* redness improvement to -0.85, and a lactic acid stinging score reduction rate to 42%, demonstrating that *Olive oleifera* leaf extract is the core anti-inflammatory and soothing component. In Comparative Example 3, the absence of *Euphorbia pulcherrima* extract resulted in a significant decrease in bacterial community ratio. The ratio of beneficial bacteria decreased to 1.25, indicating a significant weakening of the microecological regulatory capacity. In Comparative Example 4, the absence of inulin reduced the bacterial community ratio to 0.95, lower than the baseline level, confirming the crucial role of inulin in the proliferation of beneficial bacteria. In Comparative Example 5, the absence of tripeptide-32 reduced the relative expression of rhythm genes to 1.15, almost eliminating the ability to activate rhythms. In Comparative Example 6, the absence of Magnolia officinalis extract reduced the relative expression of rhythm genes to 1.45 and the tyrosinase inhibition rate to 48.5%, indicating that Magnolia officinalis extract makes a dual contribution to rhythm repair and brightening. The above data fully demonstrate that each component in the basic formula is indispensable, working together to achieve a synergistic effect of microecological balance, rhythm repair, multi-pathway brightening, and anti-inflammatory soothing.

[0088] As can be seen from Examples 1-3 and Table 1, good results can be achieved within the range of values ​​of the formulation in this application, which fully demonstrates the scientific nature and feasibility of the formulation range.

[0089] Combining Examples 1 and 4-6 with Table 1, it can be seen that the addition of arnica flower extract, bisabolol, and dipotassium glycyrrhizate to the basic formula significantly improved anti-inflammatory and soothing indicators. In Example 4, compared to Example 1, the Δa* redness improvement increased from -1.52 to -2.18, the IL-6 inhibition rate increased from 54.2% to 68.5%, and the lactic acid stinging score reduction rate increased from 62% to 78%. In Example 6, when each soothing ingredient was further increased to its upper limit, the Δa* redness improvement reached -2.45, the IL-6 inhibition rate reached 75.6%, and the lactic acid stinging score reduction rate reached 85%, indicating that the three ingredients, along with the gentian extract and olive leaf extract in the basic components, significantly enhanced the immediate redness-reducing and soothing effect of the composition.

[0090] Combining Examples 1 and 7-9 with Table 1, it can be seen that the addition of ergothioneine and evening primrose extract significantly enhanced the rhythm repair and brightening effects. In Example 7, compared to Example 1, the relative expression of the rhythm gene increased from 2.15 to 2.58, the tyrosinase inhibition rate increased from 62.5% to 68.2%, and the melanin production inhibition rate increased from 58.3% to 64.5%. In Example 9, when both were increased to their upper limits, the relative expression of the rhythm gene reached 2.85, the tyrosinase inhibition rate reached 70.5%, and the melanin production inhibition rate reached 67.8%. This indicates that the mitochondrial-targeted antioxidant effect of ergothioneine and the anti-inflammatory regulation of evening primrose extract work synergistically to improve skin rhythm disorders and dullness through both cellular energy metabolism and inflammatory response pathways.

[0091] Combining Examples 1 and 10-12 with Table 1, it can be seen that the antioxidant and anti-inflammatory effects of olive leaf extract are significantly enhanced after metal ion chelation treatment. In Example 10, the IL-6 inhibition rate increased from 54.2% to 61.5%, the lactic acid stinging score reduction rate increased from 62% to 69%, and the tyrosinase inhibition rate increased from 62.5% to 68.5% compared to Example 1. In Example 12, using more stringent chelation conditions, the IL-6 inhibition rate reached 63.5%, and the tyrosinase inhibition rate reached 70.2%, confirming that metal ion chelation treatment can effectively remove metal complexes of active polyphenolic components, improving the stability and efficacy persistence of the extract.

[0092] Combining Examples 1 and 13-15 with Table 1, it can be seen that the microecological regulation and barrier repair effects of *Echinochloa gracilis* extract were significantly enhanced after antioxidant stabilization pretreatment. In Example 13, the bacterial community ratio increased from 1.68 to 1.80 compared to Example 1, and the relative expression of rhythm genes increased from 2.15 to 2.28. In Example 15, when treated with a high concentration of antioxidants, the bacterial community ratio reached 1.85, confirming that the dual antioxidant mechanism of inert gas protection combined with natural tocopherol and ascorbate tetraisopalmitate effectively prevented the oxidative deterioration of unsaturated fatty acids and ensured the bioactivity of the extract.

[0093] Combining Examples 1 and 16-18 with Table 1, it can be seen that the prebiotic activity of inulin is significantly enhanced after enzymatic hydrolysis and modification into fructooligosaccharides. In Example 16, the bacterial community ratio increased significantly from 1.68 to 2.35 compared to Example 1; in Example 18, when the degree of enzymatic hydrolysis was increased to the upper limit, the bacterial community ratio reached 2.58, an increase of approximately 53% compared to ordinary inulin. This confirms that enzymatic hydrolysis significantly enhances the selective proliferation of beneficial bacteria after inulin is degraded into fructooligosaccharides with a degree of polymerization of 2-10.

[0094] Combining Examples 1 and 19-21 with Table 1, it can be seen that the rhythm repair and brightening effects of Magnolia officinalis extract were significantly enhanced after being prepared using a low-temperature dynamic countercurrent extraction process. In Example 19, compared to Example 1, the relative expression of the rhythm gene increased from 2.15 to 2.35, the tyrosinase inhibition rate increased from 62.5% to 70.2%, and the melanin production inhibition rate increased from 58.3% to 65.5%. In Example 21, using optimized extraction conditions, the relative expression of the rhythm gene reached 2.42, the tyrosinase inhibition rate reached 72.5%, and the melanin production inhibition rate reached 68.0%, confirming that low-temperature dynamic countercurrent extraction effectively protects the heat-sensitive active ingredients in Magnolia officinalis, allowing the brightening and rhythm repair effects to be fully realized.

[0095] 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 for restoring and brightening the microecological balance and circadian rhythm, characterized in that, Composition includes the following components by mass percentage: Licorice root extract 0.001%-1%, olive leaf extract 0.1%-5%, seaweed extract 0.1%-2%, gentian root extract 0.5%-3%, inulin 0.1%-5%, tripeptide-32 0.001%-0.01%, magnolia flower extract 0.5%-2%, balance being water.

2. The composition for microecological balance and circadian rhythm repair and brightening according to claim 1, characterized in that, Based on 100% quality, it also includes: Arnica flower extract 0.1%-2%, bisabolol 0.1%-1%, dipotassium glycyrrhizate 0.05%-0.5%.

3. The composition for microecological balance and circadian rhythm repair and brightening according to claim 1, characterized in that, The olive leaf extract is subjected to metal ion chelation treatment, and the metal ion chelation treatment method includes: Olive leaf extract was mixed with an aqueous solution of sodium phytate or disodium ethylenediaminetetraacetate at a molar concentration of 0.01%-0.1% at a mass ratio of 1:8-15, stirred at 40-50℃ for 90-110 minutes, purified by membrane separation technology and then dried.

4. The composition for microecological balance and circadian rhythm repair and brightening according to claim 1, characterized in that, The extract of *Elongatus spp.* was subjected to the following antioxidant stabilization pretreatment: Under nitrogen or argon protection, the extract of *Elongata spp.* is mixed with 3%-8% of natural tocopherol and ascorbate tetraisopalmitate at a mass ratio of 1:1-3, and stirred at 20-30°C for 20-40 minutes.

5. The composition for microecological balance and circadian rhythm repair and brightening according to claim 1, characterized in that, The inulin is enzymatically modified to obtain fructooligosaccharides with a degree of polymerization of 2-10. The enzymatic modification method includes: Inulin was prepared into an aqueous solution with a mass fraction of 10%-15%, and inulinase was added at a mass fraction of 0.2%-0.4% of the inulin. The mixture was enzymatically hydrolyzed at 53-58℃ for 3-5 hours. After enzyme inactivation, the mixture was spray-dried to obtain modified inulin with an oligofructose content ≥85%.

6. The composition for microecological balance and circadian rhythm repair and brightening according to claim 1, characterized in that, The extract of Magnolia officinalis was prepared by a low-temperature dynamic countercurrent extraction process. The extraction solvent was a 60%-75% (v / v) butanediol aqueous solution. The extraction temperature was 42-48℃, the extraction time was 2.5-3.5 hours, and the material-to-liquid ratio was 1:(8-12).

7. The composition for microecological balance and circadian rhythm repair and brightening according to claim 1, characterized in that, The composition also contains 0.001%-0.1% ergothioneine and 0.01%-0.5% evening primrose extract by weight.

8. The use of the composition for microecological balance and circadian rhythm repair and brightening as described in any one of claims 1-7 in the preparation of cosmetics for improving skin redness, dullness, and circadian rhythm disorders.