Sensitive skin micro-ecological balance-oil control and acne removal composition and application thereof

Abstract

The invention relates to a sensitive skin micro-ecological balance-oil control and acne removal composition and application thereof. The composition is prepared from the following components: lactococcus lactis / phellinus pharmacologically fermented product filtrate, lactobacillus / black mustard seed fermented product extract, citrus fruit extract and trifolium pretense flower extract. The weight ratio of the lactococcus lactis / phellinus pharmacologically fermented product filtrate to the lactobacillus / black mustard seed fermented product extract to the citrus fruit extract to the trifolium pretense flower extract is (0.1-8): (0.001-0.1): (0.001-0.2): (0.01-3), and the composition constructs a four-stage synergistic mechanism of flora regulation, barrier repair, oil control and fat suppression, anti-inflammation and soothing; the limitation of a single target spot in the traditional technology is broken through, and an efficient, safe and multi-dimensional regulation and control comprehensive solution is provided for oily sensitive skin micro-ecological balance.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of cosmetics, in particular to a sensitive skin microecological balance-oil control and acne-removing composition, a preparation method and application thereof. BACKGROUND

[0002] With the acceleration of modern life pace and the aggravation of environmental pollution, the proportion of oily sensitive skin population has increased significantly. The ecological imbalance of facial flora (such as excessive proliferation of Malassezia and reduction of Staphylococcus epidermidis) of oily sensitive skin population leads to impaired skin barrier function, and further causes complex problems such as redness, itching, excessive oil secretion, frequent acne and blackheads.

[0003] Currently, the existing technologies in the industry have significant limitations in addressing the problems of redness, itching, excessive oil secretion and acne and blackheads caused by the ecological imbalance of facial flora of oily sensitive skin population. Chemical exfoliation (such as high-concentration salicylic acid) can achieve oil control by damaging the connection between keratinocytes, but it can exacerbate barrier damage and secondary flora imbalance. Physical adsorption (such as kaolin) can only temporarily relieve oiliness and cannot regulate sebaceous gland activity. Single flora regulation products (such as probiotic supplements) are difficult to restore flora diversity and have poor stability of active ingredients. Traditional plant extraction technology has low bioavailability and high irritation risk due to organic solvent residues and molecular weight limitations. SUMMARY

[0004] The present application aims to overcome the deficiencies of the prior art and provide a sensitive skin microecological balance-oil control and acne-removing composition and its application. The composition can improve skin problems such as redness, itching, excessive oil secretion, acne and blackheads caused by the ecological imbalance of facial flora of oily sensitive skin population, and enhance the barrier protection ability of the face.

[0005] To achieve the above-mentioned purpose, the technical solution adopted by the present application is as follows:

[0006] In a first aspect, the present application provides a sensitive skin microecological balance-oil control and acne-removing composition, characterized in that the composition comprises the following components: Lactococcus lactis / fermented product filtrate of Phellinus baumii, Lactobacillus / fermented product extract of black mustard seeds, citrus fruit extract and extract of Trifolium pratense flowers; the weight ratio of the Lactococcus lactis / fermented product filtrate of Phellinus baumii, Lactobacillus / fermented product extract of black mustard seeds, citrus fruit extract and extract of Trifolium pratense flowers is (0.1-8):(0.001-0.1):(0.001-0.2):(0.01-3).

[0007] The present application realizes the overall improvement of oily sensitive skin through four-dimensional regulation of "flora-lipid-barrier-inflammation", combined with process innovation to improve the efficacy of ingredients, and provides a full-link and high-precision solution to skin problems for oily sensitive skin. The functional realization and technical advantages are as follows:

[0008] The lactococcus lactis / medicinal fomes ferments filtrate prepared based on the bidirectional fermentation technology realizes precise regulation of flora through the dual effects of "selective inhibition + prebiotic promotion": on the one hand, bacteriocins (such as lactocin) and organic acids (such as lactic acid and acetic acid) produced by lactococcus lactis can specifically inhibit the adhesion and proliferation of pathogenic bacteria such as propionibacterium acnes, and reduce the lipase activity generated by the pathogenic bacteria; on the other hand, the polysaccharide of medicinal fomes as a prebiotic can be utilized by beneficial bacteria such as staphylococcus epidermidis and lactobacillus, so as to promote the secretion of exopolysaccharide and antibacterial peptide by the beneficial bacteria, form a biological membrane protection barrier, and enhance the colonization resistance of the beneficial bacteria.

[0009] The lactobacillus / black mustard seed fermentation product extract prepared by using the natural deep eutectic solvent (NADES, choline-lactic acid system) combined with ultrasonic-assisted extraction technology, glucosinolates in black mustard seeds are hydrolyzed into isothiocyanates by lactobacillus beta-glucosinolate enzyme, which can penetrate the membrane of sebaceous gland cells, reduce the secretion of sebum by inhibiting the activity of 5 alpha-reductase, and adjust the skin pH to weak acidity to inhibit propionibacterium acnes, thereby effectively reducing the generation of comedones and blackheads.

[0010] The hesperidin and naringin in the citrus fruit extract synergistically promote the reconstruction of the desmosome structure of the stratum corneum and the synthesis of hyaluronic acid, rapidly increase the water content and barrier function of the skin, and reduce the sensitivity to external stimulation; the biochanin A and formononetin in the red clover flower extract significantly inhibit the release of pro-inflammatory factors such as IL-6 and TNF-alpha by blocking the NF-kappa B and TLR4 / MyD88 inflammatory pathways, and rapidly relieve redness, itching and irritation.

[0011] The present application ensures the activity and mildness of the components through innovative preparation processes (bidirectional fermentation to improve the stability of antibacterial peptides, NADES+ultrasonic extraction to improve the extraction rate of isothiocyanates). The bidirectional fermentation process co-cultures lactococcus lactis and medicinal fomes to form a complex of antibacterial peptides and polysaccharides, which significantly improves the stability of the antibacterial peptides, and the polysaccharide as a natural dispersing agent avoids the agglomeration of active ingredients in the filtrate; the NADES+ultrasonic extraction technology utilizes the hydrogen bond effect of choline-lactic acid to destroy the cell wall of black mustard seeds, and ultrasonic assistance further breaks the cell structure, so that the extraction rate of isothiocyanates is significantly improved.

[0012] Based on the above principles, the present application provides a safe, efficient and long-acting microecological balance-oil control and comedone solution for oily sensitive skin.

[0013] Preferably, the weight ratio of the lactococcus lactis / medicinal fomes fermentation product filtrate, the lactobacillus / black mustard seed fermentation product extract, the citrus fruit extract and the red clover flower extract is (1-5):(0.01-0.08):(0.01-0.1):(0.1-1).

[0014] More preferably, the weight ratio of Lactococcus lactis / Polyporus medicata fermentation product filtrate, Lactobacillus / black mustard seed fermentation product extract, citrus fruit extract and red clover flower extract is (2-3):(0.04-0.05):(0.04-0.05):(0.4-0.5).

[0015] Specifically, the preparation method of the fermentation product filtrate of Lactococcus lactis / Polyporus medicata includes the following steps:

[0016] (1) Activate Lactococcus lactis, inoculate the activated Lactococcus lactis into seed culture medium, and carry out anaerobic culture to obtain Lactococcus lactis seed liquid; wherein, the temperature of anaerobic culture is 35-39℃ and the time is 24-75h; the inoculation amount of Lactococcus lactis is 3-5wt% of the total mass of seed culture medium;

[0017] (2) Weigh MRS medium, medicinal strobilus powder and water, mix them in a mass ratio of (3-6):(2-6):100, adjust the pH to 5-9, sterilize, and obtain the basic fermentation medium;

[0018] (3) Inoculate the Lactococcus lactis seed culture obtained in step (1) into the basic fermentation medium obtained in step (2) for anaerobic culture. After fermentation, sterilize, centrifuge, and take the supernatant to obtain the fermentation broth. The culture temperature is 30-40℃ and the time is 48-72h. The inoculation amount of Lactococcus lactis seed culture is 10-20% of the total mass of the basic fermentation medium, and the viable count of Lactococcus lactis seed culture is (1.0-5.0)×10⁻⁶. 9 CFU / mL;

[0019] (4) The sterilized fermentation broth is subjected to primary filtration, decolorization and fine filtration to obtain the fermentation product filtrate of Lactococcus lactis / Pterocarpus medicata.

[0020] Preferably, in step (1) of preparing the fermentation product filtrate of Lactococcus lactis / Polyporus medicata, the seed culture medium is prepared by mixing MRS culture medium with water at a mass ratio of 3-6:100, autoclaving at 121°C for 15 minutes, and cooling to room temperature for later use.

[0021] Preferably, in step (2) of preparing the fermentation product filtrate of Lactococcus lactis / Polyporus medicata, the pH of the fermentation medium is 7-7.5.

[0022] Preferably, in step (4) of preparing the fermentation product filtrate of *Lactococcus lactis* / *Polyporus medicata*, the initial filtration step is as follows: filtrate 1 is obtained by filtering with a filter cloth, and then filtrate 1 is filtered again using paperboard F100 with an average pore size of 20-30 μm to obtain filtrate 2; the decolorization step is as follows: activated carbon is added to filtrate 2 at a mass of 1-3 wt%, the temperature is raised to 55-65℃, and the decolorization treatment is carried out for 0.5-1.5 h; the fine filtration step is as follows: filtrate is filtered using paperboard F100 with an average pore size of 25 μm, and filtrate 3 is collected, which is the fermentation product filtrate of *Lactococcus lactis* / *Polyporus medicata*. After initial filtration, decolorization, and fine filtration, a clear and transparent fermentation product filtrate can be obtained.

[0023] Specifically, the preparation method of the Lactobacillus / black mustard seed fermentation product extract includes the following steps:

[0024] S1. Activate Lactobacillus and inoculate the activated Lactococcus lactis into seed culture medium (MRS medium) for anaerobic culture to obtain Lactobacillus seed liquid; then weigh MRS medium and black mustard seed powder and mix them at a material-to-liquid ratio of 1g:(8-10)mL, stir and sterilize to obtain basic fermentation medium; the anaerobic culture temperature is 35-39℃ and the time is 24-75h; the inoculation amount of Lactococcus lactis is 1-3wt% of the total mass of the seed culture medium;

[0025] S2. Inoculate the Lactobacillus seed culture into the basal fermentation medium for anaerobic fermentation. After fermentation, sterilize, centrifuge, and collect the supernatant to obtain the fermentation broth. The anaerobic culture temperature is 35-39℃, the time is 48-72h, and the pH is 5.5-6.5. The inoculation amount of Lactobacillus seed culture is 4-6% of the total mass of the basal fermentation medium, and the viable count of Lactobacillus seed culture is (1.0-5.0)×10⁻⁶. 9 CFU / mL; During fermentation, pH adjustment promotes the metabolism of lactic acid bacteria to produce lactic acid and antimicrobial peptides, while inducing black mustard seeds to release glucosinolates and their degradation products (isothiocyanates). After pre-cooling the fermentation broth to 4°C, centrifuge at 7000-9000 rpm for 5-15 min, and collect the supernatant to obtain the target extract.

[0026] S3. Choline chloride and lactic acid are mixed to obtain a choline chloride-lactic acid solvent, which is effective against polar components such as lactic acid and antimicrobial peptides; choline chloride, glycerol, and sorbitol are mixed to obtain a choline chloride-glycerol-sorbitol solvent, which is effective against non-polar components such as isothiocyanates; the choline chloride-lactic acid solvent and the choline chloride-glycerol-sorbitol solvent are mixed and rotary evaporated until the mixed solvent is clear and transparent, then cooled to obtain biphase NaDES; wherein, when preparing the choline chloride-lactic acid solvent, the molar ratio of choline chloride to lactic acid is 1:(1.5-2.5); when preparing the choline chloride-glycerol-sorbitol solvent, the molar ratio of choline chloride, glycerol, and sorbitol is 1:(0.8-1.2):(0.8-1.2); the volume ratio of the choline chloride-lactic acid solvent and the choline chloride-glycerol-sorbitol solvent is 1:(0.8-1.2);

[0027] S4. The fermentation broth obtained in step S2 is mixed with the biphasic NaDES obtained in step S3 and subjected to ultrasonic extraction. After ultrasonic extraction, the mixture is centrifuged, and the supernatant is collected. The supernatant is then decolorized and desalted, and freeze-dried under vacuum to obtain the Lactobacillus / black mustard seed fermentation product extract. The volume ratio of fermentation broth to biphasic NaDES is 1:(10-20). The ultrasonic extraction power is 300-500W, the frequency is 25-35kHz, the temperature is 40-50℃, and the time is 15-25min. During ultrasonic extraction, the mixture is intermittently stirred for 10s every 5min. The Lactobacillus / black mustard seed fermentation product extract obtained by the above preparation method contains lactic acid, antimicrobial peptides, and isothiocyanates.

[0028] Preferably, in step S1 of the preparation of the Lactobacillus / black mustard seed fermentation product extract, the activation treatment of Lactobacillus is as follows: frozen Lactobacillus is inoculated into 5 mL MRS medium and cultured at 37°C and 150 rpm for 12 hours until OD600 = 0.8-1.0; the activated bacterial solution is then transferred to 100 mL MRS medium (pH 6.2-6.4) at an inoculation rate of 2% (v / v) and cultured at 37°C and 180 rpm for 18 hours until OD600 ≥ 0.8-1.0.

[0029] 1.5, to obtain activated lactococci.

[0030] Preferably, in step S1 of preparing the Lactobacillus / black mustard seed fermentation product extract, the black mustard seed powder has a particle size of 0.1-0.5 mm.

[0031] Preferably, in step S2 of the preparation of the Lactobacillus / black mustard seed fermentation product extract, the centrifugation speed is 7000-9000 rpm and the time is 5-15 min.

[0032] Preferably, in step S3 of the preparation of the Lactobacillus / black mustard seed fermentation product extract, the rotary evaporation temperature is 75-85°C.

[0033] Preferably, in step S4 of the preparation of the Lactobacillus / black mustard seed fermentation product extract, the ultrasonic extraction process involves intermittent stirring for 10 seconds every 5 minutes; the centrifugation speed is 9000-10500 rpm and the time is 5-10 minutes; the biphasic NaDEs separated by centrifugation are recovered by rotary evaporation at a temperature of 45-55℃ and a pressure of 0.05-0.2 MPa; the decolorization step involves adding 1-3 wt% of activated carbon by weight of the supernatant and decolorizing at 55-65℃ for 20-40 minutes; the desalting step involves treating the decolorized supernatant with Dowex 50WX8 ion exchange resin at a flow rate of 0.8-1.5 BV / h; and the vacuum freeze-drying temperature is -70℃ to -85℃ and the pressure is 0.05-0.2 mbar.

[0034] Specifically, the preparation method of the citrus fruit extract includes the following steps:

[0035] (1) Select fresh citrus peel, remove the pulp, seeds and white inner pulp, leaving only the outer yellow peel, vacuum dry the outer yellow peel to retain the heat-sensitive components, and then pulverize and sieve to obtain peel powder; wherein the particle size of the peel powder is 0.125-0.250 mm.

[0036] (2) Mix the fruit peel powder with a mixed solvent, perform ultrasonic extraction, centrifuge, and collect the supernatant; wherein, the mixed solvent includes anhydrous ethanol, deionized water and Tween-80 in a volume ratio of (2-4):(6-8):(0.05-0.1), and the material-to-liquid ratio of fruit peel powder to mixed solvent is 1g:15-25mL; the ultrasonic extraction power is 200-400W, the frequency is 25-35kHz, the temperature is 50±2℃, and the time is 40-50min; the function of Tween-80 is to improve the solubility of insoluble components;

[0037] (4) The supernatant was purified, concentrated and freeze-dried to obtain the citrus fruit extract.

[0038] Preferably, in step (1) of preparing the citrus fruit extract, the vacuum drying temperature is (-70)-(-85)℃, the pressure is ≤50Pa, and the time is 18-28h.

[0039] Preferably, in step (4) of the preparation of the citrus fruit extract, the purification step is as follows: the supernatant is filtered using qualitative filter paper with a pore size of 10-15 μm to remove large particulate residues, and the initial filtrate is obtained. The initial filtrate is then filtered through a 0.45 μm microporous membrane (nylon or polytetrafluoroethylene material) to further remove suspended solids, and a purified solution is obtained. The concentration step is as follows: the purified solution is concentrated using a rotary evaporator at a water bath temperature of 45-55℃, a vacuum degree of ≤0.09MPa, and a rotation speed of 70. Concentrate to 1 / 10-1 / 5 of the original volume at -120 rpm to obtain a concentrated solution; the freeze-drying steps are as follows: pre-freezing: freeze at -40℃ for 4 hours to completely solidify the sample; sublimation drying: at a vacuum degree ≤20Pa, sublimation temperature -20℃, time for 12-16h; desorption drying: desorption temperature 25℃, drying time 24-36h, to obtain a loose porous powder, which is the citrus fruit extract. Freeze-drying can retain the activity of heat-sensitive components in the citrus fruit extract.

[0040] Specifically, the preparation method of the red clover flower extract includes the following steps:

[0041] S1. Take dried red clover flowers, remove impurities, and pulverize them through a 40-60 mesh sieve to obtain red clover flower powder, in order to increase the solvent contact area and improve the extraction efficiency.

[0042] S2. Mix red clover flower powder with an ethanol aqueous solution, heat and reflux to extract, so that the intracellular components are fully dissolved, centrifuge, and collect the supernatant to obtain the extract; wherein the extraction temperature is 60-80℃ and the time is 2-5h; wherein the material-to-liquid ratio of red clover flower powder to ethanol aqueous solution is 1g:(8-13)mL, and the mass concentration of ethanol aqueous solution is 70-80wt%;

[0043] S3. The extract obtained in step S2 is concentrated, purified, dried and pulverized to obtain the red clover flower extract.

[0044] Preferably, in the preparation S3 of the red clover flower extract, the concentration step is as follows: the extract is concentrated to 1 / 10-1 / 5 of its original volume using a rotary evaporator under conditions of water bath temperature of 45-55℃, vacuum degree of ≤0.09MPa, and rotation speed of 70-120rpm, to obtain a concentrated solution; the purification step is as follows: the concentrated solution is diluted to a total solids content of 5%-10%, and then passed through AB-8 resin at a flow rate of 1-2 BV / h (bed volume / hour). First, it is washed with water at a flow rate of 2-3 BV to remove water-soluble impurities such as sugars, and then eluted with 50%-70% ethanol at a flow rate of 1-1.5 BV / h, and the extract is collected. The eluent (isoflavones enrichment section), i.e., the purified solution; drying and pulverizing steps: the purified solution is spray-dried, specifically: the feed temperature is set to 50-60℃, the inlet air temperature is controlled at 160-180℃, the outlet temperature is maintained at 80-90℃, and the atomization pressure is adjusted to 0.2-0.4MPa, while maintaining the feed rate at 10-20mL / min (in actual operation, it can be finely adjusted according to the equipment performance to ensure uniform atomization effect). After spray drying, the initial powder is obtained, which is further pulverized and sieved through a 50-70 mesh sieve to obtain an extract powder with uniform particle size distribution, which is the red clover flower extract.

[0045] In a second aspect, the present invention provides the application of the sensitive skin microecological balancing-oil-controlling and acne-removing composition of the first aspect in the preparation of cosmetics, wherein the cosmetics include at least one of toner, lotion, cream, mask, serum, gel and spray, and the sensitive skin microecological balancing-oil-controlling and acne-removing composition of the first aspect accounts for 1-15 wt% of the total mass of the cosmetics.

[0046] Thirdly, the present invention provides a gel comprising the following ingredients by weight percentage: 1%-15% of the sensitive skin microecological balance-oil control and acne-removing composition of the first aspect, 0.1%-2% thickener, 1%-15% moisturizer, 0.5%-3% preservative and 0.01%-0.3% pH adjuster, with the balance being deionized water.

[0047] Preferably, the thickener includes at least one of xanthan gum, acrylic (ester) copolymer, ammonium acryloyl dimethyl taurate / VP copolymer, hydroxyethyl cellulose, carboxymethyl cellulose, and sodium alginate.

[0048] Preferably, the moisturizer includes at least one of allantoin, sodium polyacrylate, panthenol, β-glucan, trehalose, caprylyl glycol, dipropylene glycol, sodium hyaluronate, 1,2-butanediol, glycerin, and tremella polysaccharide.

[0049] Preferably, the pH adjuster includes at least one of arginine, citric acid, NaOH, and disodium EDTA.

[0050] Preferably, the preservative includes at least one selected from 1,2-hexanediol, 1,2-pentanediol, ethylhexylglycerin, p-hydroxyacetophenone, phenoxyethanol, octanoyl hydroxamic acid, and sodium benzoate.

[0051] Fourthly, the present invention provides a method for preparing the gel described in the third aspect, comprising the following steps:

[0052] S1. Mix the humectant, thickener and deionized water, and homogenize at 75-85℃ to obtain a mixture;

[0053] S3. After the mixture cools down to 55-65℃, add the preservative and stir evenly. After the temperature drops to 35-45℃, add each component of the sensitive skin microecological balance-oil control and acne removal composition and stir evenly. Finally, add the pH adjuster to adjust the pH to obtain the gel.

[0054] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0055] Inspired by breakthroughs in microbiome research on the chain mechanism of "excessive proliferation of pathogenic bacteria - reduction of beneficial bacteria - impaired barrier function" in oily and sensitive skin, this invention combines the high selectivity of natural deep eutectic solvent (NADES) extraction technology with the synergistic effects of two-way fermentation technology. It utilizes the fermentation product filtrate of *Lactococcus lactis* / *Polyporus medicata* to regulate the balance of the microbial community, the fermentation extract of *Lactobacillus* / black mustard seed to inhibit 5α-reductase activity and reduce sebum secretion, the citrus fruit extract to strengthen the tight junctions of the stratum corneum and repair the barrier, and the red clover flower extract to block the release of inflammatory factors and alleviate sensitivity symptoms. This constructs a four-fold synergistic mechanism of "microbial regulation - barrier repair - oil control and sebum suppression - anti-inflammatory and soothing," overcoming the limitations of traditional single-target technologies and providing a comprehensive solution for efficient, safe, and multi-dimensional regulation of the microecological balance of oily and sensitive skin. Attached Figure Description Figure 1 This is a comparison image of the gel used in Example 1 before and after application. Detailed Implementation

[0056] To better illustrate the purpose, technical solution, and advantages of the present invention, the present invention will be further described below in conjunction with specific embodiments.

[0057] The sources of the raw materials used in the following examples and comparative examples are as follows:

[0058] Lactococcus lactis: Manufacturer: Guangzhou Zuoke Biotechnology Development Co., Ltd., Model: BNCC363081;

[0059] Medicinal Fomitopsis: Manufacturer is Bozhou Huozhengtang Pharmaceutical Co., Ltd., product name is Fomitopsis fissori.

[0060] Lactobacillus: Manufacturer: Jiangsu Xin Shen Ao Biotechnology Co., Ltd., Model: CCTCC M2013511;

[0061] Black mustard seeds: manufactured by Shandong Xianglongmu Seasoning Food Co., Ltd., commercially available regular black mustard seeds;

[0062] Citrus fruit: Produced by Hunan Xiangjia Juyou Agricultural Co., Ltd., commercially available regular rock sugar oranges;

[0063] Red clover flowers: The manufacturer is Shuyang Tilan Nursery Professional Cooperative, and the flowers are regular red clover flowers sold in the market;

[0064] Lactobacillus / black mustard seed fermentation product extract: Manufacturer: Beijing Dongfang Miaosen Biotechnology Co., Ltd., Model No.: 2024121292;

[0065] Unless otherwise specified, all other materials and reagents used in the examples are commercially available.

[0066] The preparation methods of each component of the sensitive skin microecological balancing-oil control and acne-removing compositions of Examples 1-7 and Comparative Examples 1-4 are as follows:

[0067] The method for preparing the fermentation product filtrate of *Lactococcus lactis* / *Polyporus medicata* includes the following steps:

[0068] (1) Lactococcus lactis is activated, and the activated Lactococcus lactis is inoculated into a seed culture medium and cultured anaerobically to obtain Lactococcus lactis seed liquid; wherein, the anaerobic culture temperature is 37℃ and the time is 36h; the inoculation amount of Lactococcus lactis is 4wt% of the total mass of the seed culture medium; the seed culture medium is prepared by mixing MRS medium and water at a mass ratio of 5:100, autoclaving at 121℃ for 15 minutes, and cooling to room temperature for later use;

[0069] (2) Weigh MRS medium, medicinal strobilus powder and water, mix them in a mass ratio of 5:4:100, adjust the pH to 7, sterilize, and obtain the basic fermentation medium.

[0070] (3) The *Lactococcus lactis* seed culture obtained in step (1) was inoculated into the basic fermentation medium obtained in step (2) for anaerobic culture. After fermentation, the culture was sterilized, centrifuged, and the supernatant was collected to obtain the fermentation broth. The culture temperature was 37℃ and the time was 54h. The inoculation amount of *Lactococcus lactis* seed culture was 15% of the total mass of the basic fermentation medium, and the viable count of *Lactococcus lactis* seed culture was 3×10⁻⁶. 9 CFU / mL;

[0071] (4) The sterilized fermentation broth is subjected to primary filtration, decolorization and fine filtration to obtain the fermentation product filtrate of Lactococcus lactis / Polyporus medicata; wherein, the primary filtration step is: filter with filter cloth to obtain filtrate 1, and then filter filtrate 1 again with paperboard F100 with an average pore size of 25μm to obtain filtrate 2; the decolorization step is: add activated carbon with a mass of 2wt% of the mass of filtrate 2 to filtrate 2, heat to 60℃ and decolorize for 1h; the fine filtration step is: filter with paperboard F100 with an average pore size of 25μm, and collect clear and transparent filtrate 3, which is the fermentation product filtrate of Lactococcus lactis / Polyporus medicata.

[0072] The preparation method of the Lactobacillus / black mustard seed fermentation product extract includes the following steps:

[0073] S1. Activate the lactobacillus and inoculate the activated lactococcus into seed culture medium (MRS medium) for anaerobic culture to obtain lactobacillus seed solution; then weigh MRS medium and black mustard seed powder, mix them at a material-to-liquid ratio of 1g:9mL, stir, and sterilize to obtain the basic fermentation medium; the anaerobic culture temperature is 37℃ and the time is 36h; the activation treatment of lactobacillus is as follows: inoculate the frozen inoculum into 5mL of MRS medium (containing 0.5% glucose and 0.2% Tween 80), and culture it at 37℃ and 150rpm for 12h until OD600=1.0 to obtain activated bacterial solution, and then transfer the activated bacterial solution to 100mL of MRS medium (pH 1.5%) at an inoculation rate of 2% (v / v). 6.2) The Lactobacillus seed culture was obtained by culturing at 37℃ and 180rpm for 18 hours until OD600≥1.5; the particle size of the black mustard seed powder was 0.3mm; the sterilization treatment was: sterilization at 115℃ for 20min;

[0074] S2. Inoculate the Lactobacillus seed culture into the basal fermentation medium for anaerobic fermentation. After fermentation, sterilize, pre-cool to 4°C, centrifuge, and collect the supernatant to obtain the fermentation broth. The anaerobic culture temperature was 37°C, the rotation speed was 100 rpm, and the time was 54 h. During the anaerobic culture, the pH was maintained at 6.0 by supplementing with 2 mol / L NaOH. The inoculation amount of Lactobacillus seed culture was 5% of the total mass of the basal fermentation medium, and the viable count of Lactobacillus seed culture was 4 × 10⁻⁶. 9 CFU / mL; centrifugation speed was 8000 rpm and time was 10 min;

[0075] S3. Choline chloride and lactic acid are mixed to obtain a choline chloride-lactic acid solvent; choline chloride, glycerol, and sorbitol are mixed to obtain a choline chloride-glycerol-sorbitol solvent; the choline chloride-lactic acid solvent and the choline chloride-glycerol-sorbitol solvent are mixed and rotary evaporated until the mixed solvent is clear and transparent, then cooled to obtain biphase NaDES; wherein, when preparing the choline chloride-lactic acid solvent, the molar ratio of choline chloride to lactic acid is 1:2; when preparing the choline chloride-glycerol-sorbitol solvent, the molar ratio of choline chloride, glycerol, and sorbitol is 1:1:1; the volume ratio of the choline chloride-lactic acid solvent and the choline chloride-glycerol-sorbitol solvent is 1:1; the rotary evaporation temperature is 80℃;

[0076] S4. The fermentation broth obtained in step S2 is mixed with the biphasic NaDES obtained in step S3, and then subjected to ultrasonic extraction. After centrifugation, the supernatant is collected, and the supernatant is decolorized and desalted. The supernatant is then freeze-dried under vacuum to obtain the Lactobacillus / black mustard seed fermentation product extract. The volume ratio of fermentation broth to biphasic NaDES is 1:15. The ultrasonic extraction power is 400W, the frequency is 30kHz, the temperature is 45℃, and the time is 20min. During ultrasonic extraction, the mixture is stirred intermittently for 10s every 5min. The centrifugation speed is 10000rpm, and the time is 8min. The biphasic NaDES separated by centrifugation is recovered by rotary evaporation at a temperature of 50℃ and a pressure of 0.1MPa. The decolorization step involves adding 2wt% activated carbon (by weight of the supernatant) and decolorizing at 60℃ for 30min. The desalting step involves using Dowex... The decolorized supernatant was treated with 50WX8 ion exchange resin at a flow rate of 1 BV / h; the vacuum freeze-drying temperature was -80℃ and the pressure was 0.1 mbar.

[0077] The method for preparing the citrus fruit extract includes the following steps:

[0078] (1) Select fresh citrus peel, remove the pulp, seeds and white inner pulp, leaving only the outer yellow peel, vacuum dry the outer yellow peel, then pulverize and sieve to obtain peel powder; wherein, the particle size of the peel powder is 0.125-0.250mm; the vacuum drying temperature is -80℃, the pressure is ≤50Pa, and the time is 24h;

[0079] (2) Mix the fruit peel powder with a mixed solvent, perform ultrasonic extraction, centrifuge, and collect the supernatant; wherein, the mixed solvent includes anhydrous ethanol, deionized water and Tween-80 in a volume ratio of 3:7:0.06, and the material-to-liquid ratio of fruit peel powder to mixed solvent is 1g:20mL; the ultrasonic extraction power is 300W, the frequency is 30kHz, the temperature is 50℃, and the time is 45min;

[0080] (4) The supernatant was purified, concentrated, and freeze-dried to obtain a loose, porous powder of citrus fruit extract. The purification steps were as follows: the supernatant was filtered using qualitative filter paper with a pore size of 15 μm to obtain the initial filtrate, which was then filtered through a 0.45 μm polytetrafluoroethylene microporous membrane to obtain the purified solution. The concentration steps were as follows: the purified solution was concentrated to 1 / 3 of its original volume using a rotary evaporator at a water bath temperature of 50℃, a vacuum degree of ≤0.09MPa, and a rotation speed of 100rpm to obtain the concentrated solution. The freeze-drying steps were as follows: pre-freezing: freezing at -40℃ for 4 hours to completely solidify the sample; sublimation drying: at a vacuum degree of ≤20Pa, a sublimation temperature of -20℃, and a time of 14h; desorption drying: at a desorption temperature of 25℃ and a drying time of 28h.

[0081] The preparation method of the red clover flower extract includes the following steps:

[0082] S1. Take dried red clover flowers, remove impurities, pulverize them, and pass them through a 50-mesh sieve to obtain red clover flower powder.

[0083] S2. Mix red clover flower powder with an ethanol aqueous solution, heat and reflux to extract, centrifuge, collect the supernatant to obtain the extract; wherein the extraction temperature is 75℃ and the time is 3h; wherein the material-to-liquid ratio of red clover flower powder to ethanol aqueous solution is 1g:10mL, and the mass concentration of ethanol aqueous solution is 75wt%.

[0084] S3. The extract obtained in step S2 is concentrated, purified, dried, and pulverized to obtain the red clover flower extract. The concentration step involves using a rotary evaporator at a water bath temperature of 50℃, a vacuum degree of ≤0.09MPa, and a rotation speed of 100rpm to concentrate the extract to 1 / 3 of its original volume, obtaining a concentrated solution. The purification step involves diluting the concentrated solution with water to a total solids content of 8%, then passing it through AB-8 resin at a flow rate of 1.5BV / h. First, it is rinsed with water at a flow rate of 2.5BV to remove water-soluble impurities such as sugars, and then... Eluent with 60% ethanol was used to elute at a flow rate of 1.3 BV / h, and the eluent (isoflavones enrichment section) was collected, which is the purified solution. Drying and pulverizing steps: The purified solution was spray-dried, specifically by setting the feed temperature to 55℃, controlling the inlet air temperature at 170℃, maintaining the outlet temperature at 85℃, adjusting the atomization pressure to 0.3 MPa, and maintaining the feed rate at 15 mL / min. After spray drying, a primary powder was obtained, which was further pulverized and sieved through a 60-mesh sieve to obtain an extract powder with a uniform particle size distribution, which is the red clover flower extract.

[0085] Example 1

[0086] A sensitive skin microecological balancing, oil-controlling, and acne-reducing composition comprises the following components: Lactococcus lactis / Polyporus medicata fermentation product filtrate, Lactobacillus / black mustard seed fermentation product extract, citrus fruit extract, and red clover flower extract; the weight ratio of the Lactococcus lactis / Polyporus medicata fermentation product filtrate, Lactobacillus / black mustard seed fermentation product extract, citrus fruit extract, and red clover flower extract is 2.6:0.044:0.048:0.43, and the total weight of the composition is 100 parts.

[0087] Example 2

[0088] A sensitive skin microecological balancing, oil-controlling, and acne-reducing composition comprises the following components: Lactococcus lactis / Polyporus medicata fermentation product filtrate, Lactobacillus / black mustard seed fermentation product extract, citrus fruit extract, and red clover flower extract; the weight ratio of the Lactococcus lactis / Polyporus medicata fermentation product filtrate, Lactobacillus / black mustard seed fermentation product extract, citrus fruit extract, and red clover flower extract is 2:0.04:0.04:0.4, and the total weight of the composition is 100 parts.

[0089] Example 3

[0090] A sensitive skin microecological balancing, oil-controlling, and acne-reducing composition comprises the following components: Lactococcus lactis / Polyporus medicata fermentation product filtrate, Lactobacillus / black mustard seed fermentation product extract, citrus fruit extract, and red clover flower extract; the weight ratio of the Lactococcus lactis / Polyporus medicata fermentation product filtrate, Lactobacillus / black mustard seed fermentation product extract, citrus fruit extract, and red clover flower extract is 3:0.05:0.05:0.5, and the total weight of the composition is 100 parts.

[0091] Example 4

[0092] A sensitive skin microecological balancing, oil-controlling, and acne-reducing composition comprises the following components: Lactococcus lactis / Polyporus medicata fermentation product filtrate, Lactobacillus / black mustard seed fermentation product extract, citrus fruit extract, and red clover flower extract; the weight ratio of the Lactococcus lactis / Polyporus medicata fermentation product filtrate, Lactobacillus / black mustard seed fermentation product extract, citrus fruit extract, and red clover flower extract is 1:0.01:0.01:0.1, and the total weight of the composition is 100 parts.

[0093] Example 5

[0094] A sensitive skin microecological balancing, oil-controlling and acne-removing composition comprises the following components: Lactococcus lactis / Polyporus medicata fermentation product filtrate, Lactobacillus / black mustard seed fermentation product extract, citrus fruit extract, and red clover flower extract; the weight ratio of the Lactococcus lactis / Polyporus medicata fermentation product filtrate, Lactobacillus / black mustard seed fermentation product extract, citrus fruit extract, and red clover flower extract is 5:0.08:0.1:1, and the total weight of the composition is 100 parts.

[0095] Example 6

[0096] A sensitive skin microecological balancing, oil-controlling, and acne-reducing composition comprises the following components: Lactococcus lactis / Polyporus medicata fermentation product filtrate, Lactobacillus / black mustard seed fermentation product extract, citrus fruit extract, and red clover flower extract; the weight ratio of the Lactococcus lactis / Polyporus medicata fermentation product filtrate, Lactobacillus / black mustard seed fermentation product extract, citrus fruit extract, and red clover flower extract is 0.1:0.001:0.001:0.01, and the total weight of the composition is 100 parts.

[0097] Example 7

[0098] A sensitive skin microecological balancing, oil-controlling and acne-removing composition comprises the following components: Lactococcus lactis / Polyporus medicata fermentation product filtrate, Lactobacillus / black mustard seed fermentation product extract, citrus fruit extract, and red clover flower extract; the weight ratio of the Lactococcus lactis / Polyporus medicata fermentation product filtrate, Lactobacillus / black mustard seed fermentation product extract, citrus fruit extract, and red clover flower extract is 8:0.1:0.2:3, and the total weight of the composition is 100 parts.

[0099] Comparative Example 1

[0100] The only difference between Comparative Example 1 and Example 1 is that the total weight of the composition remains unchanged, the fermentation product filtrate of Lactococcus lactis / Polyporus medicata is not added, and the missing amount is made up by Lactobacillus / black mustard seed fermentation product extract, citrus fruit extract and red clover flower extract in a weight ratio of 0.044:0.048:0.43.

[0101] Comparative Example 2

[0102] The only difference between Comparative Example 2 and Example 1 is that the total weight of the composition remains the same, no Lactobacillus / black mustard seed fermentation product extract is added, and the missing amount is made up by Lactococcus lactis / Polyporus medicata fermentation product filtrate, citrus fruit extract and red clover flower extract in a weight ratio of 2.6:0.048:0.43.

[0103] Comparative Example 3

[0104] The only difference between Comparative Example 3 and Example 1 is that the total weight of the composition remains unchanged, no citrus fruit extract is added, and the missing amount is made up by using Lactococcus lactis / Polyporus medicata fermentation product filtrate, Lactobacillus / black mustard seed fermentation product extract and red clover flower extract in a weight ratio of 2.6:0.044:0.43.

[0105] Comparative Example 4

[0106] The only difference between Comparative Example 4 and Example 1 is that the total weight of the composition remains the same, red clover flower extract is not added, and the missing amount is made up by Lactococcus lactis / Polyporus medicata fermentation product filtrate, Lactobacillus / black mustard seed fermentation product extract and citrus fruit extract in a weight ratio of 2.6:0.044:0.048.

[0107] Comparative Example 5

[0108] The only difference between Comparative Example 5 and Example 1 is that Comparative Example 5 contains commercially available Lactobacillus / black mustard seed fermentation product extract.

[0109] Test Example 1: Inhibition rate test of Propionibacterium acnes in each group of compositions

[0110] Experimental Methods: Liquid thioglycolate medium (FT medium, Huankai Microbiology, REF: 028033) was prepared, and 0.2 wt% Propionibacterium acnes (Guangdong Provincial Institute of Microbiology) was inoculated into the medium and cultured at 37℃ for 48 h to form a bacterial suspension. The test samples (compositions of Examples 1-7 and Comparative Examples 1-5) were mixed with DMSO to prepare a sample solution with a volume percentage of 0.01%. Then, 180 μL of bacterial suspension was added to a 96-well plate, followed by 40 μL of each sample solution to form a sample group. A control group (bacterial suspension without sample solution) was also set up. After culturing for 24 h, the OD value of each well was measured using a microplate reader. The lower the OD value, the clearer the liquid, the worse the bacterial activity, and the better the antibacterial effect. The antibacterial rate was calculated according to the following formula: Antibacterial rate (%) = (OD value of control group - OD value of sample group) / OD value of control group × 100%; specific data are shown in Table 1.

[0111] Table 1. Inhibition rate of each group of samples against Propionibacterium acnes

[0112]

[0113]

[0114] Test Example 2: Test of 5-α reductase inhibition rate of each composition

[0115] The compositions of Examples 1-7 and the compositions of Comparative Examples 1-5 were tested according to the following steps, and the results are shown in Table 2.

[0116] Experimental methods: Each composition was prepared into a 10% (v / v) sample solution using DMSO. (1) Sample tube: Add 1 mL of each example and comparative sample solution, 1 mL of enzyme solution, 1 mL of NADPH solution and 1 mL of testosterone solution to a test tube, shake gently to obtain a mixed liquid, and use a pipette to take 200 μL of the mixed liquid into a 96-well microplate. Each sample was tested in triplicate and placed in a microplate reader for detection. The absorbance was measured at 340 nm, which is the first measurement value A. 样品0 After incubating the sample at 37°C for 20 minutes, it was placed in an ELISA reader for detection. The absorbance was measured at 340 nm, which is the second measurement value A. 样品1 The enzyme solution is a 5-α reductase diluted to 50U with PBS buffer at pH 6.8; the NADPH solution is a 2mM solution of NADPH prepared with PBS at pH 6.8; and the testosterone solution is a 0.83nM solution of testosterone prepared with 75% ethanol.

[0117] (2) Enzyme tubes: Add 1 mL of PBS solution, 1 mL of enzyme solution, 1 mL of NADPH solution, and 1 mL of testosterone solution to a test tube, gently shake to obtain a mixed liquid, and use a pipette to transfer 200 μL of the mixed liquid into a 96-well microplate. Perform three replicates for each sample. Place the plate in a microplate reader and measure the absorbance at 340 nm. This value is the first measurement A. 酶0 After incubating the sample at 37°C for 20 minutes, it was placed in an ELISA reader for detection. The absorbance was measured at 340 nm, which is the second measurement value A. 酶1 Simultaneously, blank and positive control tests were performed. The blank control group consisted of replacing the sample solution in the sample tube with an equal amount of deionized water, while the positive control group consisted of 1 mL of dutasteride solution with a concentration of 25 g / L and DMSO as the solvent, following the same steps as above.

[0118] 5-α reductase inhibition rate (%) = [1-(A 样品0 -A 样品1 ) / (A 酶0 -A 酶1 The higher the 5-α reductase inhibition rate, the stronger the ability of the composition to block the conversion of testosterone to dihydrotestosterone, and the less oil secreted by the sebaceous glands.

[0119] Table 2. 5-α reductase inhibition rate of each group of samples

[0120] Group / Performance 5-alpha reductase inhibition rate / % Example 1 97.5 Example 2 97.1 Example 3 96.8 Example 4 94.6 Example 5 94.0 Example 6 90.3 Example 7 88.3 Comparative Example 1 42.7 Comparative Example 2 50.1 Comparative Example 3 73.2 Comparative Example 4 76.7 Comparative Example 5 84.0 Positive Control Group 100 Blank Control Group 0.4

[0121] As shown in Table 4, and in conjunction with the data from Examples 1-5, when the weight ratio of the Lactococcus lactis / Polyporus medicata fermentation product filtrate, Lactobacillus / black mustard seed fermentation product extract, citrus fruit extract, and red clover flower extract is (2-3):(0.04-0.05):(0.04-0.05):(0.4-0.5), the composition exhibits higher inhibition rates of Propionibacterium acnes and 5-α reductase, indicating better acne-removing, blackhead-removing, and oil-controlling effects.

[0122] Based on the data from Example 1 and Comparative Examples 1-4, it can be seen that the absence of any one component in the composition reduces the composition's acne-removing, blackhead-removing, and oil-controlling properties. This may be because the bacteriocins (such as lactobacillus) and organic acids (such as lactic acid and acetic acid) produced by the fermentation product filtrate of Lactococcus lactis / Polyporus medicamentosa can specifically inhibit the adhesion and proliferation of pathogenic bacteria such as Propionibacterium acnes, and reduce the lipase activity they produce. The glucosinolates in the Lactobacillus / black mustard seed fermentation product extract are hydrolyzed by Lactobacillus β-thioglucosidase to generate isothiocyanates, which can penetrate the sebaceous gland cell membrane, reduce sebum secretion by inhibiting 5α-reductase activity, and regulate the skin pH to a slightly acidic state to inhibit Propionibacterium acnes, effectively reducing the formation of blackheads and whiteheads. The hesperidin and naringin in the citrus fruit extract synergistically promote the reconstruction of the desmosome structure of the stratum corneum and the synthesis of hyaluronic acid, rapidly increasing the skin's moisture content and barrier function. These four components synergistically enhance the composition's oil-controlling, antibacterial, and repairing effects.

[0123] Based on the data from Example 1 and Comparative Example 5, it can be seen that the Lactobacillus / black mustard seed fermentation product extract of Example 1, through an innovative preparation process, improves the isothiocyanate extraction rate by NADES+ ultrasonic extraction, ensuring the activity and mildness of the components, and demonstrating the oil-controlling, antibacterial and repairing effects of the composition of Example 1.

[0124] Application Example 1-7 and Comparative Application Example 1-5

[0125] The compositions of Examples 1-7 and Comparative Examples 1-5 were added to the gel at a concentration of 3.6 wt% to obtain the gels of Application Examples 1-7 and Comparative Application Examples 1-5. The formulations are shown in Table 3.

[0126] The preparation methods of the gels in Application Examples 1-7 and Comparative Application Examples 1-5 include the following steps:

[0127] S1. Mix the thickener and humectant evenly, add deionized water, heat at 75℃ water bath temperature for 10 minutes, then homogenize at 4000rpm for 5 minutes, keep warm for later use, and obtain the mixture.

[0128] S2. After the mixture cools down to 60°C, add the preservative and stir evenly. When the temperature drops to 40°C, add each component of the sensitive skin microecological balance-oil control and acne removal composition and stir evenly. Finally, add the pH adjuster to adjust the pH, stop stirring, and discharge the material to obtain the gel.

[0129] Table 3. Gel formulations for Application Examples 1-7 and Comparative Application Examples 1-5.

[0130]

[0131]

[0132] Comparative Application Example 6

[0133] Compared to Application Example 6, the gel did not contain the sensitive skin microecological balance-oil control and acne-removing composition, but used an equal amount of deionized water instead of the composition, and the preparation method was the same as Application Example 1.

[0134] Test Example 3: Human Patch Test with Gel

[0135] Thirty volunteers were recruited, 15 men and 15 women, aged 20-50 years. A closed patch test method was used. Equal amounts (0.5g-0.6g) of test samples (emulsions prepared in Application Examples 1-7 and Control Examples 1-6) were placed in a specific patch applicator. The patch was then applied to the volunteers' arms with hypoallergenic adhesive tape, with six test samples applied to one arm and seven to the other. The patches were gently pressed to ensure even application to the skin and left on for 24 hours. After 24 hours, the patch applicator was removed, and skin reactions were observed and recorded at 0.5h, 24h, and 48h. The severity of adverse skin reactions is shown in Table 4 below.

[0136] Table 4. Adverse reaction levels of the skin

[0137]

[0138] After testing, the gels provided in Application Examples 1-7 and Comparative Application Examples 1-6 all showed negative reactions after human patch testing, indicating that they are safe and non-irritating to human skin.

[0139] Test Example 4: Human Efficacy Test of Gel

[0140] (1) Oil control effect test:

[0141] This cosmetic oil-control efficacy test protocol employs a randomized, double-blind, controlled trial design, recruiting 65 volunteers aged 18-40 with oily / combination skin (selection criteria: oil content on forehead and nose wings ≥150μg / cm). 2The subjects were divided into 13 groups, using the gels from Application Examples 1-7 and Control Examples 1-6, with 5 people in each group. They used the gels twice daily, morning and evening, with each application being 1g. The testing period was 28 days, including assessments of immediate effects (0.5 hours after product application) and long-term effects (after 14 and 28 days of continuous use). Before each visit, subjects uniformly cleansed their faces with a gentle cleanser and sat quietly for 30 minutes to acclimatize to the environment (temperature 22±2℃, humidity 50±5%). Subsequently, they underwent... Measure the oil levels on the forehead and nose. Measure transepidermal water loss (TEWL).

[0142] (2) Acne-removing effect test

[0143] This cosmetic efficacy testing protocol for treating acne employs a randomized, double-blind, parallel-controlled trial design. Sixty-five volunteers aged 18-35 with mild to moderate comedonal acne (selection criteria: ≥30 facial comedones without severe inflammatory papules / pustules, confirmed by a dermatologist) were recruited and divided into 13 groups. Five participants in each group used the gel from cases 1-7 (application examples) and 1-6 (control examples). The gel was applied three times daily, with each application consisting of 1g. The sample was applied to the entire face. The testing period was 28 days, including single-application tolerability observation (0.5 hours after first application) and long-term efficacy evaluation (after 14 and 28 days of continuous use). Before each visit, participants cleansed their faces with a designated gentle cleanser and sat quietly for 30 minutes to acclimatize (temperature 22±2℃, humidity 50±5%). Subsequently, a dermatologist counted the number of facial comedones (total of open and closed comedones) using a standard dermoscope (×10x) and applied... Capture facial images and perform AI-assisted analysis (acne area ratio and density).

[0144] The calculation formulas for each evaluation parameter are as follows:

[0145] Improvement rate (%) of each indicator = (mean before use - mean after use) / mean before use × 100%; data are shown in Table 5.

[0146] Table 5. Results of human efficacy tests for each group of gels.

[0147]

[0148]

[0149] As shown in Table 5, compared with the control group gel (the blank control group without the added composition), the oil control, acne removal and repair effects of each group of gels were improved.

[0150] Based on the data from Application Examples 1-7, it can be seen that when the weight ratio of the Lactococcus lactis / Polyporus medicata fermentation product filtrate, Lactobacillus / black mustard seed fermentation product extract, citrus fruit extract, and red clover flower extract is (2-3):(0.04-0.05):(0.04-0.05):(0.4-0.5), the gel has better effects in removing acne, controlling oil, and repairing skin.

[0151] Combining the data from Application Example 1 and Comparative Application Examples 1-4, it can be seen that the absence of any one component in the gel composition will reduce the oil-controlling performance of the composition. This may be because the present invention achieves the effects of microbial regulation, barrier repair, oil control and sebum inhibition, and anti-inflammatory and soothing through the synergistic effect of Lactococcus lactis / Pothomia medicinalis fermentation product filtrate, Lactobacillus / black mustard seed fermentation product extract, citrus fruit extract, and red clover flower extract, providing a safe, efficient, and long-lasting microecological balance-oil control and acne removal solution for oily and sensitive skin.

[0152] Based on the data from Application Example 1 and Comparative Application Example 5, it can be seen that the Lactobacillus / black mustard seed fermentation product extract of Application Example 1 was prepared by a natural deep eutectic solvent (NADES, choline-lactic acid system) combined with ultrasound-assisted extraction technology. Compared with commercially available Lactobacillus / black mustard seed fermentation product extract, it has better antibacterial and oil-controlling effects.

[0153] 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 sensitive skin microecological balancing, oil-controlling, and acne-removing composition, characterized in that, The composition comprises the following components: Lactococcus lactis / Fomitopsis medicata fermentation product filtrate, Lactobacillus / black mustard seed fermentation product extract, citrus fruit extract, and red clover flower extract; the weight ratio of Lactococcus lactis / Fomitopsis medicata fermentation product filtrate, Lactobacillus / black mustard seed fermentation product extract, citrus fruit extract, and red clover flower extract is (0.1-8):(0.001-0.1):(0.001-0.2):(0.01-3).

2. The sensitive skin microecological balancing-oil control and acne-removing composition as described in claim 1, characterized in that, The weight ratio of Lactococcus lactis / Polyporus medicata fermentation product filtrate, Lactobacillus / black mustard seed fermentation product extract, citrus fruit extract and red clover flower extract was (1-5):(0.01-0.08):(0.01-0.1):(0.1-1).

3. The sensitive skin microecological balancing-oil control and acne-removing composition as described in claim 1, characterized in that, The weight ratio of Lactococcus lactis / Polyporus medicata fermentation product filtrate, Lactobacillus / black mustard seed fermentation product extract, citrus fruit extract and red clover flower extract was (2-3):(0.04-0.05):(0.04-0.05):(0.4-0.5).

4. The sensitive skin microecological balancing-oil control and acne-removing composition as described in claim 1, characterized in that, The method for preparing the fermentation product filtrate of *Lactococcus lactis* / *Polyporus medicata* includes the following steps: (1) Activate Lactococcus lactis, inoculate the activated Lactococcus lactis into seed culture medium for anaerobic culture to obtain Lactococcus lactis seed liquid; wherein, the temperature of anaerobic culture is 35-39℃ and the time is 24-75h; the inoculation amount of Lactococcus lactis is 3-5wt% of the total mass of seed culture medium; (2) Weigh MRS medium, medicinal strobilus powder and water, mix them in a mass ratio of (3-6):(2-6):100, adjust the pH to 5-9, sterilize, and obtain the basic fermentation medium; (3) Inoculate the Lactococcus lactis seed culture obtained in step (1) into the basic fermentation medium obtained in step (2) for anaerobic culture. After fermentation, sterilize, centrifuge, and take the supernatant to obtain the fermentation broth. The anaerobic culture temperature is 30-40℃ and the time is 48-72h. The inoculation amount of Lactococcus lactis seed culture is 10-20% of the total mass of the basic fermentation medium, and the viable count of Lactococcus lactis seed culture is (1.0-5.0)×10⁻⁶. 9 CFU / mL; (4) The sterilized fermentation broth is subjected to primary filtration, decolorization and fine filtration to obtain the fermentation product filtrate of Lactococcus lactis / Pterocarpus medicata.

5. The sensitive skin microecological balancing-oil control and acne-removing composition as described in claim 1, characterized in that, The preparation method of the Lactobacillus / black mustard seed fermentation product extract includes the following steps: S1. Activate Lactobacillus, inoculate the activated Lactococcus lactis into seed culture medium for anaerobic culture to obtain Lactobacillus seed liquid; then weigh MRS medium and black mustard seed powder, mix them at a material-to-liquid ratio of 1g:(8-10)mL, stir, and sterilize to obtain basic fermentation medium; the anaerobic culture temperature is 35-39℃ and the time is 24-75h; the inoculation amount of Lactococcus lactis is 1-3wt% of the total mass of the seed culture medium; S2. Inoculate the Lactobacillus seed culture into the basal fermentation medium for anaerobic fermentation. After fermentation, sterilize, centrifuge, and collect the supernatant to obtain the fermentation broth. The anaerobic culture temperature is 35-39℃, the time is 48-72h, and the pH is 5.5-6.

5. The inoculation amount of Lactobacillus seed culture is 4-6% of the total mass of the basal fermentation medium, and the viable count of Lactobacillus seed culture is (1.0-5.0)×10⁻⁶. 9 CFU / mL; S3. Choline chloride and lactic acid are mixed to obtain choline chloride-lactic acid solvent; choline chloride, glycerol, and sorbitol are mixed to obtain choline chloride-glycerol-sorbitol solvent; the choline chloride-lactic acid solvent and the choline chloride-glycerol-sorbitol solvent are mixed, rotary evaporated until the mixed solvent is clear and transparent, and then cooled to obtain biphase NaDES; wherein, when preparing the choline chloride-lactic acid solvent, the molar ratio of choline chloride to lactic acid is 1:(1.5-2.5); when preparing the choline chloride-glycerol-sorbitol solvent, the molar ratio of choline chloride, glycerol, and sorbitol is 1:(0.8-1.2):(0.8-1.2); the volume ratio of the choline chloride-lactic acid solvent and the choline chloride-glycerol-sorbitol solvent is 1:(0.8-1.2); S4. The fermentation broth obtained in step S2 is mixed with the biphasic NaDES obtained in step S3 and then subjected to ultrasonic extraction. After centrifugation, the supernatant is collected, and the supernatant is decolorized and desalted. The supernatant is then freeze-dried under vacuum to obtain the Lactobacillus / black mustard seed fermentation product extract. The volume ratio of the fermentation broth to the biphasic NaDES is 1:(10-20), and the ultrasonic extraction power is 300-500W, the frequency is 25-35kHz, the temperature is 40-50℃, and the time is 15-25min.

6. The sensitive skin microecological balancing-oil control and acne-removing composition as described in claim 1, characterized in that, The method for preparing the citrus fruit extract includes the following steps: (1) Select fresh citrus peel, remove the pulp, seeds and white inner pulp, leaving only the outer yellow peel, vacuum dry the outer yellow peel, and then pulverize and sieve it to obtain peel powder; wherein the particle size of the peel powder is 0.125-0.250mm. (2) Mix the fruit peel powder with a mixed solvent, perform ultrasonic extraction, centrifuge, and collect the supernatant; wherein, the mixed solvent includes anhydrous ethanol, deionized water and Tween-80 in a volume ratio of (2-4):(6-8):(0.05-0.1), and the material-to-liquid ratio of fruit peel powder to mixed solvent is 1g:15-25mL; the ultrasonic extraction power is 200-400W, the frequency is 25-35kHz, the temperature is 50±2℃, and the time is 40-50min; (4) The supernatant was purified, concentrated and freeze-dried to obtain the citrus fruit extract.

7. The sensitive skin microecological balancing-oil control and acne-removing composition as described in claim 1, characterized in that, The preparation method of the red clover flower extract includes the following steps: S1. Take dried red clover flowers, remove impurities, pulverize them, and pass them through a 40-60 mesh sieve to obtain red clover flower powder. S2. Mix red clover flower powder with ethanol aqueous solution, heat and reflux to extract, centrifuge, collect the supernatant to obtain the extract; wherein, the extraction temperature is 60-80℃, the time is 2-5h, the material-to-liquid ratio of red clover flower powder to ethanol aqueous solution is 1g:(8-13)mL, and the mass concentration of ethanol aqueous solution is 70-80wt%. S3. The extract obtained in step S2 is concentrated, purified, dried and pulverized to obtain the red clover flower extract.

8. The application of the sensitive skin microecological balancing-oil-controlling and acne-removing composition according to any one of claims 1-6 in the preparation of cosmetics, characterized in that, The cosmetics include at least one of toner, lotion, cream, mask, serum, gel, and spray, and the sensitive skin microecological balancing-oil control and acne-removing composition accounts for 1-15 wt% of the total mass of the cosmetics.

9. A gel, characterized in that, The ingredients comprise the following ingredients by weight percentage: 1%-15% of the sensitive skin microecological balance-oil control and acne-removing composition according to any one of claims 1-6, 0.1%-2% thickener, 1%-15% moisturizer, 0.5%-3% preservative and 0.01%-0.3% pH adjuster, with the balance being deionized water.

10. The method for preparing the gel according to claim 9, characterized in that, Includes the following steps: S1. Mix the humectant, thickener and deionized water, and homogenize at 75-85℃ to obtain a mixture; S3. After the mixture cools down to 55-65℃, add the preservative and stir evenly. After the temperature drops to 35-45℃, add each component of the sensitive skin microecological balance-oil control and acne removal composition and stir evenly. Finally, add the pH adjuster to adjust the pH to obtain the gel.

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