Preparation method and application of trillium tschonoskii yeast / probiotic fermentation liquor

Abstract

The invention discloses a preparation method and application of trillium tschonoskii yeast / probiotic fermentation broth, trillium tschonoskii is fermented by adopting saccharomyces cerevisiae and compound probiotics in a combined manner, all functional components and activity of plants are effectively reserved, and loss of active components possibly caused by a traditional extraction method is avoided. The prepared trillium tschonoskii yeast / probiotic fermentation liquor does not contain chemical components, the safety of the product is ensured, and the trillium tschonoskii yeast / probiotic fermentation liquor can be directly used as a mask, essence or toner. Compared with other products on the market, the prepared trillium tschonoskii yeast / probiotic fermentation liquor is more natural and does not bring any side effect to the skin. In addition, macromolecular components are decomposed into smaller molecules in the probiotic fermentation process, so that the skin can absorb the macromolecular components more easily and fully.

Description

Technical Field

[0001] The present invention belongs to the technical field of cosmetic raw materials, and in particular relates to a preparation method of a Trillium yeast / probiotic fermentation liquid and an application thereof. Background Art

[0002] Trillium, also known as "a bead on the head" or "taro seven," belongs to the genus Trillium in the Liliaceae family. Its rhizome is commonly called "ground bead," while its fruit is commonly known as "sky bead." Both the rhizome and mature fruit of Trillium have medicinal properties and are listed in the "Compendium of Chinese Materia Medica," "National Compendium of Chinese Herbal Medicines," "Taibai Seven Medicinal Herbs," and "Flora of the Qinling Mountains." This plant has a sweet taste and mild properties, with sedative, analgesic, hemostatic, and detoxifying properties. It has demonstrated unique therapeutic benefits in treating dizziness, hypertension, neurasthenia, traumatic injuries, menstrual irregularities, post-concussion syndrome, headaches, back and leg pain, and insomnia, and is widely used, particularly among the Miao and Tujia ethnic groups. In China, Trillium grows primarily in mountain forests, valleys, and hillsides at altitudes between 1,400 and 2,200 meters, in fertile soil rich in organic matter. In recent years, with the maturity of artificial cultivation technology, the medicinal source problem of Trillium has been solved, which has increased the possibility of its application and attracted the attention of more and more scientific research teams, who are studying the efficacy and application value of Trillium.

[0003] The chemical components of Trillium spp. mainly include steroidal saponins, flavonoids, fatty acids, and polysaccharides. However, the application of Trillium spp. extracts in cosmetics has several limitations: first, organic solvents are often used in the preparation process, which may result in the extract being mixed with organic solvents; second, the polarity and water solubility of the saponins in each Trillium spp. extract vary greatly, making them difficult to use in cosmetics; third, the saponins in Trillium spp. extracts may show varying degrees of toxicity; and finally, the current extraction process only extracts a portion of the components, while the majority is discarded.

[0004] Skin care products in today's society are mainly divided into chemical skin care products and natural skin care products. Chemical skin care products usually contain chemical raw materials or chemical additives. Although they have a quick effect, they may cause side effects to users, such as "excessive lead" or "excessive mercury". In addition, due to their large molecules, chemical skin care products are often difficult to be absorbed by the skin, and the improvement effect on the skin is not ideal. Natural skin care products, including Chinese herbal skin care products and fruit and vegetable skin care products, although they are of natural origin, do not always have an ideal effect on improving the skin. If the ingredients are not properly matched, they may also cause skin allergies. Many skin care products on the market currently add some plant extracts to ordinary products and claim to have specific effects. However, due to the barrier function of the skin, the active ingredients in plant extracts, especially high molecular weight substances and water-soluble substances, are difficult to penetrate the skin and be absorbed, resulting in limited actual effects. Therefore, it has become an urgent task to develop a skin care product that can effectively improve the skin while avoiding irritation or side effects and having a soothing and moisturizing effect. Summary of the Invention

[0005] The present invention aims to overcome the defects of the prior art and provide a method for preparing a Trillium yeast / probiotic fermentation broth.

[0006] Another object of the present invention is to provide an application of the above-mentioned Trillium yeast / probiotic fermentation broth.

[0007] The technical solutions of the present invention are as follows:

[0008] A method for preparing a Trillium yeast / probiotic fermentation broth comprises the following steps:

[0009] (1) Crush the clean rhizomes of Trillium truncatum and sieve them to obtain Trillium truncatum powder, then mix the Trillium truncatum powder with deionized water, sterilize it, and cool it to obtain the Trillium truncatum inoculation solution;

[0010] (2) Saccharomyces cerevisiae, Lactobacillus plantarum, Bifidobacterium longum, Lactobacillus rhamnosus, Lactococcus lactis, and Bacillus subtilis were inoculated on a plate culture medium and cultured at a constant temperature;

[0011] (3) Pick a single colony from the plate culture medium in step (2) and inoculate it into liquid culture medium until the number of bacteria detected therein reaches 10 7-9 / mL, to obtain a seed solution; wherein, the pH of the seed solution of Saccharomyces cerevisiae is 5.5-7.0, and the pH of the seed solution of Lactobacillus plantarum, Bifidobacterium longum, Lactobacillus rhamnosus, Lactococcus and Bacillus subtilis is 4.0-6.5;

[0012] (4) inoculating the saccharomyces cerevisiae seed solution obtained in step (3) into the Trillium inoculation solution for fermentation and culturing, and obtaining the Trillium cerevisiae fermentation solution after sterilization;

[0013] (5) The seed liquids of Lactobacillus plantarum, Bifidobacterium longum, Lactobacillus rhamnosus, Lactococcus and Bacillus subtilis obtained in step (3) are mixed and inoculated into the Trillium yeast fermentation liquid obtained in step (4), and the mixture is cultured at a constant temperature of 36-38° C. for 18-24 hours. The supernatant is collected by centrifugation and filtered with a filter membrane to obtain the Trillium yeast / probiotic fermentation liquid.

[0014] In a preferred embodiment of the present invention, in step (1), the mass ratio of Trillium powder to deionized water is 2-10:100.

[0015] In a preferred embodiment of the present invention, in step (2), Saccharomyces cerevisiae is cultured at a constant temperature of 20-35°C for 48-72 hours, and Lactobacillus plantarum, Bifidobacterium longum, Lactobacillus rhamnosus, Lactococcus and Bacillus subtilis are cultured at a constant temperature of 35-37°C for 24-48 hours.

[0016] In a preferred embodiment of the present invention, in step (3), Saccharomyces cerevisiae is cultured at a constant temperature of 20-35°C for 24-72 hours, and Lactobacillus plantarum, Bifidobacterium longum, Lactobacillus rhamnosus, Lactococcus and Bacillus subtilis are cultured at a constant temperature of 36-38°C for 18-24 hours.

[0017] In a preferred embodiment of the present invention, in step (4), the seed solution of Saccharomyces cerevisiae is inoculated into the Trillium inoculation solution at a rate of 1-5%, and the fermentation temperature is 20-35° C. for 24-72 hours.

[0018] In a preferred embodiment of the present invention, in step (5), the mass ratio of the seed liquid of Lactobacillus plantarum, Bifidobacterium longum, Lactobacillus rhamnosus, Lactococcus and Bacillus subtilis is 1:1:1:1:1, and the total volume of the seed liquid of Lactobacillus plantarum, Bifidobacterium longum, Lactobacillus rhamnosus, Lactococcus and Bacillus subtilis accounts for 1-5% of the trillium yeast fermentation broth.

[0019] The trillium yeast / probiotic fermentation liquid prepared by the above preparation method is used in the preparation of a skin care composition.

[0020] In a preferred embodiment of the present invention, the skin care composition is a toner, a cream or an essence.

[0021] A skin care composition, the active ingredients of which include the trillium yeast / probiotic fermentation liquid prepared by the above preparation method.

[0022] In a preferred embodiment of the present invention, the skin care composition is a toner, a cream or an essence.

[0023] The beneficial effects of the present invention are:

[0024] 1. The present invention uses brewer's yeast and composite probiotics to jointly ferment Trillium, effectively retaining all the effective components and activities of the plant, and avoiding the loss of active ingredients that may be caused by traditional extraction methods.

[0025] 2. The present invention utilizes a combined fermentation of Saccharomyces cerevisiae and various probiotics to provide the probiotics with nutrients such as amino acids, vitamins, and pyruvate, while also providing them with energy. Furthermore, Saccharomyces cerevisiae can fully ferment and decompose the various components of Trillium, which not only reduces toxicity but also enhances the efficacy of the Trillium yeast / probiotic fermentation broth.

[0026] 3. This invention selects a specific combination of strains for compounding, and the substances secreted by different probiotics promote each other's growth. After fermentation, the beauty benefits of Trillium are significantly enhanced. It can decompose and produce a large number of small molecule nutrients that are easily absorbed by the skin, such as amino acids, small molecule compounds, oligosaccharides, and peptides. These nutrients promote skin absorption, restore skin elasticity, reduce skin sensitivity, restore cell vitality, and promote skin health.

[0027] 4. The Trillium yeast / probiotic fermentation broth produced in this invention contains no chemical ingredients, ensuring product safety and allowing it to be used directly as a facial mask, serum, or toner. Compared to other products on the market, the Trillium yeast / probiotic fermentation broth produced in this invention is more natural and does not cause any side effects on the skin. Furthermore, the probiotic fermentation process breaks down large molecules into smaller ones, making them more easily absorbed by the skin. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 (a) shows the effects of different concentrations of samples in Example 1 of the present invention on the viability of RAW264.7 cells, Figure 1 (b) shows the effects of different concentrations of samples in Example 1 of the present invention on the viability of Hacat cells.

[0029] Figure 2 The results show the effect of the Trillium yeast / probiotic fermentation broth in Example 1 of the present invention on the AQP3 content in Hacat cells.

[0030] Figure 3 The effect of the Trillium yeast / probiotic fermentation broth in Example 1 of the present invention on the morphology of RAW264.7 cells is shown.

[0031] Figure 4 The results show the effect of the Trillium yeast / probiotic fermentation broth in Example 1 of the present invention on the NO release of RAW264.7 cells.

[0032] Figure 5 Showing the results of consumer use tests of the toner in Example 2 of the present invention

[0033] Figure 6 The results of a consumer use test of the facial cream in Example 3 of the present invention are shown.

[0034] Figure 7 The results of a consumer use test of the essence in Example 4 of the present invention are shown. DETAILED DESCRIPTION

[0035] The technical solution of the present invention is further illustrated and described below through specific implementation methods in conjunction with the accompanying drawings.

[0036] Example 1

[0037] (1) Preparation of Trillium inoculation solution: Crush the clean rhizomes of Trillium and sieve through 80 mesh to obtain Trillium powder. Mix the Trillium powder and deionized water in a mass ratio of 5:100, sterilize at 121°C for 20 minutes, and cool to obtain the Trillium inoculation solution.

[0038] (2) Prepare solid culture medium, sterilize, cool, and prepare plates. In a clean bench, quickly thaw frozen Saccharomyces cerevisiae, Lactobacillus plantarum, Bifidobacterium longum, Lactobacillus rhamnosus, Lactococcus acidophilus, and Bacillus subtilis. Use an inoculation loop to dip the bacterial liquid and streak on different labeled plate culture media. Saccharomyces cerevisiae is cultured at 30°C for 72 hours, and Lactobacillus plantarum, Bifidobacterium longum, Lactobacillus rhamnosus, Lactococcus acidophilus, and Bacillus subtilis are cultured in a constant temperature incubator at 37°C for 36 hours.

[0039] (3) Prepare liquid culture medium, sterilize, and cool to room temperature. Pick individual colonies on the plate culture medium with an inoculation loop and inoculate them into liquid culture medium. Saccharomyces cerevisiae is cultured at 30°C for 36 h, while Lactobacillus plantarum, Bifidobacterium longum, Lactobacillus rhamnosus, Lactococcus, and Bacillus subtilis are cultured at 37°C for 24 h until the number of bacteria detected reaches 10 8 The pH of the seed solution of Saccharomyces cerevisiae is 6.0, and the pH of the seed solution of Lactobacillus plantarum, Bifidobacterium longum, Lactobacillus rhamnosus, Lactococcus, and Bacillus subtilis is 5.5.

[0040] (4) The seed liquid of brewer's yeast was inoculated into the Trillium inoculation liquid at a rate of 1-5%, and the mixture was fermented at 30° C. for 72 h, and then sterilized at 80° C. for 30 min to obtain the Trillium brewer's yeast fermentation liquid.

[0041] (5) The seed solutions of Lactobacillus plantarum, Bifidobacterium longum, Lactobacillus rhamnosus, Lactococcus and Bacillus subtilis obtained in step (3) are mixed in a mass ratio of 1:1:1:1:1 and inoculated into the Trillium yeast fermentation liquid obtained in step (4), wherein the total volume of the seed solutions of Lactobacillus plantarum, Bifidobacterium longum, Lactobacillus rhamnosus, Lactococcus and Bacillus subtilis accounts for 5% of the above-mentioned Trillium yeast fermentation liquid. The mixture is cultured at a constant temperature of 37° C. for 36 h, centrifuged at 5000 r / min for 30 min, the supernatant is collected and filtered with a 0.22 μm filter membrane to obtain the Trillium yeast / probiotic fermentation liquid.

[0042] The culture medium used for the aforementioned Saccharomyces cerevisiae culture was YPD medium. The preparation process involved weighing 10 g of yeast extract, 20 g of peptone, and 20 g of D-glucose using a balance, dissolving them in 1000 mL of deionized water, and adjusting the pH to 5.6. After aliquoting, sterilize the mixture with high-temperature steam at 115°C for 20 minutes. Add 20 g of agar to the solid medium.

[0043] The above-mentioned Lactobacillus plantarum, Bifidobacterium longum, Lactobacillus rhamnosus, Lactococcus, and Bacillus subtilis can all be cultured in MRS broth. Its preparation process includes taking 54g of finished product MRS broth from Guangdong Huankai Microbiological Technology Co., Ltd. (the formula of MRS broth includes the following ingredients: 10.0g peptone, 10.0g beef extract, 5.0g yeast extract, 20.0g glucose, 2.0g ammonium citrate, 5.0g sodium acetate, 0.1g magnesium sulfate, 0.05g manganese sulfate, 2.0g dipotassium hydrogen phosphate, and 1.0g Tween 80, pH 5.7), adding 1L of distilled water, stirring and boiling until completely dissolved, and sterilizing with high pressure steam at 121°C for 15min before cooling to room temperature before use. 2% agar is added to the solid medium.

[0044] To demonstrate the safety and efficacy of the Trillium yeast / probiotic fermentation broth prepared in this example, the following experiments were conducted, and the experimental results were analyzed and discussed in detail.

[0045] Cell Culture: Purchase HaCaT and RAW264.7 cells, resuscitate, and culture to an appropriate density at 37°C, 5% CO2. Discard the culture medium, wash any remaining medium with PBS, and trypsinize. Add an equal volume of FBS to the trypsin to terminate the digestion. Add two volumes of PBS, collect the cell suspension by pipetting, centrifuge at 1500 rpm for 5 minutes, and resuspend the cells in complete medium. Subculture and select cells in the logarithmic growth phase and in good condition for the next experiment.

[0046] CCK assay for cell viability: The experiment was designed with a negative control group and a sample group. Six sample concentration gradient groups were designed, with the concentrations from high to low being C1, C2, C3, C4, C5, and C6, with three parallel wells in each group. Cells in the logarithmic phase and in good condition were digested, centrifuged, and suspended in the corresponding cell culture medium. The cells were counted and a cell suspension of a certain cell concentration was prepared according to different cell sizes, culture experience, and experimental requirements. 95 μL of the cell suspension was added to each well of a 96-well plate (empirical value: 3×10 HaCaT cells). 4 RAW264.7 cells / well: 4×10 4 Each well was cultured at 37°C and 5% CO2 for 14 ± 2 h. Samples were added (5 μL of PBS was added to each well of the negative control group, and 5 μL of samples of the corresponding concentration were added to each well of the sample group). The cells were cultured at 37°C and 5% CO2 for 24 h. CCK reagent was added according to the instructions of the CCK kit, and the absorbance was detected to calculate the cell viability.

[0047] Hacat cell moisturizing activity assay: Moisturizing model - cells in the logarithmic growth phase and in good condition were digested, centrifuged, suspended in the corresponding cell culture medium, counted, and prepared into a cell suspension with a certain cell concentration. The experiment was designed with blank and sample groups, with 3 parallel wells in each group. 200 μL of cell suspension was inoculated into each well of a 48-well plate and cultured overnight at 37°C and 5% CO2 for (14±2) h. The culture medium was discarded and the cells were washed with 200 μL / well of normal saline or PBS. 190 μL of culture medium and 10 μL of PBS were added to each well of the blank group, and 190 μL of culture medium and 10 μL of sample were added to each well of the sample group. The cells were cultured at 37°C and 5% CO2 for 24 h. The cell supernatant was collected and the AQP3 content was detected and calculated according to the instructions of the ELISA kit.

[0048] RAW264.7 cells were used to detect anti-inflammatory activity: cells in the logarithmic growth phase and in good condition were digested, centrifuged, suspended in the corresponding cell culture medium, and counted to prepare a cell suspension with a certain cell concentration. The experiment was designed as a blank group (PBS group), a control group (LPS group), and a sample group, with 3 parallel wells in each group. 200 μL of cell suspension was inoculated into each well of a 48-well plate and cultured overnight at 37°C, 5% CO2 for (14±2) h. The culture medium was discarded and the cells were washed with 200 μL / well of saline or PBS. In the blank group, 195 μL of culture medium and 5 μL of PBS were added to each well. In the control group, 191 μL of culture medium, 4 μL of LPS working solution (final LPS concentration of 1 μg / mL), and 5 μL of PBS were added to each well. In the sample group, 191 μL of culture medium, 4 μL of LPS working solution (final LPS concentration of 1 μg / mL), and 5 μL of the sample to be tested were added to each well. The cells were cultured at 37°C, 5% CO2 for 24 h. The cell morphology was observed, and the cell supernatant was collected. The NO content was detected and calculated according to the kit instructions.

[0049] Hyaluronidase inhibition assay for anti-allergic activity: The hyaluronidase inhibition rate was determined using the modified Elson-Morgan method. A—sample group, B—sample control group, C—negative group, and D—negative control group were set up as shown in the table below. The experiment was performed according to the steps in Table 1. The sample was diluted 20-fold with distilled water.

[0050] Table 1 Hyaluronidase inhibition experimental process

[0051]

[0052]

[0053] Calculate the hyaluronidase inhibition rate (%) = [1-(tube A-tube B) / (tube C-tube D)]*100%

[0054] The experimental results are described as follows:

[0055] Cell viability - To ensure the reliability of the subsequent experimental data, this example tested the effect of Trillium yeast / probiotic fermentation broth at different concentrations on cell viability using CCK assay before exploring the efficacy of Trillium yeast / probiotic fermentation broth. The results are as follows: Figure 1 As shown, the fermentation broth of Trillium yeast / probiotics showed no cytotoxicity to RAW264.7 cells and HaCaT cells at different concentrations, and the cell viability was above 90%.

[0056] Moisturizing – Aquaporins (AQP3) are integral membrane proteins that penetrate biological membranes to form water channels. They are primarily responsible for transporting small molecules such as water and glycerol to different layers of skin tissue and are closely related to epidermal hydration. Numerous studies have shown that AQP3 is the most abundant AQP subtype in the skin and has the strongest correlation with skin hydration.

[0057] This example selects AQP3 as the evaluation index to explore the moisturizing effect of Trillium yeast / probiotic fermentation liquid. Figure 2 It can be seen that the fermentation broth of Trillium yeast / probiotics can effectively promote the secretion of AQP3 by HaCaT cells, increasing it from 0.98 ng / mL to 3.81 ng / mL.

[0058] Anti-inflammatory and soothing effects – Inflammation is the body's defensive response to harmful factors. This defensive process involves multiple cells and is accompanied by the release of inflammatory mediators. Nitric oxide is often used as an indicator of inflammation and a target for relieving inflammation. During the inflammatory response, activated nitric oxide synthase (iNOS) produces high concentrations of NO, activating the NF-κB signaling pathway, further inducing the release of pro-inflammatory factors and exacerbating the inflammatory response. Therefore, inhibiting the production of iNOS and NO is one way to alleviate inflammation.

[0059] This example uses LPS to induce RAW264.7 cell model and selects NO as the evaluation index to explore the anti-inflammatory activity of Trillium yeast / probiotic fermentation broth, thereby evaluating the potential soothing effect of the active ingredient. Figure 3 As shown in the figure, microscopic observation shows that LPS stimulation causes significant changes in the morphology of RAW264.7 cells, with radial tentacles extending outward, while the Trillium yeast / probiotic fermentation broth can effectively alleviate the cell changes caused by LPS stimulation. In order to further explore the anti-inflammatory mechanism of Trillium yeast / probiotic fermentation broth, this example continues to collect cell supernatant to detect the inflammatory mediator - NO. Figure 4 As shown in the LPS-activated RAW264.7 cell experiment, the NO content increased from 1.78 μM to 30.15 μM after LPS activation; after 24 hours of action of Trillium yeast / probiotic fermentation broth, the NO content decreased to 18.67 μM.

[0060] Anti-allergic and soothing - Hyaluronic acid (HA) is the main component of the extracellular matrix and intercellular matrix, and plays an important role in skin elasticity, cell adhesion, wound healing and angiogenesis. Studies have shown that HA is also closely related to allergic reactions and has a lot to do with skin sensitivity. Hyaluronidase (HAase), a proteolytic enzyme that specifically decomposes HA, is also often used in allergy-related research. Common allergic symptoms of redness, swelling, itching and pain are mainly caused by histamine, and currently common clinical medications are mainly antihistamines. Studies have shown that there is a good correlation between the inhibition of hyaluronidase activity and the inhibitory activity of mast cell release of histamine, and active ingredients that inhibit histamine often also show more obvious hyaluronidase inhibitory activity. Therefore, hyaluronidase, as a participant in type I allergic reactions, is often used to explore in vitro anti-allergic active ingredients.

[0061] This example used a hyaluronidase inhibition test to investigate the soothing efficacy of Trillium yeast / probiotic fermentation broth. Table 2 shows that the hyaluronidase inhibition rate of the Trillium yeast / probiotic fermentation broth was 79.714%, indicating that the Trillium yeast / probiotic fermentation broth can inhibit hyaluronidase activity, reduce the decomposition of hyaluronic acid, and thus reduce the release of histamine and inflammatory factors, thereby providing a certain anti-allergic and soothing effect.

[0062] Table 2 Hyaluronidase inhibition test results

[0063]

[0064] The above experiments confirmed that the Trillium yeast / probiotic fermentation broth prepared in this example can promote the production and expression of AQP3 in HaCaT cells, inhibit the release of inflammatory mediators in RAW264.7 cells, and has a significant inhibitory effect on NO. At the same time, the inhibition rate of hyaluronidase reaches 79.714%.

[0065] In summary, the Trillium yeast / probiotic fermentation liquid prepared in this example has good moisturizing and soothing effects.

[0066] Example 2

[0067] A toner comprising:

[0068] Carrier ingredients: glycerol 5%, butylene glycol 3%, deionized water 87%;

[0069] Effective ingredient: 5% of the Trillium yeast / probiotic fermentation broth prepared in Example 1.

[0070] The preparation method is as follows: weighed glycerol, butanediol and deionized water are added to a water pot and heated to 95° C., stirred and mixed, and then cooled to room temperature. Then, the Trillium yeast / probiotic fermentation liquid prepared in Example 1 is added while stirring until it is evenly stirred.

[0071] The results of the consumer use test of the toner prepared in this embodiment are as follows: Figure 5 shown.

[0072] Example 3

[0073] A facial cream comprising:

[0074] Carrier ingredients: 3 parts of 1,3-butylene glycol, 6 parts of glycerin, 74 parts of deionized water, 0.5 parts of ammonium acryloyldimethyltaurate / VP copolymer, 5 parts of caprylic / capric triglyceride, 4 parts of macadamia seed oil, 4 parts of cetearyl olivate / sorbitan olivate;

[0075] Effective ingredient: 3.5 parts of the Trillium yeast / probiotic fermentation broth prepared in Example 1.

[0076] The preparation method is as follows: weighed phase A components (glycerol, 1,3-butylene glycol, deionized water, ammonium acryloyldimethyltaurate / VP copolymer) are added to a water pot and heated to 95° C., and stirred to mix; phase B components (caprylic / capric triglyceride, macadamia seed oil, cetearyl olivate / sorbitan olivate) are heated to 80° C.; phase B components are added to phase A components and homogenized and emulsified, cooled to room temperature, and the trillium yeast / probiotic fermentation broth prepared in Example 1 is added while stirring until evenly stirred.

[0077] The results of the consumer use test of the facial cream prepared in this embodiment are as follows: Figure 6 shown.

[0078] Example 4

[0079] A serum comprising:

[0080] Carrier ingredients: 3 parts of 1,3-butanediol, 6 parts of glycerol, 86.5 parts of deionized water;

[0081] Effective ingredients: Trillium yeast / mixed probiotics combined fermentation liquid 4.5%,

[0082] The preparation method is as follows: weighed glycerol, 1,3-butanediol and deionized water are added to a water pot and heated to 95° C., stirred and mixed, cooled to room temperature, and the Trillium yeast / probiotic fermentation liquid prepared in Example 1 is added while stirring until uniformly stirred.

[0083] The results of the consumer use test of the essence prepared in this embodiment are as follows: Figure 7 shown.

[0084] The above description is merely a preferred embodiment of the present invention and therefore cannot be used to limit the scope of the present invention. In other words, equivalent changes and modifications made within the scope of the present invention and the contents of the specification should still fall within the scope of the present invention.

Claims

1. A method for preparing a Trillium yeast / probiotic fermentation broth, characterized in that: The steps include: (1) crushing the clean rhizomes of Trillium truncatum and sieving them to obtain Trillium truncatum powder, then mixing the Trillium truncatum powder with deionized water, sterilizing the mixture, and cooling the mixture to obtain a Trillium truncatum inoculation solution; (2) inoculating Saccharomyces cerevisiae, Lactobacillus plantarum, Bifidobacterium longum, Lactobacillus rhamnosus, Lactococcus aureus and Bacillus subtilis on a plate culture medium for constant temperature culture; (3) Pick a single colony from the plate culture medium in step (2) and inoculate it into a liquid culture medium for cultivation until the number of bacteria detected therein reaches 10 7-9 / mL, and the seed solution is obtained; wherein the pH of the seed solution of Saccharomyces cerevisiae is 5.5-7.0, and the pH of the seed solution of Lactobacillus plantarum, Bifidobacterium longum, Lactobacillus rhamnosus, Lactococcus and Bacillus subtilis is 4.0-6.5; (4) inoculating the saccharomyces cerevisiae seed solution obtained in step (3) into the Trillium inoculation solution for fermentation and culturing, and obtaining the Trillium saccharomyces cerevisiae fermentation solution after sterilization; (5) The seed liquids of Lactobacillus plantarum, Bifidobacterium longum, Lactobacillus rhamnosus, Lactococcus and Bacillus subtilis obtained in step (3) are mixed and inoculated into the Trillium yeast fermentation liquid obtained in step (4), and the mixture is cultured at 36-38° C. for 18-24 h. The supernatant is collected by centrifugation and filtered through a filter membrane to obtain the Trillium yeast / probiotic fermentation liquid.

2. The preparation method according to claim 1, characterized in that: In the step (1), the mass ratio of the Trillium powder to deionized water is 2-10:

100.

3. The preparation method according to claim 1, characterized in that: In the step (2), the brewer's yeast is cultured at a constant temperature of 20-35° C. for 48-72 hours, and the Lactobacillus plantarum, Bifidobacterium longum, Lactobacillus rhamnosus, Lactococcus and Bacillus subtilis are cultured at a constant temperature of 35-37° C. for 24-48 hours.

4. The preparation method according to claim 1, characterized in that: In the step (3), the brewer's yeast is cultured at a constant temperature of 20-35° C. for 24-72 hours, and the Lactobacillus plantarum, Bifidobacterium longum, Lactobacillus rhamnosus, Lactococcus and Bacillus subtilis are cultured at a constant temperature of 36-38° C. for 18-24 hours.

5. The preparation method according to claim 1, characterized in that: In the step (4), the brewer's yeast seed solution is inoculated into the Trillium inoculation solution at 1-5%, and the fermentation temperature is 20-35° C. and the fermentation time is 24-72 hours.

6. The preparation method according to claim 1, characterized in that: In the step (5), the mass ratio of the seed liquid of Lactobacillus plantarum, Bifidobacterium longum, Lactobacillus rhamnosus, Lactococcus and Bacillus subtilis is 1:1:1:1:1, and the total volume of the seed liquid of Lactobacillus plantarum, Bifidobacterium longum, Lactobacillus rhamnosus, Lactococcus and Bacillus subtilis accounts for 1-5% of the fermentation liquid of Trillium yeast.

7. Use of the Trillium yeast / probiotic fermentation liquid obtained by the preparation method according to any one of claims 1 to 6 in the preparation of a skin care composition.

8. The use according to claim 7, characterized in that: The skin care composition is toner, cream or essence.

9. A skin care composition, characterized in that: The effective ingredients thereof include the Trillium yeast / probiotic fermentation liquid prepared by the preparation method described in any one of claims 1 to 6.

10. A skin care composition according to claim 9, characterized in that: The skin care composition is toner, cream or essence.

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