Polygonatum sibiricum fermentation liquor, essence, preparation method and application

Abstract

The invention provides rhizoma polygonati fermentation liquor, essence, a preparation method and application, and particularly belongs to the technical field of skin care cosmetics. The preparation method comprises the following steps: mixing rhizoma polygonati powder with water to obtain wet-based rhizoma polygonati powder; inoculating aspergillus oryzae into the wet-based rhizoma polygonati powder for solid-state fermentation, and drying to obtain a rhizoma polygonati solid-state fermentation product; mixing the rhizoma polygonati solid-state fermentation product with water, extracting with hot water, and performing suction filtration to obtain rhizoma polygonati fermentation filtrate; and sterilizing, filtering and centrifuging the polygonatum sibiricum fermentation filtrate to obtain polygonatum sibiricum fermentation liquor (fermentation liquor for short). The prepared fermentation liquor has the effects of accelerating skin injury repair, resisting oxidation, preserving moisture, diminishing inflammation and regulating skin flora.

Description

Technical Field

[0001] The present invention belongs to the technical field of skin care cosmetics, and particularly relates to a polygonatum fermented liquid, a essence liquid, a preparation method and an application thereof. Background Art

[0002] Traditional Chinese medicine cosmetics are a class of products that, with the assistance of modern scientific and technological means, add natural Chinese herbal medicines or active ingredients of traditional Chinese medicine to cosmetics, and enhance a series of effects of cosmetics such as moisturizing, repair, and antioxidant, meeting the quality requirements of cosmetics for use. Traditional Chinese medicine has a long history of clinical application, scientific research, and application practice research for thousands of years, with a long application history and high safety. In today's society that advocates green development, traditional Chinese medicine cosmetics have become a research hotspot for many domestic and foreign cosmetics R & D institutions due to their obvious clinical therapeutic, functional, and high safety advantages. Adding Chinese herbal medicine ingredients to cosmetics can give new vitality and vigor to the cosmetics industry, and also better conform to the skin care concept of "natural, safe, and scientific".

[0003] As a traditional Chinese medicine in China, polygonatum has extremely high medicinal value and a long application history due to its effects of invigorating qi and nourishing yin, strengthening the spleen, moistening the lungs, and tonifying the kidneys. Polygonatum belongs to the genus Polygonatum of the family Liliaceae, and is a perennial herbaceous plant and a Chinese herbal medicine with both medicinal and edible uses. Its rhizome is the main medicinal part, mainly containing polysaccharides, steroidal saponins, and a small amount of flavonoids and other substances. As one of the main active ingredients of polygonatum, polygonatum polysaccharide can effectively improve the SOD activity of human cells and the body's immunity, and has various physiological functions such as improving memory, anti-dementia, anti-aging, antioxidant, lowering blood sugar and blood lipids, anti-inflammatory, anti-tumor, and enhancing immunity. At present, polygonatum has been widely studied and applied in the fields of medicine and food, but its application research in cosmetics is less. Summary of the Invention

[0004] The purpose of the present invention is to provide a polygonatum fermented liquid, a essence liquid, a preparation method and an application thereof. The polygonatum fermented liquid prepared by the present invention has the functions of accelerating skin damage repair, antioxidant, moisturizing, anti-inflammatory, and regulating skin flora.

[0005] The present invention provides a preparation method of a polygonatum fermented liquid, comprising the following steps:

[0006] Mix polygonatum powder with water according to a mass ratio of 1:(0.8 - 1.2) to obtain wet-based polygonatum powder;

[0007] Inoculate Aspergillus oryzae on the wet-based polygonatum powder for solid-state fermentation, and then dry it to obtain a polygonatum solid-state fermented product; the inoculation amount of Aspergillus oryzae is 0.05 - 1%; the conditions for the solid-state fermentation are 25 - 32°C for 24 - 72 h;

[0008] Mix the solid-state fermented product of polygonatum sibiricum with water at a mass ratio of 1:(3-5), extract with hot water, and perform suction filtration to obtain the fermented filtrate of polygonatum sibiricum; the conditions for the hot water extraction are 30-50°C and 2-5 h;

[0009] Sterilize the fermented filtrate of polygonatum sibiricum, filter, centrifuge and take the supernatant to obtain the fermented liquid of polygonatum sibiricum.

[0010] The present invention also provides the fermented liquid of polygonatum sibiricum prepared by the preparation method described in the above technical solution.

[0011] The present invention also provides the application of the fermented liquid of polygonatum sibiricum prepared by the preparation method described in the above technical solution in the preparation of products having any one or more functions of ①-⑤:

[0012] ① Accelerate the repair of skin damage; the acceleration of skin damage repair includes accelerating the repair of skin photoaging damage and / or accelerating the repair of skin cell scratch; ② Antioxidant; ③ Moisturizing; ④ Anti-inflammatory; ⑤ Regulate skin flora; the regulation of skin flora includes promoting the proliferation of staphylococcus epidermidis and / or inhibiting propionibacterium acnes.

[0013] Preferably, the product includes cosmetics.

[0014] The present invention also provides an essence containing the fermented liquid of polygonatum sibiricum prepared by the preparation method described in the above technical solution, including the following components in parts by weight:

[0015] 50-60 parts of water, 0.08-0.12 parts of dipotassium glycyrrhizinate, 0.2-0.4 parts of acrylate / C10-30 alkanol acrylate cross-linked polymer, 3-5 parts of glycerol, 0.4-0.5 parts of p-hydroxyacetophenone, 2-4 parts of butanediol, 0.02-0.04 parts of tremella heteropolysaccharide, 0.01-0.03 parts of sodium hyaluronate, 0.2-0.3 parts of arginine, 0.4-0.5 parts of 1,2-hexanediol, 1-2.5 parts of trimethylpentanediol / adipic acid / glycerol cross-linked polymer, 0.6-1 part of PEG-7 olive oil ester, and 20-30 parts of the fermented liquid of polygonatum sibiricum prepared by the preparation method described in claim 1.

[0016] The present invention also provides the preparation method of the essence described in the above technical solution, including the following steps:

[0017] Perform the first mixing of dipotassium glycyrrhizinate, acrylate / C10-30 alkanol acrylate cross-linked polymer, glycerol, p-hydroxyacetophenone, butanediol, tremella heteropolysaccharide, sodium hyaluronate and water at 75-80°C, and stir to obtain the first intermediate product;

[0018] Cool the first intermediate product to 45-50°C, perform the second mixing of the first intermediate product and arginine, and stir to obtain the second intermediate product;

[0019] The second intermediate product is cooled to 35-40°C, and the second intermediate product is mixed with polygonatum sibiricum fermented liquid, 1,2-hexanediol, trimethylpentanediol / adipic acid / glycerol cross-linked polymer and PEG-7 olive oil ester for the third time, and stirred to obtain the essence.

[0020] The present invention also provides the application of the essence described in the above technical solution or the essence prepared by the preparation method described in the above technical solution in the preparation of skin regulating and repairing cosmetics.

[0021] Preferably, the skin regulating and repairing cosmetics have good transdermal absorption effect, can promote cell vitality and can promote cell scratch healing ability.

[0022] The present invention provides a preparation method of polygonatum sibiricum fermented liquid. Aspergillus oryzae is used in the present invention for solid-state fermentation and water extraction of polygonatum cyrtonema, and the water extract is sterilized, filtered and centrifuged to prepare a fermented extract containing polygonatum oligofructose and galactan. The obtained polygonatum sibiricum fermented liquid is rich in polygonatum oligofructose and galactan. The preparation method of the present invention is more simple, energy-saving and environmentally friendly; the polygonatum sibiricum fermented liquid prepared by the present invention has a good repair effect on cell scratch and anti-photoaging damaged cells, and has a good moisturizing effect on the skin, providing new ideas for the application of natural products containing galactan and oligofructose in cosmetics.

[0023] Furthermore, the essence of the present invention is specially added with polygonatum sibiricum fermented liquid. By reasonably proportioning various active ingredients and giving full play to the synergistic skin care effects of each raw material component, natural and green skin care cosmetics can be prepared. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required to be used in the embodiments. Obviously, the following described drawings are only some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0025] Figure 1 It is the experimental flow chart of Example 1 provided by the present invention;

[0026] Figure 2 It is the result chart of the scavenging rate of polygonatum sibiricum fermented liquid on DPPH free radicals provided by the present invention;

[0027] Figure 3 It is the result chart of the scavenging rate of polygonatum sibiricum fermented liquid on ABTS free radicals provided by the present invention;

[0028] Figure 4 It is the polygonatum sibiricum fermented liquid provided by the present invention for O2 -Results graph of the scavenging rate of free radicals;

[0029] Figure 5 This is the results graph of the scavenging rate of OH free radicals by the polygonatum fermented liquid provided by the present invention; - Results graph of the scavenging rate of free radicals;

[0030] Figure 6 This is the results graph of the scavenging rate of H2O2 free radicals by the polygonatum fermented liquid provided by the present invention;

[0031] Figure 7 This is the results graph of the effect of the polygonatum fermented liquid provided by the present invention on the survival rate of HSF cells;

[0032] Figure 8 This is the results graph of the cell scratch repair effect of the polygonatum fermented liquid provided by the present invention;

[0033] Figure 9 This is the results graph of the DNA content distribution of the cell cycle analysis of the polygonatum fermented liquid provided by the present invention by flow cytometry;

[0034] Figure 10 This is the results graph of the repair of photoaging damaged cells by the polygonatum fermented liquid provided by the present invention;

[0035] Figure 11 This is the results graph of the change in skin moisture content of the polygonatum fermented liquid provided by the present invention;

[0036] Figure 12 This is the results graph of the change in the trans-epidermal water loss rate of the polygonatum fermented liquid provided by the present invention;

[0037] Figure 13 This is the results graph of the promotion rate of the polygonatum fermented liquid provided by the present invention against Staphylococcus epidermidis;

[0038] Figure 14 This is the results graph of the inhibition rate of the polygonatum fermented liquid provided by the present invention against Propionibacterium acnes;

[0039] Figure 15 This is the experimental flow chart of Example 8 provided by the present invention;

[0040] Figure 16 This is the results graph of the cumulative penetration amount per unit area of different essence liquids provided by the present invention;

[0041] Figure 17 This is the results graph of the effect of polygonatum essence liquids with different volume fractions provided by the present invention on the survival rate of HaCaT cells;

[0042] Figure 18 This is the results graph of the HaCaT cell scratch experiment of the polygonatum essence liquid provided by the present invention;

[0043] Figure 19The figure shows the results of the effects of polygonatum essence solutions with different concentrations provided by the present invention on the migration rate of HaCaT cells. Detailed implementation mode

[0044] The present invention provides a method for preparing a polygonatum fermented liquid, comprising the following steps:

[0045] Mix polygonatum powder with water according to a mass ratio of 1:(0.8 - 1.2) to obtain wet-based polygonatum powder;

[0046] Inoculate Aspergillus oryzae on the wet-based polygonatum powder for solid-state fermentation, and then dry it to obtain a polygonatum solid ferment; the inoculation amount of Aspergillus oryzae is 0.05 - 1%; the conditions for the solid-state fermentation are 25 - 32°C and 24 - 72 h;

[0047] Mix the polygonatum solid ferment with water according to a mass ratio of 1:(3 - 5), perform hot water extraction, and then suction filter to obtain a polygonatum ferment filtrate; the conditions for the hot water extraction are 30 - 50°C and 2 - 5 h;

[0048] Sterilize the polygonatum ferment filtrate, filter it, and centrifuge to take the supernatant to obtain a polygonatum fermented liquid.

[0049] The present invention mixes polygonatum powder with water according to a mass ratio of 1:(0.8 - 1.2) to obtain wet-based polygonatum powder, providing moisture for the fermentation of Aspergillus oryzae. In the present invention, the polygonatum includes Polygonatum cyrtonema Hua. In the present invention, the polygonatum powder is obtained by washing, drying, slicing, and pulverizing the rhizome of polygonatum; the drying method includes drying. In a specific embodiment, the particle size of the polygonatum powder is 60 - 100 mesh, specifically 60 mesh, 80 mesh, or 100 mesh. In the present invention, the polygonatum powder is stored sealed in a dry container. In a specific embodiment, the mass ratio is 1:0.8, 1:1, or 1:1.2. In a specific embodiment, the water includes distilled water.

[0050] After obtaining the wet-based polygonatum powder, the present invention inoculates Aspergillus oryzae on the wet-based polygonatum powder for solid-state fermentation, and then dries it to obtain a polygonatum solid ferment; the inoculation amount of Aspergillus oryzae is 0.05 - 1%; the conditions for the solid-state fermentation are 25 - 32°C and 24 - 72 h. In a specific embodiment, the temperature of the solid-state fermentation is 26 - 30°C, and the time of the solid-state fermentation is 36 - 60 h. In a specific embodiment, the drying temperature is 40 - 55°C. This step is microbial-assisted extraction, serving as a pretreatment before water extraction.

[0051] After obtaining the solid-state fermented product of polygonatum sibiricum, the present invention mixes the solid-state fermented product of polygonatum sibiricum with water at a mass ratio of 1:(3-5), performs hot water extraction, and filters by suction. The conditions for the hot water extraction are 30-50°C for 2-5 hours; a polygonatum fermented filtrate is obtained. In a specific embodiment, the conditions for the hot water extraction can be 35-45°C for 3-4 hours.

[0052] After obtaining the polygonatum fermented filtrate, the present invention sterilizes, filters, and centrifuges the polygonatum fermented filtrate to obtain the supernatant, thereby obtaining the polygonatum fermented liquid. In a specific embodiment, the temperature for sterilization is 100-120°C. In a specific embodiment, the time for sterilization is 30-60 minutes. In a specific embodiment, the conditions for filtration are 200-300 mesh. In a specific embodiment, the conditions for centrifugation are 8000-12000 r / min for 5-20 minutes.

[0053] The present invention also provides the polygonatum fermented liquid prepared by the preparation method described in the above technical solution. The polygonatum fermented liquid of the present invention has the effects of accelerating skin damage repair, antioxidant, moisturizing, anti-inflammatory, and regulating skin flora. Among them, accelerating skin damage repair includes accelerating skin photoaging damage repair and / or accelerating skin cell scratch repair; regulating skin flora includes promoting the proliferation of staphylococcus epidermidis and / or inhibiting propionibacterium acnes.

[0054] The present invention also provides the application of the polygonatum fermented liquid prepared by the preparation method described in the above technical solution in the preparation of a product having any one or more of the functions of ①-⑤: ① accelerating skin damage repair; the accelerating skin damage repair includes accelerating skin photoaging damage repair and / or accelerating skin cell scratch repair; ② antioxidant; ③ moisturizing; ④ anti-inflammatory; ⑤ regulating skin flora; the regulating skin flora includes promoting the proliferation of staphylococcus epidermidis and / or inhibiting propionibacterium acnes. In a specific embodiment, the product includes cosmetics.

[0055] The present invention also provides a serum containing the polygonatum fermented liquid prepared by the preparation method described in the above technical solution, which includes the following components in parts by weight:

[0056] 50-60 parts of water, 0.08-0.12 parts of dipotassium glycyrrhizinate, 0.2-0.4 parts of acrylate / C10-30 alkyl acrylate cross-linked polymer, 3-5 parts of glycerol, 0.4-0.5 parts of p-hydroxyacetophenone, 2-4 parts of butanediol, 0.02-0.04 parts of tremella heteropolysaccharide, 0.01-0.03 parts of sodium hyaluronate, 0.2-0.3 parts of arginine, 0.4-0.5 parts of 1,2-hexanediol, 1-2.5 parts of trimethylpentanediol / adipic acid / glycerol cross-linked polymer, 0.6-1 part of PEG-7 olive oil ester, and 20-30 parts of the polygonatum fermented liquid prepared by the preparation method described in claim 1.

[0057] The present invention does not have special limitations on the sources of the components included in the essence, and conventional commercially available products can be used.

[0058] The essence described in the present invention includes 50 - 60 parts of water. In a specific embodiment, the water includes deionized water.

[0059] The essence described in the present invention includes 0.08 - 0.12 parts of dipotassium glycyrrhizinate. In the present invention, the addition and the set addition amount of dipotassium glycyrrhizinate can bring antioxidant and anti - inflammatory effects.

[0060] The essence described in the present invention includes 0.2 - 0.4 parts of acrylate / C10 - 30 alkyl acrylate cross - linked polymer. In the present invention, the addition and the set addition amount of acrylate / C10 - 30 alkyl acrylate cross - linked polymer can bring a thickening effect.

[0061] The essence described in the present invention includes 3 - 5 parts of glycerol. In the present invention, the addition and the set addition amount of glycerol can bring moisturizing and nourishing effects.

[0062] The essence described in the present invention includes 0.4 - 0.5 parts of p - hydroxyacetophenone. In the present invention, the addition and the set addition amount of p - hydroxyacetophenone can bring antibacterial and antiseptic effects.

[0063] The essence described in the present invention includes 2 - 4 parts of butanediol. In the present invention, the addition and the set addition amount of butanediol can bring moisturizing and soothing effects.

[0064] The essence described in the present invention includes 0.02 - 0.04 parts of tremella heteropolysaccharide. In the present invention, the addition and the set addition amount of tremella heteropolysaccharide can bring moisturizing and repairing effects.

[0065] The essence described in the present invention includes 0.01 - 0.03 parts of sodium hyaluronate. In the present invention, the addition and the set addition amount of sodium hyaluronate can bring a moisturizing effect.

[0066] The essence described in the present invention includes 0.2 - 0.3 parts of arginine. In the present invention, the addition and the set addition amount of arginine can bring the effects of pH regulation and moisturizing.

[0067] The essence described in the present invention includes 0.4 - 0.5 parts of 1,2 - hexanediol. In the present invention, the addition and the set addition amount of 1,2 - hexanediol can bring a moisturizing effect.

[0068] The essence described in the present invention includes 1 - 2.5 parts of trimethylpentanediol / adipic acid / glycerol cross - linked polymer. In the present invention, the addition and the set addition amount of trimethylpentanediol / adipic acid / glycerol cross - linked polymer can bring thickening and moisturizing effects.

[0069] The essence described in the present invention comprises 0.6 - 1 part of PEG - 7 olive oil ester. In the present invention, the addition and the set amount of PEG - 7 olive oil ester can bring about the effects of emulsification and thickening, moisturizing, and repair.

[0070] The essence described in the present invention comprises 20 - 30 parts of polygonatum sibiricum fermented liquid. In the present invention, the addition and the set amount of polygonatum sibiricum fermented liquid can bring about the effects of antioxidant, repair, and antibacterial. The essence described in the present invention is specially added with polygonatum sibiricum fermented liquid rich in polygonatum sibiricum fructooligosaccharide and galactan, which has the efficacy of regulating skin flora and accelerating the repair of skin damage, and through the reasonable proportioning of various active ingredients, gives full play to the synergistic skin care efficacy of each raw material component, and produces a repair essence that is green, natural, highly safe, mild, and has little irritation.

[0071] The present invention also provides a preparation method of the essence according to the above technical solution, comprising the following steps:

[0072] Perform a first mixing of dipotassium glycyrrhizinate, acrylate / C10 - 30 alkanol acrylate cross - polymer, glycerol, p - hydroxyacetophenone, butanediol, tremella heteropolysaccharide, sodium hyaluronate and water at 75 - 80 °C, and stir to obtain a first intermediate product;

[0073] Cool the first intermediate product to 45 - 50 °C, perform a second mixing of the first intermediate product and arginine, and stir to obtain a second intermediate product;

[0074] Cool the second intermediate product to 35 - 40 °C, perform a third mixing of the second intermediate product with polygonatum sibiricum fermented liquid, 1,2 - hexanediol, trimethylpentanediol / adipic acid / glycerol cross - polymer and PEG - 7 olive oil ester, and stir to obtain the essence.

[0075] Perform a first mixing of dipotassium glycyrrhizinate, acrylate / C10 - 30 alkanol acrylate cross - polymer, glycerol, p - hydroxyacetophenone, butanediol, tremella heteropolysaccharide, sodium hyaluronate and water at 75 - 80 °C, and stir to obtain a first intermediate product. In a specific embodiment, the rotation speed of the stirring is 3000 rpm. In a specific embodiment, the stirring time is 5 - 10 min, and stir until completely dispersed and uniform. The stirring conditions hereinafter are the same as here and will not be elaborated. In a specific embodiment, dipotassium glycyrrhizinate, acrylate / C10 - 30 alkanol acrylate cross - polymer, glycerol, p - hydroxyacetophenone, butanediol, tremella heteropolysaccharide and sodium hyaluronate are added to water in sequence to make the water phase in the system fully blend.

[0076] After obtaining the first intermediate product, the present invention waits for the first intermediate product to cool to 45 - 50 °C, performs a second mixing of the first intermediate product and arginine, and stirs to obtain a second intermediate product. At this temperature, arginine can dissolve and does not lose its activity.

[0077] After obtaining the second intermediate product, the present invention cools the second intermediate product to 35-40 °C, and mixes the second intermediate product with polygonatum fermented liquid, 1,2-hexanediol, trimethylpentanediol / adipic acid / glycerol crosslinked polymer and PEG-7 olive oil ester for the third time, and stirs to obtain the essence. After obtaining the essence, the present invention samples and measures the physical and chemical properties, sends the sample for quality inspection, directly discharges the product after passing the inspection, and packs it into boxes to obtain the finished product. In a specific embodiment, the polygonatum fermented liquid, 1,2-hexanediol, trimethylpentanediol / adipic acid / glycerol crosslinked polymer and PEG-7 olive oil ester are added to the second intermediate product in sequence. At the same temperature, the aqueous phase, oil phase, active substance and preservative are mixed to make the system uniform and stable.

[0078] The present invention also provides the application of the essence described in the above technical solution or the essence prepared by the preparation method described in the above technical solution in the preparation of skin regulating and repairing cosmetics. In a specific embodiment, the skin regulating and repairing cosmetics have good transdermal absorption effect, can promote cell viability and can promote cell scratch healing ability.

[0079] In order to further illustrate the present invention, the following examples are used to describe in detail a polygonatum fermented liquid, essence and preparation method and application provided by the present invention, but they should not be construed as limiting the protection scope of the present invention.

[0080] Example 1

[0081] Preparation of polygonatum fermented liquid by Aspergillus oryzae fermentation

[0082] 1. Fermentation method

[0083] The first step of Aspergillus oryzae fermentation: Weigh 20 g of pulverized and sieved polygonatum sibiricum powder through 80 meshes, add distilled water according to the mass ratio of 1:

[0084] 1.2, add 0.5% Aspergillus oryzae at 30 °C, and statically culture for 48 h. After the fermentation is completed, obtain the solid polygonatum fermented product, and dry it at 45 °C for later use.

[0085] The second step of water extraction: According to the mass of the starting material, add distilled water to the dried solid polygonatum fermented product until the mass ratio is 1:20. At a temperature of 45 °C, extract for 3 h to obtain the hot water extract of polygonatum. Vacuum filter the hot water extract of polygonatum, and the supernatant is the fermented filtrate of polygonatum. After sterilizing the fermented filtrate of polygonatum at 120 °C for 30 min, filter it with a 250-mesh filter cloth, and centrifuge the filtrate at 12,000 r / min for 15 min to obtain the fermented liquid of polygonatum.

[0086] 2. Experimental flow chart

[0087] The experimental flow chart is shown in Figure 1 .

[0088] Example 2

[0089] Component Analysis of Polygonatum sibiricum Reduced Liquid

[0090] 1. Experimental Procedure

[0091] 1.1 Polysaccharide Fractionation and Alcohol Precipitation of Polygonatum sibiricum Reduced Liquid

[0092] Add absolute ethanol to the Polygonatum sibiricum reduced liquid to make the ethanol volume 50% (v / v) and 85% (v / v) of the total system volume respectively. Let it stand for 24 h, centrifuge at 6000 xg for 15 min, and measure the polysaccharide content of the precipitate and supernatant respectively. Remove the supernatant, and the precipitate is the crude polysaccharide sample. Redissolve the alcohol-precipitated precipitate and freeze-dry it under vacuum at -80 °C (0 - 5 Pa) to obtain the Polygonatum sibiricum polysaccharide sample.

[0093] 1.2 Determination of Polysaccharide Molecular Weight

[0094] The high performance gel permeation chromatography (HPGPC) method was used to determine the molecular weight. The weight-average molecular weights of the Polygonatum sibiricum polysaccharides precipitated with 50% and 85% ethanol were 12.2 kDa and 3.688 kDa respectively.

[0095] 1.3 Analysis of Monosaccharide Composition of Polysaccharide

[0096] The high performance anion exchange chromatography was used to determine the monosaccharide composition of the obtained sample.

[0097] 2. Experimental Related Data

[0098] 2.1 The polysaccharide yield of Polygonatum sibiricum reduced liquid is shown in Table 1.

[0099] Table 1 Polysaccharide Yield of Polygonatum sibiricum Reduced Liquid

[0100] Alcohol precipitation concentration % (v / v) Yield of precipitated polysaccharide (%) Yield of polysaccharide in supernatant (%) Yield of total sugar (%) 50 29.91±0.11 22.20±0.03 52.11 85 26.51±0.08 31.55％±0.06 58.06

[0101] 2.2 The monosaccharide composition of the 50% alcohol-precipitated polysaccharide of Polygonatum sibiricum reduced liquid is shown in Table 2.

[0102] Table 2 Monosaccharide Composition

[0103]

[0104] Result Analysis: Combining the results of molecular weight and monosaccharide composition can prove that the Polygonatum sibiricum reduced liquid of the present invention is rich in galactan and fructooligosaccharide.

[0105] Example 3

[0106] Determination of Antioxidant Activity of Polygonatum sibiricum Reduced Liquid

[0107] 1. Experimental Procedure

[0108] 1.1 Scavenging Rate of DPPH Free Radical by Polygonatum sibiricum Reduced Liquid

[0109] Scavenging rate of polygonatum sibiricum fermented liquid on DPPH free radicals: After gradient dilution of the polygonatum sibiricum fermented liquid, it was reacted with the DPPH solution, and the absorbance was measured and the inhibition rate was calculated.

[0110] Calculation formula for DPPH free radical inhibition rate: DPPH inhibition rate (%) = (B + C - A) / B; where A is the absorbance of tube A, B is the absorbance of tube B, and C is the absorbance of tube C.

[0111] 1.2 Scavenging rate of polygonatum sibiricum fermented liquid on ABTS free radicals

[0112] After diluting the polygonatum sibiricum fermented liquid, it was reacted with the ABTS working solution, and the absorbance at 734 nm was measured and the inhibition rate was calculated.

[0113] Calculation formula for ABTS free radical inhibition rate: ABTS inhibition rate (%) = [(A0 - A) / A0] * 100%; where A0 is the absorbance of tube A0 and A is the absorbance of the test solution.

[0114] 1.3 Scavenging rate of polygonatum sibiricum fermented liquid on superoxide anions

[0115] The luminescence peak value of the diluted polygonatum sibiricum fermented liquid was measured using the pyrogallol - luminol system, and the scavenging rate was calculated.

[0116] Calculation formula for superoxide anion scavenging rate: Superoxide anion scavenging rate (%) = [(A0 - A1) / A0] * 100%

[0117] 1.4 Scavenging rate of polygonatum sibiricum fermented liquid on hydroxyl free radicals

[0118] The luminescence peak value of the diluted polygonatum sibiricum fermented liquid was measured through the H2O2 - ferrous chloride - luminol system, and the scavenging rate was calculated.

[0119] Calculation formula for hydroxyl free radical scavenging rate: Hydroxyl free radical scavenging rate (%) = [(A0 - A1) / A0] * 100%

[0120] 1.5 Scavenging rate of polygonatum sibiricum fermented liquid on hydrogen peroxide

[0121] The luminescence peak value of the diluted polygonatum sibiricum fermented liquid was measured using the H2O2 - luminol system, and the scavenging rate was calculated.

[0122] Calculation formula for hydrogen peroxide scavenging rate: Hydrogen peroxide scavenging rate (%) = [(A0 - A1) / A0] * 100%

[0123] 2. Experimental related data

[0124] 2.1 The DPPH free radical scavenging rate of polygonatum sibiricum fermented liquid is shown in Figure 2 and Table 3.

[0125] Table 3 DPPH free radical scavenging rate of polygonatum sibiricum fermented liquid

[0126]

[0127] Result analysis: From Figure 2 and Table 3, it can be seen that the fermented liquid of Polygonatum sibiricum in the present invention can significantly inhibit DPPH free radicals at a dilution multiple of less than 10, and the inhibition rate can reach more than 50%.

[0128] 2.2 The ABTS free radical scavenging rate of the fermented liquid of Polygonatum sibiricum is shown in Figure 3 and Table 4.

[0129] Table 4 ABTS free radical scavenging rate of the fermented liquid of Polygonatum sibiricum

[0130]

[0131]

[0132] Result analysis: From Figure 3 and Table 4, it can be seen that the inhibition rate of the ABTS free radicals of the fermented liquid of Polygonatum sibiricum in the present invention can reach more than 20% at a dilution multiple of less than 10.

[0133] 2.3 The superoxide anion scavenging rate of the fermented liquid of Polygonatum sibiricum is shown in Figure 4 and Table 5.

[0134] Table 5 Superoxide anion scavenging rate of the fermented liquid of Polygonatum sibiricum

[0135] Original solution of Polygonatum sibiricum Delar. ex Redoute fermentation broth Diluted 5 times Diluted 10 times Diluted 20 times Diluted 40 times Diluted 80 times Diluted 160 times 97.72％ 75.91％ 52.55％ 31.11％ 18.95％ 12.33％ 5.71％

[0136] Result analysis: From Figure 4 and Table 5, it can be seen that the superoxide anion scavenging rate of the fermented liquid of Polygonatum sibiricum in the present invention can reach more than 50% at a dilution multiple of less than 10.

[0137] 2.4 The hydroxyl radical scavenging rate of the fermented liquid of Polygonatum sibiricum is shown in Figure 5 and Table 6.

[0138] Table 6 Hydroxyl radical scavenging rate of the fermented liquid of Polygonatum sibiricum

[0139] Original solution of Polygonatum sibiricum Delar. ex Redoute fermentation broth Diluted 5 times Diluted 10 times Diluted 20 times Diluted 40 times Diluted 80 times Diluted 160 times 71.45％ 29.96％ 19.18％ 14.57％ 12.39％ 5.96％ 4.25％

[0140] Result analysis: From Figure 5 and Table 6, it can be seen that the superoxide anion scavenging rate of the fermented liquid of Polygonatum sibiricum in the present invention can reach more than 19% at a dilution multiple of less than 10.

[0141] 2.5 The hydrogen peroxide scavenging rate of the fermented liquid of Polygonatum sibiricum is shown in Figure 6 and Table 7.

[0142] Table 7 Hydrogen peroxide scavenging rate of the fermented liquid of Polygonatum sibiricum

[0143] Original solution of Polygonatum sibiricum Delar. ex Redoute fermentation broth Diluted 5 times Diluted 10 times Diluted 20 times Diluted 40 times Diluted 80 times Diluted 160 times 95.76％ 92.43％ 88.63％ 81.31％ 67.76％ 23.58％ 0

[0144] Result analysis: As can be seen from Figure 6 and Table 7, the superoxide anion scavenging rate of the fermented liquid of Polygonatum sibiricum in this invention can reach more than 60% when the dilution multiple is below 40.

[0145] Example 4

[0146] Cytotoxicity and repair ability of the fermented liquid of Polygonatum sibiricum on HSF cells

[0147] 1. Experimental process

[0148] The promoting effects on the viability, migration rate and ultraviolet damage repair of HSF cells were evaluated by MTT method and scratch experiment.

[0149] 2. Experimental related data

[0150] 2.1 Cytotoxicity results of the fermented liquid of Polygonatum sibiricum

[0151] The cytotoxic effects of the fermented liquid of Polygonatum sibiricum at different volume fractions (the fermented liquid of Polygonatum sibiricum was diluted to different concentrations with PBS: 0.20%, 0.39%, 0.78%, 1.56%, 3.13%, 6.25%, 12.5%, 25%, 50%) on normal HSF cells were detected by MTT method, and the results are as Figure 7 shown in Table 8.

[0152] Table 8 Cytotoxicity results

[0153]

[0154] Result analysis: When the concentration of the fermented liquid of Polygonatum sibiricum is lower than 1.56%, it has a certain promoting effect on the viability of HSF cells. As the concentration of the fermented liquid of Polygonatum sibiricum increases, the survival rate of HSF cells gradually decreases and shows a dose-dependence. When the concentration of the fermented liquid of Polygonatum sibiricum is 50%, the cell survival rate is above 80%. The concentration of 50% fermented liquid of Polygonatum sibiricum was selected for further study on the ability to promote the scratch healing of HSF cells.

[0155] 2.2 The results of the cell scratch repair ability of the fermented liquid of Polygonatum sibiricum are shown in Table 9 and Figure 8 .

[0156] Table 9 Results of the cell scratch repair ability of the fermented liquid of Polygonatum sibiricum

[0157] Before repair μm Repaired for 24 h μm Migration rate (%) Polygonatum sibiricum Delar. ex Redoute fermentation broth (A3) 626.6 509.1 0.187519949 Blank (A1) 482.7 423.6 0.122436296

[0158] Result analysis: The cell scratch repair results of the fermented liquid of Polygonatum sibiricum ( Figure 8 ), the promoting migration rate of the 50% fermented liquid of Polygonatum sibiricum on the scratch of human skin fibroblasts is 0.19%, which is higher than 0.12% of the blank group. It shows that the fermented liquid containing fructooligosaccharides and galactan of Polygonatum sibiricum can promote the proliferation and migration of fibroblasts, and thus promote the repair of skin wounds.

[0159] 2.3 The ability of the polygonatum fermented liquid to repair photoaging damage to cells is shown in Figure 9 and Figure 10 .

[0160] Result analysis: The results of repairing photoaging damage to cells by the polygonatum fermented liquid ( Figure 9 and Figure 10 ). By comparing the model group and the 50% polygonatum fermented liquid group, it was found that the G0 / G1 phase of the model group was significantly prolonged, proving that UVB caused damage to the cells and induced cell cycle arrest, indicating that the cell model was successfully established. Compared with the blank group, there was no significant change in the peak value of the G0 / G1 phase after treatment with 50% polygonatum fermented liquid, that is, the fermented liquid containing fructooligosaccharide and galactan of polygonatum had a certain repairing effect on the photoaging damage of human skin fibroblasts.

[0161] Example 5

[0162] Anti-inflammatory activity of the polygonatum fermented liquid

[0163] 1. Experimental procedure

[0164] The inhibition rate was determined by the respiratory burst experiment of RAW264.7 cells.

[0165] The formula for calculating the inhibition rate of the fermented liquid on the respiratory burst is:

[0166]

[0167] 2. Experimental relevant data

[0168] The inhibition rate of the fermented liquid on the respiratory burst was 38.76%.

[0169] Example 6

[0170] Moisturizing properties of the polygonatum fermented liquid containing fructooligosaccharide and galactan of polygonatum

[0171] 1. Experimental procedure

[0172] The polygonatum fermented liquid was applied to the arms of 10 subjects aged 20 - 24 years old, and the skin water content and transepidermal water loss rate were measured.

[0173] 2. Experimental relevant data

[0174] 2.1 Changes in the skin water content after applying the fermented liquid are shown in Figure 11 .

[0175] 2.2 Changes in the transepidermal water loss rate after applying the fermented liquid are shown in Figure 12 .

[0176] Result analysis: Within 1 hour after application, the skin moisture content was the highest when applying 20% polygonatum fermented liquid. Compared with the blank group (water), the skin moisture content was significantly increased when applying 10% and 20% polygonatum fermented liquid, with increases of 9.24% and 26.7% respectively. After 5 hours of application, the skin moisture content of applying 10% and 20% polygonatum fermented liquid was still increased by 7.33% and 10.56% respectively compared with the blank group, that is, the polygonatum fermented liquid had a certain moisturizing effect. At 2 hours after application, the improvement of transepidermal water loss was the most obvious, and the water loss rates after applying 10% and 20% polygonatum fermented liquid were reduced by 9.71% and 7.64% respectively compared with the blank group.

[0177] Example 7

[0178] Effect of polygonatum fermented liquid on specific skin flora

[0179] 1. Experimental process

[0180] The proliferation promotion rate of Staphylococcus epidermidis and the inhibition rate of Propionibacterium acnes were determined through the bacterial suspension culture experiment.

[0181]

[0182] 2. Experimental related data

[0183] 2.1 The effect of the fermented liquid on Staphylococcus epidermidis is shown in Figure 13 .

[0184] 2.2 The effect of the fermented liquid on Propionibacterium acnes is shown in Figure 14 .

[0185] The results showed that the fermented liquid could promote the proliferation of Staphylococcus epidermidis ( Figure 13 ), compared with the blank group, 25% of the fermented liquid could promote the proliferation by 31.98%. And Staphylococcus epidermidis could affect the growth of conditional pathogenic bacteria on the skin and indirectly improve the skin hydration. That is, the polygonatum fermented liquid containing fructooligosaccharide and galactan could promote the proliferation of the skin symbiotic bacterium Staphylococcus epidermidis.

[0186] The fermented liquid had a certain inhibitory effect on Propionibacterium acnes ( Figure 14 ), and the inhibition rate of 25% of the fermented liquid on Propionibacterium acnes was 17.19%. That is, the polygonatum fermented liquid containing fructooligosaccharide and galactan could regulate specific skin flora to a certain extent and could be applied to cosmetics.

[0187] Example 8

[0188] Preparation of polygonatum ferment repair essence

[0189] 1. Process flow

[0190] In the first step, heat water to about 80 °C, and successively add dipotassium glycyrrhizinate, acrylate / C10-30 alkyl acrylate cross-linked polymer, glycerol, p-hydroxyacetophenone, butanediol, Tremella fuciformis fruit body extract, sodium hyaluronate (wherein butanediol, Tremella fuciformis fruit body extract and sodium hyaluronate need to be dispersed evenly in advance), and homogenize and stir at 3000 rpm for 5 min until completely dispersed evenly;

[0191] In the second step, cool the above liquid to 45 °C, add arginine, and stir until completely dispersed evenly;

[0192] In the third step, wait for the liquid to cool to 40 °C, successively add the polygonatum fermented liquid, 1,2-hexanediol, trimethylpentanediol / adipic acid / glycerol cross-linked polymer, and PEG-7 olive oil ester, and stir until completely dispersed evenly, then take a sample to measure physical and chemical properties;

[0193] In the fourth step, send the prepared sample for quality inspection, and discharge it after passing the quality inspection; carry out packaging into boxes to obtain the finished product.

[0194] 2. Experimental related data

[0195] 2.1 The formula of the polygonatum fermented repair essence is shown in Table 10.

[0196] Table 10 Formula table of polygonatum essence

[0197]

[0198]

[0199] By weight, the usage amounts of various raw materials for 500 g of the sample are shown in Table 10.

[0200] At the same time, replace the polygonatum fermented liquid in the raw materials with distilled water, and prepare a blank essence solution group under the same process conditions as a control.

[0201] 2.2 The process flow chart of the preparation of the polygonatum fermented repair essence (polygonatum essence) is shown in Figure 15 .

[0202] The sensory and physical and chemical indexes of the polygonatum essence are shown in Table 11. According to Table 11, the obtained polygonatum essence has good cosmetic properties.

[0203] Table 11 Sensory and physical and chemical indexes of polygonatum essence

[0204]

[0205] Example 9

[0206] In vitro percutaneous absorption experiment of polygonatum fermented repair essence

[0207] 1. Experimental procedure

[0208] The cumulative permeation amount and permeation kinetic parameters of polysaccharides in porcine skin were determined using a Franz diffusion cell.

[0209] 2. Experimental related data

[0210] 2.1 The changes in the cumulative permeation amount of the fermented polygonatum cirrhifolium repair essence (hereinafter referred to as polygonatum essence) at different time points are shown in Figure 16 .

[0211] 2.2 The in vitro permeation kinetic parameters of the fermented polygonatum cirrhifolium repair essence are shown in Table 12.

[0212] Table 12 In vitro permeation kinetic parameters of polygonatum essence

[0213] Sample Jss (μg / h·cm2) Kp (cm / h) Tlag / h Polygonatum sibiricum Delar. ex Redoute essence 0.378 0.00004834 6.439153439 Blank essence 0.236 0.00003018 14.69491525

[0214] Result analysis: After adding the fermented polygonatum cirrhifolium liquid, the cumulative permeation amount, steady-state transdermal rate, and permeability coefficient of the fermented polygonatum cirrhifolium repair essence of the present invention are all increased compared with the blank fermented liquid. The cumulative permeation amount of the polygonatum essence reaches 11.75 mg / mL at 24 h.

[0215] Example 10

[0216] Effect of polygonatum essence on the viability of HaCaT cells and determination of its ability to promote cell scratch healing

[0217] 1. Experimental procedure

[0218] Its effects on the viability and migration rate of HaCaT cells were evaluated by MTT assay and scratch assay.

[0219] 2. Experimental related data

[0220] 2.1 The cytotoxic effects of polygonatum essence at different volume fractions (the polygonatum essence was diluted to different concentrations with PBS: 0.39%, 0.78%, 1.56%, 3.13%, 6.25%, 12.5%, 25%) on normal HaCaT cells were detected by MTT assay, and the results are as Figure 17 shown in and Table 13.

[0221] Table 13 Cytotoxicity results

[0222]

[0223] Results: When the concentration of Polygonatum sibiricum essence was lower than 3.13%, it had a certain promoting effect on the viability of HaCaT cells. With the increase of the concentration of Polygonatum sibiricum essence, the survival rate of HaCaT cells gradually decreased and was dose-dependent. When the concentration of Polygonatum sibiricum essence was between 3.13% and 12.5%, the cell survival rate was between 90% and 100%. The concentration of Polygonatum sibiricum essence with a cell survival rate of more than 90% was selected to further study the ability of promoting HaCaT cell scratch healing.

[0224] 2.2 Results of HaCaT cell scratch test with Polygonatum sibiricum essence Figure 18 As shown in Table 14.

[0225] Table 14 Results of HaCaT cell scratch test with Polygonatum sibiricum essence

[0226]

[0227]

[0228] 2.3 Effects of different concentrations of Polygonatum sibiricum extract on the migration rate of HaCaT cells Figure 19 shown.

[0229] Result analysis: According to the results of the HaCaT cell scratch test of the polygonatum sibiricum essence, the polygonatum sibiricum essence of the present invention can promote cell migration and achieve the effect of repairing scratches.

[0230] Embodiment 11

[0231] Safety evaluation of Polygonatum sibiricum essence——Human patch test

[0232] 1. Experimental Procedure

[0233] A total of 46 subjects who met the volunteer selection criteria were selected, with an age distribution of 18 to 58 years old. After the volunteers were stabilized under constant temperature and humidity conditions for half an hour, the spot tester was attached to the curved side of the volunteer's forearm, 0.02g of 30% Polygonatum sibiricum essence was weighed into the spot tester, and gently pressed to make it stick to the skin. After 24 hours, the spot tester was removed, and the experimental results were observed and recorded after stabilization for 30 minutes. The results were recorded according to the skin reaction grading standards in the "Cosmetic Safety Technical Specifications 2015 Edition".

[0234] 2. Experimental related data

[0235] The safety of Polygonatum sibiricum essence was evaluated through patch tests. The experimental results showed that none of the 46 test subjects showed symptoms such as light red spots, erythema, edematous erythema, and significant edema, indicating that the essence is safe and has no obvious irritation to human skin.

[0236] Although the above embodiments have described the present invention in detail, they are only a part of the embodiments of the present invention, rather than all of them. People can also obtain other embodiments based on these embodiments without creative efforts, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A method for preparing a polygonatum fermented liquid, characterized in that, It includes the following steps: Mix polygonatum sibiricum powder with water at a mass ratio of 1:(0.8 - 1.2) to obtain wet-based polygonatum sibiricum powder; Inoculate aspergillus oryzae into the wet-based polygonatum sibiricum powder for solid-state fermentation, and then dry it to obtain solid-state fermented polygonatum sibiricum; the inoculation amount of aspergillus oryzae is 0.05 - 1%; the conditions for solid-state fermentation are 25 - 32°C for 24 - 72 h; Mix the solid-state fermented polygonatum sibiricum with water at a mass ratio of 1:(3 - 5), extract with hot water, and filter by suction to obtain polygonatum fermented filtrate; the conditions for hot water extraction are 30 - 50°C for 2 - 5 h; Sterilize the polygonatum fermented filtrate, filter it, and centrifuge to take the supernatant to obtain polygonatum fermented liquid.

2. The polygonatum fermented liquid prepared by the preparation method described in claim 1.

3. The application of the polygonatum fermented liquid prepared by the preparation method described in claim 1 in the preparation of a product having any one or more functions of ① - ⑤: ① Accelerate skin injury repair; the acceleration of skin injury repair includes accelerating the repair of skin photoaging injury and / or accelerating the repair of skin cell scratch; ② Antioxidation; ③ Moisturizing; ④ Anti-inflammatory; ⑤ Regulate skin flora; the regulation of skin flora includes promoting the proliferation of staphylococcus epidermidis and / or inhibiting propionibacterium acnes.

4. The application according to claim 3, characterized in that, The product includes cosmetics.

5. An essence containing the polygonatum fermented liquid prepared by the preparation method described in claim 1, characterized in that, It includes the following components in parts by weight: 50 - 60 parts of water, 0.08 - 0.12 parts of dipotassium glycyrrhizinate, 0.2 - 0.4 parts of acrylate / C10 - 30 alkyl acrylate cross-linked polymer, 3 - 5 parts of glycerol, 0.4 - 0.5 parts of p-hydroxyacetophenone, 2 - 4 parts of butanediol, 0.02 - 0.04 parts of tremella heteropolysaccharide, 0.01 - 0.03 parts of sodium hyaluronate, 0.2 - 0.3 parts of arginine, 0.4 - 0.5 parts of 1,2 - hexanediol, 1 - 2.5 parts of trimethylpentanediol / adipic acid / glycerol cross-linked polymer, 0.6 - 1 part of PEG - 7 olive oil ester, and 20 - 30 parts of the polygonatum fermented liquid prepared by the preparation method described in claim 1.

6. The preparation method of the essence according to claim 5, characterized in that, It includes the following steps: First mix dipotassium glycyrrhizinate, acrylate / C10 - 30 alkyl acrylate cross-linked polymer, glycerol, p-hydroxyacetophenone, butanediol, tremella heteropolysaccharide, sodium hyaluronate and water at 75 - 80°C, and stir to obtain a first intermediate product; Cool the first intermediate product to 45 - 50°C, and second mix the first intermediate product with arginine and stir to obtain a second intermediate product; Cool the second intermediate product to 35 - 40°C, and third mix the second intermediate product with the polygonatum fermented liquid, 1,2 - hexanediol, trimethylpentanediol / adipic acid / glycerol cross-linked polymer and PEG - 7 olive oil ester and stir to obtain the essence.

7. The application of the essence described in claim 5 or the essence prepared by the preparation method described in claim 6 in the preparation of skin-regulating and repairing cosmetics.

8. The application according to claim 7, characterized in that, The skin-regulating and repairing cosmetics have good transdermal absorption effect, can promote cell vitality and can promote the ability of cell scratch healing.

Images