A Yunnan Paris polyphylla extract with scalp inflammatory symptom repairing function and its application

Abstract

The present invention belongs to the technical field of external skin preparations, and specifically relates to a Yunnan Paris extract with a function of repairing inflammatory symptoms of the scalp and its application. The present invention crushes and extracts dried Yunnan Paris, and then undergoes one-stage enzymolysis, two-stage enzymolysis, and three-stage enzymolysis. The enzymolysis product is inoculated with Aspergillus oryzae CICC 40188 and Bacillus subtilis GDMCC NO: 1.1750 for fermentation, and then ethanol is added to adjust the concentration for extraction, and filtered and ultrafiltered using an ultrafiltration membrane with a molecular weight cutoff of less than 2000Da, so that the types and concentrations of anti-inflammatory active ingredients in the Yunnan Paris extract are improved, and the obtained extract has good anti-inflammatory and maintenance and repair effects on scalp inflammation. At the same time, the prepared Yunnan Paris extract is matched with a kelp extract, which can effectively remove free radicals and has good antioxidant effects.

Description

Technical Field

[0001] The invention belongs to the technical field of external skin preparations, and specifically relates to a Paris polyphylla extract with a function of repairing scalp inflammatory symptoms and an application thereof. Background Art

[0002] Scalp inflammation is mainly caused by seborrheic dermatitis, which is a chronic inflammatory skin disease. Seborrheic dermatitis often begins to appear after puberty and recurs repeatedly. The clinical manifestations are mainly greasy scales and erythema on the scalp, accompanied by itching. The cause of seborrheic dermatitis is still unclear, but several major factors related to the onset are known: Malassezia colonization, excessive sebum secretion, genetic susceptibility and immune factors.

[0003] Yunnan Paris polyphylla Paris polyphylla yunnanensis ) is also called Yunnan Paris, Caoheche, and Single-legged Lotus. The nutritional components of Yunnan Paris are very rich, and contain a variety of steroidal saponins. Yunnan Paris has the effects of clearing away heat and detoxifying, reducing swelling and relieving pain, cooling the liver and calming the nerves, and has a good effect on diseases such as carbuncle, sore throat, snake bites, and falls. The invention patent with publication number CN114191359A discloses a shower gel containing Yunnan Paris extract and a preparation method thereof, which records that adding Yunnan Paris extract to the formula can make the shower gel have good antibacterial and disinfection effects.

[0004] As people pay more attention to the concept of environmental protection and desire to return to nature, plant herbal care has attracted people's attention, and the skin topical preparations or cosmetics industry are becoming more and more natural, effective, mild and safe. Therefore, developing a Yunnan Paris polyphylla plant extract with scalp inflammation repair as a raw material to deal with the problems caused by scalp inflammation and to realize its application in various scalp care products is a technical problem that needs to be solved at present. Summary of the invention

[0005] The present invention aims at the above-mentioned technical problems and provides a solution of the present invention. The solution of the present invention relates to the technical field of skin external preparations, and specifically to a Yunnan Paris polyphylla extract having the function of repairing scalp inflammatory symptoms and its application.

[0006] Specifically, the Paris polyphylla extract is prepared by the following steps:

[0007] S1: Crush the dried rhizome of Paris polyphylla to obtain rhizome powder, add water 15-30 times the mass of the rhizome powder, heat and stir for 4-8 hours to obtain a stirred material; cool and keep warm, add 0.5-5% cellulase and 0.4-4.5% pectinase by mass of the rhizome powder to perform enzymolysis for 3-6 hours to obtain enzymolyzate 1.

[0008] S2: inactivate the enzyme in the enzymolysate 1 to obtain the inactivated material 1, add 0.8-3.5% of the weight of the rhizome powder with a neutral protease, perform enzymolysis for 2.5-5 hours, and obtain the enzymolysate 2.

[0009] S3: The enzyme of the enzymolyzate 2 is inactivated to obtain the inactivated material 2, and then 0.7-4% of papain by weight of the rhizome powder is added, and the enzymolysis is performed for 3-5.5 hours to obtain the enzymolyzate 3.

[0010] S4: inactivate the enzyme in the hydrolyzate 3 to obtain material A, add 3-10% carbon source by weight of the rhizome powder and inoculate fermentation bacteria to obtain material B, and ferment at a temperature of 25-35° C. for 24-60 hours to obtain a fermented material.

[0011] S5: The fermentation material is sterilized, and then anhydrous ethanol is added, heated to 40-55° C., stirred for 4-8 hours, and filtered to obtain filtrate 1.

[0012] S6: Filter the filtrate 1 using an ultrafiltration membrane with a molecular weight cutoff lower than 2000Da to obtain a filtrate 2, which is then concentrated and dried to obtain the Paris polyphylla extract.

[0013] Furthermore, the inoculated bacteria are Aspergillus oryzae CICC 40188 and Bacillus subtilis GDMCC NO: 1.1750.

[0014] Furthermore, the concentration of the inoculum in material B is 2.5×10 8 -9.7×10 8 cfu / g.

[0015] Furthermore, the bacterial quantity ratio of Aspergillus oryzae CICC 40188 and Bacillus subtilis GDMCC NO: 1.1750 in material B is 1:0.3-0.8.

[0016] Furthermore, in S5, the amount of anhydrous ethanol added is 0.5-1 times the mass of water added in S1.

[0017] Furthermore, in S1, the powder is crushed to a particle size less than 1 mm.

[0018] Furthermore, in said S1, the temperature is heated to 45-65°C.

[0019] Furthermore, in S1, the stirring speed is 40-400 rpm.

[0020] Furthermore, in S1, the temperature is cooled to 35-42°C.

[0021] Furthermore, in said S1, the temperature of enzymatic hydrolysis is 35-50°C.

[0022] Furthermore, in S1, the enzymatic hydrolysis process is carried out at a rotation speed of 40-400 rpm.

[0023] Furthermore, in S2, enzymolysis is performed at a temperature of 35-42°C.

[0024] Furthermore, in S2, the enzymatic hydrolysis process is carried out at a rotation speed of 40-400 rpm.

[0025] Furthermore, in S3, enzymolysis is performed at a temperature of 38-45°C.

[0026] Furthermore, in S3, the enzymatic hydrolysis process is carried out at a rotation speed of 40-400 rpm.

[0027] Furthermore, in S4, the carbon source includes at least one of sucrose, glucose, fructose and maltose.

[0028] Furthermore, in S5, the filtration is performed using a filter membrane with a pore size of 0.5-10 microns.

[0029] Furthermore, in S6, filtration is performed using an ultrafiltration membrane with a molecular weight cutoff of 1200 Da.

[0030] Furthermore, in S6, the concentration is performed under reduced pressure, the concentration temperature is 58-62° C., and the concentration is performed to 10-20% of the mass of the filtrate 2.

[0031] Furthermore, in S6, the drying is freeze drying, the freeze drying temperature is -40°C to -28°C, and the freeze drying is performed until the water content is less than 5.5wt%.

[0032] On the other hand, the present invention also provides an application of the Paris polyphylla extract, including using the Paris polyphylla extract to prepare a preparation having anti-inflammatory and free radical scavenging functions.

[0033] Furthermore, the preparation is a skin external medicine or a cosmetic type preparation.

[0034] Furthermore, the preparation consists of the above-mentioned Paris polyphylla extract, Laminaria japonica extract, ethylene glycol, glycerin, a preservative and water.

[0035] Furthermore, the preparation is composed of the following components in terms of mass: 0.1-1 parts of the above-mentioned Paris polyphylla extract, 0.05-0.6 parts of Laminaria japonica extract, 2-5 parts of ethylene glycol, 3-7 parts of glycerol, 0.05-0.3 parts of preservatives and 30-50 parts of water.

[0036] Furthermore, the mass ratio of the Paris polyphylla extract to the Laminaria lappa extract is 1:0.4-0.7.

[0037] Furthermore, the preparation method of the kelp extract comprises the following steps:

[0038] Step 1: Grind the dried kelp and pass it through a 50-mesh sieve to obtain kelp powder;

[0039] Step 2: placing kelp powder and ethanol aqueous solution with a mass concentration of 20-40% in a container at a mass ratio of 1:12-40, heating to 35-55° C., and stirring for 6-12 hours to obtain a mixture;

[0040] Step 3: Filter the mixture using a 3-micron pore size filter membrane to obtain filtrate 1, and then filter the filtrate 1 using an ultrafiltration membrane with a molecular weight cutoff of 1200Da to obtain filtrate 2. Concentrate filtrate 2 and freeze-dry it until the water content is less than 6.8%, thereby obtaining the kelp extract.

[0041] The preparation method of the preparation comprises the steps of mixing and sterilizing the raw materials.

[0042] The beneficial effects of the present invention are as follows:

[0043] The invention crushes and extracts dried parsley from Yunnan and then performs one-stage enzymolysis, two-stage enzymolysis and three-stage enzymolysis, inoculates the enzymolysis product with Aspergillus oryzae CICC 40188 and Bacillus subtilis GDMCC NO: 1.1750 for fermentation, adds ethanol to adjust the concentration for extraction, and performs filtering and ultrafiltration using an ultrafiltration membrane with a molecular weight cutoff lower than 2000 Da, so that the types and concentrations of anti-inflammatory active ingredients in the parsley from Yunnan are improved, and the obtained extract has good anti-inflammatory and maintenance and repair effects on scalp inflammation.

[0044] The Paris polyphylla extract of the present invention can reduce the TNF-α content in the serum of sensitized mice. The Paris polyphylla extract prepared by the present invention has a good anti-inflammatory effect and has a protective function on inflammatory symptoms of the scalp.

[0045] The fermentation process of the present invention can improve the anti-inflammatory effect of the extract. Furthermore, by using Aspergillus oryzae CICC 40188 and Bacillus subtilis GDMCC NO: 1.1750 for joint fermentation, the types and contents of anti-inflammatory active ingredients are improved, thereby improving the anti-inflammatory effect of the extract. In the fermentation process of the present invention, the combination of the two bacteria has a synergistic effect in promoting the anti-inflammatory effect of the extract. And in the preparation system of the present invention, using a ratio of 1:0.3-0.8 of the number of the two bacteria can bring better anti-inflammatory effects.

[0046] When preparing the Paris polyphylla extract of Yunnan, the present invention uses ethanol to adjust the ethanol concentration in the extraction solvent, and uses an ultrafiltration membrane with a molecular weight cutoff of 1200Da for filtration, both of which can increase the anti-inflammatory effect of the extract.

[0047] The preparation prepared by the combination of the Yunnan Paris polyphylla extract prepared by the present invention with a kelp extract, a polyol component, etc. has a good free radical scavenging effect; after the preparation is diluted to a concentration of 25wt%, it can scavenge 92.78% of DPPH free radicals and 86.17% of superoxide free radicals, has a good effect of eliminating human free radicals, and imparts an anti-aging effect to the skin.

[0048] The present invention uses an ethanol aqueous solution with a mass concentration of 20-40% to extract kelp, and the obtained extraction can better improve the free radical scavenging effect of the preparation and give the skin a better anti-aging function; and when extracting the kelp, an ultrafiltration membrane with a molecular weight cutoff of 1200Da is used for treatment, which can further improve the efficiency of the preparation in scavenging DPPH free radicals and superoxide free radicals.

[0049] When the preparation is prepared by the present invention, the Paris polyphylla extract and the Laminaria lappa extract prepared by the present invention are used to mutually promote the improvement of the antioxidant effect of the preparation, and the combination of the two has a synergistic effect in increasing the effect of the preparation in eliminating DPPH free radicals and superoxide free radicals.

[0050] The preparation prepared by the invention has excellent stability and good heat resistance and cold resistance effects.

[0051] The preparation method of the invention is simple, green and environmentally friendly, and the prepared product has good performance and good application prospects. DETAILED DESCRIPTION

[0052] The technical solution of the present invention will be described below in conjunction with specific implementation methods, and the effects of the technical solution of the present invention will be made clearer and more specific.

[0053] 1. Yunnan Paris polyphylla extract and its effect test

[0054] 1. Preparation of Paris polyphylla extract

[0055] Cellulase: 20,000 U / g, Hubei Chengfeng Chemical Co., Ltd.

[0056] Pectinase: 20,000 U / g, Shandong Fangchang Biotechnology Co., Ltd.

[0057] Neutral protease: 100,000 U / g, Shandong Xinxiong Biotechnology Co., Ltd.

[0058] Papain: 100,000 U / g, Xi'an Tianzhixiang Biotechnology Co., Ltd.

[0059] Aspergillus oryzae CICC 40188: The strain number is CICC 40188, and its Latin name is Aspergillus oryzae , purchased from China Food Fermentation Industries Research Institute Co., Ltd.

[0060] Bacillus subtilis GDMCC NO: 1.1750: The strain number is GDMCC 1.1750, and its Latin name is Bacillus subtilis subsp. Subtilis , purchased from Guangdong Microbiological Culture Collection Center.

[0061] Yunnan Paris Extract I:

[0062] The preparation method is as follows:

[0063] S1: Crush the dried rhizomes of Paris polyphylla to a particle size of less than 1 mm to obtain rhizome powder, add water 20 times the mass of the rhizome powder, heat to 50°C and stir at a speed of 75 rpm for 6 hours to obtain a stirred material; cool to 37°C and keep warm, add 2.4% of cellulase and 3.2% of pectinase by mass of the rhizome powder, and perform enzymolysis at a temperature of 37°C and a stirring speed of 65 rpm for 4.5 hours to obtain an enzymolyzate 1;

[0064] S2: Inactivate the enzyme in the enzymatic hydrolysate 1 to obtain the inactivated material 1, add 2.7% of the weight of the rhizome powder neutral protease, and perform enzymolysis for 4 hours at a temperature of 38°C and a rotation speed of 60 rpm to obtain the enzymatic hydrolysate 2;

[0065] S3: inactivate the enzyme in the hydrolysate 2 to obtain the inactivated material 2, then add 2.8% papain by weight of the rhizome powder, and perform enzymolysis for 4.5 hours at a temperature of 40°C and a rotation speed of 75 rpm to obtain the hydrolysate 3;

[0066] S4: inactivate the enzyme in the hydrolyzate 3 to obtain material A, add 6.2% of the carbon source glucose by weight of the rhizome powder and inoculate fermentation bacteria to obtain material B, and ferment at 30°C for 48 hours to obtain a fermented material;

[0067] S5: sterilize the fermentation material, then add anhydrous ethanol, heat to 45°C and stir for 7 hours, filter using a filter membrane with a pore size of 2 microns to obtain filtrate 1;

[0068] S6: Filter the filtrate 1 using an ultrafiltration membrane with a molecular weight cutoff of 1200Da to obtain filtrate 2, and then concentrate under reduced pressure at a concentration temperature of 60°C to 15.4% of the mass of the filtrate 2, and freeze-dry at a temperature of -35°C to a water content of 4.92wt%, to obtain the Paris polyphylla extract I.

[0069] Wherein, the inoculated bacteria are composed of Aspergillus oryzae CICC 40188 and Bacillus subtilis GDMCC NO: 1.1750 in a bacterial quantity ratio of 1:0.5.

[0070] The concentration of the inoculum in material B is 7.2×10 8 cfu / g.

[0071] In S5, the amount of anhydrous ethanol added is 0.8 times the mass of water added in S1.

[0072] Yunnan Paris Extract II:

[0073] The preparation method is as follows:

[0074] S1: Crush the dried rhizome of Paris polyphylla to a particle size of less than 1 mm to obtain rhizome powder, add water 25 times the mass of the rhizome powder, heat to 45°C and stir at a speed of 80 rpm for 4.5 hours to obtain a stirred material; cool to 37°C and keep warm, add 4.5% of cellulase and 2.5% of pectinase by mass of the rhizome powder, perform enzymolysis at a temperature of 37°C and a stirring speed of 70 rpm for 5.5 hours to obtain an enzymolysate 1;

[0075] S2: Inactivate the enzyme in the enzymatic hydrolysate 1 to obtain the inactivated material 1, add 1.2% of the weight of the rhizome powder neutral protease, and perform enzymolysis for 3 hours at a temperature of 40°C and a rotation speed of 80 rpm to obtain the enzymatic hydrolysate 2;

[0076] S3: inactivate the enzyme in the hydrolysate 2 to obtain the inactivated material 2, then add 3.5% papain by weight of the rhizome powder, and perform enzymolysis for 3.8 hours at a temperature of 42° C. and a rotation speed of 80 rpm to obtain the hydrolysate 3;

[0077] S4: inactivate the enzyme in the hydrolyzate 3 to obtain material A, add 7.2% of the carbon source glucose by weight of the rhizome powder and inoculate fermentation bacteria to obtain material B, and ferment at 32°C for 50 hours to obtain a fermented material;

[0078] S5: sterilize the fermentation material, then add anhydrous ethanol, heat to 50°C and stir for 5.5 hours, filter using a filter membrane with a pore size of 2 microns to obtain filtrate 1;

[0079] S6: Filter the filtrate 1 using an ultrafiltration membrane with a molecular weight cutoff of 1200Da to obtain filtrate 2, and then concentrate under reduced pressure at a concentration temperature of 60°C to 13.7% of the mass of the filtrate 2, and freeze-dry at a temperature of -40°C to a water content of 5.07wt%, to obtain the Paris polyphylla extract II.

[0080] Wherein, the inoculated bacteria are composed of Aspergillus oryzae CICC 40188 and Bacillus subtilis GDMCC NO: 1.1750 in a bacterial quantity ratio of 1:0.3.

[0081] The concentration of the inoculum in material B is 7.28×10 8 cfu / g.

[0082] In S5, the amount of anhydrous ethanol added is 0.5 times the mass of water added in S1.

[0083] Yunnan Paris polyphylla extract III: The difference from Yunnan Paris polyphylla extract I is that fermentation bacteria are not inoculated in S4, and the rest are the same as Yunnan Paris polyphylla extract I.

[0084] Yunnan Paris polyphylla extract IV: The difference from Yunnan Paris polyphylla extract I is that an equal amount of water is used to replace anhydrous ethanol in S5, and the rest is the same as Yunnan Paris polyphylla extract I.

[0085] Yunnan Paris polyphylla extract V: The difference from Yunnan Paris polyphylla extract I is that no ultrafiltration membrane filtration is performed in S6, and the rest is the same as Yunnan Paris polyphylla extract I.

[0086] Yunnan Paris polyphylla extract VI: The difference from Yunnan Paris polyphylla extract I is that the amount of anhydrous ethanol used in S5 is twice the mass of water added in S1, and the rest is the same as Yunnan Paris polyphylla extract I.

[0087] Yunnan Paris polyphylla extract VII: The difference from Yunnan Paris polyphylla extract I is that the bacteria inoculated in S4 is only Aspergillus oryzae CICC 40188 and the concentration in material B is 7.2×10 8 cfu / g, and the others were the same as those of Paris polyphylla extract I.

[0088] Yunnan Paris polyphylla extract VIII: The difference from Yunnan Paris polyphylla extract I is that the bacteria inoculated in S4 is only Bacillus subtilis GDMCC NO: 1.1750 and the concentration in material B is 7.2×10 8 cfu / g, and the others were the same as those of Paris polyphylla extract I.

[0089] Yunnan Paris polyphylla extract IX: The difference from Yunnan Paris polyphylla extract I is that the inoculum is composed of Aspergillus oryzae CICC 40188 and Bacillus subtilis GDMCC NO: 1.1750 in a bacterial ratio of 1:0.1 and the concentration of the inoculum in material B is 7.2×10 8 cfu / g, and the others were the same as those of Paris polyphylla extract I.

[0090] Yunnan Paris polyphylla extract X: The difference from Yunnan Paris polyphylla extract I is that the inoculum is composed of Aspergillus oryzae CICC 40188 and Bacillus subtilis GDMCC NO: 1.1750 in a bacterial ratio of 1:1.5 and the concentration of the inoculum in material B is 7.2×10 8 cfu / g, and the others were the same as those of Paris polyphylla extract I.

[0091] 2. Effect test of Yunnan Paris polyphylla extract

[0092] TNF-α is a pro-inflammatory factor, and the generation of inflammation will cause an increase in the concentration of TNF-α; the present invention tests the concentration of TNF-α in mouse serum to verify the effect of the Yunnan Paris polyphylla extract of the present invention in inhibiting inflammation.

[0093] 130 healthy BALB / c mice weighing 18-22 grams were selected, half of which were male and half were female, and randomly divided into 13 groups according to their weight, namely, Yunnan Paris extract groups I to X, blank control group, positive drug group and model group, a total of 13 groups, 10 mice in each group, half of which were male and half were female. The same test area of ​​3cm×3cm on the abdomen of each group of mice was shaved. The test area of ​​each mouse in the Yunnan Paris extract groups I to X, the positive drug group and the model group was smeared with a 2,4-dinitrochlorophenylacetone solution with a concentration of 7wt% for sensitization. On the 5th day, the inside and outside of the right ear of each mouse were smeared with a 2,4-dinitrochlorophenylacetone solution with a mass concentration of 0.5% for stimulation, and the stimulation was performed at a frequency of 3 days / time, for a total of 4 times; the blank control group was treated according to the above operation, except that the 2,4-dinitrochlorophenylacetone solution was replaced with saline.

[0094] 2 hours after the first challenge, the mice in the Paris polyphylla extract groups I to X were evenly smeared with the Paris polyphylla extract (diluted with water to a solution with a mass concentration of 10%) on the front and back of the right ear of the mice at a dosage of 0.5 g solution / mouse, and the administration frequency was twice a day; the positive drug group was smeared with 0.5 g of an aqueous solution with a mass concentration of 1% hydrocortisone cream (produced by Beijing Shuangji Pharmaceutical Co., Ltd.); the blank control group and the model group were smeared with an equal amount of normal saline in the same manner; the above administration was continued until the last challenge, and 24 hours after the last challenge, the mouse eyeballs were removed for blood sampling, and the TNF-α value in the serum was detected. The results are shown in Table 1.

[0095] Table 1: TNF-α values ​​in serum

[0096] test TNF-α / pg / mL Blank control group 56.29±6.07 Model Group 60.83±5.51 Positive drug group 55.94±4.98 Paridis yunnanensis extract I 57.48±5.24 Yunnan Paris Extract II 57.57±5.93 Yunnan Paris Extract III 58.65±5.51 Yunnan Paris Extract IV 58.29±6.19 Yunnan Paris polyphylla extract V 58.17±5.04 Yunnan Paris polyphylla extract VI 58.06±5.15 Yunnan Paris Extract VII 57.81±5.76 Yunnan Paris Extract VIII 57.95±5.38 Yunnan Paris Extract IX 57.79±5.29 Yunnan Paris ExtractX 57.64±6.45

[0097] From the test results in Table 1, it can be seen that the mice in the model group were sensitized and stimulated with 2,4-dinitrochlorophenylpropionone solution, and no drug application was used for treatment. The test showed that the TNF-α content in the serum of the model group was high; while the mice in the blank control group were not sensitized and stimulated with 2,4-dinitrochlorophenylpropionone solution, and no drug application was used for treatment, and the serum TNF-α content was maintained at a normal level; the mice in the Yunnan Paris extract group were sensitized and stimulated with 2,4-dinitrochlorophenylpropionone solution, and the Yunnan Paris extract was used for treatment, and the TNF-α content in their serum was maintained close to the level of TNF-α content in the serum of the blank control group. It can be seen that the Yunnan Paris extract of the present invention can reduce the TNF-α content in the serum of mice after sensitization, thereby indicating that the Yunnan Paris extract prepared by the present invention has a good anti-inflammatory effect, and it has a maintenance and repair function for scalp inflammatory symptoms.

[0098] By comparing extracts I and III, it can be seen that the fermentation process of the present invention can better improve the anti-inflammatory effect of the extract. Furthermore, by using Aspergillus oryzae CICC 40188 and Bacillus subtilis GDMCC NO: 1.1750 for joint fermentation, the types and contents of anti-inflammatory active ingredients are improved, and the anti-inflammatory effect of the extract is improved. By comparing extracts I, VII, and VIII, it can be obtained that the two have a synergistic effect in promoting the anti-inflammatory effect of the extract. And in the preparation system of the present invention, using a ratio of 1:0.3-0.8 of the number of bacteria of the two can bring better anti-inflammatory effects, which can be obtained by comparing extracts I, IX, and X.

[0099] When preparing the Yunnan Paris polyphylla extract, the present invention uses ethanol to adjust the ethanol concentration in the extraction solvent, and uses an ultrafiltration membrane with a molecular weight cutoff of 1200Da for filtration, both of which can increase the anti-inflammatory effect of the extract. The above content can be obtained by comparing extracts I, IV, V, and VI.

[0100] 2. Preparation of the preparation and testing of its effect

[0101] The above-mentioned extract of Paris polyphylla was used to prepare a preparation, and then the effect test was performed. The preparation method of the specific preparation is as follows.

[0102] Preparation 1:

[0103] Raw material composition: by weight, 0.4 parts of Yunnan Paris extract I, 0.2 parts of Laminaria extract, 4.2 parts of ethylene glycol, 6 parts of glycerol, 0.17 parts of phenoxyethanol and 45 parts of water; the mass ratio of Yunnan Paris extract to Laminaria extract is 1:0.5;

[0104] The preparation method of kelp extract is as follows:

[0105] Step 1: Grind the dried kelp and pass it through a 50-mesh sieve to obtain kelp powder;

[0106] Step 2: placing kelp powder and 40% ethanol aqueous solution in a container at a mass ratio of 1:22, heating to 48° C., and stirring for 10 hours to obtain a mixture;

[0107] Step 3: filtering the mixture through a 3-micron pore size filter membrane to obtain filtrate 1, and then filtering the filtrate 1 through an ultrafiltration membrane with a molecular weight cutoff of 1200 Da to obtain filtrate 2, concentrating the filtrate 2, and freeze-drying it to a water content of about 4.25% to obtain the kelp extract;

[0108] The preparation method of the preparation is as follows: the raw materials are stirred and mixed at a rotation speed of 60 rpm for 0.5 hour, and then sterilized to obtain the preparation 1.

[0109] Preparation 2:

[0110] Raw material composition: by weight, 0.42 parts of Yunnan Paris extract II, 0.18 parts of Laminaria extract, 4.8 parts of ethylene glycol, 5.4 parts of glycerol, 0.16 parts of phenoxyethanol and 45 parts of water; the mass ratio of Yunnan Paris extract to Laminaria extract is 1:0.43;

[0111] The preparation method of kelp extract is as follows:

[0112] Step 1: Grind the dried kelp and pass it through a 50-mesh sieve to obtain kelp powder;

[0113] Step 2: placing kelp powder and ethanol aqueous solution with a mass concentration of 20% in a container at a mass ratio of 1:35, heating to 50° C., and stirring for 9.5 hours to obtain a mixture;

[0114] Step 3: filtering the mixture through a 3-micron pore size filter membrane to obtain filtrate 1, and then filtering the filtrate 1 through an ultrafiltration membrane with a molecular weight cutoff of 1200 Da to obtain filtrate 2, concentrating the filtrate 2, and freeze-drying it to a water content of about 4.39% to obtain the kelp extract;

[0115] The preparation method of the preparation is as follows: the raw materials are stirred and mixed at a rotation speed of 60 rpm for 0.5 hour, and then sterilized to obtain the preparation 2.

[0116] Preparation 3:

[0117] Raw material composition: by mass, 0.6 parts of Paris polyphylla extract I, 4.2 parts of ethylene glycol, 6 parts of glycerol, 0.17 parts of phenoxyethanol and 45 parts of water; the rest are the same as Preparation 1.

[0118] Preparation 4:

[0119] Raw material composition: by mass, 0.6 parts of kelp extract, 4.2 parts of ethylene glycol, 6 parts of glycerol, 0.17 parts of phenoxyethanol and 45 parts of water; the rest are the same as Preparation 1.

[0120] Preparation 5:

[0121] The preparation method of kelp extract is as follows:

[0122] Step 1: Grind the dried kelp and pass it through a 50-mesh sieve to obtain kelp powder;

[0123] Step 2: Place kelp powder and water in a container at a mass ratio of 1:22, heat to 48° C., and stir for 10 hours to obtain a mixture;

[0124] Step 3: filtering the mixture through a 3-micron pore size filter membrane to obtain filtrate 1, and then filtering the filtrate 1 through an ultrafiltration membrane with a molecular weight cutoff of 1200 Da to obtain filtrate 2, concentrating the filtrate 2, and freeze-drying it to a water content of about 4.25% to obtain the kelp extract;

[0125] The rest are the same as Preparation 1.

[0126] Preparation 6:

[0127] The preparation method of kelp extract is as follows:

[0128] Step 1: Grind the dried kelp and pass it through a 50-mesh sieve to obtain kelp powder;

[0129] Step 2: placing kelp powder and ethanol aqueous solution with a mass concentration of 80% in a container at a mass ratio of 1:22, heating to 48° C., and stirring for 10 hours to obtain a mixture;

[0130] Step 3: filtering the mixture through a 3-micron pore size filter membrane to obtain filtrate 1, and then filtering the filtrate 1 through an ultrafiltration membrane with a molecular weight cutoff of 1200 Da to obtain filtrate 2, concentrating the filtrate 2, and freeze-drying it to a water content of about 4.25% to obtain the kelp extract;

[0131] The rest are the same as Preparation 1.

[0132] Preparation 7:

[0133] The preparation method of kelp extract is as follows:

[0134] Step 1: Grind the dried kelp and pass it through a 50-mesh sieve to obtain kelp powder;

[0135] Step 2: placing kelp powder and 40% ethanol aqueous solution in a container at a mass ratio of 1:22, heating to 48° C., and stirring for 10 hours to obtain a mixture;

[0136] Step 3: filtering the mixture through a 3-micron pore size filter membrane to obtain filtrate 1, concentrating the filtrate 1, and freeze-drying it to a water content of about 4.25% to obtain the kelp extract;

[0137] The rest are the same as Preparation 1.

[0138] The above preparations 1-7 were tested for their effects, as follows:

[0139] (1) Test on the effect of scavenging DPPH free radicals

[0140] Preparations 1-7 were diluted with deionized water to a concentration of 25 wt % as test samples.

[0141] Take 1.4 mL of sample and 2 mL of 0.05 mg / mL DPPH ethanol solution and mix them. Incubate at 25°C in the dark for half an hour. Measure the absorbance at 519 nm and record it as A1.

[0142] Take 1.4 mL of deionized water and 2 mL of 0.05 mg / mL DPPH ethanol solution and mix them. Incubate at 25°C in the dark for half an hour. Measure the absorbance at 519 nm and record it as A0.

[0143] Take 1.4 mL of sample and mix with 2 mL of ethanol, react at 25°C in the dark for half an hour, and measure the absorbance at 519 nm, which is recorded as A2.

[0144] Then the DPPH free radical scavenging rate was calculated according to the formula: DPPH free radical scavenging rate = 100% × [1-(A1-A2) / A0]. The results are shown in Table 2.

[0145] Table 2: Test on the effect of scavenging DPPH free radicals

[0146] test DPPH free radical scavenging rate / % Preparation 1 92.78 Preparation 2 91.26 Preparation 3 90.05 Preparation 4 87.73 Preparation 5 86.14 Preparation 6 83.97 Preparation 7 79.69

[0147] (2) Test on the effect of scavenging superoxide free radicals

[0148] Preparations 1-7 were diluted with deionized water to a concentration of 25 wt % as test samples.

[0149] Take 2 mL of sample, 4.5 mL of 50 mmol / L Tri-HCl (pH 8.2) buffer and 0.4 mL of 25 mmol / L pyrogallol aqueous solution, mix them, keep them warm at 25°C for 5 minutes, then add 1 mL of 8 mmol / L HCl solution, mix well, and measure the absorbance at 325 nm, which is recorded as B1.

[0150] Take 2 mL of deionized water, 4.5 mL of 50 mmol / L Tri-HCl (pH 8.2) buffer and 0.4 mL of 25 mmol / L pyrogallol aqueous solution, mix them, keep them warm at 25°C for 5 minutes, then add 1 mL of 8 mmol / L HCl solution, mix well, and measure the absorbance at 325 nm, which is recorded as B0.

[0151] Take 2 mL of sample, 4.5 mL of 50 mmol / L Tri-HCl (pH 8.2) buffer and 0.4 mL of deionized water, mix them, keep them at 25°C for 5 minutes, then add 1 mL of 8 mmol / L HCl solution, mix well, and measure the absorbance at 325 nm, which is recorded as B2.

[0152] Then the superoxide free radical scavenging rate was calculated according to the formula: superoxide free radical scavenging rate = 100% × [1-(B1-B2) / B0]. The results are shown in Table 3.

[0153] Table 3: Test of the effect of scavenging superoxide free radicals

[0154] test Superoxide free radical scavenging rate / % Preparation 1 86.21 Preparation 2 84.54 Preparation 3 83.25 Preparation 4 80.67 Preparation 5 79.06 Preparation 6 76.74 Preparation 7 72.18

[0155] Free radicals can cause cells to be oxidized, leading to skin aging and other phenomena; therefore, the present invention uses the free radical scavenging effect to verify the anti-aging effect of the preparation. The test results in Tables 2-3 show that the preparation prepared by the Yunnan Paris extract prepared by the present invention, combined with kelp extract, polyol components, etc., has a good free radical scavenging effect; after the preparation is diluted to a concentration of 25wt%, it can scavenge 92.78% of DPPH free radicals and 86.17% of superoxide free radicals, and has a good effect of eliminating human free radicals, giving the skin anti-aging effects and the like.

[0156] By comparing preparations 1, 5-6, it can be seen that the present invention uses an ethanol aqueous solution with a mass concentration of 20-40% to extract kelp, and the obtained extraction can better enhance the free radical scavenging effect of the preparation, giving the skin better anti-aging function; and when extracting kelp, an ultrafiltration membrane with a molecular weight cutoff of 1200Da is used for treatment, which can further enhance the efficiency of the preparation in scavenging DPPH free radicals and superoxide free radicals.

[0157] From the comparison results of preparations 3 and 4 with preparation 1 in Table 2-3, it can be seen that the use of the Paris polyphylla extract and the Laminaria elata extract prepared by the present invention in the preparations can mutually promote the antioxidant effect of the preparations, and the combination of the two has a synergistic effect in increasing the effect of the preparations in eliminating DPPH free radicals and superoxide free radicals.

[0158] (3) Stability test

[0159] Preparations 1-7 were subjected to the following stability effect tests:

[0160] Heat stability: Place the preparation in a thermostat at 50°C for one week, take it out, cool it naturally to room temperature, and observe the appearance of the preparation for precipitation or stratification.

[0161] Cold stability: Place the preparation in a -20℃ constant temperature box for one week, take it out, naturally heat it to room temperature, and observe whether there is precipitation or stratification on the appearance of the preparation.

[0162] The test results are shown in Table 4.

[0163] Table 4: Stability test of formulations

[0164] test Heat stability Cold resistance and stability Preparation 1 No stratification, no precipitation No stratification, no precipitation Preparation 2 No stratification, no precipitation No stratification, no precipitation Preparation 3 No stratification, no precipitation No stratification, no precipitation Preparation 4 No stratification, no precipitation No stratification, no precipitation Preparation 5 No stratification, no precipitation No stratification, no precipitation Preparation 6 No stratification, no precipitation No stratification, no precipitation Preparation 7 No stratification, no precipitation No stratification, no precipitation

[0165] The test results in Table 4 show that the preparation prepared by the present invention has excellent stability and good heat and cold resistance.

[0166] Those skilled in the art may make various changes and modifications to the present invention. If these changes and modifications fall within the scope of the claims of the present invention and their equivalents, the present invention is intended to include these changes and modifications.

Claims

1. A Yunnan Paris polyphylla extract having the function of repairing scalp inflammatory symptoms, characterized in that: The Paris polyphylla extract is prepared by the following steps: S1: crushing the dried rhizome of Paris polyphylla to obtain rhizome powder, adding water 15-30 times the weight of the rhizome powder, heating and stirring for 4-8 hours to obtain a stirred material; cooling and keeping warm, adding 0.5-5% of cellulase and 0.4-4.5% of pectinase by weight of the rhizome powder for enzymolysis for 3-6 hours to obtain an enzymolysate 1; S2: inactivate the enzyme in the enzymatic hydrolysate 1 to obtain the inactivated material 1, add 0.8-3.5% of the weight of the rhizome powder with a neutral protease, and perform enzymolysis for 2.5-5 hours to obtain the enzymatic hydrolysate 2; S3: inactivate the enzyme in the hydrolysate 2 to obtain the inactivated material 2, then add 0.7-4% papain by weight of the rhizome powder, and perform enzymolysis for 3-5.5 hours to obtain the hydrolysate 3; S4: inactivate the enzyme in the hydrolyzate 3 to obtain material A, add 3-10% of the mass of the rhizome powder to the carbon source and inoculate the fermentation bacteria to obtain material B, and ferment at a temperature of 25-35° C. for 24-60 hours to obtain a fermented material; S5: sterilize the fermentation material, then add anhydrous ethanol, heat to 40-55°C, stir for 4-8 hours, filter, and obtain filtrate 1; S6: Filter the filtrate 1 using an ultrafiltration membrane with a molecular weight cutoff lower than 2000Da to obtain a filtrate 2, which is then concentrated and dried to obtain the Paris polyphylla extract.

2. The extract according to claim 1, characterized in that The concentration of the fermentation bacteria in material B is 2.5×10 8 -9.7×10 8 cfu / g.

3. The extract according to claim 1, characterized in that In S4, the fermentation bacteria are Aspergillus oryzae CICC 40188 and Bacillus subtilis GDMCC NO: 1.1750, and the bacterial quantity ratio of Aspergillus oryzae CICC 40188 and Bacillus subtilis GDMCC NO: 1.1750 in material B is 1:0.3-0.

8.

4. The extract according to claim 1, characterized in that In S5, the amount of anhydrous ethanol added is 0.5-1 times the mass of water added in S1.

5. The extract according to claim 1, characterized in that In S1, the temperature of enzymatic hydrolysis is 35-50°C.

6. The extract according to claim 1, characterized in that In S2, enzymolysis is performed at a temperature of 35-42°C.

7. The extract according to claim 1, characterized in that In S3, enzymatic hydrolysis is performed at a temperature of 38-45°C.

8. Use of the Paris polyphylla extract according to any one of claims 1 to 7 in the preparation of a preparation having anti-inflammatory and free radical scavenging functions.

9. The use according to claim 8, characterized in that: The preparation having anti-inflammatory and free radical scavenging functions is composed of the following components in parts by mass: 0.1-1 parts of the Polygonum multiflorum extract of any one of claims 1-7, 0.05-0.6 parts of kelp extract, 2-5 parts of ethylene glycol, 3-7 parts of glycerol, 0.05-0.3 parts of preservatives and 30-50 parts of water; the mass ratio of the Polygonum multiflorum extract to the kelp extract is 1:0.4-0.

7.

10. The use according to claim 9, characterized in that: The preparation method of the kelp extract comprises the following steps: Step 1: Grind the dried kelp and pass it through a 50-mesh sieve to obtain kelp powder; Step 2: placing kelp powder and ethanol aqueous solution with a mass concentration of 20-40% in a container at a mass ratio of 1:12-40, heating to 35-55° C., and stirring for 6-12 hours to obtain a mixture; Step 3: Filter the mixture using a 3-micron pore size filter membrane to obtain filtrate 1, and then filter the filtrate 1 using an ultrafiltration membrane with a molecular weight cutoff of 1200Da to obtain filtrate 2. Concentrate filtrate 2 and freeze-dry it until the water content is less than 6.8%, thereby obtaining the kelp extract.