CN114869831A - A method for preparing cosmetic composition containing natural plants - Google Patents
A method for preparing cosmetic composition containing natural plants Download PDFInfo
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- CN114869831A CN114869831A CN202210580056.3A CN202210580056A CN114869831A CN 114869831 A CN114869831 A CN 114869831A CN 202210580056 A CN202210580056 A CN 202210580056A CN 114869831 A CN114869831 A CN 114869831A
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Images
Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9794—Liliopsida [monocotyledons]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9728—Fungi, e.g. yeasts
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
- A61K2800/59—Mixtures
- A61K2800/592—Mixtures of compounds complementing their respective functions
- A61K2800/5922—At least two compounds being classified in the same subclass of A61K8/18
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/805—Corresponding aspects not provided for by any of codes A61K2800/81 - A61K2800/95
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/82—Preparation or application process involves sonication or ultrasonication
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/84—Products or compounds obtained by lyophilisation, freeze-drying
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/85—Products or compounds obtained by fermentation, e.g. yoghurt, beer, wine
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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Birds (AREA)
- Epidemiology (AREA)
- Botany (AREA)
- Biotechnology (AREA)
- Engineering & Computer Science (AREA)
- Dermatology (AREA)
- Gerontology & Geriatric Medicine (AREA)
- Cosmetics (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention provides a preparation method of a cosmetic compound containing natural plants, which relates to the technical field of plant extraction and purification, fermentation and cosmetics and comprises the following steps: the method comprises the following steps: respectively extracting ginseng, angelica, glycyrrhiza glabra, white truffle and poria cocos, and purifying to obtain related extracts; step two: preparing rice fermentation liquor step by step according to a set sequence, firstly adding protease, amylase and cellulase for aerobic fermentation, and then adding rumen fluid for anaerobic fermentation; step three: according to the invention, multiple plant extracts are compounded and then compounded, and rice fermentation liquor is added, so that the defects of poor effect and incomplete exertion of a single extract are effectively overcome, effective components are converted into micromolecules which can be better absorbed through fermentation, and then multiple plant extracts are compounded, so that the active components and active components of various plants are high, and the effects of whitening, moisturizing, resisting oxidation and resisting aging are achieved.
Description
Technical Field
The invention provides a preparation method of a cosmetic compound containing natural plants, and relates to the technical field of plant extraction and purification, fermentation and cosmetics.
Background
With the development of science and technology, human skin tends to become aged gradually due to factors such as age, ultraviolet radiation, diet, environment and the like. The current research suggests that skin aging mainly includes two categories, natural aging and photoaging. There are various theories as to the cause thereof, such as a genetic theory, an immunological theory, a free radical theory, etc., and among them, the free radical theory is becoming more and more popular. The theory of free radicals states that skin changes with age are caused by the side effects of free radicals. Normally, the production and disappearance of free radicals in the body are in a state of dynamic equilibrium, and when the body is aged, the ability of the body to scavenge free radicals appears to be acutely or chronically diminished. The surplus free radicals can generate destructive reaction on the chemical structure of the biomacromolecule forming the tissue chest, so that the integrity of the normal tissue form and function is damaged, and therefore, the development of corresponding skin care products aiming at mechanisms of causing skin aging and the like has wide development space.
In recent years, natural cosmetics having anti-aging, whitening, etc. effects are being relied upon by consumers. The active ingredients in natural cosmetics mainly come from plants, animals and microorganisms, wherein the application of the plant active ingredients is the most extensive, and the plant active ingredients are successfully applied to the fields of skin care, whitening, aging resistance and the like. Compared with conventional cosmetics, the plant active ingredient is natural substance extracted from plant, has smaller molecules, is easier to be absorbed and digested by skin, and does not generate deposition in vivo. Meanwhile, the plant extract cosmetics also have the advantages of obvious curative effect, strong pertinence and no side effect or side effect after long-term use. The extraction method of the plant active ingredients is researched and applied to a cosmetic formula for human skin experiments, so that the traditional Chinese medicine whitening anti-aging composition is obtained, and a research thought is provided for developing new whitening raw materials for the cosmetic industry and developing natural plant raw materials.
Disclosure of Invention
In order to obtain a cosmetic raw material with multiple effects and multiple combined processes, which can whiten and tender skin, resist oxidation, aging and the like, the preparation method of the cosmetic compound containing natural plants is provided, and comprises the following steps: the method comprises the following steps: respectively extracting ginseng, angelica, glycyrrhiza glabra, white truffle and poria cocos, and purifying to obtain related extracts; step two: preparing rice fermentation liquor step by step according to a set sequence, firstly adding protease, amylase and cellulase for aerobic fermentation, and then adding rumen fluid for anaerobic fermentation; step three: mixing the related extracts according to a set mass fraction, adding a preservative, sterilizing and filtering to obtain the product.
The product obtained by adopting the technical scheme is natural, free of stimulation, rich in nutrition, strong in efficacy, easy to absorb, safe and safe, can be used safely, and is suitable for the fields of skin care, beauty treatment and the like.
Further, crushing white ginseng by using a crusher to obtain white ginseng powder, adding white rot fungi laccase, cellulase and mannase into the white ginseng powder, adding water into the white ginseng powder in a thermostatic water bath at 40 ℃ for 2 hours, wherein the mass ratio of the white ginseng powder to the white rot fungi laccase, the cellulase and the mannase is 1:0.05: 0.01, extracting the white ginseng powder by using a 60-75% ethanol solution, the mass ratio of the white ginseng powder to the ethanol solution is 1: 15-1: 25, the ultrasonic temperature is 45-55 ℃, the ultrasonic frequency is 40-50kHz, the ultrasonic extraction time is 0.5-1.5 hours, heating the white ginseng powder to 65-75 ℃, performing reflux extraction, the extraction time is 3-6 hours, and filtering and collecting filtrate; carrying out secondary extraction on the filter residue, wherein the mass ratio of the filter residue to the ethanol solution is 1: 10-1: 20, heating and refluxing for 3-6 hours for extraction, and filtering and collecting filtrate; and combining the two filtrates, adsorbing the combined filtrate by macroporous resin, and heating and concentrating the filtrate at 75-85 ℃ until no alcohol exists, thereby obtaining the ginseng extract.
By adopting the technical scheme, as the ginseng contains effective components such as ginsenoside, ginseng polysaccharide, ginseng polypeptide, vitamins, trace elements and the like, the invention mainly extracts and purifies the ginsenoside, so that according to the principle of similarity and intermiscibility, the white ginseng which is grown for 5 years is selected firstly, and then the white ginseng is extracted by compounding a plurality of processes, the extraction yield is effectively improved, and the active substances can be effectively enriched compared with the active substances extracted by a single process.
Further, crushing angelica sinensis, sieving with a 24-mesh sieve, enabling the powder sieved with a 60-mesh sieve to be not more than 40%, soaking and mixing with 65-75% methanol in an amount which is 15-25 times of the angelica sinensis powder, carrying out ultrasonic extraction for three times, wherein the total ultrasonic extraction time is 0.5-1.0 h, the ultrasonic extraction temperature is 35-55 ℃, combining filtrates extracted for three times, firstly heating and concentrating under reduced pressure to half volume of the filtrate, precipitating with 95% ethanol in an amount which is 3 times of the mass of the concentrate after the concentrate is cooled, centrifugally leaching supernatant, collecting centrifugal precipitate, collecting the supernatant through a 0.45-micron microporous filter membrane, filtering and precipitating, mixing the centrifugal precipitate with the filtered precipitate, and drying to obtain the angelica sinensis extract.
By adopting the technical scheme, the angelica is crushed into coarse angelica powder or medium angelica powder, so that the extraction is convenient, and the influence of the fineness of the angelica powder on the filtering effect is avoided; multiple times of ultrasonic extraction and purification are carried out, the purity of the extract is improved, the extraction pollution is reduced, and the economic benefit and the environmental benefit are high.
Further, crushing poria cocos to obtain poria cocos powder, sieving the poria cocos powder with a 60-mesh sieve, adding distilled water with the mass being 20-30 times of that of the poria cocos powder to perform microwave extraction, wherein the microwave power is 450-550W, the radiation time is 5-9min, performing suction filtration, repeatedly extracting filter residues once again, combining two extracting solutions, performing vacuum concentration on the extracting solutions by using a rotary evaporator to obtain a paste, and drying the paste in a drying oven at 65-75 ℃ for 18-24 hours to obtain the white poria cocos extract.
Further, cleaning fresh white pine dew, then carrying out vacuum freeze drying, then sending the vacuum dried white pine dew into a grinder for grinding to obtain white pine dew powder, soaking and mixing 40% -60% ethanol 15-20 times of the white pine dew powder, carrying out ultrasonic-assisted extraction for 1-2 h with the ultrasonic frequency of 3-6kHz, filtering after the ultrasonic-assisted extraction to obtain a crude extract, purifying the crude extract by nonpolar macroporous resin (styrene-divinylbenzene polymer), carrying out pretreatment on the macroporous resin, fully swelling the macroporous resin by absolute ethanol, then placing the macroporous resin into a chromatographic column, carrying out flow cleaning by absolute ethanol until the liquid after cleaning is not mixed by adding equivalent distilled water, then stopping washing the macroporous resin by using a large amount of distilled water until no ethanol smell exists, carrying out sampling on the crude extract with the flow rate of 3ml/min, adsorbing the crude extract by the macroporous resin, then eluting by using 40% -60% ethanol solution, eluting at flow rate of 2ml/min, heating the eluate to 65-75 deg.C, and concentrating the filtrate under reduced pressure to obtain Gymnolenium giganteum extract.
By adopting the technical scheme, the physical, chemical and biological states of the white truffle are basically unchanged by using the vacuum freeze drying technology, so that the nutrient substances and effective components of the white truffle are maximally reserved, the damage to volatile components and heat-sensitive substances in the white truffle in the drying process is reduced, and the reduction of the polysaccharide content in the white truffle during extraction is avoided.
Further, adding 60-75% of ethanol solvent into glycyrrhiza glabra to extract for 3 times, wherein the first material-liquid ratio is 1: 8-1: 10, the second material-liquid ratio is 1: 5-1: 8, the third material-liquid ratio is 1: 5-1: 8, the extraction temperature is 70-80 ℃, the extraction time is 4-6 hours, filtering after extraction, combining the three filtrates to obtain a crude extract, concentrating the crude extract at 60-75 ℃ under reduced pressure until the extract is basically free of ethanol to obtain a residual solution, adding dichloromethane with the mass of 2-3 times of that of the crude extract into the residual solution to extract, collecting a dichloromethane layer, repeating the extraction operation for 2-3 times, combining dichloromethane filtrates of each time, concentrating the dichloromethane filtrate at 50-60 ℃ under reduced pressure until the total content is 30-50%, purifying the concentrated solution by a silica gel column, adsorbing the concentrated solution, adding methanol to elute, wherein the mass ratio of the dichloromethane to the methanol is 90:1, eluting at 1-3 drops/s, collecting eluate, concentrating the eluate at 50-60 deg.C under reduced pressure until no liquid flows out, and freeze drying the concentrated solid to obtain Glycyrrhiza glabra root extract.
Further, crushing dried rice to obtain rice powder, sieving the rice powder with a 80-mesh sieve, adding water, cooking to obtain rice powder slurry, wherein the mass ratio of the rice powder to the water is 1: 10-1: 30, adding 0.1-0.8% of protease, 0.1-1.2% of amylase and 0.1-0.65% of cellulase into the rice powder slurry in sequence, continuously stirring and dissolving, heating to 55-60 ℃, fermenting for 6-10h, wherein the fermentation pH is 7.0-7.3, and filtering sterile air oxygen is continuously introduced during the fermentation process, and the air flow rate is 2-4L/min to obtain the rice fermentation broth primary product.
By adopting the technical scheme, digestive enzymes such as protease, amylase, cellulase and the like are added firstly, so that the anti-nutritional factors of the rice can be eliminated, macromolecular proteins are degraded, and the aerobic fermentation of the rice is realized firstly.
Further, placing rumen fluid in a separating funnel, incubating at 35-45 ℃ for 1-3h, then, reserving the middle-upper layer rumen fluid, adding 0.5-1.5% rumen fluid into the rice fermentation liquor primary product, stirring for 2-3min, then fermenting, setting the fermentation temperature at 36-42 ℃, the fermentation pH value at 6.6-6.8, continuously introducing carbon dioxide in the fermentation process, mixing and stirring gas and liquid for 1min, standing for 9min, fermenting for 8-12h, sterilizing the fermentation liquor after fermentation is completed, filtering and collecting the filtrate, thus obtaining the rice fermentation liquor.
By adopting the technical scheme, the anaerobic fermentation is carried out by utilizing rumen microorganisms after the rice is subjected to aerobic fermentation, so that the fiber and protein can be effectively degraded, the rice is degraded into small molecular substances which are harmless and more suitable for being absorbed by a human body, active substances are enriched, the product mildness is improved, the toxic and side effects are reduced, and a plurality of new active substances can be generated with plant extracts. By simulating the digestion of food in the stomach in vitro and by adopting a bionic multiple fermentation process, rice is fermented into active ingredients which are better absorbed and more safely obtained.
Further, 10-20% of ginseng root extract, 8-16% of angelica sinensis extract, 8-16% of poria cocos extract, 8-16% of glycyrrhiza glabra root extract, 8-16% of chrysosporium extract, 0.5-2% of 1, 2-hexanediol and 0.2-1% of p-hydroxyacetophenone are taken, and the balance is supplemented to 100% by rice fermentation liquor, and the product is obtained by stirring uniformly, sterilizing and filtering.
Drawings
FIG. 1 is a process flow diagram of a method for preparing a cosmetic composition containing natural plants;
FIG. 2 is a graph of DPPH inhibition by a cosmetic composition containing natural plants;
FIG. 3 is a concentration profile of a tyrosinase activity assay using a cosmetic composition containing natural plants.
Detailed Description
The present invention will be described in further detail with reference to the following drawings and examples. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
Example 1
A method for preparing cosmetic composition containing natural plants, as shown in figure 1, comprises the following steps:
1. the ginseng root extract is prepared by selecting 5-year-old white ginseng, crushing the selected white ginseng by using a crusher, adding enzyme water bath, extracting by using 70% ethanol as a solvent at the temperature of 45 ℃ for 1 hour at 70 ℃ under the ultrasonic assistance with the solid-liquid ratio of 1:20, heating and refluxing for 5 hours, filtering and collecting filtrate; extracting the filter residue twice with a solid-to-liquid ratio of 1:12 under heating and refluxing for 4.5 hours, filtering and collecting the filtrate. Mixing the two filtrates, and concentrating at 80 deg.C until no alcohol exists to obtain Ginseng radix extract.
2. The angelica root extract is prepared by crushing and sieving angelica, carrying out ultrasonic extraction on angelica powder for 0.6h by using 70% methanol in an amount which is 1:20 times of the weight of the angelica powder, carrying out ultrasonic extraction for three times at 40 ℃, combining filtrate, carrying out vacuum concentration on the filtrate to half, cooling the concentrated solution, then carrying out precipitation by using 95% ethanol in an amount which is 3 times of the weight of the angelica powder, carrying out centrifugal leaching on supernatant, collecting centrifugal precipitation, filtering the supernatant, collecting filtered precipitation, mixing the centrifugal precipitation with the filtered precipitation, and drying the mixture to obtain the angelica extract.
3. Poria extract is prepared by pulverizing Poria to obtain Poria powder, adding distilled water, extracting Poria with microwave assistance at solid-to-liquid ratio of 1:20, radiating for 7min at microwave power of 520W, vacuum filtering, extracting the residue for 1 time, mixing the two extractive solutions, vacuum concentrating with rotary evaporator to obtain paste, and drying to obtain Poria extract.
4. The preparation method comprises the following steps of carrying out vacuum freeze drying on the white truffle extract and fresh white truffle, then crushing the white truffle, carrying out ultrasonic assisted extraction for 1 hour by using 50% ethanol 15 times of the white truffle powder, carrying out ultrasonic frequency of 5kHz to obtain a crude extract, carrying out adsorption elution on the crude extract by using macroporous resin to obtain an eluent, and carrying out reduced pressure concentration on the eluent at 70 ℃ until no alcohol exists to obtain the white truffle extract.
5. Crushing liquorice into powder, extracting for 3 times by using 70% ethanol as a solvent, wherein the ratio of the first material to the liquid is 1:8, the ratio of the second material to the liquid is 1:6, the ratio of the third material to the liquid is 1:6, the extraction temperature is 70 ℃, the extraction time is 4 hours, filtering after extraction, combining the three filtrates to obtain a crude extract, concentrating the crude extract at 70 ℃ to remove ethanol to obtain a residual liquid, adding two times of dichloromethane into the residual liquid for mixing, standing for layering after uniform mixing, collecting a dichloromethane layer and removing an extraction reagent, and repeatedly extracting for 3 times to ensure the completeness of an extraction effect. And (3) adsorbing and eluting the concentrated solution after extraction through a silica gel column to collect eluent, concentrating the eluent at 60 ℃ under reduced pressure, and freeze-drying the concentrated solid to obtain the glycyrrhiza glabra extract.
6. Adding 0.5% protease, 0.7% amylase and 0.35% cellulase into rice fermentation filtrate, performing aerobic fermentation at 58 deg.C and pH of 7.1 for 8 hr to obtain rice fermentation broth primary product; adding 1% rumen fluid into the rice fermentation broth, performing anaerobic fermentation at 37 deg.C and pH6.7 for 10 hr, sterilizing the fermentation broth after fermentation, filtering, and collecting filtrate to obtain rice fermentation broth.
7. After the plant extracts are obtained, 15% of ginseng root extract, 10% of angelica extract, 10% of tuckahoe extract, 10% of glycyrrhiza glabra extract, 10% of truffle extract, 43.8% of yeast/rice fermentation product filtrate, 1, 2-hexanediol and 0.2% of p-hydroxyacetophenone are mixed, sterilized and filtered to obtain the product.
Example 2
A method for preparing cosmetic composition containing natural plants, as shown in figure 1, comprises the following steps:
1. the ginseng root extract is prepared by selecting 5-year-old white ginseng, crushing the selected white ginseng by using a crusher, adding enzyme water bath, extracting by using 70% ethanol as a solvent at 45 ℃ for 1.5 hours at a solid-liquid ratio of 1:18 under the ultrasonic assistance of 1.5 hours and a temperature of 65 ℃, heating and refluxing for 5.5 hours, filtering and collecting filtrate; and carrying out secondary extraction on the filter residue with the solid-liquid ratio of 1:15, heating and refluxing for 5 hours, filtering and collecting filtrate. Mixing the two filtrates, and concentrating at 75 deg.C until there is no alcohol to obtain Ginseng radix extract.
2. Pulverizing radix Angelicae sinensis, sieving, ultrasonically extracting radix Angelicae sinensis powder with 1:18 times of 70% methanol for 0.8 hr, ultrasonically extracting for three times at 45 deg.C, mixing filtrates, concentrating the filtrate under reduced pressure to half, cooling the concentrated solution, precipitating with 3 times of 95% ethanol, centrifuging to obtain supernatant, collecting supernatant, centrifuging, filtering, collecting filtrate, mixing the supernatant with the filtrate, and drying to obtain radix Angelicae sinensis extract.
3. Poria extract is prepared by pulverizing Poria, adding distilled water, extracting Poria with microwave at solid-to-liquid ratio of 1:25 under microwave power of 520W for 10min, vacuum filtering, extracting the residue under the same conditions for 1 time, mixing extractive solutions, vacuum concentrating with rotary evaporator to obtain paste, and drying.
4. The preparation method comprises the following steps of carrying out vacuum freeze drying on the white truffle extract and fresh white truffle, then crushing the white truffle, carrying out ultrasonic assisted extraction for 1.5 hours by using 45% ethanol 15 times of the white truffle powder, carrying out ultrasonic frequency of 6kHz to obtain a crude extract, carrying out adsorption elution on the crude extract by using macroporous resin to obtain an eluent, and concentrating the eluent at 70 ℃ until no alcohol exists to obtain the white truffle extract.
5. A glycyrrhiza glabra root extract is prepared by crushing glycyrrhiza glabra into powder, extracting for 3 times by using 75% ethanol as a solvent, wherein the ratio of the first material to the liquid is 1:9, the ratio of the second material to the liquid is 1:8, the ratio of the third material to the liquid is 1:8, the extraction temperature is 70 ℃, the extraction time is 5.5 hours, filtering after extraction, combining the three filtrates to obtain a crude extract, concentrating the crude extract at 70 ℃ to remove ethanol to obtain a residual liquid, adding two times of dichloromethane into the residual liquid, mixing uniformly, standing for layering, collecting a dichloromethane layer, removing an extraction reagent, and repeating the extraction for 3 times to ensure the completeness of the extraction effect. And (3) adsorbing and eluting the concentrated solution after extraction through a silica gel column to collect eluent, concentrating the eluent at 60 ℃ under reduced pressure, and freeze-drying the concentrated solid to obtain the glycyrrhiza glabra extract.
6. Adding 0.5% protease, 0.7% amylase and 0.35% cellulase into rice fermentation filtrate, performing aerobic fermentation at 58 deg.C and pH7.1 for 8 hr to obtain rice fermentation broth primary product; adding 1% rumen fluid into the rice fermentation broth, performing anaerobic fermentation at 37 deg.C and pH6.7 for 10 hr, sterilizing the fermentation broth after fermentation, filtering, and collecting filtrate to obtain rice fermentation broth.
7. After the plant extracts are obtained, 15 percent of ginseng root extract, 15 percent of angelica extract, 15 percent of tuckahoe extract, 15 percent of glabrous greenbrier rhizome extract, 15 percent of white truffle extract, 23.5 percent of yeast/rice fermentation product filtrate, 1, 2-hexanediol and 0.5 percent of p-hydroxyacetophenone are mixed, sterilized and filtered to obtain the product.
Effect example 1:
DPPH radical scavenging test
First, reagent preparation
(1)3.913mgDPPH 95% ethanol to 50mL
(2)8mgTrolox is fixed to 100mL by 95% ethanol
(3) Random example 1 composition (hereinafter referred to as example 1)
Second, the experimental procedure
(1) UV zero calibration, 95% ethanol as blank
(2) Preliminary experiments (DPPH concentration selection)
Shaking 500 μ L DPPH +1000 μ L95% ethanol for 10s, standing for 10min, and measuring absorbance (absorbance at 520nm of 0.45-0.55, can be used as DPPH solution for the following experiment)
(3) Determination of Trolox
| Sample | Trolox(μL) | 95% ethanol (μ L) | DPPH(μL) |
| |
100 | 900 | 500 |
| |
100 | 900+500 | 0 |
| |
0 | 100+900 | 500 |
| |
0 | 100+900+500 | 0 |
Adding DPPH or 95% ethanol, shaking for 10s, standing for 10min, and measuring absorbance
(4) Experimental procedure
Adding DPPH or 95% ethanol, shaking for 10s, and standing for 10min
(5) The absorbance values were measured at 520nm, tabulated and plotted to obtain IC50 values.
Calculating the formula: inhibition rate (%) [1- (S-SB)/(C-CB) ]. 100
Third, experimental results
The experimental results are as follows: example 1 has the effect of scavenging DPPH free radicals, the higher the concentration of the product, the higher the efficiency of scavenging DPPH free radicals, and as can be seen from FIG. 2, the IC50 value is about 27. mu.L/mL.
Effect example 2:
and (3) detecting the moisturizing effect:
1. procedure of experiment
The instrument comprises the following steps: hornemeter CMB25 (Germany CK corporation)
And (3) testing a sample: no. 1 (pure water), No. 2 (random example sample)
2. Conditions of the experiment
(I) indoor Environment
② no strong sunlight or direct lighting
③ the ambient temperature is 20 +/-2 ℃ and the ambient humidity is 40-60 percent
3. Subject Condition
Firstly, the normal subjects with age of 18-60 years can be selected from male and female, and female has priority
② patients without serious systemic diseases, immunodeficiency or autoimmune diseases
③ patients with inactive allergic diseases
Fourthly, no history of allergy to skin care cosmetics
Hormone medicine and immunosuppressant have not been used all the time within one month
Sixthly, those who do not participate in other clinical experiments
Seventhly, volunteers add ginseng and can finish the specified contents according to the experimental requirements
4. Skin moisture content detection working principle
The moisture content of the skin can be measured by adopting a worldwide accepted Cornemeter-capacitance method, the principle of which is based on that the dielectric constant (< 7) of water (81) and other substances is changed greatly, and a properly-shaped measuring capacitor changes along with the change of the capacitance of the skin according to the difference of water content, and the capacitance of the skin is in a measuring range. The capacitance measuring method is superior to other methods in that the test result is practically not affected by polarization effect and ionic conductivity since the skin to be tested is not in unnatural contact with the test probe and almost no current flows through the skin to be tested. The instrument probe and the water in the skin have no inertia in the process of establishing balance, and can realize quick measurement, thereby eliminating the influence of active skin on the measurement result.
5. Test procedure
Testing an area: inner side of forearm
② preparation before testing
The skin whitening products (cosmetics, external medicines or internal health products) cannot be used 2-3 days before the tested part, before the test, the testee needs to uniformly clean the inner side of the forearm, the forearm is wiped clean by dry facial tissue, after the cleaning, the inner side of the forearm of the testee is marked with an area, before formal test, the testee needs to sit still in a room meeting the standard for at least 30min, the forearm is exposed, and the testee is placed in a test state to keep relaxed.
Testing process
Forearm medial marking 3 x 4cm in the experiment 2 In the experimental area, the same arm can mark a plurality of areas at the same time, and the intervals of the areas are 2 cm. Test products and placebo were randomly distributed on the forearmIn the above, the blank value 30min after cleaning of each test area was measured, and then the sample/cm was measured as 2.0. + -. 0.1mg 2 The dosage of the test sample is uniformly coated in the experimental area by using a latex finger cot, and the skin moisture content of the test area and the control area is distributed and measured for 0h, 1h, 2h and 4h after coating. The measurements of the same volunteer were performed by the same measurement staff.
6. Measurement index
Measuring water content of skin twice, and taking average value
7. Control format
Performing different treatment controls by itself
8. Results and analysis
Participating in the measurement of the tester
As a result: as can be seen from the table, the skin moisture content of the blank group was substantially unchanged for different test persons within 4 hours.
In the water group, the skin moisture content slightly increased within 1 hour after the skin test site was applied with pure water, but immediately returned to the previous level at the later time, indicating that the application of pure water did not increase the skin moisture content.
In the sample group, after the skin test part is smeared with the sample, the moisture content of the skin is obviously increased, and the moisture content is stable within 2 hours, so that the composition has the moisturizing effect.
Effect example 3
1. The purpose is as follows: semi-inhibitory effect of cosmetic composition containing natural plants on tyrosinase
2. Materials: sample preparation: cosmetic composition containing natural plants (composition of example 1)
Buffer solution: k2HPO4(8.71g/500ml) + KH2PO4(6.805g/500ml) → pH 6.80(± 0.02) spectrophotometer: uv, 480nm
3. The method comprises the following steps:
(1) enzyme concentration selection:
taking a small amount of enzyme, dissolving the enzyme by using a buffer solution, and obtaining brown color; adding 500. mu.L of (L-tyrosine (0.3mg/ml), 500. mu.L of buffer solution, 450. mu.L of water and 50. mu.L of enzyme) into a 1.5ml centrifuge tube; before adding enzyme, at 37 deg.C for 20 min; after the addition of the enzyme, the absorbance was measured at a wavelength of 480nm in a spectrophotometer at 37 ℃ for 10min (enzyme addition-test). The enzyme concentration was adjusted to give an Absorbance (ABS) of between 0.45 and 0.55. The enzyme solution at this concentration was used as an experimental enzyme solution.
(2) Experimental addition
(3) The absorbance at 480nm was measured, and a table was prepared. Plotting to obtain IC 50 .
Calculating the formula: barrier rate (%) [1- (S-SB)/(C-CB) ]. 100
Note that: before adding enzyme, at 37 deg.C for 20 min; after the addition of the enzyme, the absorbance was measured at a wavelength of 480nm in a spectrophotometer at 37 ℃ for 10min (enzyme addition-test). Without enzyme at 37 deg.C for 20 min.
S：Sample SB：Sample Blank C：Control CB：Control Blank
4. Results
The results of the tyrosinase activity inhibition of the cosmetic composition containing natural plants are shown in the table:
5. and (4) conclusion: the cosmetic composition containing natural plants has effect in inhibiting tyrosinase activity, and the higher the concentration of the composition, the stronger the inhibition effect on tyrosinase activity, and as can be seen from FIG. 3, the IC50 value of the composition is about 4 μ L/mL.
Claims (9)
1. A method for preparing a cosmetic composition containing natural plants, comprising: the method comprises the following steps:
the method comprises the following steps: respectively extracting ginseng, angelica, glycyrrhiza glabra, white truffle and poria cocos, and purifying to obtain related extracts;
step two: preparing rice fermentation liquor step by step according to a set sequence, firstly adding protease, amylase and cellulase for aerobic fermentation, and then adding rumen fluid for anaerobic fermentation;
step three: mixing the related extracts according to a set mass fraction, adding a preservative, sterilizing and filtering to obtain the product.
2. The method for preparing a cosmetic composition comprising natural plants according to claim 1, wherein: crushing white ginseng by using a crusher to obtain white ginseng powder, adding white rot fungi laccase, cellulase and mannase into the white ginseng powder, adding water into the white ginseng powder, performing thermostatic waterbath at 40 ℃ for 2 hours, wherein the mass ratio of the white ginseng powder to the white rot fungi laccase to the cellulase to the mannase is 1:0.05: 0.01, then extracting by using a 60-75% ethanol solution, the mass ratio of the white ginseng powder to the ethanol solution is 1: 15-1: 25, the ultrasonic temperature is 45-55 ℃, the ultrasonic frequency is 40-50kHz, the ultrasonic extraction time is 0.5-1.5 hours, heating to 65-75 ℃, performing reflux extraction, the extraction time is 3-6 hours, and then filtering and collecting filtrate; carrying out secondary extraction on the filter residue, wherein the mass ratio of the filter residue to the ethanol solution is 1: 10-1: 20, heating and refluxing for 3-6 hours for extraction, and filtering and collecting filtrate; and combining the two filtrates, heating the filtrate at 75-85 ℃, and concentrating the filtrate until no alcohol exists to obtain the ginseng root extract.
3. The method for preparing a cosmetic composition comprising natural plants according to claim 1, wherein: crushing angelica, sieving with a 24-mesh sieve, enabling powder sieved with a 60-mesh sieve to be not more than 40%, soaking and mixing with 65-75% methanol in an amount which is 15-25 times of the amount of the angelica powder, carrying out ultrasonic extraction for three times, enabling the total ultrasonic extraction time to be 0.5-1.0 h, enabling the ultrasonic extraction temperature to be 35-55 ℃, combining filtrates obtained by three times of extraction, heating and concentrating under reduced pressure to half volume of the filtrate, cooling the concentrated solution, precipitating with 95% ethanol in an amount which is 3 times of the mass of the concentrated solution, carrying out centrifugal leaching on supernatant, collecting centrifugal precipitate, collecting the supernatant through a 0.45-micrometer microporous filter membrane, filtering the precipitate, mixing the centrifugal precipitate with the filtered precipitate, and drying to obtain the angelica extract.
4. The method for preparing a cosmetic composition comprising natural plants according to claim 1, wherein: pulverizing poria cocos to obtain poria cocos powder, sieving the poria cocos powder with a 60-mesh sieve, adding distilled water with the mass being 20-30 times of that of the poria cocos powder, performing microwave extraction with the microwave power being 450-550W and the radiation time being 5-9min, performing suction filtration, repeatedly extracting filter residues once, combining two extracting solutions, performing vacuum concentration on the extracting solutions by using a rotary evaporator to obtain a paste, and drying the paste in a drying oven at the temperature of 65-75 ℃ for 18-24 hours to obtain the poria cocos extract.
5. The method for preparing a cosmetic composition comprising natural plants according to claim 1, wherein: cleaning fresh white truffle, then carrying out vacuum freeze drying, then sending the vacuum dried white truffle into a grinder for grinding to obtain white truffle powder, soaking and mixing the white truffle powder with 40-60% ethanol which is 15-20 times of the white truffle powder, carrying out ultrasonic auxiliary extraction for 1-2 h with the ultrasonic frequency of 3-6kHz, filtering after the ultrasonic auxiliary extraction to obtain crude extract, purifying the crude extract by nonpolar macroporous resin (styrene-divinylbenzene polymer), carrying out pretreatment on the macroporous resin, fully swelling the macroporous resin by absolute ethanol, then placing the macroporous resin into a chromatographic column, carrying out flow cleaning by the absolute ethanol until the liquid after cleaning is not mixed by adding equivalent distilled water, then washing the macroporous resin by a large amount of distilled water until the smell of the ethanol does not exist, carrying out sampling by controlling the flow rate of the crude extract to be 3ml/min, adsorbing the crude extract by the macroporous resin, then eluting by 40-60% ethanol solution, eluting at flow rate of 2ml/min, heating the eluate to 65-75 deg.C, and concentrating the filtrate under reduced pressure to obtain Gymnolenium giganteum extract.
6. The method for preparing a cosmetic composition comprising natural plants according to claim 1, wherein: adding 60-75% of ethanol solvent into glycyrrhiza glabra to extract for 3 times, wherein the ratio of the first material to the liquid is 1: 8-1: 10, the ratio of the second material to the liquid is 1: 5-1: 8, the ratio of the third material to the liquid is 1: 5-1: 8, the extraction temperature is 70-80 ℃, the extraction time is 4-6 h, filtering after extraction, combining the three filtrates to obtain a crude extract, concentrating the crude extract at 60-75 ℃ under reduced pressure to an ethanol-free state to obtain a residual solution, adding 2-3 times of dichloromethane to the residual solution to extract, collecting dichloromethane layers, repeating the extraction operation for 2-3 times, combining the dichloromethane filtrates of each time, concentrating the dichloromethane filtrate at 50-60 ℃ under reduced pressure to 30-50% of the total content, purifying the concentrated solution by a silica gel column, adsorbing the concentrated solution, adding methanol to elute, and controlling the mass ratio of the dichloromethane to the methanol to be 90:1, eluting at 1-3 drops/s, collecting eluate, concentrating the eluate at 50-60 deg.C under reduced pressure until no liquid flows out, and freeze drying the concentrated solid to obtain Glycyrrhiza glabra root extract.
7. The method for preparing a cosmetic composition comprising natural plants according to claim 1, wherein: crushing dried rice to obtain rice powder, sieving the rice powder with a 80-mesh sieve, adding water, cooking to obtain rice flour slurry, wherein the mass ratio of the rice powder to the water is 1: 10-1: 30, adding 0.1-0.8% of protease, 0.1-1.2% of amylase and 0.1-0.65% of cellulase into the rice flour slurry in sequence, continuously stirring and dissolving, heating to 55-60 ℃, fermenting at the pH value of 7.0-7.3 for 6-10h, continuously introducing filtered sterile air oxygen during the fermentation process, and allowing the air flow rate to be 2-4L/min to obtain a rice fermentation liquid primary product.
8. The method for preparing a cosmetic composition comprising natural plants according to claim 7, wherein: putting rumen fluid in a separating funnel, incubating at 35-45 deg.C for 1-3h, collecting middle and upper layer rumen fluid, adding 0.5-1.5% rumen fluid into the rice fermentation liquid, stirring for 2-3min, fermenting, setting fermentation temperature at 36-42 deg.C, fermentation pH at 6.6-6.8, introducing carbon dioxide continuously during fermentation, mixing gas and liquid, stirring for 1min, standing for 9min, fermenting for 8-12h, sterilizing the fermentation liquid, filtering, and collecting filtrate to obtain rice fermentation liquid.
9. The method for preparing a cosmetic composition comprising natural plants according to claim 1, wherein: taking 10-20% of ginseng root extract, 8-16% of angelica extract, 8-16% of poria extract, 8-16% of glycyrrhiza glabra root extract, 8-16% of chrysosporium extract, 0.5-2% of 1, 2-hexanediol and 0.2-1% of p-hydroxyacetophenone, and supplementing the balance to 100% by rice fermentation liquor, uniformly stirring, sterilizing and filtering to obtain the product.
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