CN114931537B - Composite ferment capable of delaying aging and preparation method and application thereof - Google Patents
Composite ferment capable of delaying aging and preparation method and application thereof Download PDFInfo
- Publication number
- CN114931537B CN114931537B CN202210483369.7A CN202210483369A CN114931537B CN 114931537 B CN114931537 B CN 114931537B CN 202210483369 A CN202210483369 A CN 202210483369A CN 114931537 B CN114931537 B CN 114931537B
- Authority
- CN
- China
- Prior art keywords
- composite
- fermentation
- stock solution
- ginseng
- raw materials
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- 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]
-
- 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
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
- A61K8/34—Alcohols
- A61K8/345—Alcohols containing more than one hydroxy group
-
- 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
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/55—Phosphorus compounds
- A61K8/553—Phospholipids, e.g. lecithin
-
- 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
-
- 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]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- 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
-
- 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
- A61K2800/74—Biological properties of particular ingredients
- A61K2800/78—Enzyme modulators, e.g. Enzyme agonists
- A61K2800/782—Enzyme inhibitors; Enzyme antagonists
-
- 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
-
- 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
- A61K2800/85—Products or compounds obtained by fermentation, e.g. yoghurt, beer, wine
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/54—Improvements relating to the production of bulk chemicals using solvents, e.g. supercritical solvents or ionic liquids
Abstract
The invention belongs to the technical field of cosmetics, and discloses a composite ferment capable of delaying senescence, a preparation method and application thereof. The preparation method of the composite ferment comprises the following steps: mixing the raw materials with lecithin and glycerol, and homogenizing under high pressure to obtain a composite stock solution; mixing the composite stock solution with a fermentation strain, and fermenting to obtain a composite fermentation product; the raw materials include two or more of Ginseng radix, ganoderma, tricholoma matsutake, herba Dendrobii, saviae Miltiorrhizae radix, rehmanniae radix, and fructus Lycii; the fermentation strain comprises one or more of bacillus subtilis, monascus and saccharomyces cerevisiae. The invention combines high-pressure homogenization and directional microorganism fermentation technology organically, lecithin and glycerin are assisted in the high-pressure homogenization process, so that plant cells can be fully crushed and fully released, the utilization efficiency of raw materials is improved, the composite stock solution obtained by high-pressure homogenization is used as a subsequent fermentation substrate, and the composite fermentation product with obvious aging delaying effect can be prepared through the bioconversion capability of microorganisms.
Description
Technical Field
The invention belongs to the technical field of cosmetics, and particularly relates to a composite ferment capable of delaying senescence, and a preparation method and application thereof.
Background
In recent years, people have increasingly strong demands for delaying skin aging, and research on anti-skin aging is gradually in progress, and various anti-aging active substances are continuously developed aiming at skin aging mechanisms. Along with the continuous development of science, active ingredients in natural plants and the efficacy thereof are continuously excavated. At present, the active ingredients in natural plants are obtained by adopting a water extraction method or an alcohol extraction method, the traditional extraction methods have the problems of low yield and high energy consumption, and the obtained extract has no obvious effect; some manufacturers use supercritical CO 2 Extraction is carried out, but equipment requirements are high, and the cost is high.
Disclosure of Invention
The present invention aims to solve at least one of the technical problems in the prior art described above. Therefore, the invention provides a composite ferment capable of delaying senescence, a preparation method and application thereof, lecithin and glycerol are assisted in a high-pressure homogenizing process, so that cells are fully crushed, the dissolution rate of active ingredients is improved, and the composite ferment is organically combined with a microbial fermentation technology, so that the composite ferment has a remarkable senescence delaying effect.
The first aspect of the invention provides a method for preparing a composite ferment, comprising the following steps:
mixing the raw materials with lecithin and glycerol, and homogenizing under high pressure to obtain a composite stock solution;
mixing the composite stock solution with a fermentation strain, and fermenting to obtain the composite fermentation product;
the raw materials comprise two or more of ginseng, ganoderma lucidum, tricholoma matsutake, dendrobium, red sage root, rehmannia root and medlar;
the pressure of the high-pressure homogenization is 20-100MPa;
the fermentation strain comprises one or more of bacillus subtilis, monascus and saccharomyces cerevisiae.
Compared with the traditional extraction technology, the development of the modern microbial fermentation technology brings new revelation for plant extraction. The plant active ingredient extraction technology formed by combining modern fermentation engineering can develop raw materials with the dual characteristics of plants and microorganisms. Research has shown that microorganisms can produce many bioactive components including various enzymes, small molecule compounds, etc. during growth, which are excellent skin care materials. After the plants are subjected to microbial fermentation treatment, the products of the plants contain more excellent and more effective skin protection effects; the products are compounded and applied by a scientific method, so that the activity of skin tissue cells can be enhanced, the metabolism of the skin tissue cells can be promoted, the microcirculation of the skin tissue can be improved, and the skin can be in a healthy state. The invention takes two or more of ginseng, ganoderma lucidum, tricholoma matsutake, dendrobium, red sage root, rehmannia root and medlar as raw materials, organically combines high-pressure homogenization and directional microorganism fermentation technologies, and fully breaks plant cells by lecithin and glycerin in the high-pressure homogenization process, so that various components in the cells are quickly and fully released, the utilization efficiency of the raw materials is improved, the composite stock solution obtained by high-pressure homogenization is used as a subsequent fermentation substrate, and the composite fermentation product with obvious aging delaying effect can be prepared through the bioconversion capability of microorganisms.
The raw materials used in the invention have the following effects:
the ginseng has the effects of whitening skin, removing speckle, removing wrinkle, resisting aging, moistening skin and retaining youthful looks, and mainly contains ginsenoside and ginseng polysaccharide.
Ganoderma has effects of enhancing qi color, removing wrinkle and resisting aging, and mainly contains triterpenes, polysaccharides, polypeptides and volatile oil.
The medlar has the effects of nourishing Rong Zengyan, removing spots and moisturizing skin, and mainly contains flavonoids, alkaloids, polysaccharide and vitamins.
The black tricholoma matsutake and the dendrobium contain rich polysaccharide and alkaloid components, and have good free radical removal capability and moisturizing effect.
The red sage root contains a large amount of phenolic acid components and has strong effects of resisting oxidation and promoting blood circulation.
The rehmannia contains iridoid components and has good anti-inflammatory effect.
Preferably, the temperature of the high-pressure homogenization is 25-50 ℃, and the time of the high-pressure homogenization is 5-40min. More preferably, the temperature of the high-pressure homogenization is 30-45 ℃, the pressure of the high-pressure homogenization is 40-100MPa, and the time of the high-pressure homogenization is 10-30min.
Preferably, the preparation method of the composite ferment comprises the following steps:
mixing Ginseng radix, ganoderma, tricholoma matsutake, herba Dendrobii, saviae Miltiorrhizae radix, rehmanniae radix, fructus Lycii with lecithin, glycerol, and water, homogenizing under high pressure to obtain composite stock solution;
mixing the composite stock solution with a fermentation strain, and fermenting to obtain the composite fermentation product;
the pressure of the high-pressure homogenization is 20-100MPa;
the fermentation strain comprises one or more of bacillus subtilis, monascus and saccharomyces cerevisiae;
the dosage ratio of the ginseng, the lucid ganoderma, the tricholoma matsutake, the dendrobium, the root of red-rooted salvia, the rehmannia, the medlar, the lecithin, the glycerol and the ionized water is (0.01-50 g): (0.01-50 g): (0.01-50 g): (0.01-50 g): (0.01-50 g): (0.01-50 g): (0.01-50 g): (0.01-50 g): 1g: (1-1000 g). More preferably, the dosage ratio of ginseng, ganoderma lucidum, tricholoma matsutake, dendrobium, red sage root, rehmannia root, medlar, lecithin, glycerol and ionized water is (0.01-1 g): (0.01-1 g): (0.01-1 g): (0.01-1 g): (0.01-1 g): (0.01-1 g): (0.01-1 g): (0.01-1 g): 1g: (1-100 g).
Preferably, the medium and water are also added before the fermentation is performed.
Preferably, the ratio of the amounts of the complex stock solution, the fermentation strain, the medium and the water is (1-500 g): (0.01-10 mL): 1g: (1-500 g).
Preferably, the concentration of the fermentation strain is 10 5 -10 9 CFU/mL。
Preferably, the temperature of the fermentation is 20-45 ℃, and the time of the fermentation is 12-120h. More preferably, the temperature of the fermentation is 25-40 ℃, and the time of the fermentation is 12-84h.
The second aspect of the invention provides a composite ferment prepared by the preparation method of the invention.
In a third aspect, the invention provides the use of a composite ferment according to the invention for the preparation of a cosmetic.
Based on the application, the invention also provides a cosmetic which comprises the composite ferment and auxiliary materials acceptable in the cosmetic, wherein the composite ferment accounts for 0.01-80% of the mass of the cosmetic.
Preferably, the cosmetically acceptable adjuvants include one or more of emulsifying agent, humectant, thickener, and antiseptic.
Preferably, the composite ferment accounts for 0.05-20% of the mass of the cosmetic.
Compared with the prior art, the invention has the following beneficial effects:
the invention organically combines high-pressure homogenization and directional microorganism fermentation technologies, takes two or more of ginseng, ganoderma lucidum, tricholoma matsutake, dendrobium, red sage root, rehmannia root and medlar as raw materials, and takes lecithin and glycerin as auxiliary materials in the high-pressure homogenization process, so that plant cells can be fully crushed, various components in the cells can be quickly and fully released, the utilization efficiency of the raw materials is improved, the composite stock solution obtained by the high-pressure homogenization is used as a subsequent fermentation substrate, and the composite fermentation product with obvious aging delaying effect can be prepared through the bioconversion capability of microorganisms.
The composite ferment prepared by the invention has high DPPH free radical clearance (up to 87.52%), can reduce the quantity of aging cells in human skin fibroblasts (HDF), reduce the relative expression quantity of matrix metalloproteinase-1 (MMP-1) mRNA in the HDF, improve the relative expression quantity of I-type collagen (COL-I) mRNA, has a protective effect on human immortalized keratinocytes (HaCat) after UV irradiation, promotes cell growth, has obvious anti-aging effect, and can reduce the content of inflammatory factors IL-1 beta and TNF-alpha generated by the HaCat cells after UV irradiation.
Detailed Description
In order to make the technical solutions of the present invention more apparent to those skilled in the art, the following examples will be presented. It should be noted that the following examples do not limit the scope of the invention.
The starting materials, reagents or apparatus used in the following examples are available from conventional commercial sources or may be obtained by methods known in the art unless otherwise specified.
Example 1
A method for preparing a composite ferment, comprising the following steps:
mixing 10g of ginseng, 10g of ganoderma lucidum, 5g of tricholoma matsutake, 5g of dendrobium, 5g of red sage root, 5g of rehmannia root, 5g of medlar, 5g of lecithin, 40g of glycerol and 310g of deionized water, adding into a high-pressure homogenizer, homogenizing at 40 ℃ under 80MPa for 20min to obtain a composite stock solution;
100g of the composite stock solution and 2mL of bacillus subtilis bacterial liquid (concentration is 10) 7 CFU/mL), 4g of LB medium and 100g of deionized water, fermenting at 30 ℃ for 48h, sterilizing the fermentation broth at 121 ℃ for 30min after the fermentation is finished, centrifuging for 30min at a rotation speed of 5000rpm/min, and taking the supernatant after centrifugation to obtain a composite fermentation product.
Example 2
A method for preparing a composite ferment, comprising the following steps:
mixing 10g of ginseng, 10g of lucid ganoderma, 10g of dendrobium, 5g of red sage root, 5g of rehmannia root, 5g of medlar, 5g of lecithin, 40g of glycerol and 310g of deionized water, adding into a high-pressure homogenizer, homogenizing at 40 ℃ under 80MPa for 20min to obtain a composite stock solution;
100g of the composite stock solution and 2mL of bacillus subtilis bacterial liquid (concentration is 10) 7 CFU/mL), 4g of LB medium and 100g of deionized water, fermenting at 30 ℃ for 48h, sterilizing the fermentation broth at 121 ℃ for 30min after the fermentation is finished, centrifuging for 30min at a rotation speed of 5000rpm/min, and taking the supernatant after centrifugation to obtain a composite fermentation product.
Example 3
A method for preparing a composite ferment, comprising the following steps:
mixing 10g of ginseng, 10g of lucid ganoderma, 10g of dendrobium, 5g of red sage root, 10g of rehmannia root, 5g of lecithin, 40g of glycerol and 310g of deionized water, adding into a high-pressure homogenizer, homogenizing at 40 ℃ under 80MPa for 20min to obtain a composite stock solution;
100g of the composite stock solution and 2mL of bacillus subtilis bacterial liquid (concentration is 10) 7 CFU/mL), 4g of LB medium and 100g of deionized water, fermenting at 30 ℃ for 48h, sterilizing the fermentation broth at 121 ℃ for 30min after the fermentation is finished, centrifuging for 30min at a rotation speed of 5000rpm/min, and taking the supernatant after centrifugation to obtain a composite fermentation product.
Example 4
A method for preparing a composite ferment, comprising the following steps:
mixing 10g of ginseng, 10g of lucid ganoderma, 10g of dendrobium, 5g of red sage root, 10g of medlar, 5g of lecithin, 40g of glycerol and 310g of deionized water, adding into a high-pressure homogenizer, homogenizing at 40 ℃ under 80MPa for 20min to obtain a composite stock solution;
100g of the composite stock solution and 2mL of bacillus subtilis bacterial liquid (concentration is 10) 7 CFU/mL), 4g of LB medium and 100g of deionized water, fermenting at 30 ℃ for 48h, sterilizing the fermentation broth at 121 ℃ for 30min after the fermentation is finished, centrifuging for 30min at a rotation speed of 5000rpm/min, and taking the supernatant after centrifugation to obtain a composite fermentation product.
Example 5
A method for preparing a composite ferment, comprising the following steps:
mixing 15g of ginseng, 10g of lucid ganoderma, 10g of dendrobium, 10g of medlar, 10g of lecithin, 35g of glycerol and 310g of deionized water, and then adding into a high-pressure homogenizer at a homogenizing temperature of 40 ℃ and a homogenizing pressure of 100MPa for 15min to obtain a composite stock solution;
100g of the composite stock solution and 2mL of bacillus subtilis bacterial liquid (concentration is 10) 7 CFU/mL), 4g of LB medium and 100g of deionized water, fermenting at 30 ℃ for 48h, sterilizing the fermentation broth at 121 ℃ for 30min after the fermentation is finished, centrifuging for 30min at a rotation speed of 5000rpm/min, and taking the supernatant after centrifugation to obtain a composite fermentation product.
Example 6
A method for preparing a composite ferment, comprising the following steps:
mixing 15g of ginseng, 15g of lucid ganoderma, 10g of dendrobium, 5g of lecithin, 55g of glycerol and 300g of deionized water, and adding into a high-pressure homogenizer at a homogenizing temperature of 40 ℃ and a homogenizing pressure of 100MPa for 15min to obtain a composite stock solution;
100g of the composite stock solution and 2mL of bacillus subtilis bacterial liquid (concentration is 10) 7 CFU/mL), 4g LB culture medium and 100g deionized water, fermenting at 30deg.C for 48h, sterilizing the fermentation broth at 121deg.C for 30min, and rotating at rotation speedCentrifuging at 5000rpm/min for 30min, and collecting supernatant to obtain composite fermentation product.
Example 7
A method for preparing a composite ferment, comprising the following steps:
mixing 20g of ginseng, 20g of ganoderma lucidum, 5g of lecithin, 55g of glycerol and 300g of deionized water, and then adding into a high-pressure homogenizer at a homogenizing temperature of 40 ℃ and a homogenizing pressure of 80MPa for 25min to obtain a composite stock solution;
100g of the composite stock solution and 2mL of bacillus subtilis bacterial liquid (concentration is 10) 7 CFU/mL), 4g of LB medium and 100g of deionized water, fermenting at 30 ℃ for 48h, sterilizing the fermentation broth at 121 ℃ for 30min after the fermentation is finished, centrifuging for 30min at a rotation speed of 5000rpm/min, and taking the supernatant after centrifugation to obtain a composite fermentation product.
Example 8
A method for preparing a composite ferment, comprising the following steps:
mixing 10g of ginseng, 10g of ganoderma lucidum, 5g of tricholoma matsutake, 5g of dendrobium, 5g of red sage root, 5g of rehmannia root, 5g of medlar, 5g of lecithin, 40g of glycerol and 310g of deionized water, adding into a high-pressure homogenizer, homogenizing at 40 ℃ under 80MPa for 20min to obtain a composite stock solution;
100g of the composite stock solution and 1mL of monascus bacterial solution (concentration is 10) 7 CFU/mL), 4g of PDB medium and 100g of deionized water, fermenting at 30 ℃ for 30 hours, sterilizing the fermentation broth at 121 ℃ for 30 minutes after the fermentation is finished, centrifuging for 30 minutes at a rotation speed of 5000rpm/min, and taking the supernatant after centrifugation to obtain a composite fermentation product.
Example 9
A method for preparing a composite ferment, comprising the following steps:
mixing 10g of ginseng, 10g of lucid ganoderma, 10g of dendrobium, 5g of red sage root, 5g of rehmannia root, 5g of medlar, 5g of lecithin, 40g of glycerol and 310g of deionized water, adding into a high-pressure homogenizer, homogenizing at 40 ℃ under 80MPa for 20min to obtain a composite stock solution;
100g of composite stock solution and 1mL of red riceMould liquid (concentration 10) 7 CFU/mL), 4g of PDB medium and 100g of deionized water, fermenting at 30 ℃ for 30 hours, sterilizing the fermentation broth at 121 ℃ for 30 minutes after the fermentation is finished, centrifuging for 30 minutes at a rotation speed of 5000rpm/min, and taking the supernatant after centrifugation to obtain a composite fermentation product.
Example 10
A method for preparing a composite ferment, comprising the following steps:
mixing 10g of ginseng, 10g of lucid ganoderma, 10g of dendrobium, 5g of red sage root, 10g of rehmannia root, 5g of lecithin, 40g of glycerol and 310g of deionized water, adding into a high-pressure homogenizer, homogenizing at 40 ℃ under 80MPa for 20min to obtain a composite stock solution;
100g of the composite stock solution and 1mL of monascus bacterial solution (concentration is 10) 7 CFU/mL), 4g of PDB medium and 100g of deionized water, fermenting at 30 ℃ for 30 hours, sterilizing the fermentation broth at 121 ℃ for 30 minutes after the fermentation is finished, centrifuging for 30 minutes at a rotation speed of 5000rpm/min, and taking the supernatant after centrifugation to obtain a composite fermentation product.
Example 11
A method for preparing a composite ferment, comprising the following steps:
mixing 10g of ginseng, 10g of lucid ganoderma, 10g of dendrobium, 5g of red sage root, 10g of medlar, 5g of lecithin, 40g of glycerol and 310g of deionized water, adding into a high-pressure homogenizer, homogenizing at 40 ℃ under 80MPa for 20min to obtain a composite stock solution;
100g of the composite stock solution and 1mL of monascus bacterial solution (concentration is 10) 7 CFU/mL), 4g of PDB medium and 100g of deionized water, fermenting at 30 ℃ for 30 hours, sterilizing the fermentation broth at 121 ℃ for 30 minutes after the fermentation is finished, centrifuging for 30 minutes at a rotation speed of 5000rpm/min, and taking the supernatant after centrifugation to obtain a composite fermentation product.
Example 12
A method for preparing a composite ferment, comprising the following steps:
mixing 15g of ginseng, 10g of lucid ganoderma, 10g of dendrobium, 10g of medlar, 10g of lecithin, 35g of glycerol and 310g of deionized water, and then adding into a high-pressure homogenizer at a homogenizing temperature of 40 ℃ and a homogenizing pressure of 100MPa for 15min to obtain a composite stock solution;
100g of the composite stock solution and 1mL of monascus bacterial solution (concentration is 10) 7 CFU/mL), 4g of PDB medium and 100g of deionized water, fermenting at 30 ℃ for 30 hours, sterilizing the fermentation broth at 121 ℃ for 30 minutes after the fermentation is finished, centrifuging for 30 minutes at a rotation speed of 5000rpm/min, and taking the supernatant after centrifugation to obtain a composite fermentation product.
Example 13
A method for preparing a composite ferment, comprising the following steps:
mixing 15g of ginseng, 15g of lucid ganoderma, 10g of dendrobium, 5g of lecithin, 55g of glycerol and 300g of deionized water, and adding into a high-pressure homogenizer at a homogenizing temperature of 40 ℃ and a homogenizing pressure of 100MPa for 15min to obtain a composite stock solution;
100g of the composite stock solution and 1mL of monascus bacterial solution (concentration is 10) 7 CFU/mL), 4g of PDB medium and 100g of deionized water, fermenting at 30 ℃ for 30 hours, sterilizing the fermentation broth at 121 ℃ for 30 minutes after the fermentation is finished, centrifuging for 30 minutes at a rotation speed of 5000rpm/min, and taking the supernatant after centrifugation to obtain a composite fermentation product.
Example 14
A method for preparing a composite ferment, comprising the following steps:
mixing 20g of ginseng, 20g of ganoderma lucidum, 5g of lecithin, 55g of glycerol and 300g of deionized water, and then adding into a high-pressure homogenizer at a homogenizing temperature of 40 ℃ and a homogenizing pressure of 80MPa for 25min to obtain a composite stock solution;
100g of the composite stock solution and 1mL of monascus bacterial solution (concentration is 10) 7 CFU/mL), 4g of PDB medium and 100g of deionized water, fermenting at 30 ℃ for 30 hours, sterilizing the fermentation broth at 121 ℃ for 30 minutes after the fermentation is finished, centrifuging for 30 minutes at a rotation speed of 5000rpm/min, and taking the supernatant after centrifugation to obtain a composite fermentation product.
Example 15
A method for preparing a composite ferment, comprising the following steps:
mixing 10g of ginseng, 10g of ganoderma lucidum, 5g of tricholoma matsutake, 5g of dendrobium, 5g of red sage root, 5g of rehmannia root, 5g of medlar, 5g of lecithin, 40g of glycerol and 310g of deionized water, adding into a high-pressure homogenizer, homogenizing at 40 ℃ under 80MPa for 20min to obtain a composite stock solution;
100g of the composite stock solution and 1mL of Saccharomyces cerevisiae solution (concentration 10) 7 CFU/mL), 2g of PDB medium and 100g of deionized water, fermenting at 32 ℃ for 72 hours, sterilizing the fermentation liquor at 121 ℃ for 30 minutes after the fermentation is finished, centrifuging for 30 minutes at a rotation speed of 5000rpm/min, and taking the supernatant after centrifugation to obtain a composite fermentation product.
Example 16
A method for preparing a composite ferment, comprising the following steps:
mixing 10g of ginseng, 10g of lucid ganoderma, 10g of dendrobium, 5g of red sage root, 5g of rehmannia root, 5g of medlar, 5g of lecithin, 40g of glycerol and 310g of deionized water, adding into a high-pressure homogenizer, homogenizing at 40 ℃ under 80MPa for 20min to obtain a composite stock solution;
100g of the composite stock solution and 1mL of Saccharomyces cerevisiae solution (concentration 10) 7 CFU/mL), 2g of PDB medium and 100g of deionized water, fermenting at 32 ℃ for 72 hours, sterilizing the fermentation liquor at 121 ℃ for 30 minutes after the fermentation is finished, centrifuging for 30 minutes at a rotation speed of 5000rpm/min, and taking the supernatant after centrifugation to obtain a composite fermentation product.
Example 17
A method for preparing a composite ferment, comprising the following steps:
mixing 10g of ginseng, 10g of lucid ganoderma, 10g of dendrobium, 5g of red sage root, 10g of rehmannia root, 5g of lecithin, 40g of glycerol and 310g of deionized water, adding into a high-pressure homogenizer, homogenizing at 40 ℃ under 80MPa for 20min to obtain a composite stock solution;
100g of the composite stock solution and 1mL of Saccharomyces cerevisiae solution (concentration 10) 7 CFU/mL), 2g PDB culture medium and 100g deionized water, fermenting at 32deg.C for 72 hr, sterilizing the fermentation broth at 121deg.C for 30min after fermentation, centrifuging at 5000rpm/min for 30min, collecting the supernatant, and collecting the composite fermentationAnd (3) an object.
Example 18
A method for preparing a composite ferment, comprising the following steps:
mixing 10g of ginseng, 10g of lucid ganoderma, 10g of dendrobium, 5g of red sage root, 10g of medlar, 5g of lecithin, 40g of glycerol and 310g of deionized water, adding into a high-pressure homogenizer, homogenizing at 40 ℃ under 80MPa for 20min to obtain a composite stock solution;
100g of the composite stock solution and 1mL of Saccharomyces cerevisiae solution (concentration 10) 7 CFU/mL), 2g of PDB medium and 100g of deionized water, fermenting at 32 ℃ for 72 hours, sterilizing the fermentation liquor at 121 ℃ for 30 minutes after the fermentation is finished, centrifuging for 30 minutes at a rotation speed of 5000rpm/min, and taking the supernatant after centrifugation to obtain a composite fermentation product.
Example 19
A method for preparing a composite ferment, comprising the following steps:
mixing 15g of ginseng, 10g of lucid ganoderma, 10g of dendrobium, 10g of medlar, 10g of lecithin, 35g of glycerol and 310g of deionized water, and then adding into a high-pressure homogenizer at a homogenizing temperature of 40 ℃ and a homogenizing pressure of 100MPa for 15min to obtain a composite stock solution;
100g of the composite stock solution and 1mL of Saccharomyces cerevisiae solution (concentration 10) 7 CFU/mL), 2g of PDB medium and 100g of deionized water, fermenting at 32 ℃ for 72 hours, sterilizing the fermentation liquor at 121 ℃ for 30 minutes after the fermentation is finished, centrifuging for 30 minutes at a rotation speed of 5000rpm/min, and taking the supernatant after centrifugation to obtain a composite fermentation product.
Example 20
A method for preparing a composite ferment, comprising the following steps:
mixing 15g of ginseng, 15g of lucid ganoderma, 10g of dendrobium, 5g of lecithin, 55g of glycerol and 300g of deionized water, and adding into a high-pressure homogenizer at a homogenizing temperature of 40 ℃ and a homogenizing pressure of 100MPa for 15min to obtain a composite stock solution;
100g of the composite stock solution and 1mL of Saccharomyces cerevisiae solution (concentration 10) 7 CFU/mL), 2g PDB medium and 100g deionized water, and fermenting at 32deg.C for 72hSterilizing the fermentation liquor at 121 ℃ for 30min after fermentation is finished, centrifuging for 30min at the rotation speed of 5000rpm/min, and taking the supernatant after centrifugation to obtain the composite fermentation product.
Example 21
A method for preparing a composite ferment, comprising the following steps:
mixing 20g of ginseng, 20g of ganoderma lucidum, 5g of lecithin, 55g of glycerol and 300g of deionized water, and then adding into a high-pressure homogenizer at a homogenizing temperature of 40 ℃ and a homogenizing pressure of 80MPa for 25min to obtain a composite stock solution;
100g of the composite stock solution and 1mL of Saccharomyces cerevisiae solution (concentration 10) 7 CFU/mL), 2g of PDB medium and 100g of deionized water, fermenting at 32 ℃ for 72 hours, sterilizing the fermentation liquor at 121 ℃ for 30 minutes after the fermentation is finished, centrifuging for 30 minutes at a rotation speed of 5000rpm/min, and taking the supernatant after centrifugation to obtain a composite fermentation product.
Comparative example 1 (differing from example 1 in that conventional water decoction was used)
A preparation method of the compound extract comprises the following steps:
mixing 10g of ginseng, 10g of ganoderma lucidum, 5g of tricholoma matsutake, 5g of dendrobium, 5g of red sage root, 5g of rehmannia root, 5g of medlar, 5g of lecithin, 40g of glycerol and 310g of deionized water, heating and extracting for 2 hours at 100 ℃, filtering, and collecting filtrate to obtain a compound extract.
Comparative example 2 (the difference from example 1 is that the Bacillus subtilis liquid was replaced with deionized water)
This comparative example was compared with example 1, except that the bacillus subtilis liquid was replaced with deionized water, and the addition amounts were the same, and other components, addition amounts, and preparation methods were the same as in example 1.
Comparative example 3 (differing from example 1 in that no lecithin or glycerin was added)
This comparative example is compared to example 1, except that equal amounts of deionized water are used instead of lecithin and glycerin, and other components, amounts of addition, and preparation methods are the same as in example 1.
Comparative example 4 (differs from example 1 in that lecithin was replaced with soybean oil)
This comparative example was compared with example 1, except that lecithin was replaced with soybean oil and the addition amount was the same, and other components, addition amounts, and preparation method were the same as in example 1.
Comparative example 5 (differs from example 1 in that glycerol is replaced by butanediol)
This comparative example was compared with example 1, except that glycerin was replaced with butanediol, and the addition amount was the same, and other components, addition amounts, and preparation methods were the same as example 1.
Comparative example 6 (differing from example 1 in the use of a single raw material)
A method for preparing a composite ferment, comprising the following steps:
mixing 45g of ginseng, 5g of lecithin, 40g of glycerol and 310g of deionized water, adding into a high-pressure homogenizer, homogenizing at 40 ℃ under 80MPa for 20min to obtain a composite stock solution;
100g of the composite stock solution and 2mL of bacillus subtilis bacterial liquid (concentration is 10) 7 CFU/mL), 4g of LB medium and 100g of deionized water, fermenting at 30 ℃ for 48h, sterilizing the fermentation broth at 121 ℃ for 30min after the fermentation is finished, centrifuging for 30min at a rotation speed of 5000rpm/min, and taking the supernatant after centrifugation to obtain a composite fermentation product.
Comparative example 7 (differing from example 8 in the use of a single starting material)
Mixing 45g of medlar, 5g of lecithin, 40g of glycerol and 310g of deionized water, and then adding into a high-pressure homogenizer at a homogenizing temperature of 40 ℃ and a homogenizing pressure of 80MPa for 20min to obtain a composite stock solution;
100g of the composite stock solution and 1mL of monascus bacterial solution (concentration is 10) 7 CFU/mL), 4g of pdb medium and 100g of deionized water, fermenting at 30 ℃ for 48 hours, sterilizing the fermentation broth at 121 ℃ for 30 minutes after the fermentation is finished, centrifuging for 30 minutes at a rotation speed of 5000rpm/min, and taking the supernatant after centrifugation to obtain a composite fermentation product.
Comparative example 8 (differs from example 15 in the use of a single starting material)
45g of lucid ganoderma, 5g of lecithin, 40g of glycerin and 310g of deionized water are mixed and then added into a high-pressure homogenizer, the homogenizing temperature is 40 ℃, the homogenizing pressure is 80MPa, and the homogenizing time is 20min, so that a composite stock solution is obtained;
100g of the composite stock solution and 1mL of Saccharomyces cerevisiae solution (concentration 10) 7 CFU/mL), 2g of pdb medium and 100g of deionized water, fermenting at 30 ℃ for 48 hours, sterilizing the fermentation broth at 121 ℃ for 30 minutes after the fermentation is finished, centrifuging for 30 minutes at a rotation speed of 5000rpm/min, and taking the supernatant after centrifugation to obtain a composite fermentation product.
Comparative example 9 (the difference from example 1 is that the order of addition of lecithin and glycerin is different)
A method for preparing a composite ferment, comprising the following steps:
mixing 10g of ginseng, 10g of ganoderma lucidum, 5g of tricholoma matsutake, 5g of dendrobium, 5g of red sage root, 5g of rehmannia root, 5g of medlar and 310g of deionized water, adding into a high-pressure homogenizer, homogenizing at 40 ℃ under 80MPa for 20min to obtain a composite stock solution;
100g of the composite stock solution and 2mL of bacillus subtilis bacterial liquid (concentration is 10) 7 CFU/mL), 4g of LB medium, 5g of lecithin, 40g of glycerol and 100g of deionized water, fermenting at 30 ℃ for 48h, sterilizing the fermentation broth at 121 ℃ for 30min after the fermentation is finished, centrifuging for 30min at a rotating speed of 5000rpm/min, and taking the supernatant after centrifugation to obtain the composite fermentation product.
Test example 1: erythrocyte hemolysis evaluation
The basic principle of the erythrocyte hemolysis experiment is to evaluate the damage of chemicals to ocular tissue cells by measuring the leaching amount and the denaturation degree of hemoglobin.
The samples prepared in examples 1-21 and comparative examples 1-9 were subjected to a erythrocyte hemolysis test in accordance with the RBC test method and fractionation criteria of European alternative validation center (ECVAM). Where HD50 is the concentration of the sample at which 50% of the erythrocytes are hemolyzed, DI is the protein denaturation index, and L/D is the ratio of HD50 to DI. The evaluation criteria are shown in Table 1; the evaluation results are shown in Table 2.
TABLE 1
| L/D | Grading |
| L/D>100 | No irritation |
| 10<L/D≤100 | Microstimulation |
| 1<L/D≤10 | Mild irritation |
| 0.1<L/D≤1 | Toxicity irritation |
| L/D≤0.1 | Severe irritation |
TABLE 2
| Sample of | Grading | Sample of | Grading |
| Example 1 | No irritation | Example 16 | No irritation |
| Example 2 | No irritation | Example 17 | No irritation |
| Example 3 | No irritation | Example 18 | No irritation |
| Example 4 | No irritation | Example 19 | No irritation |
| Example 5 | No irritation | Example 20 | No irritation |
| Example 6 | No irritation | Example 21 | No irritation |
| Example 7 | No irritation | Comparative example 1 | Microstimulation |
| Implementation of the embodimentsExample 8 | No irritation | Comparative example 2 | No irritation |
| Example 9 | No irritation | Comparative example 3 | No irritation |
| Example 10 | No irritation | Comparative example 4 | No irritation |
| Example 11 | No irritation | Comparative example 5 | No irritation |
| Example 12 | No irritation | Comparative example 6 | No irritation |
| Example 13 | No irritation | Comparative example 7 | No irritation |
| Example 14 | No irritation | Comparative example 8 | No irritation |
| Example 15 | No irritation | Comparative example 9 | No irritation |
As can be seen from Table 2, the samples prepared in examples 1-21 and comparative examples 2-9 were not stimulatory to red blood cells except for comparative example 1.
Test example 2: DPPH radical scavenging ability evaluation
The DPPH free radical scavenging ability is to some extent the antioxidant capacity of the reactive species. The higher the free radical scavenging rate, the stronger the antioxidant capacity and the stronger the anti-aging capacity. Therefore, the anti-aging effect of the DPPH free radical scavenging ability can be judged by the research of the sample. The following test was carried out on the samples prepared in examples 1 to 21 and comparative examples 1 to 9, and the results are shown in Table 3.
Adding 2.0mL of DPPH test solution with the mass concentration of 45.8mg/L and 0.05mL of sample into a test tube, supplementing the total volume to 3mL by 50% ethanol (v/v), shaking uniformly, carrying out light-shielding reaction for 30min, and measuring absorbance at 517nm wavelength by using A1 cm cuvette, and marking as A1; adding 2mL of absolute ethyl alcohol and a sample to be detected with a corresponding volume into a test tube, supplementing the total volume to 3mL by using 50% ethanol (v/v), and recording the measured absorbance as A2; to the test tube, 2mL of DPPH test solution and 1mL of 50% ethanol (v/v) were added, and the absorbance was measured and designated as A3. Calculating the clearance rate (Y) of the liquid to be tested to DPPH free radicals according to the following formula;
the calculation formula of the DPPH free radical clearance is as follows: y (%) = [1- (A1-A2)/A3 ] ×100;
wherein A1 is the absorbance value of the system after the blank is cleared of DPPH, A2 is the absorbance value of the system after the sample is cleared of DPPH, and A3 is the absorbance value of the reaction system before the medicine is added.
TABLE 3 Table 3
The results in Table 3 show that the complex ferments prepared in examples 1-21 have a stronger DPPH radical scavenging ability than those prepared in comparative examples 1-9.
Test example 3: senescence-associated beta-galactosidase (SA-beta-gal) assay
Human skin fibroblasts (HDF) were 1×10 per well in 6-well plates 4 The density of individual cells was at 37℃with a mass fraction of 5% CO 2 Culturing for 24 hours. After 24 hours of treatment of HDF with the sample (5%), the HDF was washed twice with Phosphate Buffered Saline (PBS) at 30J/cm 2 Is irradiated with UVB. Then, PBS was removed and the HDF sample (5%) was subjected to a mass fraction of 5% CO at 37% 2 Culturing for 24 hours. Wash with 1ml PBS, then add 0.5ml fixative, then leave at room temperature for 15 minutes to fix. To the immobilized HDF, 0.5mL of a mixed staining solution (470. Mu.L of a staining solution, 5. Mu.L of a staining replenishment solution, 25. Mu.L of dimethylformamide containing 20mg/mL of X-gal) was added, and the cells were stained by culturing at 37℃for 24 hours. The stained cells were then washed with PBS, and 500 fibroblast microscopes were randomly selected for each sample plate to take photographs and the SA-. Beta. -Gal stained fibroblasts were counted. Relative ratio of senescent cells = number of senescent cells in sample/number of senescent cells in model group x 100; the results are shown in Table 4.
TABLE 4 Table 4
Table 4 shows that the composite ferments prepared in examples 1-21 have an effect of delaying aging by reducing the number of aging cells in HDF after treating HDF with the composite ferments prepared in examples 1-21, compared with comparative examples 1-9.
Test example 4: human type I collagen and matrix metalloproteinase analysis
HDF was 2×10 per well in 6-well plates 4 The density of individual cells was at 37℃with a mass fraction of 5% CO 2 Culturing for 24 hours. After 24 hours of treatment of HDF with the sample (5%), the HDF was washed twice with PBS at 30J/cm 2 Is irradiated with UVB. Then, PBS was removed and the HDF sample (5%) was subjected to a mass fraction of 5% CO at 37% 2 Culturing for 24 hours. After the completion of the culture, the contents of human type I collagen (COL-I) and matrix metalloproteinase-1 (MMP-1) were determined according to the ELISA kit procedure.
Purification of the total RNA extract was performed according to the kit protocol. RNA (3. Mu.L) concentration and purity were measured by a ratio of absorbance at 260/280nm and the remaining RNA solution was stored at-80℃for reverse transcription. The total RNA was converted to first strand complementary DNA (cDNA) by the following reverse transcription system: 4. Mu.L of 5x prime Script RT Master MIX, 0.5. Mu.g of total RNA and 20. Mu.L of RNase-free water in the reaction. The cDNA was used in real-time PCR and the reaction system contained 10. Mu. L SYBR Premix EX Taq (2X), 1. Mu.L forward primer (10. Mu.M), 1. Mu.L reverse primer (10. Mu.M) and 8. Mu.L cDNA, which was amplified in Applied Biosystems7500Fast Real time PCR System v version 2.3 according to the following conditions: 50.0deg.C for 3 min, 95.0deg.C for 3 min, then 95.0deg.C for 10 sec and 60.0deg.C for 30 sec for 40 cycles. Threshold cycle (Ct) was analyzed by instrument software and according to the comparative Ct method (2 -ΔΔCt ) Fold change in mRNA expression was calculated. The forward and reverse primer sequences of the genes are shown in Table 5; the test results are shown in tables 6 and 7.
TABLE 5
TABLE 6
TABLE 7
The results in tables 6 and 7 show that the complex ferments prepared in examples 1-21 are better able to reduce MMP-1 content and increase COL-I content than those prepared in comparative examples 1-9. In addition, the complex ferment prepared in examples 1-21 can reduce the relative expression amount of MMP-1mRNA in HDF and improve the relative expression of COL-I mRNA. As the improvement of MMP-1 and the reduction of COL-I can cause skin aging and wrinkle, the composite fermentation products prepared in the embodiments 1-21 of the invention can improve the content of COL-I while reducing the content of MMP-1, which shows that the composite fermentation products in the experimental examples 1-21 have stronger anti-aging effect.
Test example 5: effect on human immortalized keratinocyte (HaCat) cell viability following UV irradiation
HaCat cells were seeded into 6-well plates at 37℃and 5% CO 2 Is incubated overnight. When the cell plating rate in the 6-hole plate reaches 50% -60%, adding cell culture medium into blank control group and model group, adding cell culture medium containing sample (5%) with corresponding concentration into sample group, pre-treating for 1 hr, and performing UVB irradiation (30 mJ/cm 2 ) And 40min. At 37℃and 5% CO 2 After continuous culture for 24 hours, the culture medium was aspirated, washed 2-3 times with PBS, and the cell viability was tested by MTT method, cell relative viability (%) =sample group cell viability/model group cell viability×100, and the test results are shown in table 8.
TABLE 8
| Sample of | Relative cell viability (%) | Sample of | Relative cell viability (%) |
| Example 1 | 170.3 | Example 16 | 158.8 |
| Example 2 | 157.6 | Example 17 | 161.3 |
| Example 3 | 160.9 | Example 18 | 155.5 |
| Example 4 | 148.2 | Example 19 | 154.0 |
| Example 5 | 140.1 | Example 20 | 144.2 |
| Example 6 | 143.5 | Example 21 | 141.3 |
| Example 7 | 138.2 | Comparative example 1 | 109.7 |
| Example 8 | 166.8 | Comparative example 2 | 111.0 |
| Example 9 | 160.2 | Comparative example 3 | 117.2 |
| Example 10 | 161.3 | Comparative example 4 | 122.1 |
| Example 11 | 154.2 | Comparative example 5 | 119.4 |
| Example 12 | 146.3 | Comparative example 6 | 125.6 |
| Example 13 | 137.8 | Comparative example 7 | 120.3 |
| Example 14 | 140.3 | Comparative example 8 | 121.5 |
| Example 15 | 168.0 | Comparative example 9 | 124.4 |
The results in Table 8 show that the complex fermentates prepared in examples 1-21 have better protection against UV-irradiated HaCat cells and promote cell growth compared to comparative examples 1-9.
Test example 6: effect on IL-1 beta and TNF-alpha content in HaCaT cells after UV irradiation
HaCat cells were seeded into 6-well plates at 37℃and 5% CO 2 Is incubated overnight. When the cell plating rate in the 6-hole plate reaches 50% -60%, adding cell culture medium into blank control group and model group, adding cell culture medium containing sample (5%) of corresponding concentration of sample into sample group, pretreating for 1 hr, and performing UVB irradiation (30 mJ/cm 2 ) And 40min. At 37℃and 5% CO 2 The culture was continued for 24 hours. After the completion of the incubation, the IL-1β and TNF- α contents were measured according to the ELISA kit procedure, and the test results are shown in Table 9.
TABLE 9
The results in Table 9 show that the complex ferments prepared in examples 1-21 can reduce the content of inflammatory factors IL-1. Beta. And TNF-alpha. Produced by HaCat cells after UV irradiation, and exhibit anti-inflammatory activity, as compared with comparative examples 1-9.
Application example 1
The application example 1 provides an essence containing the composite fermentation product prepared in the embodiment 1, the specific formula of the essence is shown in table 10, and the preparation method is prepared according to the conventional preparation method of the essence.
Table 10
Comparative application example 1
This comparative application example 1 is different from application example 1 in that the complex fermented product obtained in example 1 is replaced with the complex extract obtained in comparative example 1, and the addition amounts are the same, and other components, addition amounts, and preparation methods are the same as those of application example 1.
Evaluation of effects of essence of application example 1 and comparative application example 1
1. The subjects selected 30 volunteers, aged 20-45 years, male and female halves. The essence of application example 1 was used on the left side, the essence of comparative application example 1 was used on the right side, once each day in the morning and evening, each time 1g of the product was applied to the face, and the face was massaged for 2 minutes. And filling out a trial evaluation table according to the sensory evaluation. Statistics were performed once for 7 days, 14 days, and 28 days. The volunteers evaluate the effect of skin elasticity (0-5 min) and compactness (0-5 min) on the samples during use, wherein the highest score is 5, which indicates remarkable effect, and the lowest score is 0, which indicates no effect. The results are shown in Table 11.
TABLE 11
2. The subjects selected 30 volunteers and were female aged 30-45 years. Prior to testing, photographs were taken using a VISIA facial image analyzer to analyze the appearance of volunteer eye fine lines. The volunteers were asked to use the corresponding group of products after cleansing, wherein the left eye was using the essence of application example 1, the right eye was using the essence of comparative application example 1, and photographing and analysis were performed again after 1 minute as the instant wrinkle-lightening effect of the samples. The volunteer was then asked to continuously test the sample to be tested, 2 times daily, 0.5g each time, for continuous trial 28d, and follow-up on days 7 and 28, take a facial photograph and analyze as a long-lasting moire-free effect of the sample. The detection of the same volunteer is done by the same inspector. And calculating the change rate of the eye fine lines through an analysis system arranged in the detection instrument, and counting the result. The results are shown in Table 12.
Table 12
The results in tables 11 and 12 show that volunteers considered a significant improvement in skin elasticity, firmness, and eye fine wrinkles after using the concentrate prepared in application example 1, which is related to the ability of the complex ferment to increase COL-I content and decrease MMP-1 content over the concentrate prepared in comparative application example 1. The application of the composite ferment of the invention to cosmetics is proved to have remarkable anti-aging effect.
While the preferred embodiments of the present invention have been illustrated and described, the present invention is not limited to the embodiments, and various equivalent modifications and substitutions can be made by one skilled in the art without departing from the spirit of the present invention, and these are intended to be included in the scope of the present invention as defined in the appended claims.
Claims (10)
1. A method for preparing a composite ferment, which is characterized by comprising the following steps:
mixing the raw materials with lecithin and glycerol, and homogenizing under high pressure to obtain a composite stock solution;
mixing the composite stock solution with a fermentation strain, and fermenting to obtain the composite fermentation product;
the raw materials are ginseng and ganoderma lucidum;
or the raw materials are selected from ginseng, ganoderma lucidum and dendrobium;
or the raw materials are selected from ginseng, ganoderma lucidum, dendrobium and medlar;
or the raw materials are selected from ginseng, ganoderma lucidum, dendrobium, red sage root and medlar;
or the raw materials are selected from ginseng, ganoderma lucidum, dendrobium, red sage root and rehmannia root;
or the raw materials are selected from ginseng, ganoderma lucidum, dendrobium, red sage root, rehmannia root and medlar;
or the raw materials are selected from ginseng, ganoderma lucidum, tricholoma matsutake, dendrobium, red sage root, rehmannia root and medlar;
the pressure of the high-pressure homogenization is 20-100MPa;
the fermentation strain is composed of one or more of bacillus subtilis, monascus and saccharomyces cerevisiae.
2. The method according to claim 1, wherein the high pressure homogenization temperature is 25-50 ℃ and the high pressure homogenization time is 5-40min.
3. The method of claim 1, wherein a medium and water are also added prior to the fermentation.
4. The method according to claim 3, wherein the ratio of the amounts of the complex stock solution, the fermentation strain, the medium and the water is (1-500 g): (0.01-10 mL): 1g: (1-500 g).
5. The method according to claim 1, wherein the concentration of the fermentation strain is 10 5 -10 9 CFU/mL。
6. The method according to claim 1, wherein the fermentation is carried out at a temperature of 20 to 45 ℃ for a period of 12 to 120 hours.
7. A composite ferment prepared by the method of any one of claims 1 to 6.
8. Use of the complex ferment of claim 7 in the preparation of cosmetics.
9. A cosmetic, characterized by comprising the composite ferment as claimed in claim 7 and acceptable auxiliary materials in the cosmetic, wherein the composite ferment accounts for 0.01-80% of the mass of the cosmetic.
10. The cosmetic according to claim 9, wherein the composite ferment is 0.05-20% by mass of the cosmetic.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210483369.7A CN114931537B (en) | 2022-05-05 | 2022-05-05 | Composite ferment capable of delaying aging and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210483369.7A CN114931537B (en) | 2022-05-05 | 2022-05-05 | Composite ferment capable of delaying aging and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114931537A CN114931537A (en) | 2022-08-23 |
| CN114931537B true CN114931537B (en) | 2023-05-02 |
Family
ID=82863714
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210483369.7A Active CN114931537B (en) | 2022-05-05 | 2022-05-05 | Composite ferment capable of delaying aging and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114931537B (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106344510A (en) * | 2016-08-26 | 2017-01-25 | 晨光生物科技集团股份有限公司 | Industrial production method of lycopene microemulsion |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109431899A (en) * | 2018-11-27 | 2019-03-08 | 北京工商大学 | One kind 5 joins circle proferment pulp cosmetic and the preparation method and application thereof |
| CN111840384A (en) * | 2020-08-01 | 2020-10-30 | 武汉真福医药股份有限公司 | Anti-senile plaque composition and preparation method thereof |
| CN113520942B (en) * | 2021-07-19 | 2022-12-06 | 广州环亚化妆品科技股份有限公司 | Anti-oxidation and anti-aging five-ginseng fermentation product and preparation method and application thereof |
-
2022
- 2022-05-05 CN CN202210483369.7A patent/CN114931537B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106344510A (en) * | 2016-08-26 | 2017-01-25 | 晨光生物科技集团股份有限公司 | Industrial production method of lycopene microemulsion |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114931537A (en) | 2022-08-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111297791B (en) | Anti-aging and repairing skin care composition containing symbiotic bacteria combined fermentation product, essence milk and preparation method and application of essence milk | |
| CN112494406A (en) | Whitening and freckle-removing cosmetic and preparation method thereof | |
| CN111743851B (en) | Whitening and spot-fading composition and application thereof | |
| CN108815072B (en) | Fresh plant composition fermented raw pulp with moisturizing and relieving effects and preparation method and application thereof | |
| CN113520942B (en) | Anti-oxidation and anti-aging five-ginseng fermentation product and preparation method and application thereof | |
| CN115678805B (en) | Preparation method and application of tricholoma matsutake yeast fermentation liquor with repairing and anti-aging effects | |
| CN114588061B (en) | Anti-aging wrinkle-removing composition, preparation method thereof and skin care product | |
| CN104434760A (en) | Facial cream containing ingredient of cordyceps militaris and preparation method thereof | |
| CN113244127A (en) | Moisture-preserving and antioxidant cordyceps sinensis mask emulsion and preparation method thereof | |
| CN114931537B (en) | Composite ferment capable of delaying aging and preparation method and application thereof | |
| CN114632055B (en) | Whitening and relieving plant fermentation product and preparation method and application thereof | |
| CN111437243A (en) | Wrinkle-removing and anti-aging composition, formula of anti-aging cosmetic and preparation method of anti-aging cosmetic | |
| CN111374935A (en) | Polypeptide-containing composition and application thereof | |
| CN111419733B (en) | Ceramide polypeptide compound Anmyo honey | |
| CN111568831B (en) | Composition of Chinese herbal medicine extract for skin cosmetics and preparation method thereof | |
| CN115414289A (en) | Cordyceps flower cosmetics and preparation method thereof | |
| CN111603404B (en) | Skin care essence containing cell extracting solution and preparation method thereof | |
| CN112137939A (en) | Composition with functions of reducing fine lines and increasing skin elasticity, preparation method and application thereof | |
| CN112915060A (en) | Emulsion containing anti-aging composition and preparation method thereof | |
| CN114224800B (en) | Rosa plant extract and preparation method, composition and application thereof | |
| CN113440457A (en) | Bletilla striata formula and preparation method thereof | |
| CN115006328B (en) | Whitening and freckle-removing composition, and preparation method and application thereof | |
| CN115317410B (en) | Fermentation oil for relieving, preserving moisture and enhancing cell viability and preparation method and application thereof | |
| CN116421525B (en) | Paeonia root extract and preparation method thereof | |
| CN117338626B (en) | Anti-aging whitening and moisturizing composition and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |