CN113876616A - Skin care composition with wrinkle removing effect and preparation method thereof - Google Patents
Skin care composition with wrinkle removing effect and preparation method thereof Download PDFInfo
- Publication number
- CN113876616A CN113876616A CN202111196356.3A CN202111196356A CN113876616A CN 113876616 A CN113876616 A CN 113876616A CN 202111196356 A CN202111196356 A CN 202111196356A CN 113876616 A CN113876616 A CN 113876616A
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- yeast
- water
- skin care
- care composition
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- 238000002360 preparation method Methods 0.000 title claims abstract description 31
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims abstract description 61
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 54
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- 238000000855 fermentation Methods 0.000 claims abstract description 46
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- PRAKJMSDJKAYCZ-UHFFFAOYSA-N squalane Chemical compound CC(C)CCCC(C)CCCC(C)CCCCC(C)CCCC(C)CCCC(C)C PRAKJMSDJKAYCZ-UHFFFAOYSA-N 0.000 claims abstract description 42
- 229940041514 candida albicans extract Drugs 0.000 claims abstract description 32
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- HIQIXEFWDLTDED-UHFFFAOYSA-N 4-hydroxy-1-piperidin-4-ylpyrrolidin-2-one Chemical compound O=C1CC(O)CN1C1CCNCC1 HIQIXEFWDLTDED-UHFFFAOYSA-N 0.000 claims abstract description 21
- JXTPJDDICSTXJX-UHFFFAOYSA-N n-Triacontane Natural products CCCCCCCCCCCCCCCCCCCCCCCCCCCCCC JXTPJDDICSTXJX-UHFFFAOYSA-N 0.000 claims abstract description 21
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- CDQSJQSWAWPGKG-UHFFFAOYSA-N butane-1,1-diol Chemical compound CCCC(O)O CDQSJQSWAWPGKG-UHFFFAOYSA-N 0.000 claims abstract description 13
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- LXAHHHIGZXPRKQ-UHFFFAOYSA-N 5-fluoro-2-methylpyridine Chemical compound CC1=CC=C(F)C=N1 LXAHHHIGZXPRKQ-UHFFFAOYSA-N 0.000 claims abstract description 5
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- 239000003109 Disodium ethylene diamine tetraacetate Substances 0.000 description 18
- ZGTMUACCHSMWAC-UHFFFAOYSA-L EDTA disodium salt (anhydrous) Chemical compound [Na+].[Na+].OC(=O)CN(CC([O-])=O)CCN(CC(O)=O)CC([O-])=O ZGTMUACCHSMWAC-UHFFFAOYSA-L 0.000 description 18
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- NHBKXEKEPDILRR-UHFFFAOYSA-N 2,3-bis(butanoylsulfanyl)propyl butanoate Chemical compound CCCC(=O)OCC(SC(=O)CCC)CSC(=O)CCC NHBKXEKEPDILRR-UHFFFAOYSA-N 0.000 description 2
- HVBSAKJJOYLTQU-UHFFFAOYSA-N 4-aminobenzenesulfonic acid Chemical compound NC1=CC=C(S(O)(=O)=O)C=C1 HVBSAKJJOYLTQU-UHFFFAOYSA-N 0.000 description 2
- 101500025419 Homo sapiens Epidermal growth factor Proteins 0.000 description 2
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- GVUGOAYIVIDWIO-UFWWTJHBSA-N nepidermin Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)NC(=O)CNC(=O)[C@@H](NC(=O)[C@@H](NC(=O)[C@H](CS)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CS)NC(=O)[C@H](C)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](C)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CCSC)NC(=O)[C@H](CS)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CS)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CC(C)C)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CS)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O)C(C)C)[C@@H](C)CC)C(C)C)C(C)C)C1=CC=C(O)C=C1 GVUGOAYIVIDWIO-UFWWTJHBSA-N 0.000 description 2
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- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 1
- 235000020777 polyunsaturated fatty acids Nutrition 0.000 description 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 1
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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
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
-
- 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
- A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
- A61K8/36—Carboxylic acids; Salts or anhydrides thereof
- A61K8/365—Hydroxycarboxylic acids; Ketocarboxylic acids
-
- 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
- A61K8/40—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing nitrogen
- A61K8/44—Aminocarboxylic acids or derivatives thereof, e.g. aminocarboxylic acids containing sulfur; Salts; Esters or N-acylated derivatives thereof
-
- 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]
-
- 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
- A61K8/99—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from microorganisms other than algae or fungi, e.g. protozoa or bacteria
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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
-
- 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
- A61K2800/51—Chelating agents
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Birds (AREA)
- Mycology (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Engineering & Computer Science (AREA)
- Botany (AREA)
- Emergency Medicine (AREA)
- Tropical Medicine & Parasitology (AREA)
- Gerontology & Geriatric Medicine (AREA)
- Dermatology (AREA)
- Cosmetics (AREA)
Abstract
The invention discloses a skin care composition with wrinkle removing effect and a preparation method thereof, wherein the skin care composition is prepared from the following raw materials in percentage by weight: 11-13% of squalane, 0.01-0.03% of EDTA disodium, 4-6% of glycerol, 4.4-4.5% of butanediol, 0.2-0.4% of sodium lactate, 0.9-1.1% of isopropyl myristate, 1-5% of recombinant human elastin, 0.3-0.6% of scutellaria and tricholoma matsutake compound fermentation product, 1.5-2.0% of yeast extract, 1-5% of dry cell exosome, 0.01-0.03% of essence and the balance of water. The preparation method provided by the invention is reasonable in design, safe and stable, free of side effect and wide in application prospect.
Description
Technical Field
The invention relates to the technical field of skin care products, in particular to a skin care composition with a wrinkle removing effect and a preparation method thereof.
Background
Wrinkles on the face, deep or shallow, are marks left to us all the year round, but many beauty MM do not allow any wrinkles to appear on the face, and the wrinkles are formed by reducing elastic fibers for maintaining normal skin tension, weakening sebaceous gland secretion, reducing subcutaneous fat, and enabling the skin to be too loose and folded with deep tissues. Facial wrinkles are particularly common, and with the increase of age, the metabolism of the organism is weakened, and aging wrinkles are gradually formed.
Most skin care products with the wrinkle removing effect cannot solve the problems from the inside, and not only do not have the wrinkle removing effect, but also have the stimulation effect on the skin. Skin care products are not always safe, and some have various chemical components added to them, which, once they reach the surrounding of deep living cells through damaged, sensitive skin barriers, can cause irritation, but rather more serious skin problems.
Therefore, it is very meaningful to develop a cosmetic having a remarkable wrinkle-removing effect.
Disclosure of Invention
The invention aims to provide a skin care composition which is easier to absorb by skin and has remarkable skin wrinkle removing effect.
The above purpose is achieved by the following scheme:
the skin care composition with the wrinkle removing effect is characterized by being prepared from the following raw materials in percentage by weight: 11-13% of squalane, 0.01-0.03% of EDTA disodium, 4-6% of glycerol, 4.4-4.5% of butanediol, 0.2-0.4% of sodium lactate, 0.9-1.1% of isopropyl myristate, 1-5% of recombinant human elastin, 0.3-0.6% of scutellaria and tricholoma matsutake compound fermentation product, 1.5-2.0% of yeast extract, 1-5% of dry cell exosome, 0.01-0.03% of essence and the balance of water.
The skin care composition with the wrinkle removing effect is characterized in that the scutellaria baicalensis and tricholoma matsutake compound fermentation product is prepared from the following raw materials in parts by weight: 10-20 parts of scutellaria baicalensis, 3-5 parts of tricholoma matsutake, 0.3-0.6 part of lactic acid, 0.3-0.6 part of yeast powder, 0.8-1 part of peptone and 65-85 parts of water.
The preparation method of the skin care composition with the wrinkle removing effect is characterized by comprising the following steps: the method comprises the following steps:
(1) adding appropriate amount of water into yeast extract, and stirring to obtain yeast extract water solution;
(2) adding isopropyl myristate and recombinant human elastin into yeast extract water solution, centrifuging, removing supernatant, and drying to obtain compound material;
(3) adding squalane, EDTA disodium, glycerol and appropriate amount of water into a stirring pot, heating to 70-80 deg.C, adding yeast lysate extract, stem cell exosome, compound material and other raw materials, stirring at 20-30 deg.C, cooling, and discharging.
The preparation method of the skin care composition with the wrinkle removing effect is characterized by comprising the following steps: the preparation method of the scutellaria and tricholoma matsutake compound fermentation product comprises the following steps:
(1) cleaning Scutellariae radix and Tricholoma matsutake, mixing, stirring, extracting with ethanol, and concentrating under reduced pressure to obtain compound extract;
(2) inoculating yeast to a culture medium, performing activation culture to obtain activated yeast, inoculating the activated yeast to a liquid culture medium, and performing fermentation culture to obtain a yeast liquid;
(3) adding lactic acid, yeast powder, peptone, water, compound extract and yeast liquid into a fermentation tank according to the weight parts, and fermenting at 30-35 ℃ for 12-24h to obtain fermentation liquor;
(4) adding the obtained fermentation liquor into absolute ethyl alcohol, carrying out ultrasonic treatment, centrifuging, filtering and drying to obtain the product.
The invention has the beneficial effects that:
the skin care product formula containing the recombinant human-like elastin and the stem cell exosomes, provided by the invention, supplements collagen and simultaneously forms elastic fibers, effectively prevents collagen from losing and eliminates wrinkles, and compared with other products, the skin care product formula can lighten lines, tighten, resist aging, eliminate wrinkles and stimulate collagen regeneration through the recombinant human-like elastin; the stem cell exosome is a cell secretion (vesicle with the diameter of 40-100 nm) from stem cells, carries bioactive substances such as protein, mRNA and microRNA of the stem cells, has some functions of the stem cells, can help to promote the repair and regeneration of damaged epidermis, promote the growth capacity of collagen and repair the elasticity of aged and fractured collagen; the disodium EDTA can chelate metal ions, so that the damage of the metal ions to the skin is reduced; the butanediol is a micromolecular moisturizing component, can retain water in the horny layer, has a good moisturizing effect and has small irritation to the skin; the yeast extract contains various active ingredients, can promote the metabolism of skin, and has good effects of removing wrinkles, resisting aging, whitening and moisturizing. Elastin is the main component of elastic fiber, the elastic fiber and collagen fiber coexist to endow the tissue with elasticity and expansion capability, the elastic fiber is called as 'human rubber', and the lactic acid compound extract can provide better functions of moisture retention and anti-aging. The preparation method provided by the invention is reasonable in design, safe and stable, free of side effect and wide in application prospect.
Detailed Description
Examples 1,
A skin care composition with wrinkle removing effect is prepared from the following raw materials in percentage by weight: the balance of water, 12% of squalane, 0.02% of disodium EDTA, 5% of glycerol, 4.46% of butanediol, 0.3% of sodium lactate, 1% of isopropyl myristate, 1% of recombinant human elastin, 0.3% of scutellaria baicalensis and tricholoma matsutake compound fermentation product, 1.5% of yeast extract, 5% of stem cell exosome and 0.02% of essence.
The scutellaria and tricholoma matsutake compound fermentation product is prepared from the following raw materials in parts by weight: 10 parts of scutellaria baicalensis, 5 parts of tricholoma matsutake, 0.5 part of lactic acid, 0.5 part of yeast powder, 1 part of peptone and 75 parts of water.
The preparation method of the skin care composition with the wrinkle removing effect comprises the following steps:
(1) adding appropriate amount of water into yeast extract, and stirring to obtain yeast extract water solution;
(2) adding isopropyl myristate and recombinant human elastin into yeast extract water solution, centrifuging, removing supernatant, and drying to obtain compound material;
(3) adding squalane, disodium EDTA, glycerol and appropriate amount of water into a stirring pot, heating to 80 deg.C, adding yeast lysate extract, stem cell exosome, compound material and other raw materials, stirring at 26 deg.C, cooling, and discharging.
The preparation method of the scutellaria and tricholoma matsutake compound fermentation product comprises the following steps:
(1) cleaning Scutellariae radix and Tricholoma matsutake, mixing, stirring, extracting with ethanol, and concentrating under reduced pressure to obtain compound extract;
(2) inoculating yeast to a culture medium, performing activation culture to obtain activated yeast, inoculating the activated yeast to a liquid culture medium, and performing fermentation culture to obtain a yeast liquid;
(3) adding lactic acid, yeast powder, peptone, water, a compound extract and a yeast liquid into a fermentation tank according to the weight parts, and fermenting for 24 hours at 35 ℃ to obtain a fermentation liquid;
(4) adding the obtained fermentation liquor into absolute ethyl alcohol, carrying out ultrasonic treatment, centrifuging, filtering and drying to obtain the product.
Examples 2,
A skin care composition with wrinkle removing effect is prepared from the following raw materials in percentage by weight: the balance of water, 12% of squalane, 0.02% of disodium EDTA, 5% of glycerol, 4.46% of butanediol, 0.3% of sodium lactate, 1% of isopropyl myristate, 2% of recombinant human elastin, 0.35% of scutellaria baicalensis and tricholoma matsutake compound fermentation product, 1.5% of yeast extract, 5% of stem cell exosome and 0.02% of essence. The preparation method is conventional in the field, and the raw materials are uniformly mixed.
The preparation method of the skin care composition with the wrinkle removing effect comprises the following steps:
(1) adding appropriate amount of water into yeast extract, and stirring to obtain yeast extract water solution;
(2) adding isopropyl myristate and recombinant human elastin into yeast extract water solution, centrifuging, removing supernatant, and drying to obtain compound material;
(3) adding squalane, disodium EDTA, glycerol and appropriate amount of water into a stirring pot, heating to 80 deg.C, adding yeast lysate extract, stem cell exosome, compound material and other raw materials, stirring at 26 deg.C, cooling, and discharging.
The preparation method of the scutellaria and tricholoma matsutake compound fermentation product comprises the following steps:
(1) cleaning Scutellariae radix and Tricholoma matsutake, mixing, stirring, extracting with ethanol, and concentrating under reduced pressure to obtain compound extract;
(2) inoculating yeast to a culture medium, performing activation culture to obtain activated yeast, inoculating the activated yeast to a liquid culture medium, and performing fermentation culture to obtain a yeast liquid;
(3) adding lactic acid, yeast powder, peptone, water, a compound extract and a yeast liquid into a fermentation tank according to the weight parts, and fermenting for 24 hours at 35 ℃ to obtain a fermentation liquid;
(4) adding the obtained fermentation liquor into absolute ethyl alcohol, carrying out ultrasonic treatment, centrifuging, filtering and drying to obtain the product.
Examples 3,
A skin care composition with wrinkle removing effect is prepared from the following raw materials in percentage by weight: the balance of water, 12% of squalane, 0.02% of disodium EDTA, 5% of glycerol, 4.46% of butanediol, 0.3% of sodium lactate, 1% of isopropyl myristate, 5% of recombinant human elastin, 0.4% of scutellaria baicalensis and tricholoma matsutake compound fermentation product, 1.5% of yeast extract, 2% of stem cell exosome and 0.02% of essence. The preparation method is conventional in the field, and the raw materials are uniformly mixed.
The preparation method of the skin care composition with the wrinkle removing effect comprises the following steps:
(1) adding appropriate amount of water into yeast extract, and stirring to obtain yeast extract water solution;
(2) adding isopropyl myristate and recombinant human elastin into yeast extract water solution, centrifuging, removing supernatant, and drying to obtain compound material;
(3) adding squalane, disodium EDTA, glycerol and appropriate amount of water into a stirring pot, heating to 80 deg.C, adding yeast lysate extract, stem cell exosome, compound material and other raw materials, stirring at 26 deg.C, cooling, and discharging.
The preparation method of the scutellaria and tricholoma matsutake compound fermentation product comprises the following steps:
(1) cleaning Scutellariae radix and Tricholoma matsutake, mixing, stirring, extracting with ethanol, and concentrating under reduced pressure to obtain compound extract;
(2) inoculating yeast to a culture medium, performing activation culture to obtain activated yeast, inoculating the activated yeast to a liquid culture medium, and performing fermentation culture to obtain a yeast liquid;
(3) adding lactic acid, yeast powder, peptone, water, a compound extract and a yeast liquid into a fermentation tank according to the weight parts, and fermenting for 24 hours at 35 ℃ to obtain a fermentation liquid;
(4) adding the obtained fermentation liquor into absolute ethyl alcohol, carrying out ultrasonic treatment, centrifuging, filtering and drying to obtain the product.
Examples 4,
A skin care composition with wrinkle removing effect is prepared from the following raw materials in percentage by weight: the balance of water, 12% of squalane, 0.02% of disodium EDTA, 5% of glycerol, 4.46% of butanediol, 0.3% of sodium lactate, 1% of isopropyl myristate, 5% of recombinant human elastin, 0.5% of scutellaria baicalensis and tricholoma matsutake compound fermentation product, 1.5% of yeast extract, 5% of stem cell exosome and 0.02% of essence. The preparation method is conventional in the field, and the raw materials are uniformly mixed.
The preparation method of the skin care composition with the wrinkle removing effect comprises the following steps:
(1) adding appropriate amount of water into yeast extract, and stirring to obtain yeast extract water solution;
(2) adding isopropyl myristate and recombinant human elastin into yeast extract water solution, centrifuging, removing supernatant, and drying to obtain compound material;
(3) adding squalane, disodium EDTA, glycerol and appropriate amount of water into a stirring pot, heating to 80 deg.C, adding yeast lysate extract, stem cell exosome, compound material and other raw materials, stirring at 26 deg.C, cooling, and discharging.
The preparation method of the scutellaria and tricholoma matsutake compound fermentation product comprises the following steps:
(1) cleaning Scutellariae radix and Tricholoma matsutake, mixing, stirring, extracting with ethanol, and concentrating under reduced pressure to obtain compound extract;
(2) inoculating yeast to a culture medium, performing activation culture to obtain activated yeast, inoculating the activated yeast to a liquid culture medium, and performing fermentation culture to obtain a yeast liquid;
(3) adding lactic acid, yeast powder, peptone, water, a compound extract and a yeast liquid into a fermentation tank according to the weight parts, and fermenting for 24 hours at 35 ℃ to obtain a fermentation liquid;
(4) adding the obtained fermentation liquor into absolute ethyl alcohol, carrying out ultrasonic treatment, centrifuging, filtering and drying to obtain the product.
Examples 5,
A skin care composition with wrinkle removing effect is prepared from the following raw materials in percentage by weight: the balance of water, 12% of squalane, 0.02% of disodium EDTA, 5% of glycerol, 4.46% of butanediol, 0.3% of sodium lactate, 1% of isopropyl myristate, 2% of recombinant human elastin, 0.6% of scutellaria baicalensis and tricholoma matsutake compound fermentation product, 2.0% of yeast extract, 5% of stem cell exosome and 0.02% of essence. The preparation method is conventional in the field, and the raw materials are uniformly mixed.
The preparation method of the skin care composition with the wrinkle removing effect comprises the following steps:
(1) adding appropriate amount of water into yeast extract, and stirring to obtain yeast extract water solution;
(2) adding isopropyl myristate and recombinant human elastin into yeast extract water solution, centrifuging, removing supernatant, and drying to obtain compound material;
(3) adding squalane, disodium EDTA, glycerol and appropriate amount of water into a stirring pot, heating to 80 deg.C, adding yeast lysate extract, stem cell exosome, compound material and other raw materials, stirring at 26 deg.C, cooling, and discharging.
The preparation method of the scutellaria and tricholoma matsutake compound fermentation product comprises the following steps:
(1) cleaning Scutellariae radix and Tricholoma matsutake, mixing, stirring, extracting with ethanol, and concentrating under reduced pressure to obtain compound extract;
(2) inoculating yeast to a culture medium, performing activation culture to obtain activated yeast, inoculating the activated yeast to a liquid culture medium, and performing fermentation culture to obtain a yeast liquid;
(3) adding lactic acid, yeast powder, peptone, water, a compound extract and a yeast liquid into a fermentation tank according to the weight parts, and fermenting for 24 hours at 35 ℃ to obtain a fermentation liquid;
(4) adding the obtained fermentation liquor into absolute ethyl alcohol, carrying out ultrasonic treatment, centrifuging, filtering and drying to obtain the product.
Comparative examples 1,
A skin care composition with wrinkle removing effect is prepared from the following raw materials in percentage by weight: the balance of water, 12% of squalane, 0.02% of disodium EDTA, 5% of glycerol, 4.46% of butanediol, 0.3% of sodium lactate, 1% of isopropyl myristate and 0.02% of essence.
The preparation method is according to the conventional process.
Comparative examples 2,
A skin care composition with wrinkle removing effect is prepared from the following raw materials in percentage by weight: the balance of water, squalane 12%, disodium EDTA 0.02%, glycerin 5%, butylene glycol 4.46%, sodium lactate 0.3%, isopropyl myristate 1%, yeast extract 1.5%, and essence 0.02%.
The preparation method is according to the conventional process.
Comparative examples 3,
A skin care composition with wrinkle removing effect is prepared from the following raw materials in percentage by weight: the balance of water, squalane 12%, disodium EDTA 0.02%, glycerin 5%, butylene glycol 4.46%, sodium lactate 0.3%, isopropyl myristate 1%, lactic acid 0.4%, and essence 0.02%.
The preparation method is according to the conventional process.
Comparative examples 4,
A skin care composition with wrinkle removing effect is prepared from the following raw materials in percentage by weight: the balance of water, 12% of squalane, 0.02% of disodium EDTA, 5% of glycerol, 4.46% of butanediol, 0.3% of sodium lactate, 1% of isopropyl myristate, 5% of recombinant human elastin and 0.02% of essence.
The preparation method is according to the conventional process.
Comparative examples 5,
A skin care composition with wrinkle removing effect is prepared from the following raw materials in percentage by weight: the balance of water, 12% of squalane, 0.02% of disodium EDTA, 5% of glycerol, 4.46% of butanediol, 0.3% of sodium lactate, 1% of isopropyl myristate, 5% of stem cell exosomes and 0.02% of essence.
The preparation method is according to the conventional process.
1. Safety test
The number of tested persons: 50 people in total, half of men and women, age 22-50 years.
Groups were randomized into 5 groups of 5 males and 5 females. The face is used as a tested part, and any product (cosmetics, external medicines or internal health care products) cannot be used in the tested part within 15 days before the test. Before the test, the subject washes the face and sits still for 30min in a constant temperature and humidity room with the temperature of 22 +/-1 ℃ and the humidity of 50 +/-5% 2h after washing, and keeps a relaxed state, and the evaluation test of the lactic acid stimulation efficacy of the face is carried out:
in the test, a half-face test was performed on a random scale, one side was the example product and the other side was the other comparative product, and 50. mu.L of a 10% lactic acid solution was dropped on a single-layer filter paper having a diameter of 0.8cm, attached to the test site, and the subject was asked to feel stinging after 3min, 4min and 5min, respectively, and a score was recorded as the side lactic acid stimulation score (D0). The product is applied as required 2 times a day, and the application is continued for 14 days, and the test is required to be revisited on the 7 th (D7) and 14 th (D14) days.
The test results are shown in the following table
From the results of the table, it can be seen that: the examples and the comparative examples have the effect of preventing skin irritation, namely, have a certain repairing effect on facial wrinkles, wherein the better effects of the examples 3, 4 and 5 show that the squalane, disodium EDTA, glycerol, butanediol, sodium lactate, isopropyl myristate, recombinant human elastin, lactic acid, yeast extract, stem cell exosome and essence composition have better wrinkle removing effects. In conclusion, the composition has a good fine line repairing effect and a lactic acid stimulation reducing effect.
2. Superoxide radical clearance Activity assay
The experimental principle is as follows: superoxide anion free radicals are used as free radicals generated in the metabolic process of organisms, can attack biological macromolecules, such as lipids, proteins, nucleic acids, polyunsaturated fatty acids and the like, cause cross-linking or breakage of the biological macromolecules, cause damage to cell structures and functions, have close relation with aging and pathological changes of organisms, and have attracted extensive attention on research on removal of superoxide anion free radicals.
The AP-TEMED system produces superoxide anion, which reacts with hydroxylamine hydrochloride to form NO2-，NO2-The azo compound which generates red color under the action of sulfanilic acid and alpha-naphthylamine has a characteristic absorption peak at 530nm, and the scavenging capacity of the sample to superoxide anions is negatively correlated with the absorbance value at 530 nm.
Procedure for the preparation of the
The reagents in the kit were added in the following order.
Calculating the formula: superoxide anion clearance I% (a control tube-a assay tube)/a control tube × 100%
As a result:
the experiment result shows that the composition has superoxide radical scavenging activity.
3. Recombinant human epidermal growth factor bioactivity test
Reagent preparation
Measuring the complete culture solution to 100ml of calf serum, and adding the culture solution to a constant volume of 1000 ml.
The maintenance culture solution is measured by 4ml of calf serum, and the culture solution is added to a constant volume of 1000 ml.
Digestion solution: weighing 0.2g of disodium ethylene diamine tetraacetate, 8g of sodium chloride, 0.2g of potassium chloride, 1.152g of disodium hydrogen phosphate, 0.2g of monopotassium phosphate and 2.5g of trypsin, adding purified water to dissolve, fixing the volume to 1000ml, filtering and sterilizing.
Thiazole blue (MTT) solution: 0.10g of MTT powder was weighed, dissolved in PBS20ml, and sterilized by filtration through a 0.22 μm filter. Storing at 4 ℃ in the dark.
PBS: weighing 8.0g of sodium chloride, 0.20g of potassium chloride, 1.44g of disodium hydrogen phosphate and 0.24g of potassium dihydrogen phosphate, adding purified water to dissolve, fixing the volume to 1000ml, and sterilizing at 121 ℃ for 15 minutes.
Cell lines: BALB/C3T3 cells.
rhEGF national standard: 6800IU per branch, lyophilized powder;
test procedure
Preparing a standard substance: mu.l of the standard was diluted 10-fold in 450. mu.l of complete medium and the procedure was repeated until the dilution was 100-fold. Mu.l of 100-fold diluted standard was diluted 1.36-fold in 265. mu.l of complete medium. Taking 136-fold diluent in a 96-hole cell culture plate, sequentially carrying out 4-fold incremental gradient dilution from 1: 4 (the specific operation steps are firstly adding 150 mu l of complete culture solution in the 96-hole cell culture plate, adding 50 mu l of 1000-fold diluted sample solution in the 1 st column in the 96-hole plate, carrying out blow-on dilution, after full dilution, extracting 50 mu l in the 2 nd column in the 1 st column, carrying out blow-on dilution, extracting 50 mu l in the 3 rd column in the 2 nd column after full dilution, carrying out blow-on dilution, repeating the operation till the 10 th column, carrying out 10 dilutions (1-10 holes) totally, and simultaneously setting a duplicate hole for each dilution.
Preparing a recombinant human epidermal growth factor test sample: mu.l of the sample was diluted 10-fold in 900. mu.l of complete medium and the procedure was repeated until the dilution was 1000-fold. Taking 1000-time dilution liquid in a 96-hole cell culture plate, sequentially carrying out 4-time incremental gradient dilution from 1: 4 (the specific operation steps are that firstly, 150 mu l of complete culture liquid is added in the 96-hole cell culture plate, 50 mu l of 1000-time diluted sample solution is added in the 1 st column of the 96-hole plate, blow-on dilution is carried out, after full dilution, 50 mu l of sample solution is extracted in the 2 nd column of the 96-hole plate, blow-on dilution is carried out, the operation is repeated until the 10 th column, 10 dilution degrees (1-10 holes) are carried out totally, and each dilution degree is provided with a compound hole at the same time.
Note: the sample pre-dilution factor was adjusted according to its activity value.
Measurement method
Firstly, BALB/C3T3 cell line was cultured with 5% carbon dioxide at 37 ℃ using the whole culture medium, and the cell concentration was controlled to 1.0X 10 per 1ml5-5.0×105And (4) carrying out biological activity determination on the individual cells 24-36 hours after passage.
② discarding the culture solution in the culture flask, digesting and collecting cells and preparing the complete culture solution containing 5.0X 10 per 1ml4～8.0×104Of individual cellsThe cell suspension is inoculated in a 96-well plate, the cell suspension is continuously shaken up in the inoculation process, the inoculation number of each well is kept to be the same, each well is 100 mu l, and the cell suspension is cultured under the conditions of 37 ℃ and 5% carbon dioxide.
And replacing the culture medium with maintaining culture medium after 24 hours. Incubated at 37 ℃ for 24 hours with 5% carbon dioxide.
Fourthly, removing maintenance liquid from the prepared cell culture plate, and adding 100 mul of standard solution and test solution into each hole. Culturing the cells at 37 ℃ in 5% carbon dioxide for 64-72 hours.
MTT colorimetric method: mu.l of MTT solution was added to each well, and the mixture was incubated at 37 ℃ with 5% carbon dioxide for 5 hours. The above operations are carried out under aseptic conditions. After discarding the liquid in the culture plate, adding 100 μ l DMSO into each well, mixing well, measuring absorbance on a microplate reader with 630nm as reference wavelength and 570nm as test wavelength, and recording the measurement result.
Data processing
And performing four-parameter fitting on the OD value data of each sample and each standard.
Calculation of results
The test data is processed by a computer program or a four-parameter regression calculation method.
Calculating the working titer of the sample with the formula U ═ pre-dilution multiple 4C (C is the C value in the four-parameter equation)
The test article is corrected for potency and the result is calculated according to the following formula:
corrected titer (IU/ml) ═ Pr Ds/Dr Er
In the formula, Pr is the biological activity of a standard substance, IU/ml;
ds is the pre-dilution multiple of the test supply;
es is the dilution multiple of the half-effect number of the test sample equivalent to the standard sample;
dr is the pre-dilution multiple of the standard substance;
er is the dilution multiple of half effective amount of the standard substance;
the experiment result shows that the composition has the cell growth promoting activity.
The applicant states that the present invention is illustrated by the above examples of the skin care composition having wrinkle removing effect of the present invention, but the present invention is not limited to the above examples, i.e., it does not mean that the present invention must be implemented by means of the above examples. The equivalent replacement of each raw material of the product, the addition of auxiliary components, the selection of a specific mode and any other improvement are all within the protection scope and the disclosure scope of the invention.
It should be noted that the various technical features described in the above embodiments can be combined in any suitable manner without contradiction, and the invention is not described in any way for the possible combinations in order to avoid unnecessary repetition.
4. Cell migration Rate test
The experimental principle is as follows: the method is based on artificially creating a blank area, called a "scratch", on the fused monolayer of cells as they grow until they fuse into a single layer. The cells at the scratch edge will gradually enter the blank area to heal the scratch. Images were captured initially and periodically during cell migration, and the rate of cell migration was determined by comparing the images.
Test materials
Cell lines: MDBK bovine kidney cells;
other materials and equipment: newborn bovine serum and DMEM medium;
96-well cell culture plates; 200 mul, 1ml sterile tip; penicillin-streptomycin solution;
200 μ l, 1ml pipette, ruler, etc.
Carbon dioxide incubator, clean bench, refrigerator, etc.
Test method
Firstly, a marker pen is used behind a 6-hole plate, a ruler is used for aligning, transverse lines are uniformly drawn, and the transverse lines cross through holes approximately every 0.5-1 cm.
② adding 5 multiplied by 10 of concentration into 6-hole plate holes52ml of MDBK cell suspension per ml.
And thirdly, observing that cells in the 6-hole plate grow all over a single layer on the next day, scratching the cells perpendicular to a transverse line at the back of the plate by using the gun head compared with the straight ruler as much as possible, and scratching 2 channels in each hole.
And fourthly, washing the cells for 3 times by using PBS, and washing off the scraped suspension cells.
Adding 1.8ml of corresponding culture solution into the wells according to groups, then adding 200 mul of test sample, and adding the same amount of PBS into the cell control wells. The test wells and cell control wells in the 12-well plate are shown in the table below. Because MDBK cells are good in viability, only a serum-free group is arranged.
Sixthly, adding 5 percent CO at 37 DEG C2And (5) an incubator for culture. And taking a picture at 0 moment, and recording the picture taking position in each hole. And observing and photographing the fixed position during subsequent observation.
And calculating the migration rate of the cells by comparing the migration numbers of the cells at the same scratch at different times to obtain a conclusion.
Claims (4)
1. The skin care composition with the wrinkle removing effect is characterized by being prepared from the following raw materials in percentage by weight: 11-13% of squalane, 0.01-0.03% of EDTA disodium, 4-6% of glycerol, 4.4-4.5% of butanediol, 0.2-0.4% of sodium lactate, 0.9-1.1% of isopropyl myristate, 1-5% of recombinant human elastin, 0.3-0.6% of scutellaria and tricholoma matsutake compound fermentation product, 1.5-2.0% of yeast extract, 1-5% of dry cell exosome, 0.01-0.03% of essence and the balance of water.
2. The skin care composition with the wrinkle removing effect according to claim 1, wherein the scutellaria baicalensis and tricholoma matsutake compound fermentation product is prepared from the following raw materials in parts by weight: 10-20 parts of scutellaria baicalensis, 3-5 parts of tricholoma matsutake, 0.3-0.6 part of lactic acid, 0.3-0.6 part of yeast powder, 0.8-1 part of peptone and 65-85 parts of water.
3. A method of preparing a skin care composition having wrinkle removing effect according to claim 1, wherein: the method comprises the following steps:
(1) adding appropriate amount of water into yeast extract, and stirring to obtain yeast extract water solution;
(2) adding isopropyl myristate and recombinant human elastin into yeast extract water solution, centrifuging, removing supernatant, and drying to obtain compound material;
(3) adding squalane, EDTA disodium, glycerol and appropriate amount of water into a stirring pot, heating to 70-80 deg.C, adding yeast lysate extract, stem cell exosome, compound material and other raw materials, stirring at 20-30 deg.C, cooling, and discharging.
4. A method of preparing a skin care composition with wrinkle removing effect according to claim 3, wherein: the preparation method of the scutellaria and tricholoma matsutake compound fermentation product comprises the following steps:
(1) cleaning Scutellariae radix and Tricholoma matsutake, mixing, stirring, extracting with ethanol, and concentrating under reduced pressure to obtain compound extract;
(2) inoculating yeast to a culture medium, performing activation culture to obtain activated yeast, inoculating the activated yeast to a liquid culture medium, and performing fermentation culture to obtain a yeast liquid;
(3) adding lactic acid, yeast powder, peptone, water, compound extract and yeast liquid into a fermentation tank according to the weight parts, and fermenting at 30-35 ℃ for 12-24h to obtain fermentation liquor;
(4) adding the obtained fermentation liquor into absolute ethyl alcohol, carrying out ultrasonic treatment, centrifuging, filtering and drying to obtain the product.
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