CN118319830B

CN118319830B - Skin tightening and anti-wrinkle essence and preparation method thereof

Info

Publication number: CN118319830B
Application number: CN202410485729.6A
Authority: CN
Inventors: 唐晋雯; 杜育娟; 周广练; 涂世平
Original assignee: Guangzhou B&c Cosmetics Co ltd
Current assignee: Guangzhou B&c Cosmetics Co ltd
Priority date: 2024-04-22
Filing date: 2024-04-22
Publication date: 2025-01-28
Anticipated expiration: 2044-04-22
Also published as: CN118319830A

Abstract

本发明提出了一种紧致皮肤抗皱精华液及其制备方法，属于化妆品技术领域。由以下原料按重量份制备而成：活性提取物7‑10份、多肽组合物5‑7份、促透剂组合物0.5‑2份、DNA钠0.1‑0.5份、可溶性胶原0.3‑0.5份、羟丙基四氢吡喃三醇5‑10份、润肤剂2‑8份、保湿剂5‑10份、乳化剂0.5‑2份、增稠剂0.2‑0.5份、水蛭素0.1‑0.5份、EDTA二钠0.02‑0.05份、去离子水50‑70份。本发明制得的紧致皮肤抗皱精华液具有美白、抗氧化、抗衰老、抗光老化、紧致皮肤、提高皮肤弹性、淡化皮肤皱纹等功效，多组分协同作用，效果更明显，且成本较低，制备方法简单，具有广阔的应用前景。The present invention proposes a skin tightening and anti-wrinkle essence and a preparation method thereof, and belongs to the field of cosmetic technology. It is prepared by the following raw materials by weight: 7-10 parts of active extracts, 5-7 parts of polypeptide compositions, 0.5-2 parts of penetration enhancer compositions, 0.1-0.5 parts of DNA sodium, 0.3-0.5 parts of soluble collagen, 5-10 parts of hydroxypropyl tetrahydropyrantriol, 2-8 parts of emollients, 5-10 parts of moisturizers, 0.5-2 parts of emulsifiers, 0.2-0.5 parts of thickeners, 0.1-0.5 parts of hirudin, 0.02-0.05 parts of disodium EDTA, and 50-70 parts of deionized water. The skin tightening and anti-wrinkle essence prepared by the present invention has the effects of whitening, anti-oxidation, anti-aging, anti-photoaging, tightening skin, improving skin elasticity, and fading skin wrinkles, and the multi-component synergistic effect is more obvious, and the cost is low, the preparation method is simple, and it has broad application prospects.

Description

Tightening skin anti-wrinkle essence and preparation method thereof

Technical Field

The invention relates to the technical field of cosmetics, in particular to a compact skin anti-wrinkle essence and a preparation method thereof.

Background

The biological aging process is the process that cells continuously dry up and die, and most of skin cells are water like other cells. When the water is sufficient, the skin cells are full of vigor, and the ability of resisting severe environment and repairing themselves is strong. In the current society, the air environment pollution is serious, and various bacteria, viruses, radiation and haze can cause the skin to lose moisture more quickly and the skin can age quickly. Skin dryness, tightness, roughness, desquamation, loss of elasticity and large pores are all manifestations of skin water deficiency, and all need to be immediately moisturized. The skin moisture mainly comes from the inside of the body, the protection effect of the sebum membrane is weakened or destroyed along with the aging and the change of the external environment, the natural moisturizing factors of the horny layer are lost, at the moment, the horny layer of the skin is easy to be in a water-deficient state, the phenomenon that the horny metabolism is bad, the intracellular impurities and the metabolites cannot be removed in time is easy to occur, the phenomenon of thick and thick horny layer is generated, the skin color is influenced, the skin looks dark and dull, and the penetration of the maintenance product is also hindered. Even if whitening care is performed, the effect is limited.

Because modern people have a fast pace of life and are affected by various environments, skin is often dry and dull, and cosmetics are becoming daily necessities and are widely used by people. The essence water has the characteristics of high nutrient content, small skin irritation, capability of regulating skin cell activity and the like, and is more taken as a necessity for basic skin care by mass females.

The Chinese patent application CN111150701A discloses an anti-wrinkle tightening essence and a preparation method thereof, wherein the anti-wrinkle tightening composition comprises, by weight, 0.5-3 parts of spirulina maxima extract, 0.005-0.1 part of sodium hyaluronate, 0.01-0.05 part of cyclodextrin, 0.01-0.05 part of ubiquinone, 0.1-1 part of avenanthramide, 1-5 parts of yeast extract, 0.1-1 part of fucoidan, 0.01-0.1 part of humic acid, 0.1-3 parts of dipyridamole extract, 0.05-0.3 part of decapeptides, 0.05-0.3 part of glycoprotein and 0.005-0.05 part of rose oil. The invention can quickly permeate to deep skin to supplement water, smooth skin, smooth small lines and wrinkles, endow skin with elasticity, tighten and delay skin aging, promote regeneration of young cells and make skin appear bright and glossy, but the invention directly extracts small molecular polypeptide obtained by protease hydrolysis, which is easy to have the problems of poor stability, poor anti-wrinkle effect and the like.

Disclosure of Invention

The invention aims to provide a compact skin anti-wrinkle essence and a preparation method thereof, which have the effects of whitening, resisting oxidation, resisting aging, resisting photoaging, compacting skin, improving skin elasticity, reducing skin wrinkles and the like, have more obvious effect, lower cost and simple preparation method, and have wide application prospect.

The technical scheme of the invention is realized as follows:

The invention provides a compact skin anti-wrinkle essence which is prepared from the following raw materials, by weight, 7-10 parts of an active extract, 5-7 parts of a polypeptide composition, 0.5-2 parts of a penetration enhancer composition, 0.1-0.5 part of DNA sodium, 0.3-0.5 part of soluble collagen, 5-10 parts of hydroxypropyl tetrahydropyran triol, 2-8 parts of an emollient, 5-10 parts of a humectant, 0.5-2 parts of an emulsifier, 0.2-0.5 part of a thickener, 0.1-0.5 part of hirudin, 0.02-0.05 part of EDTA disodium and 50-70 parts of deionized water.

As a further improvement of the invention, the permeation enhancer composition comprises ethoxydiglycol and a permeation enhancing compound, wherein the mass ratio of the ethoxydiglycol to the permeation enhancing compound is 4-7:3, and the structural formula of the permeation enhancing compound is shown as formula I:

Wherein r=c6-C18 alkyl chain.

The synthesis method of the permeation promoting compound comprises the steps of dissolving dihaloalkane in dichloromethane, dropwise adding dichloromethane solution of caprolactam and triethylamine, heating and refluxing, stirring and reacting for a first time period, then dropwise adding dichloromethane solution of menthol, heating and refluxing, stirring and reacting for a second time period, removing solvent under reduced pressure, and separating and purifying by column chromatography to obtain the permeation promoting compound.

As a further improvement of the invention, the molar ratio of dihaloalkane, caprolactam, menthol and triethylamine is 1:0.9-1:0.9-1:4-6, the first time period is 3-5h, and the second time period is 4-6h.

As a further improvement of the invention, the polypeptide composition is a composition of palmitoyl tripeptide-5, hexapeptide-11, hexapeptide-9, acetyl hexapeptide-8 and arginine/lysine polypeptide, and the mass ratio is 4-6:1-3:2-4:0.5-1:3-5.

As a further improvement of the present invention, the active extract is prepared as follows:

S1, supercritical fluid extraction, namely drying, crushing and extracting sticktight and dendrobium candidum by using a supercritical fluid to prepare an oil extract, and reserving the rest materials;

s2, deep eutectic solvent enzymolysis, namely adding the rest materials in the step S1 into an aqueous solution containing the deep eutectic solvent, adding complex enzyme for enzymolysis, and inactivating enzyme to obtain an enzymolysis product;

S3, fermenting, namely mixing the enzymolysis product obtained in the step S2 with water, sterilizing, inoculating bifidobacterium longum and bifidobacterium breve seed liquid, fermenting, culturing, filtering, washing solids, collecting eluent, concentrating to obtain bacterial liquid, and reserving filtrate;

S4, ethanol precipitation, namely adding ethanol into the filtrate obtained in the step S3 to precipitate, centrifuging, washing and drying to obtain a polysaccharide composition, recovering ethanol from supernatant fluid, and reserving the residual liquid;

s5, purifying the residual liquid in the step S4 by macroporous resin, removing the solvent by decompression, purifying the filtrate by decompression, diluting and loading the filtrate by 65-75wt% ethanol, loading the filtrate into a column by a wet method, collecting eluent with the concentration of 65-75wt% ethanol, removing the ethanol by decompression, and drying to obtain an active mixture;

S6, preparing a yeast lysate, namely freezing the bacterial liquid prepared in the step S3 by liquid nitrogen, dissolving at room temperature, centrifuging, collecting supernatant, and freeze-drying to prepare the yeast lysate;

s7, preparing an active extract, namely uniformly mixing the oil extract prepared in the step S1, the polysaccharide composition prepared in the step S4, the active mixture prepared in the step S5, the yeast lysate prepared in the step S6 and the vitamin C to prepare the active extract.

According to the invention, the mass ratio of spanishneedles herb to dendrobium candidum in the step S1 is 7-10:3-5, the supercritical fluid extraction condition is that the pressure is 27-35MPa, the temperature is 32-37 ℃ and the time is 70-90min, the mass ratio of the rest material to the aqueous solution containing the deep eutectic solvent in the step S2 is 10:35-60, the water content of the aqueous solution containing the deep eutectic solvent is 40-60wt%, the deep eutectic solvent is prepared by mixing choline chloride and glucose according to the molar ratio of 1:1, the compound enzyme is formed by mixing cellulase and pectase, the mass ratio is 10-15:3-5, the addition amount of the compound enzyme is 1-2wt% of the total mass of the system, the enzymolysis temperature is 40-50 ℃ and the time is 2-4h, the mass ratio of the enzymolysis product to the water in the step S3 is 1:1.5-3, the inoculation amount of bifidobacterium longum seed and bifidobacterium breve seed liquid is 2-4 v/3 v/1-3 u/v/the bacterial seed liquid is 5736-150 mL/v/150-150 mL/v/the bacterial strain is cultured under the condition of no fermentation condition.

As a further improvement of the invention, the ethanol is added to 60-70wt% of the ethanol content of the system in the step S4, the precipitation time is 3-5 hours, the macroporous resin is selected from at least one of XDA-2, NKA-9, SP-825, D101, HPD600, X-5, LSA-10, NKI-9 and AB-8 in the step S5, the ethanol is diluted to a concentration of 0.8-1.2mg/mL by 65-75wt%, and the mass ratio of the oil extract, the polysaccharide composition, the active mixture, the split yeast lysate and the vitamin C in the step S7 is 5-7:12-15:10-17:3-5:1-2.

As a further improvement of the invention, the emollient is at least one selected from the group consisting of glycerol tri (ethyl caproate), isononyl isononanoate, polydimethylsiloxane, caprylic/capric triglyceride and butter tree (BUTYROSPERMUM PARKII) fruit fat, the humectant is at least one selected from the group consisting of glycerol, butanediol, propylene glycol, hexylene glycol, sodium hyaluronate, betaine and trehalose, the emulsifier is at least one selected from the group consisting of glycerol stearate, PEG-100 stearate, cetostearyl oleate, sorbitan olive oleate, C12-20 alkyl glucoside and polyglycerol-2 stearate, and the thickener is at least one selected from the group consisting of carbomer, acrylic acid (esters) or C10-30 alcohol acrylate cross-linked polymer, sodium polyacryl dimethyl taurate and xanthan gum.

The invention further provides a preparation method of the tightening skin anti-wrinkle essence, which comprises the following steps:

(1) Adding active extract, polypeptide composition, soluble collagen, DNA sodium, hydroxypropyl tetrahydropyran triol, hirudin, humectant, thickener, EDTA disodium into deionized water, heating to 70-80deg.C, and stirring at 300-500r/min for 10-20min to obtain phase A

(2) Adding the penetration enhancer composition into emollient and emulsifier, heating to 70-80deg.C, and stirring at 300-500r/min for 10-20min to obtain phase B;

(3) Adding phase B into phase A, stirring for 5-15min at 300-500r/min, emulsifying for 10-20min at 7000-10000r/min, maintaining the temperature for 10-20min, cooling, and discharging to obtain compact skin anti-wrinkle essence.

The invention has the following beneficial effects:

The penetration-promoting compound is prepared by connecting menthol and caprolactam through alkyl chains, overcomes the defect that menthol is easy to volatilize due to smaller molecular weight, is simple in preparation method, mild in reaction condition and higher in yield, has better effects of promoting active substances to penetrate into skin, and playing better effects of tightening, improving skin elasticity and resisting wrinkles, wherein a caprolactam part can interact with a stratum corneum lipid bilayer to increase fluidity, and a menthol part can damage conformational sequence of the stratum corneum lipid bilayer and reduce phase transition temperature of stratum corneum lipid. The prepared penetration-promoting compound and the ethoxydiglycol are mixed for use, and the penetration-promoting compound and the ethoxydiglycol have a synergistic effect.

The DNA sodium is added into the compact skin anti-wrinkle essence, and the DNA nucleotide fragment has remarkable effects in increasing the basic values of moisture retention, elasticity and skin thickness and reducing skin wrinkles, and simultaneously, the DNA sodium stimulates cell repair, reduces inflammation symptoms, accelerates the healing of skin micro-lesions, has regeneration and photoprotection effects on keratinocytes and fibroblasts and has protection effects on ultraviolet-induced fibroblast injury.

Hydroxypropyl tetrahydropyran triol is the core component in Pro-XylaneTM (vitrein). Hydroxypropyl tetrahydropyran triol can promote the production of glycosaminoglycans (GAGs) and Proteoglycans (PGs) in the skin.

The polypeptide composition comprises palmitoyl tripeptide-5, hexapeptide-11, hexapeptide-9, acetyl hexapeptide-8 and arginine/lysine polypeptide, wherein the palmitoyl tripeptide-5 can stimulate the synthesis of collagen and glycosaminoglycan, strengthen the skin firmness and elasticity, play a role in resisting wrinkle and aging, the hexapeptide-11 can improve the water holding capacity, reduce wrinkles, resist aging and enable the skin to return to a tender and compact state, the hexapeptide-9 can increase the type I collagen, the type IV collagen, the laminin-5 and the integrin synthesis, promote the differentiation and maturation of epidermal cells and the regeneration of the skin, reduce wrinkles, the acetyl hexapeptide-8 has the effects of scavenging free radicals, promoting the synthesis of collagen, moisturizing and reducing fine lines and wrinkles, and the arginine/lysine polypeptide can improve the skin defense function, improve the skin sensitivity and regulate the water-oil balance of the skin. Several protein peptides act synergistically to play a good role in tightening skin, resisting wrinkle, keeping moisture and improving skin elasticity.

The raw materials of the active extract prepared by the invention are sticktight extract and dendrobium candidum, and the oil extract extracted by the supercritical carbon dioxide extraction technology has unique pharmacological characteristics, such as antibacterial, anti-inflammatory and antiallergic effects, detoxification and detumescence effects, heat-clearing and pain-relieving effects, blood circulation promoting and stasis removing effects, qi regulating and food retention removing effects and the like.

The invention uses enzyme as catalyst to hydrolyze plant cell wall, to promote the effective component to diffuse from cell to extraction medium, but uses deep eutectic solvent as reaction medium, to improve the extraction of compound insoluble in water, and has the advantages of low toxicity, green property, dynamic property, incombustibility and strong solubility. Glucose is used as one of the raw materials in the deep eutectic solvent, and a carbon source is also provided for subsequent fermentation.

The invention ferments by bifidobacteria, and the product of the bifidobacteria fermentation contains abundant vitamins, amino acids, mineral substances of trophoblasts and the like, accelerates skin metabolism and repairs damaged skin, and can improve the dissolution of active components in raw materials, so that the fermentation product contains abundant substances such as flavone, organic acid, terpene alcohol, polysaccharide and the like.

The polysaccharide composition obtained by alcohol precipitation of the fermentation product has very excellent moisturizing effect and antioxidant property. The active mixture obtained by macroporous resin separation and purification contains rich flavonoid compounds such as isorhamnoside, apigenin and the like, has strong antioxidation, can help skin resist free radical damage, slow aging, contains phytanic acid, can promote the generation and repair of collagen and elastin by stimulating the activity of skin cells, thereby being beneficial to improving the elasticity and the compactness of the skin and leading the skin to be younger and healthier, contains diterpenoid alcohol similar to the diterpenoid alcohol A, can be combined with retinoic acid receptor and retinol X receptor after being converted into phytanic acid in vivo, thereby regulating gene expression and protein production and promoting epidermal metabolism.

On the one hand, the fermentation process generates abundant fermentation byproducts, simultaneously promotes the proliferation of bifidobacteria, and after liquid nitrogen freezing wall breaking and bifidobacteria wall breaking, the content is dissolved out, the prepared yeast lysate can regulate the expression of the deacetylase-6 gene, is beneficial to prolonging the service life of cells, delays aging, and has the effects of whitening, resisting oxidation and resisting aging.

The compact skin anti-wrinkle essence prepared by the invention has the effects of whitening, resisting oxidation, resisting aging, resisting photoaging, compacting skin, improving skin elasticity, reducing skin wrinkles and the like, has more obvious effect and lower cost due to multi-component synergistic effect, and has a wide application prospect due to simple preparation method.

Detailed Description

The following description of the technical solutions in the embodiments of the present invention will be clear and complete, and it is obvious that the described embodiments are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.

Cellulase, 1.1U/g, pectase, 6U/g, purchased from Xia Cheng (Beijing) Biotechnology development Co., ltd. Bifidobacterium longum, 100 hundred million cfu/g, purchased from Guangdong, hongdou biotechnology Co., ltd, and Bifidobacterium breve, 100 hundred million cfu/g, purchased from Shaanxi, seen biotechnology Co., ltd.

The preparation method of the strain seed liquid comprises inoculating strain into Gao's medium, activating and culturing at 37deg.C and 100r/min for 24 hr to obtain strain seed liquid with a strain content of 10 ⁸-10⁹ cfu/mL.

PREPARATION EXAMPLE 1 Synthesis of penetration enhancing Compounds

The synthetic route is as follows:

The method comprises the steps of dissolving 0.1mol of 1, 12-dichlorododecane in 200mL of dichloromethane, dropwise adding 50mL of dichloromethane solution containing 0.09mol of caprolactam and 0.5mol of triethylamine, heating and refluxing, stirring and reacting for 4h, then dropwise adding 20mL of dichloromethane solution containing 0.09mol of menthol, heating and refluxing, stirring and reacting for 5h, decompressing and removing the solvent, and separating and purifying by column chromatography (petroleum ether: ethyl acetate volume ratio=20:1), so as to obtain the permeation-promoting compound with the yield of 82.5%. ESI-MS calculated, C ₂₈H₅₄NO₂(M+H)⁺ 436.41, found 436.4.

Nuclear magnetic results ：¹H NMR(300MHz,CDCl₃)δ3.37(t,2H),3.2(m,4H),2.78(m,1H),2.17(t,2H),1.80(m,1H),1.73(m,1H),160(m,1H),1.55-1.59(m,8H),1.45(m,2H),1.40(m,4H),1.27-1.30(m,18H),1.04(s,3H),1.01(d,6H).

Preparation example 2 preparation of active extract

The method comprises the following steps:

s1, supercritical fluid extraction, namely drying and crushing 7 parts by weight of sticktight and 3 parts by weight of dendrobium candidum, and extracting by using a supercritical fluid to prepare an oil extract, wherein the rest materials are left for use;

the supercritical fluid extraction condition is that the pressure is 27MPa, the temperature is 32 ℃ and the time is 70min;

S2, deep eutectic solvent enzymolysis, namely adding 10 parts by weight of the rest materials in the step S1 into 35 parts by weight of aqueous solution containing the deep eutectic solvent, adding complex enzyme, wherein the addition amount of the complex enzyme is 1wt% of the total mass of the system, carrying out enzymolysis for 2 hours at 40 ℃, and inactivating enzyme to obtain an enzymolysis product;

the water content of the aqueous solution containing the deep eutectic solvent is 40wt%, and the deep eutectic solvent is prepared by mixing choline chloride and glucose according to a molar ratio of 1:1;

the compound enzyme is formed by mixing cellulase and pectase, and the mass ratio is 10:3;

S3, fermenting, namely mixing 10 parts by weight of the enzymolysis product prepared in the step S2 with 15 parts by weight of water, sterilizing, inoculating bifidobacterium longum and bifidobacterium breve seed liquid, wherein the inoculum sizes of the bifidobacterium longum and the bifidobacterium breve seed liquid are respectively 2v/v% and 1v/v%, fermenting and culturing for 24 hours at 36 ℃ under the anaerobic condition, filtering, washing solids, collecting eluent, concentrating to obtain bacterial liquid, and reserving filtrate;

S4, alcohol precipitation, namely adding ethanol into the filtrate obtained in the step S3 until the content of the ethanol in the system is 60wt%, precipitating for 3 hours, centrifuging, washing, drying to obtain a polysaccharide composition, recovering the ethanol from supernatant fluid, and reserving the residual liquid;

S5, purifying the residual liquid in the step S4 by using macroporous resin NKA-9, removing the solvent from the filtrate by decompression, diluting the filtrate with 65wt% ethanol to a concentration of 0.8mg/mL, loading the sample into a column by a wet method, ensuring that the specification of the chromatographic column is 15mm multiplied by 500mm, ensuring that the column volume is 50mL, collecting eluent with the concentration of 65wt% ethanol, removing the ethanol by decompression, and drying to obtain an active mixture;

S7, preparing an active extract, namely stirring and mixing 5 parts by weight of the oil extract prepared in the step S1, 12 parts by weight of the polysaccharide composition prepared in the step S4, 10 parts by weight of the active mixture prepared in the step S5, 3 parts by weight of the yeast lysate prepared in the step S6 and 1 part by weight of vitamin C for 20 minutes to prepare the active extract.

Preparation example 3 preparation of active extract

The method comprises the following steps:

S1, supercritical fluid extraction, namely drying and crushing 10 parts by weight of sticktight and 5 parts by weight of dendrobium candidum, and extracting by using a supercritical fluid to prepare an oil extract, wherein the rest materials are left for use;

the supercritical fluid extraction condition is that the pressure is 35MPa, the temperature is 37 ℃ and the time is 90min;

s2, deep eutectic solvent enzymolysis, namely adding 10 parts by weight of the rest materials in the step S1 into 60 parts by weight of aqueous solution containing the deep eutectic solvent, adding complex enzyme, wherein the addition amount of the complex enzyme is 2wt% of the total mass of the system, carrying out enzymolysis for 4 hours at 50 ℃, and inactivating the enzyme to obtain an enzymolysis product;

the water content of the aqueous solution containing the deep eutectic solvent is 60wt%, and the deep eutectic solvent is prepared by mixing choline chloride and glucose according to a molar ratio of 1:1;

the compound enzyme is formed by mixing cellulase and pectase, and the mass ratio is 15:5;

S3, fermenting, namely mixing 10 parts by weight of the enzymolysis product prepared in the step S2 with 30 parts by weight of water, sterilizing, inoculating bifidobacterium longum and bifidobacterium breve seed liquid, wherein the inoculum sizes of the bifidobacterium longum and the bifidobacterium breve seed liquid are respectively 4v/v% and 3v/v%, fermenting and culturing for 48 hours at 39 ℃ under the anaerobic condition, filtering, washing solids, collecting eluent, concentrating to obtain bacterial liquid, and reserving filtrate;

S4, alcohol precipitation, namely adding ethanol into the filtrate obtained in the step S3 until the ethanol content of the system reaches 70wt%, precipitating for 5 hours, centrifuging, washing, drying to obtain a polysaccharide composition, recovering ethanol from supernatant fluid, and reserving the residual liquid;

S5, purifying the residual liquid in the step S4 by using macroporous resin NKA-9, removing the solvent from the filtrate by decompression, diluting the filtrate with 75wt% ethanol to a concentration of 1.2mg/mL, loading the sample into a column by a wet method, ensuring that the specification of the chromatographic column is 15mm multiplied by 500mm, ensuring that the column volume is 50mL, collecting eluent with the concentration of 75wt% ethanol, removing the ethanol by decompression, and drying to obtain an active mixture;

S7, preparing an active extract, namely stirring and mixing 7 parts by weight of the oil extract prepared in the step S1, 15 parts by weight of the polysaccharide composition prepared in the step S4, 17 parts by weight of the active mixture prepared in the step S5, 5 parts by weight of the yeast lysate prepared in the step S6 and 2 parts by weight of vitamin C for 20 minutes to prepare the active extract.

Preparation example 4 preparation of active extract

The method comprises the following steps:

S1, supercritical fluid extraction, namely drying 8 parts by weight of sticktight and 4 parts by weight of dendrobium candidum, crushing, and extracting by using a supercritical fluid to prepare an oil extract, wherein the rest materials are left for use;

The supercritical fluid extraction condition is that the pressure is 30MPa, the temperature is 35 ℃ and the time is 85min;

S2, deep eutectic solvent enzymolysis, namely adding 10 parts by weight of the rest materials in the step S1 into 45 parts by weight of an aqueous solution containing the deep eutectic solvent, adding complex enzyme, wherein the addition amount of the complex enzyme is 1.5 weight percent of the total mass of the system, carrying out enzymolysis for 3 hours at 45 ℃, and inactivating enzyme to obtain an enzymolysis product;

the water content of the aqueous solution containing the deep eutectic solvent is 50wt%, and the deep eutectic solvent is prepared by mixing choline chloride and glucose according to a molar ratio of 1:1;

The compound enzyme is formed by mixing cellulase and pectase, and the mass ratio is 12:4;

S3, fermenting, namely mixing 10 parts by weight of the enzymolysis product prepared in the step S2 with 22 parts by weight of water, sterilizing, inoculating bifidobacterium longum and bifidobacterium breve seed liquid, wherein the inoculum sizes of the bifidobacterium longum and the bifidobacterium breve seed liquid are respectively 3v/v% and 2v/v%, fermenting and culturing for 36h under the anaerobic condition at 37 ℃ and 170r/min, filtering, washing solids, collecting eluent, concentrating to obtain bacterial liquid, and reserving filtrate;

s4, alcohol precipitation, namely adding ethanol into the filtrate obtained in the step S3 until the ethanol content of the system reaches 65wt%, precipitating for 4 hours, centrifuging, washing, drying to obtain a polysaccharide composition, recovering ethanol from supernatant fluid, and reserving the residual liquid;

S5, purifying the residual liquid in the step S4 by using macroporous resin NKA-9, removing the solvent from the filtrate by decompression, diluting the filtrate with 70wt% ethanol to a concentration of 1mg/mL, loading the sample into a column by a wet method, ensuring that the specification of the chromatographic column is 15mm multiplied by 500mm, ensuring that the column volume is 50mL, collecting eluent with the concentration of 70wt% ethanol, removing the ethanol by decompression, and drying to obtain an active mixture;

S7, preparing an active extract, namely stirring and mixing 6 parts by weight of the oil extract prepared in the step S1, 13 parts by weight of the polysaccharide composition prepared in the step S4, 14 parts by weight of the active mixture prepared in the step S5, 4 parts by weight of the yeast lysate prepared in the step S6 and 1.5 parts by weight of vitamin C for 20 minutes to prepare the active extract.

Preparation example 5

The difference from preparation example 4 is that the complex enzyme is a single cellulase.

Preparation example 6

The difference compared to preparation example 4 is that the complex enzyme is a single pectase.

Comparative preparation example 1

Compared with preparation example 4, the difference is that no Bidens pilosa is added in step S1.

Comparative preparation example 2

Compared with preparation example 4, the difference is that no Dendrobium officinale was added in step S1.

Comparative preparation example 3

In comparison with preparation example 4, the difference is that no complex enzyme was added in step S2.

Comparative preparation example 4

The difference from preparation example 4 is that the seed solution of Bifidobacterium longum strain was not inoculated in step S3.

Comparative preparation example 5

The difference from preparation example 4 is that the seed solution of bifidobacterium breve was not inoculated in step S3.

Comparative preparation example 6

In comparison with preparation example 4, the difference is that step S3 is not performed.

Comparative preparation example 7

The difference from preparation example 4 is that no oil extract was added in step S7.

Comparative preparation example 8

The difference from preparation example 4 is that the polysaccharide composition was not added in step S7.

Comparative preparation example 9

In comparison with preparation example 4, the difference is that no active mixture is added in step S7.

Comparative preparation example 10

In comparison with preparation example 4, the difference is that no yeast lysate was added in step S7.

Example 1

The embodiment provides a compact skin anti-wrinkle essence, and the preparation method comprises the following steps:

(1) Uniformly mixing ethoxydiglycol and the permeation-promoting compound prepared in preparation example 1 according to a mass ratio of 4:3 to prepare a permeation-promoting agent composition;

(2) Uniformly mixing palmitoyl tripeptide-5, hexapeptide-11, hexapeptide-9, acetyl hexapeptide-8 and arginine/lysine polypeptide according to the mass ratio of 4:1:2:0.5:3 to prepare a polypeptide composition;

(3) Adding 7 parts by weight of the active extract prepared in preparation example 2,5 parts by weight of the polypeptide composition, 0.3 part by weight of soluble collagen, 0.1 part by weight of DNA sodium, 5 parts by weight of hydroxypropyl tetrahydropyran triol, 0.1 part by weight of hirudin, 5 parts by weight of butanediol, 0.2 part by weight of carbomer and 0.02 part by weight of EDTA disodium into 50 parts by weight of deionized water, heating to 70 ℃, stirring for 10min at 300r/min to obtain phase A

(4) Adding 0.5 weight part of the permeation enhancer composition into 2 weight parts of capric triglyceride and 0.5 weight part of sorbitan olive oleate, heating to 70 ℃, and stirring for 10min at 300r/min to obtain phase B;

(5) Adding phase B into phase A, stirring for 5min at 300r/min, emulsifying for 10min at 7000r/min, maintaining the temperature for 10min, cooling, and discharging to obtain compact skin anti-wrinkle essence.

Example 2

(1) Uniformly mixing ethoxydiglycol and the permeation-promoting compound prepared in preparation example 1 according to the mass ratio of 7:3 to prepare a permeation-promoting agent composition;

(2) Uniformly mixing palmitoyl tripeptide-5, hexapeptide-11, hexapeptide-9, acetyl hexapeptide-8 and arginine/lysine polypeptides according to the mass ratio of 6:3:4:1:5 to prepare a polypeptide composition;

(3) Adding 10 parts by weight of the active extract prepared in preparation example 3, 7 parts by weight of the polypeptide composition, 0.5 part by weight of soluble collagen, 0.5 part by weight of DNA sodium, 10 parts by weight of hydroxypropyl tetrahydropyran triol, 0.5 part by weight of hirudin, 10 parts by weight of trehalose, 0.5 part by weight of polyethylene glycol 2000 and 0.05 part by weight of disodium EDTA into 70 parts by weight of deionized water, heating to 80 ℃, stirring for 20min at 500r/min to obtain phase A

(4) Adding 2 parts by weight of a permeation enhancer composition to 8 parts by weight of polydimethylsiloxane and 2 parts by weight of cetostearyl alcohol olive oleate, heating to 80 ℃, and stirring for 20min at 500r/min to obtain a phase B;

(5) Adding phase B into phase A, stirring for 15min at 500r/min, emulsifying for 20min at 10000r/min, maintaining the temperature for 20min, cooling, and discharging to obtain compact skin anti-wrinkle essence.

Example 3

(1) Uniformly mixing ethoxydiglycol and the permeation-promoting compound prepared in preparation example 1 according to a mass ratio of 5:3 to prepare a permeation-promoting agent composition;

(2) Uniformly mixing palmitoyl tripeptide-5, hexapeptide-11, hexapeptide-9, acetyl hexapeptide-8 and arginine/lysine polypeptide according to the mass ratio of 5:2:3:0.7:4 to prepare a polypeptide composition;

(3) 8 parts by weight of the active extract obtained in preparation example 4, 6 parts by weight of the polypeptide composition, 0.4 part by weight of soluble collagen, 0.3 part by weight of DNA sodium, 7 parts by weight of hydroxypropyl tetrahydropyran triol, 0.3 part by weight of hirudin, 7 parts by weight of sodium hyaluronate, 0.35 part by weight of xanthan gum and 0.035 part by weight of disodium EDTA were added to 60 parts by weight of deionized water, heated to 75 ℃, and stirred for 15min at 400r/min to obtain phase A

(4) Adding 1 part by weight of the penetration enhancer composition into 5 parts by weight of triglyceride (ethylhexanoic acid) and 1 part by weight of glycerol stearate, heating to 75 ℃, and stirring for 15min at 400r/min to obtain a phase B;

(5) Adding phase B into phase A, stirring for 10min at 400r/min, emulsifying for 15min at 8000r/min, maintaining the temperature for 15min, cooling, and discharging to obtain compact skin anti-wrinkle essence.

Example 4

The difference compared to example 3 is that the permeation enhancer composition is replaced by a single ethoxydiglycol.

Example 5

The difference compared to example 3 is that the permeation enhancer composition is replaced by a single permeation enhancer compound prepared in preparation example 1.

Example 6

The difference compared to example 3 is that the active extract from preparation 4 is replaced by the active extract from preparation 5.

Example 7

The difference compared to example 3 is that the active extract from preparation 4 is replaced by the active extract from preparation 6.

Comparative example 1

The difference compared to example 3 is that the active extract obtained in preparation 4 is replaced by the active extract obtained in comparative preparation 1.

Comparative example 2

The difference compared to example 3 is that the active extract from preparation 4 is replaced by the active extract from comparative preparation 2.

Comparative example 3

The difference compared to example 3 is that the active extract obtained in preparation 4 is replaced by the active extract obtained in comparative preparation 3.

Comparative example 4

The difference compared to example 3 is that the active extract prepared in preparation 4 is replaced by the active extract prepared in comparative preparation 4.

Comparative example 5

The difference compared to example 3 is that the active extract from preparation 4 is replaced by the active extract from comparative preparation 5.

Comparative example 6

The difference compared to example 3 is that the active extract from preparation 4 is replaced by the active extract from comparative preparation 6.

Comparative example 7

The difference compared to example 3 is that the active extract from preparation 4 is replaced by the active extract from comparative preparation 7.

Comparative example 8

The difference compared to example 3 is that the active extract from preparation 4 is replaced by the active extract from comparative preparation 8.

Comparative example 9

The difference compared to example 3 is that the active extract from preparation 4 is replaced by the active extract from comparative preparation 9.

Comparative example 10

The difference compared to example 3 is that the active extract obtained in preparation example 4 is replaced by the active extract obtained in comparative preparation example 10.

Comparative example 11

In comparison with example 3, the difference is that no permeation enhancer composition was added.

Comparative example 12

In comparison with example 3, the polypeptide composition was not added.

Comparative example 13

The difference compared to example 3 is that no active extract was added.

Test example 1 safety evaluation

The tightening skin anti-wrinkle essence prepared in examples 1 to 7 and comparative examples 1 to 13 of the present invention were subjected to safety evaluation. The results were as follows:

the standard of sanitary chemistry and microbiology has lead content less than 10mg/kg, mercury less than 0.05mg/kg, arsenic less than 1mg/kg, total bacteria less than 10cfu/mL, and no detection of fecal coliform, staphylococcus aureus and pseudomonas aeruginosa. The indexes meet the specification of national 'hygienic Standard for cosmetics' GB 7916-87.

The acute oral toxicity test of mice shows that the acute oral toxicity LD ₅₀ values of female and male mice are both larger than 5000mg/kg body weight, and the acute oral toxicity test belongs to the actual non-toxic grade according to GB7919-87 chemical acute toxicity grading standards.

Skin multiple stimulation test, skin stimulation integral is greater than 0.4, less than 0.5, pathological histology integral <4. The skin irritation intensity evaluation standard according to the chemical substances GB7916-87 meets the national standard of cosmetic hygiene, and belongs to the light irritation substances.

Test example 2 clearance to DPPH

The compact dermatological anti-wrinkle essence prepared in examples 1 to 7 and comparative examples 1 to 13 was diluted with deionized water to 1mg/mL as a sample solution. Vitamin C was used as a control group.

Respectively taking 2.00 multiplied by 10 ^-4 mol/L DPPH absolute ethyl alcohol solution and 3mL of sample solution, adding into a test tube, shaking uniformly, placing for 30min in a dark place, measuring absorbance A _i of the mixed solution at 517nm, simultaneously measuring absorbance A ₀ of 3mL of pure water mixed with 3mL of DPPH absolute ethyl alcohol solution and absorbance A _j of 3mL of sample solution mixed with 3mL of absolute ethyl alcohol, and calculating the clearance (%) of DPPH according to a formula. The results are shown in Table 1.

DPPH clearance (%) = [1- (A _i-A_j)/A₀ ]. Times.100%)

TABLE 1

As shown in the table above, the compact anti-skin wrinkle essence prepared in the examples 1-3 of the invention has better antioxidant activity.

Test example 3

Face was selected as the test site, and subjects meeting the conditions were selected to be divided into 20 groups of 10 persons each of examples 1 to 7 and comparative examples 1 to 13, respectively. The subjects used the compact dermatological anti-wrinkle essence prepared in examples 1 to 7 and comparative examples 1 to 13 as test products according to the use requirements, collected skin data at the corresponding test sites of the subjects, and analyzed the change of the skin data before and after the use of the samples in the test areas.

Improvement rate (%) = | (last data-initial data) |/initial data×100%

The improvement rate is absolute value, the improvement rate is guaranteed to be always positive, and when the essence is used in actual test, the moisture content of the skin cuticle is improved, the skin elasticity R2 value is improved, and the skin compactness F4 value is reduced.

The specific test process is as follows:

test conditions subjects on the day of the test were allowed to sit still for 30min in a laboratory at a temperature of (25.+ -. 1) °C and humidity of 45%.+ -. 5% RH with the designated facial samples for facial cleansing.

Test method Corneometer test skin stratum corneum moisture content, cutometer test elasticity R2 value, and compactibility F4 value. The results are shown in Table 2.

(1) Test subject skin initiation data prior to the start of the experiment.

(2) After the initial data acquisition is completed, samples are dispensed to the subjects, full face smears are carried out once a day for 1 week, and the last data of the skin of the subjects are tested under the same test conditions after 1 week.

TABLE 2

Group of	Moisture content improvement Rate of skin horny layer (%)	Improvement rate of skin elasticity R2 value (%)	Skin firmness F4 value improvement rate (%)
				Example 1	52.2	23.1	33.2
Example 2	51.9	23.8	32.9
				Example 3	53.1	24.2	33.7
Example 4	44.1	19.7	28.9
				Example 5	46.9	21.0	30.4
Example 6	50.2	21.4	31.2
				Example 7	49.8	21.9	31.5
Comparative example 1	45.2	16.7	27.5
				Comparative example 2	46.9	17.2	26.4
Comparative example 3	47.9	20.2	30.6
				Comparative example 4	48.6	20.6	27.1
Comparative example 5	48.2	19.7	28.3
				Comparative example 6	46.7	16.8	25.9
Comparative example 7	49.1	16.1	24.4
				Comparative example 8	44.9	20.2	28.7
Comparative example 9	46.1	18.4	28.4
				Comparative example 10	47.6	17.5	27.6
Comparative example 11	43.0	18.2	27.1
				Comparative example 12	48.8	15.7	25.1
Comparative example 13	42.5	14.9	22.7

From the above table, the tightening skin anti-wrinkle essence prepared in the embodiments 1-3 has better effects of moisturizing, tightening and improving skin elasticity.

The foregoing description of the preferred embodiments of the invention is not intended to be limiting, but rather is intended to cover all modifications, equivalents, alternatives, and improvements that fall within the spirit and scope of the invention.

Claims

1. A skin tightening and anti-wrinkle essence, characterized in that it is prepared from the following raw materials by weight: 7-10 parts of active extract, 5-7 parts of polypeptide composition, 0.5-2 parts of penetration enhancer composition, 0.1-0.5 parts of DNA sodium, 0.3-0.5 parts of soluble collagen, 5-10 parts of hydroxypropyl tetrahydropyrantriol, 2-8 parts of emollient, 5-10 parts of humectant, 0.5-2 parts of emulsifier, 0.2-0.5 parts of thickener, 0.1-0.5 parts of hirudin, 0.02-0.05 parts of disodium EDTA, and 50-70 parts of deionized water;

The penetration enhancer composition comprises ethoxydiglycol and a penetration enhancing compound, and the structural formula of the penetration enhancing compound is shown in Formula I:

Wherein, R = C6-C18 alkyl chain;

The preparation method of the active extract is as follows:

S1. Supercritical fluid extraction: The Bidens pilosa and Dendrobium candidum were dried, crushed, and subjected to supercritical fluid extraction to obtain an oil extract, and the remaining material was retained;

S2. deep eutectic solvent enzymatic hydrolysis: adding the remaining material in step S1 to an aqueous solution containing a deep eutectic solvent, adding a composite enzyme for enzymatic hydrolysis, inactivating the enzyme, and obtaining an enzymatic hydrolysis product; the deep eutectic solvent is prepared by mixing choline chloride and glucose, and the composite enzyme is composed of a mixture of cellulase and pectinase;

S3 fermentation: the enzymatic product obtained in step S2 was mixed with water, sterilized, inoculated with long Bifidobacterium and breve Bifidobacterium species seed solution, fermented, filtered, washed with solids, the eluate was collected and concentrated to obtain a bacterial solution, and the filtrate was retained;

S4. Alcohol precipitation: adding ethanol to the filtrate in step S3, centrifuging, washing, drying, to obtain a polysaccharide composition, recovering ethanol from the supernatant, and retaining the remaining liquid;

S5. Purification by macroporous resin: The solvent of the remaining liquid in step S4 is removed under reduced pressure, and the filtrate is purified by macroporous resin, and the solvent is removed under reduced pressure, and the sample is diluted with 65-75wt% ethanol, and the column is wet loaded, and the column volume is 45-55mL, and the eluate with 65-75wt% ethanol concentration is collected, and the ethanol is removed under reduced pressure and dried to obtain an active mixture;

S6. Preparation of bifid yeast lysate: freezing the bacterial solution obtained in step S3 with liquid nitrogen, thawing it at room temperature, centrifuging it, collecting the supernatant, and freeze-drying it to obtain bifid yeast lysate;

S7. Preparation of active extract: the oil extract prepared in step S1, the polysaccharide composition prepared in step S4, the active mixture prepared in step S5, the bifida yeast lysate prepared in step S6 and vitamin C are uniformly mixed to prepare the active extract.

2. The skin tightening and anti-wrinkle essence according to claim 1, characterized in that the mass ratio of the ethoxydiglycol to the penetration-promoting compound is 4-7:3.

3. The skin tightening and anti-wrinkle essence according to claim 1 is characterized in that the synthesis method of the penetration-promoting compound is as follows: dissolving 1,12-dichlorododecane in dichloromethane, adding a dichloromethane solution of caprolactam and triethylamine dropwise, heating to reflux, stirring to react for a first period of time, then adding a dichloromethane solution of menthol dropwise, heating to reflux, stirring to react for a second period of time, removing the solvent under reduced pressure, separating and purifying by column chromatography, and obtaining the penetration-promoting compound.

4. The skin tightening and anti-wrinkle essence according to claim 3, characterized in that the molar ratio of 1,12-dichlorododecane, caprolactam, menthol and triethylamine is 1:0.9-1:0.9-1:4-6, the first time period is 3-5 hours, and the second time period is 4-6 hours.

5. The skin tightening and anti-wrinkle essence according to claim 1, characterized in that the polypeptide composition is a composition of palmitoyl tripeptide-5, hexapeptide-11, hexapeptide-9, acetyl hexapeptide-8, and arginine/lysine polypeptides, with a mass ratio of 4-6:1-3:2-4:0.5-1:3-5.

6. The skin tightening and anti-wrinkle essence according to claim 1, characterized in that the mass ratio of Bidens pilosa and Dendrobium officinale in step S1 is 7-10:3-5, the conditions of the supercritical fluid extraction are pressure 27-35 MPa, temperature 32-37° C., and time 70-90 min; the mass ratio of the residual material and the aqueous solution containing the deep eutectic solvent in step S2 is 10:35-60, the water content in the aqueous solution containing the deep eutectic solvent is 40-60wt%, and the The molar ratio of choline chloride to glucose is 1:1, the mass ratio of the cellulase to the pectinase is 10-15:3-5, the added amount of the complex enzyme is 1-2wt% of the total mass of the system, the temperature of the enzymolysis is 40-50°C, and the time is 2-4h; the mass ratio of the enzymolysis product to water in step S3 is 1:1.5-3, the inoculation amounts of the long bifidobacterium and the breve bifidobacterium seed liquid are 2-4v/v% and 1-3v ^/ v%, respectively, the bacterial content of the seed liquid is ^108-109 cfu/mL, and the fermentation culture conditions are anaerobic conditions, 36-39°C, 150-200r/min, and the fermentation culture is 24-48h.

7. The skin tightening and anti-wrinkle essence according to claim 1, characterized in that, in step S4, ethanol is added to 60-70wt% of the ethanol content of the system, and the precipitation time is 3-5h; in step S5, the macroporous resin is selected from at least one of XDA-2, NKA-9, SP-825, D101, HPD600, X-5, LSA-10, NKI-9 and AB-8, and is diluted with 65-75wt% ethanol to a concentration of 0.8-1.2mg/mL; in step S7, the mass ratio of the oil extract, the polysaccharide composition, the active mixture, the bifida yeast lysate and the vitamin C is 5-7:12-15:10-17:3-5:1-2.

8. The skin tightening and anti-wrinkle essence according to claim 1, characterized in that the emollient is selected from at least one of triethylhexanoin, isononyl isononanoate, dimethicone, caprylic/capric triglyceride, and avocado tree (BUTYROSPERMUM PARKII) fruit fat; the humectant is selected from at least one of glycerol, butylene glycol, propylene glycol, hexylene glycol, sodium hyaluronate, betaine, and trehalose; the emulsifier is selected from at least one of glyceryl stearate, PEG-100 stearate, cetearyl olive oil ester, sorbitan olive oil ester, C12-20 alkyl glucoside, and polyglyceryl-2 stearate; the thickener is selected from at least one of carbomer, acrylic acid (ester)/C10-30 alcohol acrylate crosspolymer, sodium polyacryloyldimethyl taurate, and xanthan gum.

9. A method for preparing the skin tightening and anti-wrinkle essence according to any one of claims 1 to 8, characterized in that it comprises the following steps:

(1) Add the active extract, polypeptide composition, soluble collagen, DNA sodium, hydroxypropyl tetrahydropyrantriol, hirudin, moisturizer, thickener, and disodium EDTA into deionized water, heat to 70-80°C, and stir at 300-500 r/min for 10-20 min to prepare phase A.

(2) adding the penetration enhancer composition to the emollient and emulsifier, heating to 70-80° C., and stirring at 300-500 r/min for 10-20 min to obtain phase B;

(3) Add phase B into phase A, stir at 300-500 r/min for 5-15 min, emulsify at 7000-10000 r/min for 10-20 min, keep warm for 10-20 min, cool down, discharge, and obtain a firming skin anti-wrinkle essence.