CN111388570B - Yellow tea compound extract and application thereof in skin external preparation - Google Patents
Yellow tea compound extract and application thereof in skin external preparation Download PDFInfo
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- CN111388570B CN111388570B CN202010424984.1A CN202010424984A CN111388570B CN 111388570 B CN111388570 B CN 111388570B CN 202010424984 A CN202010424984 A CN 202010424984A CN 111388570 B CN111388570 B CN 111388570B
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/23—Apiaceae or Umbelliferae (Carrot family), e.g. dill, chervil, coriander or cumin
- A61K36/232—Angelica
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
- A61K36/288—Taraxacum (dandelion)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/35—Caprifoliaceae (Honeysuckle family)
- A61K36/355—Lonicera (honeysuckle)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/82—Theaceae (Tea family), e.g. camellia
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
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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
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
Abstract
The invention discloses a yellow tea compound extract, which is prepared by the following method: (a) the honeysuckle flower with the weight ratio of 1-2:1-2:1-2: 1: dandelion: chinese angelica: extracting honeysuckle stem with a solvent to obtain an extracting solution A; (b) carrying out ultrasonic extraction on the yellow tea to obtain an extracting solution B; (c) mixing the extracting solution A and the extracting solution B according to the weight ratio of 1:0.8-2.5 to obtain the final yellow tea compound extract. The invention also discloses application of the yellow tea compound extract in skin moisturizing and/or skin barrier maintaining functions and application in preparing a skin external preparation for skin moisturizing and/or skin barrier maintaining functions.
Description
Technical Field
The invention aims to provide a yellow tea compound extract with moisturizing effect, which can promote the expression of keratinocyte tight junction protein (Claudin-1), can be used as an effect additive to be added into cosmetics and used for skin care.
Background
Skin refers to the tissue of the body surface outside the muscles, is the largest organ of the human body, and mainly plays a role in protecting the body, removing sweat, feeling cold and heat, pressure and the like. The skin covers the whole body, protects tissues and organs in the human body from being invaded by external substances such as pathogenic microorganisms and the like and mechanical injuries such as physics and the like, and effectively resists external stimulation; the skin also keeps the loss of water, electrolytes, nutrient substances and the like of the human body, effectively maintains the stability of the internal environment of the human body and is a natural barrier for protecting the human body.
Healthy skin is soft, smooth, and elastic, largely due to the abundant moisture in the skin, which is high and accounts for 18-20% of the total body mass. When the moisture content of the skin is lower than 10%, the skin becomes dry, rough and even chapped, and the skin loses water due to dry environment, unbalanced water and oil of the skin, excessive cleaning and the like. When the skin loses water excessively, the skin barrier is broken, so that the skin barrier cannot play a role in sufficiently retaining water. The skin does not have sufficient moisture for a long time, so that the skin is dry, the skin aging is accelerated, and various skin problems are caused; in addition, the sufficient water content of the skin is a prerequisite for improving the physiological environment of the skin and promoting the metabolism of the skin. Therefore, maintaining skin moisturization is particularly important in daily skin care.
Currently, there are two approaches to skin moisturization, one to prevent water loss and one to increase water absorption. Under normal conditions, the skin must first absorb sufficient water and then retain it. Therefore, moisture absorption is a prerequisite for skin moisturization. Moisture absorption of the skin is mainly accomplished by the combination of moisture absorption factors and water molecules in the skin interior and external skin care products. Strong binding force with water molecules, large water absorption capacity and good moisture retention effect.
Skin barrier function generally refers to the physical barrier structure of the epidermis, especially the stratum corneum. From the viewpoint of cell differentiation and tissue formation, the physical barrier function of the skin depends not only on the stratum corneum but also on the entire structure of the epidermis. From the aspects of biochemical composition and functional effect, the physical barrier structure of epidermis is not only related to lipid of epidermis, but also closely related to protein, water, inorganic salt and other metabolites of epidermis, and the abnormality of these components may affect the barrier function of skin, thereby causing a series of skin problems and even skin diseases.
Keratinocytes and tight junctions are two important structures in the physical barrier of the skin. While the Claudin-1 protein is one of the important transmembrane proteins in this structure, it maintains the skin barrier function and barrier function together with keratinocytes. If the expression is abnormal, the epithelial permeability barrier is damaged, the polarity of the cells is lost, and the adhesion between the cells is reduced. For example, in the Furute and Hata article, "important role of Claudin-based tight junctions for mammalian epidermal barrier" (Claudin-based light junctions are small for the mammalian epidermal barrier), Claudin-1 deficiency is reported to directly result in skin shrinkage, severe water deficit, increased transdermal water loss (TEWL) and decreased skin adhesion in mice. Therefore, promoting the expression of Claudin-1 can improve a series of skin problems caused by dryness and impaired skin barrier to a certain extent.
The flos Lonicerae is dried bud or flower with initial bloom of Lonicera japonica Thunb of Caprifoliaceae. Most areas in China produce the most of the Chinese herbs, the Shandong yield is the largest, and the Henan yield is better in quality. The main components of the medicine are chlorogenic acid, luteolin and the like, and the medicine mainly has the effects of resisting inflammation, relieving fever, resisting bacteria and viruses and the like.
The herba Taraxaci is dried whole plant of Compositae herba Taraxaci (Taraxacum mongolicum hand-Mazz.), herba Taraxaci (Taraxacum sinicum Kitag), or plants of the same genus. The distribution is distributed in most areas of the country. The whole herb contains dandelin, taraxol, inulin, choline and other components. Has antibacterial, antiinflammatory, and oxygen free radical resisting effects.
Caulis Lonicerae is dried stem and branch of Lonicera japonica (Lonicera japonica Thunb.) of Caprifoliaceae. Mainly produced in Zhejiang, Sichuan, Jiangsu and Henan provinces. Mainly contains chlorogenic acid, isochlorogenic acid and other components. Mainly has antibacterial and anti-inflammatory effects.
The Latin name (Camellia sinensis O. Ktze.) of the original yellow tea plant is a unique tea in China. The basic processing technology comprises spreading out, deactivating enzyme, stewing to yellow, rolling and drying. Mainly contains tea polyphenols, catechin, etc. Mainly has the functions of resisting oxidation, resisting allergy, resisting 5 alpha reductase and the like.
Angelica sinensis is the dried root of Angelica sinensis (Oliv.) Diels of Umbelliferae, and is divided into Gansu, Sichuan and Yunnan provinces. Contains ligustilide, ferulic acid, etc. Has effects of resisting oxidation and whitening skin.
The invention unexpectedly discovers that the compound extract containing the yellow tea, the honeysuckle, the dandelion, the honeysuckle stem and the angelica has the moisturizing effect of increasing the water absorption and/or the effect of promoting the expression of the keratinocyte tight junction protein (Claudin-1).
Therefore, the application takes the compound extract of the yellow tea, the honeysuckle, the dandelion, the honeysuckle stem and the angelica as a research object for the first time, and inspects the moisturizing effect and/or the skin barrier maintenance effect of the compound extract when the compound extract is externally applied to the skin.
Disclosure of Invention
The inventors of the present invention have found, after extensive and intensive studies: the compound extract of the yellow tea, the honeysuckle, the dandelion, the honeysuckle stem and the angelica has the moisturizing effect of increasing water absorption and/or has the effect of promoting the expression of keratinocyte tight junction protein (Claudin-1). Therefore, the compound extract can be used as an efficacy additive to be added into skin external preparations, especially cosmetics, and is used for moisturizing skin.
In one aspect, the invention provides a yellow tea compound extract, which is prepared by the following method:
(a) the honeysuckle flower with the weight ratio of 1-2:1-2:1-2: 1: dandelion: chinese angelica: extracting honeysuckle stem with a solvent to obtain an extracting solution A;
(b) carrying out ultrasonic extraction on the yellow tea to obtain an extracting solution B; and
(c) mixing the extracting solution A and the extracting solution B according to the weight ratio of 1:0.8-2.5 to obtain the final yellow tea compound extract.
In a preferred embodiment, the solvent in step (a) is selected from: deionized water, methanol, ethanol, acetone, propylene glycol, butylene glycol, glycerol, or combinations thereof.
In a preferred embodiment, step (b) comprises ultrasonic extraction with ethanol. In a preferred embodiment, step (b) comprises extraction with ethanol at a concentration of 70% by volume.
On the other hand, the invention also provides application of the yellow tea compound extract prepared according to the method in skin moisturizing and/or skin barrier function maintaining.
In a preferred embodiment, skin moisturization is achieved as a result of increased moisture absorption.
In a preferred embodiment, maintaining skin barrier function is achieved by promoting expression of keratinocyte tight junction protein.
In another aspect, the invention also provides application of the yellow tea compound extract prepared according to the method in preparing a skin external preparation for moisturizing skin and/or maintaining skin barrier function.
In a preferred embodiment, the skin moisturizing of the external skin preparation is achieved as a result of increasing moisture absorption.
In a preferred embodiment, the skin barrier function maintenance of the external preparation for skin is achieved by promoting the expression of keratinocyte tight junction protein.
In a preferred embodiment, the external preparation for skin comprises 0.0001 to 20% by weight, preferably 0.001 to 10% by weight, more preferably 0.001 to 5% by weight, most preferably 0.01 to 5% by weight of the yellow tea compound extract.
Drawings
FIG. 1 shows the results of the effect of the extracts described herein on the expression of epidermal keratinocyte tight junction protein (Claudin-1). Wherein, FIG. 1A shows the results of the negative control; figure 1B shows keratinocyte claudin expression following treatment with 0.0016 wt% of the yellow tea compound extract of example 5; figure 1C shows keratinocyte tight junction protein expression following treatment with 0.2 wt.% of the extract of example 7; figure 1D shows keratinocyte claudin expression after treatment with 0.04 wt.% of the extract of example 8.
Detailed Description
The invention discovers for the first time that the yellow tea compound extract has the moisturizing effect of increasing water absorption, and the compound extract has the capability of promoting the expression of keratinocyte tight junction protein (Claudin-1). Therefore, the yellow tea compound extract can be used as an efficacy additive to be added into a skin external agent (such as a skin care product) to assist in improving skin problems such as skin dryness, skin barrier damage and the like related to Claudin-1.
The invention provides a yellow tea compound extract, which is prepared by the following steps: (a) the honeysuckle flower with the weight ratio of 1-2:1-2:1-2: 1: dandelion: chinese angelica: extracting honeysuckle stem with a solvent to obtain an extracting solution A; (b) carrying out ultrasonic extraction on the yellow tea to obtain an extracting solution B; and (c) mixing the extracting solution A and the extracting solution B according to the weight ratio of 1:0.8-2.5 to obtain the final yellow tea compound extract.
In some preferred embodiments of the present invention, the ratio of honeysuckle: dandelion: chinese angelica: the weight ratio of the honeysuckle stem is 2:2:2: 1.
The yellow tea compound extract can be applied to skin external preparations, especially cosmetics, and different dosage is added according to different types of preparations.
The yellow tea compound extract can be used as an additive with a moisturizing effect to be applied to a skin external preparation. In a specific embodiment, the yellow tea compound extract can be applied to cosmetics as an additive with moisturizing effect. In a particular embodiment, the cosmetic is selected from: face cleaning lotion, cosmetic water, lotion, cream, jelly and facial mask. Different amounts are added according to different types of preparations.
The yellow tea compound extract can be used as an additive with the function of maintaining skin barrier to be applied to skin external preparations. In a specific embodiment, the yellow tea compound extract can be applied to cosmetics as an additive with the function of maintaining the skin barrier. In a particular embodiment, the cosmetic is selected from: face cleaning lotion, cosmetic water, lotion, cream, jelly and facial mask. Different amounts are added according to different types of preparations.
In some preferred embodiments, the amount of the yellow tea compound extract contained in the external preparation for skin may be 0.001% to 20% (w/w). Preferably 0.01-20% (w/w). More preferably 0.01% to 10% (w/w). More preferably 0.1% to 5% (w/w). In some preferred embodiments of the present invention, the amount of the yellow tea compound extract contained in the external preparation for skin may be 0.001% -10% (w/w), 0.002% -10% (w/w), 0.003% -10% (w/w), 0.01% -10% (w/w), 0.02% -10% (w/w), 0.03% -10% (w/w), 0.1% -10% (w/w), 0.2% -10% (w/w), 0.3% -10% (w/w). In some preferred embodiments of the present invention, the amount of the yellow tea compound extract contained in the external preparation for skin may be 0.001% to 1% (w/w), for example, 0.002% to 1% (w/w), 0.003% to 1% (w/w), 0.01% to 1% (w/w), 0.02% to 1% (w/w), 0.03% to 1% (w/w), 0.1% to 1% (w/w), 0.2% to 1% (w/w), 0.3% to 1% (w/w).
In some embodiments, the yellow tea compound extract for skin moisturizing and/or maintaining skin barrier function is used in an amount of 0.001-5 wt%. In some embodiments, the yellow tea compound extract for skin moisturizing and/or skin barrier function maintenance is used in an amount of 0.001-1 wt%. In some embodiments, the yellow tea compound extract for skin moisturizing and/or skin barrier function maintenance is used in an amount of 0.001-0.2 wt%. In some embodiments, the yellow tea compound extract for skin moisturizing and/or skin barrier function maintenance is used in an amount of 0.001 to 0.04 wt%. In some embodiments, the yellow tea compound extract for skin moisturizing and/or skin barrier function maintenance is used in an amount of 0.001 to 0.008 wt%. In some embodiments, the yellow tea compound extract for skin moisturizing and/or skin barrier function maintenance is used in an amount of 0.001 to 0.002 wt%.
In another aspect of the present invention, there is provided a skin external preparation having skin moisturizing and/or skin barrier maintaining functions, comprising the compound extract of camellia sinensis prepared according to the method of the present invention and a cosmetically acceptable excipient.
The external preparation for skin is a general concept of all ingredients generally used for the external skin, and may be, for example, a cosmetic composition or a pharmaceutical composition. The cosmetic composition may be a basic cosmetic, a face makeup cosmetic, a body cosmetic, a hair care cosmetic, etc., and the formulation thereof is not particularly limited and may be appropriately selected depending on the purpose.
The cosmetic composition also contains different cosmetically acceptable media or matrix excipients according to different formulations and purposes.
The cosmetically, dermatologically or pharmaceutically acceptable vehicle that can be used in the composition for external application to skin of the present invention is in the form of a water phase, an oil phase, a gel, a wax-in-water emulsion, an oil-in-water emulsion or a water-in-oil emulsion. The aqueous phase is a mixture of one or more water-soluble or dispersible components, which may be liquid, semi-solid, or solid at room temperature (25 ℃). The vehicle includes or may be in the form of a suspension, dispersion or solution in an aqueous or hydro-alcoholic vehicle, which may contain a thickening or gelling agent. The person skilled in the art can select suitable product forms, the components contained therein, based on the knowledge of the person skilled in the art.
The composition may comprise an aqueous phase which may contain water or a mixture of water and at least one hydrophilic organic solvent such as an alcohol, in particular a linear or branched lower monohydric alcohol containing from 2 to 5 carbon atoms, such as ethanol or propanol; polyols, such as propylene glycol, sorbitol, glycerol, panthenol or polyethylene glycols and mixtures thereof.
When the composition of the invention is in the form of an emulsion, the composition may also optionally comprise a surfactant.
The composition may also comprise film-forming polymers such as polyurethanes, polyacrylic acid homo-or copolymers, polyesters, hydrocarbon-based resins and/or silicone resins. The polymer may be dissolved or dispersed in a cosmetically acceptable vehicle and optionally combined with a plasticizer.
The compositions of the present invention may also comprise an oil phase containing oil-soluble or oil-dispersible components that are liquid at room temperature (25 ℃) and/or substances that are oily or waxy at room temperature, such as waxes, semisolids, gums, and mixtures thereof. The oil phase may also contain an organic solvent.
Typically liquid at room temperature, suitable oily substances include: hydrocarbon-based oils of animal origin, such as perhydrosqualene; hydrocarbon-based vegetable oils, such as liquid triglycerides of C4-10 fatty acids, e.g. heptanoic acid or octanoic acid triglycerides, or oils, e.g. sunflower oil, corn oil, soybean oil, grapeseed oil, castor oil, avocado oil, octanoic/decanoic acid triglycerides, jojoba oil; linear or branched hydrocarbons of mineral or synthetic origin, such as liquid paraffin and its derivatives, vaseline; synthetic esters and ethers, in particular esters of fatty alcohols, such as isopropyl myristate, 2-ethylhexyl palmitate, 2-octyldodecyl stearate, isostearyl isostearate; hydroxylated esters, such as isostearyl lactate, octyl hydroxystearate, octyl dodecyl hydroxystearate, heptanoates, octanoates and decanoates of fatty alcohols; polyol esters such as propylene glycol dicaprylate, neopentyl glycol diheptanoate, diethylene glycol diisononanoate, and pentaerythritol esters; c12-26-containing fatty alcohols, such as octyldodecanol, 2-butyloctanol, 2-hexyldecanol, 2-undecylpentadecanol, oleyl alcohol; fluoro and/or fluorosilicone oils based in part on hydrocarbons, silicone oils, volatile or non-volatile linear or cyclic polymethylsiloxanes which are liquid or semi-solid at room temperature, such as cyclic polydimethylsiloxanes and polydimethylsiloxanes, optionally containing phenyl groups, such as phenyltrimethicones, silicones and mixtures thereof.
The composition of the present invention may further comprise any component commonly used in the cosmetic field. These components include preservatives, aqueous phase thickeners (extract biopolymers, synthetic polymers) and fatty phase thickeners, fragrances, hydrophilic and lipophilic active agents and mixtures thereof.
The compositions of the invention may also comprise an additional particulate phase, which may be a pigment and/or a pearlescent agent and/or a filler used in cosmetic compositions.
Pigments may be present in the composition, suitable inorganic pigments include titanium oxide, zirconium oxide and cerium oxide as well as zinc oxide, iron oxide and ferric blue; suitable organic pigments include barium, strontium, calcium and aluminum lakes and carbon black.
Pearling agents may be present in the composition, suitable pearling agents include mica coated with titanium oxide, iron oxide or natural pigments.
Fillers may be present in the composition, suitable fillers include talc, silica, zinc stearate, mica, kaolin, nylon powder, polyethylene powder, teflon, starch, boron nitride, copolymer microspheres, such as silicone resin microbeads.
The oil phase of the compositions of the present invention may comprise one or more waxes, gums or mixtures thereof. Waxes include hydrocarbon-based waxes, fluoro waxes, and/or silicone waxes, and may be derived from vegetable, mineral, animal, and/or synthetic sources. Suitable waxes include beeswax, carnauba wax, candelilla wax, paraffin wax, microcrystalline wax, ozokerite; synthetic waxes include polyethylene waxes, silicone waxes containing C16-45. Gums are generally polydimethylsiloxanes or sodium carboxymethylcellulose or extracts, and semisolid substances are generally hydrocarbon-based compounds, such as lanolin and its derivatives.
The compositions of the present invention may be formulated into any suitable product form. Such product forms include, but are not limited to, aerosol sprays, creams, lotions, solids, liquids, dispersions, foams, gels, lotions, mousses, ointments, powders, patches, pomades, solutions, hand pump sprays, sticks, masks and towelettes. The compositions of the present invention may be conveniently used to prepare or as cosmetic, dermatological or pharmaceutical topical products by various methods well known in the art.
The composition for external skin preparations of the present invention may include one or more of the following ingredients: anti-allergic agents, antimicrobial agents, antioxidants, chelating agents, colorant depigmenting agents, emollients, emulsifiers, exfoliants, film formers, fragrances, humectants, insect repellents, lubricants, pharmaceutically active agents, moisturizers, light stabilizers, preservatives, skin protectants, skin penetration enhancers, sunscreens, stabilizers, surfactants, thickeners, viscosity modifiers, vitamins, or any combination thereof.
The invention is further illustrated below with reference to specific examples. It is to be understood, however, that these examples are illustrative only and are not to be construed as limiting the scope of the present invention. Test methods in which specific conditions are not specified in the following examples are generally carried out under conventional conditions or under conditions recommended by the manufacturer. All percentages and parts are by weight unless otherwise indicated.
Yellow tea, black tea and black tea adopted in the following examples are purchased from Fujian Asia Tong green garden tea industry Co Ltd; the adopted honeysuckle, dandelion, angelica and honeysuckle stem are all purchased from Shanghai medicine Huayu pharmaceutical Co Ltd.
Example 1: preparation of yellow tea extract
Weighing 100g of yellow tea, pulverizing into coarse powder (type of pulverizer: Dade medicine machine DFY-600C), performing ultrasonic extraction with 10 times of 70% ethanol (v/v) for 45 minutes (ultrasonic instrument: TGCXN-2B of Beijing Hongxianglong biotechnology Co., Ltd.), filtering with filter paper (specification of filter paper: 60 x60 cm rapid filter paper), concentrating the filtrate at 55 deg.C, standing overnight at 4 deg.C, taking out, centrifuging at 3 deg.C and 3500rpm for 15 minutes, filtering with filter paper (specification of filter paper: 60 x60 cm rapid filter paper), and making into 1g/ml solution.
Example 2: preparation of yellow tea compound extract
Weighing honeysuckle: dandelion: chinese angelica: 200g of honeysuckle stem (1:1:1:1), extracting with 10 times of deionized water for 1h and 2 times, filtering cotton, combining filtrates, adding 95% ethanol (v/v) for alcohol precipitation, uniformly stirring, standing overnight, filtering with filter paper (specification of filter paper: 60 x60 cm rapid filter paper), concentrating the filtrate to 1 time of crude drug amount, standing overnight at 4 ℃, centrifuging for 15 minutes at 3500rpm of 3 ℃, filtering with filter paper (specification of filter paper: 60 x60 cm rapid filter paper), and finally preparing into 1g/ml solution, namely extracting solution A.
Weighing 200g of yellow tea, pulverizing into coarse powder (powder grinding machine model: Dade medicine machine DFY-600C), performing ultrasonic extraction with 10 times of 70% ethanol (v/v) for 45 minutes (ultrasonic instrument: TGCXN-2B of Beijing Honghong biotechnology, Inc.), filtering with filter paper (filter paper specification: 60 x60 cm rapid filter paper), concentrating the filtrate at 55 deg.C, standing overnight at 4 deg.C, centrifuging at 3 deg.C and 3500rpm for 15 minutes, filtering with filter paper (filter paper specification: 60 x60 cm rapid filter paper), and making into 1g/ml solution, i.e. extractive solution B.
Mixing the extract A and the extract B (3:1), and finally preparing 1g/ml yellow tea compound extract.
Example 3: preparation of yellow tea compound extract
Weighing honeysuckle: dandelion: chinese angelica: 100g of honeysuckle stem (2:2:1:1), extracting with 10 times of deionized water for 1h and 2 times, filtering cotton, combining filtrates, adding 95% ethanol (v/v) for alcohol precipitation, uniformly stirring, standing overnight, filtering with filter paper (specification of filter paper: 60 x60 cm rapid filter paper), concentrating the filtrate to 1 time of crude drug amount, standing overnight at 4 ℃, centrifuging for 15 minutes at 3500rpm of 3 ℃, filtering with filter paper (specification of filter paper: 60 x60 cm rapid filter paper), and finally preparing into 1g/ml solution, namely extracting solution A.
Weighing 100g of yellow tea, pulverizing into coarse powder (powder grinding machine model: Dade medicine machine DFY-600C), performing ultrasonic extraction with 10 times of 70% ethanol (v/v) for 45 minutes (ultrasonic instrument: TGCXN-2B of Beijing Honghong biotechnology, Inc.), filtering with filter paper (filter paper specification: 60 x60 cm rapid filter paper), concentrating the filtrate at 55 deg.C, standing overnight at 4 deg.C, centrifuging at 3 deg.C and 3500rpm for 15 minutes, filtering with filter paper (filter paper specification: 60 x60 cm rapid filter paper), and making into 1g/ml solution, i.e. extractive solution B.
Mixing the extract A and the extract B (1:1), and finally preparing 1g/ml yellow tea compound extract.
Example 4: preparation of yellow tea compound extract
Weighing honeysuckle: dandelion: chinese angelica: 150g of honeysuckle stem (2:2:1:1), extracting with 10 times of deionized water for 1h and 2 times, filtering cotton, combining filtrates, adding 95% ethanol (v/v) for alcohol precipitation, uniformly stirring, standing overnight, filtering with filter paper (specification of filter paper: 60 x60 cm rapid filter paper), concentrating the filtrate to 1 time of crude drug amount, standing overnight at 4 ℃, centrifuging for 15 minutes at 3500rpm of 3 ℃, filtering with filter paper (specification of filter paper: 60 x60 cm rapid filter paper), and finally preparing into 1g/ml solution, namely extracting solution A.
Weighing 150g of yellow tea, pulverizing into coarse powder (powder pulverizing machine model: Dade medicine machine DFY-600C), extracting with 10 times of 70% ethanol (v/v) by ultrasonic extraction for 45 minutes (ultrasonic instrument: TGCXN-2B of Beijing Honghong biotechnology, Inc.), filtering with filter paper (filter paper specification: 60 x60 cm rapid filter paper), concentrating the filtrate at 55 deg.C, standing overnight at 4 deg.C, centrifuging at 3 deg.C and 3500rpm for 15 minutes, filtering with filter paper (filter paper specification: 60 x60 cm rapid filter paper), and making into 1g/ml solution to obtain extract B.
Mixing the extractive solution A and the extractive solution B (1:3), and making into 1g/ml compound extractive solution.
Example 5: preparation of yellow tea compound extract
Weighing honeysuckle: dandelion: chinese angelica: 140g of honeysuckle stem (2:2:2:1), extracting with 10 times of deionized water for 1h and 2 times, filtering cotton, combining filtrates, adding 95% ethanol (v/v) for alcohol precipitation, uniformly stirring, standing overnight, filtering with filter paper (specification of filter paper: 60 x60 cm rapid filter paper), concentrating the filtrate to 1 time of crude drug amount, standing overnight at 4 ℃, centrifuging for 15 minutes at 3500rpm of 3 ℃, filtering with filter paper (specification of filter paper: 60 x60 cm rapid filter paper), and finally preparing into 1g/ml solution, namely extracting solution A.
Weighing 140g of yellow tea, pulverizing into coarse powder (powder pulverizing machine model: Dade medicine machine DFY-600C), performing ultrasonic extraction with 10 times of 70% ethanol (v/v) for 45 minutes (ultrasonic instrument: TGCXN-2B of Beijing Honghong biotechnology, Inc.), filtering with filter paper (filter paper specification: 60 x60 cm rapid filter paper), concentrating the filtrate at 55 deg.C, standing overnight at 4 deg.C, centrifuging at 3 deg.C and 3500rpm for 15 minutes, filtering with filter paper (filter paper specification: 60 x60 cm rapid filter paper), and making into 1g/ml solution to obtain extract B.
Mixing the extractive solution A and the extractive solution B (1:0.8), and making into 1g/ml compound extractive solution.
Example 6: preparation of yellow tea compound extract
Weighing honeysuckle: dandelion: chinese angelica: 70g of honeysuckle stem (2:2:2:1), extracting with 10 times of deionized water for 1h and 2 times, filtering cotton, combining filtrates, adding 95% ethanol (v/v) for alcohol precipitation, uniformly stirring, standing overnight, filtering with filter paper (specification of filter paper: 60 x60 cm rapid filter paper), concentrating the filtrate to 1 time of crude drug amount, standing overnight at 4 ℃, centrifuging for 15 minutes at 3500rpm of 3 ℃, filtering with filter paper (specification of filter paper: 60 x60 cm rapid filter paper), and finally preparing into 1g/ml solution, namely extracting solution A.
Weighing 70g of yellow tea, pulverizing into coarse powder (powder pulverizing machine model: Dade medicine machine DFY-600C), performing ultrasonic extraction with 10 times of 70% ethanol (v/v) for 45 minutes (ultrasonic instrument: TGCXN-2B of Beijing Honghong biotechnology, Inc.), filtering with filter paper (filter paper specification: 60 x60 cm rapid filter paper), concentrating the filtrate at 55 deg.C, standing overnight at 4 deg.C, centrifuging at 3 deg.C and 3500rpm for 15 minutes, filtering with filter paper (filter paper specification: 60 x60 cm rapid filter paper), and making into 1g/ml solution to obtain extract B.
Mixing the extract A and the extract B (1:2.5), and finally preparing 1g/ml yellow tea compound extract.
Example 7: preparation of black tea compound extract
Weighing honeysuckle: dandelion: chinese angelica: 100g of honeysuckle stem (2:2:2:1), extracting with 10 times of deionized water for 1h and 2 times, filtering cotton, combining filtrates, adding 95% ethanol (v/v) for alcohol precipitation, uniformly stirring, standing overnight, filtering with filter paper (specification of filter paper: 60 x60 cm rapid filter paper), concentrating the filtrate to 1 time of crude drug amount, standing overnight at 4 ℃, centrifuging for 15 minutes at 3500rpm of 3 ℃, filtering with filter paper (specification of filter paper: 60 x60 cm rapid filter paper), and finally preparing into 1g/ml solution, namely extracting solution A.
Weighing 100g of black tea, pulverizing into coarse powder (powder pulverizing machine model: Dade medicine machine DFY-600C), performing ultrasonic extraction with 10 times of 70% ethanol (v/v) for 45 minutes (ultrasonic instrument: TGCXN-2B of Beijing Honghong biotechnology, Inc.), filtering with filter paper (filter paper specification: 60 x60 cm rapid filter paper), concentrating the filtrate at 55 deg.C, standing overnight at 4 deg.C, centrifuging at 3 deg.C and 3500rpm for 15 minutes, filtering with filter paper (filter paper specification: 60 x60 cm rapid filter paper), and making into 1g/ml solution to obtain extract B.
Mixing the extractive solution A and the extractive solution B (1:0.8), and making into 1g/ml compound extractive solution.
Example 8: preparation of black tea compound extract
Weighing honeysuckle: dandelion: chinese angelica: 100g of honeysuckle stem (2:2:2:1), extracting with 10 times of deionized water for 1h and 2 times, filtering cotton, combining filtrates, adding 95% ethanol (v/v) for alcohol precipitation, uniformly stirring, standing overnight, filtering with filter paper (specification of filter paper: 60 x60 cm rapid filter paper), concentrating the filtrate to 1 time of crude drug amount, standing overnight at 4 ℃, centrifuging for 15 minutes at 3500rpm of 3 ℃, filtering with filter paper (specification of filter paper: 60 x60 cm rapid filter paper), and finally preparing into 1g/ml solution, namely extracting solution A.
Weighing black tea 100g, pulverizing into coarse powder (powder grinding machine model: Dade medicine machine DFY-600C), extracting with 10 times of 70% ethanol (v/v) by ultrasonic extraction for 45 min (ultrasonic instrument: TGCXN-2B of Beijing Honghong biotechnology Co., Ltd.), filtering with filter paper (filter paper specification: 60 x60 cm rapid filter paper), concentrating the filtrate at 55 deg.C, standing overnight at 4 deg.C, centrifuging at 3 deg.C and 3500rpm for 15 min, filtering with filter paper (filter paper specification: 60 x60 cm rapid filter paper), and making into 1g/ml solution to obtain extract B.
Mixing the extractive solution A and the extractive solution B (1:0.8), and making into 1g/ml compound extractive solution.
Example 9: preparation of yellow tea compound extract
Weighing honeysuckle: dandelion: 280g of honeysuckle stem (2:2:1), extracting with 10 times of deionized water for 1h and 2 times, filtering cotton, combining filtrates, adding 95% ethanol (v/v) for alcohol precipitation, uniformly stirring, standing overnight, filtering with filter paper (the specification of the filter paper is 60 x60 cm rapid filter paper), concentrating the filtrate to 1 time of crude drug amount, standing overnight at 4 ℃, centrifuging for 15 minutes at the temperature of 3 ℃ and 3500rpm, filtering with filter paper (the specification of the filter paper is 60 x60 cm rapid filter paper), and finally preparing into 1g/ml solution, namely extracting solution A.
Weighing 100g of yellow tea, pulverizing into coarse powder (powder grinding machine model: Dade medicine machine DFY-600C), performing ultrasonic extraction with 10 times of 70% ethanol (v/v) for 45 minutes (ultrasonic instrument: TGCXN-2B of Beijing Honghong biotechnology, Inc.), filtering with filter paper (filter paper specification: 60 x60 cm rapid filter paper), concentrating the filtrate at 55 deg.C, standing overnight at 4 deg.C, centrifuging at 3 deg.C and 3500rpm for 15 minutes, filtering with filter paper (filter paper specification: 60 x60 cm rapid filter paper), and making into 1g/ml solution, i.e. extractive solution B.
Mixing the extract A and the extract B (1:2.5), and finally preparing 1g/ml yellow tea compound extract.
Example 10: evaluation of moisture Retention Property
According to the characteristic that saturated salt solution in a closed space can provide a constant humidity value, a sample aqueous solution with a certain concentration is placed in a dryer containing potassium acetate saturated solution (the relative humidity is 60%) for a period of time, the water loss condition of the sample solution along with the change of time is calculated, the water retention rate is compared to measure the water loss prevention capability of the sample under a dry environment, and the higher the water retention rate is, the better the moisture retention effect is.
0.1g (1mg/ml) of the prepared extracts of examples 1-9 was placed in small plates, respectively, placed in a desiccator containing a saturated solution of potassium acetate (relative humidity 60%), weighed for 3 times at times of 0, 24, 36, 72, 96 and 120 hours, respectively, and the water loss of the sample solution as a function of time was calculated and the ability of the sample to prevent water loss in a dry environment was measured by comparing the water retention. The capacity of the sample for absorbing outside moisture is compared according to the water absorption, and the higher the water absorption is, the better the moisture absorption effect is. The results are shown in Table 1.
Moisture absorption rate (%) - (M2-M1)/M1X 100%
M1-sample Dry weight before moisture absorption (g)
M2-sample weight after moisture absorption (g)
TABLE 1
Note: in the table ". indicates that the value was significantly different from the water loss value of water at the same time (negative control) (. p.p.0.05;. p.0.01;. p.0.001).
Moisture absorption rate in vitro experiment results show that the yellow tea compound has different degrees of absorption on water under different moisture absorption time. Under the same moisture absorption time, the moisture absorption rate of the yellow tea compound in example 6 is the highest, and the moisture absorption rate of the yellow tea compound in example 1 only and the moisture absorption rate of the yellow tea compound in example 9 without angelica sinensis are not as high as that of example 6 at each time, which shows that the yellow tea compound has the moisture retention effect (p is less than 0.05) under a specific ratio.
Example 11: effect on expression of epidermal keratinocyte tight junction protein (Claudin-1)
1. The experimental method comprises the following steps:
(1) cell culture: HaCaT cells (immortalized human skin keratinocytes (HaCaT, ATCC) were inoculated into a culture flask containing DK-SFM medium (DK-SFM medium, Gibco) and placed at 37 ℃ in CO2(volume fraction is 5%) for 5 days in a constant-temperature incubator until the cells are fused, and changing the culture solution every other day. For the experiment, the cell culture flask was taken out, and HaCaT was trypsinized (Sigma Co.) to give a fine powderDigesting the cells for 4 minutes, then inoculating the cells into a 96-well plate, incubating for 24 hours, adding 200 mu l of example extracting solution diluted by culture solution into each well of the medicine adding group, directly adding 200 mu l of culture solution into each well of the control group, and culturing for 48-72 hours in an incubator. The test of cytotoxicity and the capability of synthesizing the moisturizing related protein is carried out.
(2) The compound extracts of example 5, example 7 and example 8 were prepared into compound solutions of corresponding final concentrations (5%, 1%, 0.2%, 0.04%, 0.008%, 0.0016% by weight), and tested for cytotoxicity by XTT.
(3) According to the results of the cytotoxicity test, after the end of the drug-adding treatment at the mass concentrations which did not affect the cell activities in examples 5, 7 and 8, respectively, the cell culture solution in the 96-well plate was removed, fixed with acetone, and then 200. mu.l of a 1% aqueous Triton X-100 solution was added thereto and allowed to stand at room temperature for 1 hour. After adding the primary antibody Claudin-1 (Invitrogen) overnight at 4 deg.C, the corresponding secondary antibody (Invitrogen) was added and incubated l h away from light at room temperature. DAPI (Sigma) was stained, and finally, the blocking piece was added and mounted, and observed and photographed under a fluorescent microscope (OLYMPUS IX50 inverted microscope and OLYMPUSB X60-32 fluorescent microscope, OLYMPUS, Inc., magnification 400 times).
2. The experimental results are as follows:
as can be seen from the results of FIGS. 1A to 1D, 0.0016 wt% of the extract solution of example 5, i.e., the yellow tea infusion, was most effective in promoting the expression of epidermal keratinocyte Claudin-1 in the tea formulations of the respective examples, relative to 0.2 wt% of example 7 and 0.04 wt% of example 8.
The compound extracts prepared in examples 2 to 6 and example 9 were used for the preparation of external preparations for skin. The skin external preparation is preferably a cosmetic composition such as a lotion, essence, cream, etc. The weight percentage of the compound extract in the skin external preparation is 0.0001-20% (w/w). Preferably 0.001-10% (w/w). More preferably 0.001-5% (w/w). Most preferably 0.01% to 5% (w/w).
The following are examples of specific applications of the compound extract in skin external preparations, and formulations and preparation methods of these preparations. In the tables, "-" indicates no addition.
Example 12: preparation of face cream
Example 13: preparation of the emulsion
Example 14: preparation of jelly
Example 15: preparation of astringent
Example 16: preparation of essence
Example 17: preparation of facial mask
Example 18: preparation of eye cream
Example 19: preparation of an aerosol (cleaning foam)
Example 20: preparation of the spray
Example 21: preparation of shower gel
Example 22: preparation of facial cleanser
Claims (7)
1. A yellow tea compound extract for moisturizing skin and/or maintaining skin barrier function is prepared by the following steps:
(a) mixing the honeysuckle flower with the weight ratio of 2:2:2: 1: dandelion: chinese angelica: extracting honeysuckle stem with a solvent to obtain an extracting solution A;
(b) carrying out ultrasonic extraction on the yellow tea to obtain an extracting solution B; and
(c) mixing the extract A and the extract B according to the weight ratio of 1:0.8-2.5 to obtain the final yellow tea compound extract,
wherein the solvent in step (a) is deionized water,
wherein the step (b) comprises ultrasonic extraction with ethanol.
2. Use of the yellow tea compound extract according to claim 1 for skin moisturizing and/or skin barrier function maintenance for non-therapeutic purposes.
3. Use of the yellow tea compound extract according to claim 1 for preparing a skin external preparation for moisturizing skin and/or maintaining skin barrier function.
4. The use as claimed in claim 3, wherein the external preparation for skin comprises 0.0001-20 wt% of the yellow tea complex extract.
5. The use as claimed in claim 4, wherein the external preparation for skin comprises 0.001-10 wt% of the yellow tea complex extract.
6. The use as claimed in claim 5, wherein the external preparation for skin comprises 0.001-5 wt% of the yellow tea complex extract.
7. The use as claimed in claim 6, wherein the external preparation for skin comprises 0.01 to 5% by weight of the yellow tea complex extract.
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