CN106726898A - A kind of Ramulus et Folium Mussaendae Pubescentis extract and preparation method and application - Google Patents
A kind of Ramulus et Folium Mussaendae Pubescentis extract and preparation method and application Download PDFInfo
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- CN106726898A CN106726898A CN201611116033.8A CN201611116033A CN106726898A CN 106726898 A CN106726898 A CN 106726898A CN 201611116033 A CN201611116033 A CN 201611116033A CN 106726898 A CN106726898 A CN 106726898A
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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
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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
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/805—Corresponding aspects not provided for by any of codes A61K2800/81 - A61K2800/95
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Abstract
The invention provides a kind of Ramulus et Folium Mussaendae Pubescentis extract, its preparation method is：Raw material white tea pulverizes and sieves, plus ethanol water carries out normal temperature homogenate extraction, centrifugation, collects supernatant and is homogenate extraction liquid, and filter residue is standby；Filter residue is taken, is added water, neutral proteinase, digested, boiling water bath goes out enzyme afterwards, is cooled to room temperature, is centrifuged, and collects supernatant and is enzyme-extracting solution；Homogenate extraction liquid is merged with enzyme-extracting solution, ultrafiltration is carried out, filtered solution vacuum concentration is taken, vacuum freeze drying obtains described Ramulus et Folium Mussaendae Pubescentis extract；Gained Ramulus et Folium Mussaendae Pubescentis extract of the invention, in the case of without various chemical addition agents, can be prepared into the natural white tea function toner that a kind of suitable various adult's skin quality are used, and with moisturizing, whitening, moist, anti-ageing various functions of waiting for a long time.
Description
(1) technical field
Natural work(is being prepared the present invention relates to a kind of Ramulus et Folium Mussaendae Pubescentis extract and preparation method thereof, and described Ramulus et Folium Mussaendae Pubescentis extract
Application in energy toner.
(2) background technology
With the development of science and technology and society, the raising that people are required cosmetics and skincare product promotes the fast of cosmetic industry
Speed development.Various functions, the cosmetics of formulation are come out one after another, and Cosmetic Market is a feast for the eyes, colourful.In numerous cosmetics
Among kind class toner with its it is exclusive it is salubrious moisten, natural permeable the features such as occupy increasingly consequence, its product
Yield and sales volume are just rising year by year and development prospect is considerable.Toner is that a kind of liquid type with water as primary raw material is made up
Product, mainly there is transparent type and the major class of emulsion-type two in the market.It can make the clean quiet wetting of skin, appropriateness protection skin and promotion
The physiological action of skin.To numerous consumer groups' common need be typically used for kept the skin wet to the cutin of skin and moisturizing into
Point or supply skin nourishment composition, make skin emolliency, it is plump, meet skin physiology state, keep skin health and have when using
Salubrious sense.
The product category of current toner is a lot, can be such as classified as by function：Spatter property toner, convergence are made up
Water, flexibility toner, moisturizing astringent, whitening lotion and specific function toner etc..Wherein moisturizing, nutrition, skin-active,
Whitening lotion is accounted for more than more than half.Common moisturizing (moisturizing) Essence stresses replenishing water and preserving moisture, and facial-nourishing dew stresses nutrition, smoothing toner
Stress to make epidermis soft, skin activating water stresses active cell, and whitening water stresses whitening function.
But in toner of the prior art, except a small number of plant extract beyond the region of objective existences, base starting material is generally containing from mineral stone
The hydro carbons of oil, alcohols etc., coordinate raw material to be chemical products preservative, colouring agent and odorant.Such as the glycerine in NMF, the third two
The polyalcohols such as alcohol, sorbierite；Salicylic acid, hydroquinones in whitening agent, fluorescent material；Nipagin esters, benzene oxygen in preservative
Base ethanol, to hydroxyl this formic acid esters, ethanol (while there is astriction)；Resorcinol, benzoperoxide sulphur in acne removing agent
Sulphur, allantoin etc.；Human body can be penetrated into by skin during these composition long-term uses, by its different degrees of absorption, to the skin of people
Skin mucous membrane produces harmful effect, or has certain stimulation to skin.
Therefore in current cosmetics production, raw material naturalization receives much attention, it has been recognized that natural raw material is than closing
It is more healthy into chemical composition.Skin-protection product increasingly tends to naturalization, and the exploitation of natural active matter have turned into cosmetic
The important directions of aquatic products exploitation.
Although the patent on toner has kind more than 200 at present in addition, compounding pattern is generally used, and contain mostly
There are various artificial synthesized chemical compositions.Such as " a kind of toner and preparation method thereof ", application number 201010299313.3, mandate public affairs
The B of announcement CN 101966135, the invention toner is included：Surfactant, polyalcohol, emollient, NMF, preservative and
Other additives." cosmetics water for women ", application number 200410018024.6, Authorization Notice No. CN 1248678C, although invention
Contain absolute alcohol in planting active ingredient, but formula using natural aromatics such as thing Lavender, styrax essential oil, rose waters.
In other functional cosmetic water, moisturizing class toner, such as " a kind of moisturizing astringent " (application number 201510489660.5), formula
In comprising glycerine, DPG, alcohol, Sodium Hyaluronate, sodium hyaluronate sodium, disodium succinate, methyl ether, butanedioic acid, pungent
The Multiple components such as glycol；Whitening spot-removing class toner, such as " a kind of freckle removing and whitening toner and preparation method thereof " (application number
201410621660.1), raw material components water containing cation, lipoic acid, potassium sorbate, polysorbate, essence, the red sage root are extracted
Thing, propane diols Multiple components；Anti-aging moisture retention water, such as " a kind of toner with anti-ageing anti-wrinkle and moisturizing moisturizing effect and its
Preparation method ", described toner is formed by the various ingredients such as alcohol phase, water phase and additive compounding；Anti-acne class toner, such as
" a kind of anti-acne toner and preparation method thereof ", the toner by perilla leaf, capsicum leaf, cushaw stem, kochia scoparia, folium lycii,
The extracts such as Aloe mesophyll, felwort, Chinese violet, dandelion and papain, vitamin E, vitamin B6Part, distilled water etc.
Various ingredients compounding is formed.
The artificial synthesized chemical composition generally contained in existing patent or cosmetic aquatic products, deposits to consumer health and appearance
In potential hazard；In addition Multiple components compounding, not only needs more purchase cost, and need multiple fine to determine in production
Amount and dissolving married operation, overall manufacturing are relatively costly.
White tea is one of six big teas of China, is also the peculiar teas in Fujian Province, (belongs to green different from Anji white tea
Tea).Main product in Fujian Fuding, have stable political situation, the ground such as Songxi and Jianyang.Compared with green tea, traditional preparation method of white tea is unique, no
Need frying to knead and finish, it is only necessary to wither naturally and dry, belong to slight fermented tea.Into tea qualitative characteristics for appearance completely drapes over one's shoulders white
In the least, color and luster silvery white is grayish green, therefore claims " white tea ", and into tea shoot, the tender even, milli in complete, leaf bottom is fragrant clear fresh in the least, and the yellowish green limpid, flavour of soup look is clear
Light time sweet.Its main designs and varieties has silver tip pekoe, white peony, tribute eyebrow, Shou Mei.Since ancient times, white tea have " 1 year tea, three
The good reputation of year medicine, 7 years treasured ", causes the favor of more and more consumers in recent years..
White tea due to its natural manufacture craft, remain abundant active enzyme, Polyphenols, catechin, flavonoids,
Polysaccharide, theanine class, coffee bases, vitamins, chlorophyll etc. beneficial to health bioactivator, wherein mainly
(in terms of dry weight) is that (10%-20% is left comprising tea polyphenols (15%-35% or so is as many as 3 times of green tea), catechin
It is right), flavonoids (1% or so) in interior aldehydes matter and amino acid (5%-10% or so), with outstanding pharmacology and health care
Effect.Research shows that white tea has regulation blood fat, prevention cardiovascular and cerebrovascular disease, anti-oxidant, anti-aging, anti-mutation, hypoglycemic, anti-
The many-sides such as tumour, regulation immunologic function, radioresistance, protection protect liver, antibacterial, eliminating inflammation and expelling toxin are respectively provided with significant effect.White tea
It is in the traditional export trade teas of China, to be widely believed that most one of tealeaves of health-care efficacy, phase is occupied in the international market
Work as consequence.Additionally, also contain the nutritional ingredients such as soluble sugar (3% or so), protein (20% or so) in white tea, but
Mostly water-insoluble protein.
The current utilization of document and patent to Fujian white tea is concentrated mainly on edible aspect, such as " instant radix pseudostellariae white tea and
Its preparation technology " (application number 201110458421.5, the B of Authorization Notice No. CN 102488040), a kind of " white tea beverage processing
Technique " (application number 201310281684.2, the B of Authorization Notice No. CN 103283878), " a kind of white tea instant powder and its preparation
Method " (application number 201610279715.4), " a kind of active component composition containing Ramulus et Folium Mussaendae Pubescentis extract and preparation method thereof and should
With " (application number 201610428974.9), " a kind of method that anti-oxidation active substance is extracted in old white tea from golden flower " is " a kind of
Solid tea containing Ramulus et Folium Mussaendae Pubescentis extract " (application number 201210552533.1), a kind of " processing method of cold instant white tea powder " (application number
200710140565.X, Authorization Notice No. CN 101366427 B), protrude the " a kind of containing white tea of skin-care functional
Skin beautifying tea bag " (application number 201110268973.X, Authorization Notice No. CN 102349585 B) is also to be taken in leading
On the way.Above patent some or white tea is formed with other compoundings, is not the single raw material of white tea.So far rarely seen useful Fujian white tea is
Raw material is used for the document report in terms of cosmetics, especially with white tea for single raw material prepares natural function cosmetic water.
The aging of skin, fold, dimness are caused by inside and outside many factors, but the main reason of skin senescence is body
Interior free radical destroys normal cell behavior, accelerates cell senescence.Can be prevented if it can remove or suppress free radical or
Alleviate the aging of skin.In terms of beautifying face and moistering lotion skin care, the aldehydes matter such as Polyphenols, catechin, flavonoids in white tea, tool
There are the effects such as significant anti-oxidant, anti-aging, uvioresistant, whitening, antibacterial, anti-inflammatory.
Aldehydes matter in white tea is very easy to oxidation in the environment of high temperature, aerobic, high light are present.Traditional high temperature
Extraction can effectively improve the production efficiency and recovery rate of phenols, but high-temp extracting is easily caused the brown stain of extract solution color and luster and local flavor in vexed
Ripe taste deterioration, although conventional low temperature extraction is not oxidizable, extraction time is long, and recovery rate is low.
The present invention quickly effectively extracts the aldehydes matter in white tea using homogenate extraction method low temperature.Homogenate extraction is (also referred to as
Dodge formulation) it is a kind of Novel extraction method developed in recent years, according to the principle that historrhexis extracts, flash extracter leads to
Crossing historrhexis makes the fully exposed composition that is extracted be divided by solvent under the various external force effects such as negative pressure, shearing, high velocity impact
Attached bag is enclosed, dissociates, is dissolved, and moment reaches the balance of solvent strength, so as to reach the purpose of rapid extraction, reduces active ingredient
Degraded or loss.There is the functions such as high-speed stirred, superpower vibration, negative pressure diafiltration to reach extraction purpose simultaneously.Due to once
Extracting can complete typically within the several seconds to a few minutes, and its extraction rate is more than hundred times of conventional method, therefore is referred to as
It is " homogenate extraction ", it is possible to effectively extracted in room temperature, does not destroy heat-sensitive ingredients.Extraction process has efficient, quick, section
The various features such as energy, low consumption, environmental protection.
The present invention has carried out appropriate enzymolysis to the water-insoluble protein in white tea, and not only water-soluble is good, and skin care work(
Can be stronger.Small active peptides are strong to the permeability of skin, can pass through cuticula and combined with skin epithelial cell, participate in and improve
The metabolism of Skin Cell, strengthens blood circulation, reaches the purpose of skin care.Simultaneously through the polypeptide after enzymolysis, contain in molecule
Substantial amounts of hydrophilic group exposure, with preferably keeping skin to moisten and moisture-keeping efficacy, while the amino acid in white tea is also nutritious
Skin care is acted on, and the carbohydrate content of polyhydroxy has moisture-keeping function in white tea.
The present invention uses homogenate extraction aldehydes matter, enzymolysis and extraction white tea polypeptide, ultrafiltration, freeze-drying etc. to Fujian white tea
Technological means has carried out white tea feature extract and prepared by white tea functional cosmetic water, and functional evaluation finds the extract and change
Adornment water has preferable antioxidation in vitro, suppression tyrosinase isoreactivity, and has good moisture-keeping function to human body skin.
(3) content of the invention
It is an object of the invention to provide a kind of Ramulus et Folium Mussaendae Pubescentis extract and preparation method and application.
, with Fujian white tea as raw material, using homogenate extraction aldehydes matter, enzymolysis and extraction white tea polypeptide is ultrafiltration, cold for the present invention
Freezing the technological means such as dry has carried out the preparation of white tea feature extract.Obtained white tea feature extract of the invention can be answered
For preparing the natural white tea functional cosmetic water that suitable human body skin is used.The toner have moisturizing, whitening, moisten, it is anti-
Various functions such as aging, while the various skin quality of adult can be adapted to using.Functional evaluation finds Ramulus et Folium Mussaendae Pubescentis extract of the present invention and change
Adornment water has preferable antioxidation in vitro, suppression tyrosinase isoreactivity, and has good moisture-keeping function to human body skin.
The present invention is adopted the following technical scheme that：
A kind of Ramulus et Folium Mussaendae Pubescentis extract, is prepared as follows obtaining：
(1) raw material white tea pulverizes and sieves (to 40~60 mesh), by feed liquid mass ratio 1：15~30 is 60% with volume fraction
Supernatant is collected in~80% ethanol water mixing, normal temperature 2~4min of homogenate extraction, centrifugation, and filter cake is flash to be carried 1~3 time again
Filter residue is collected afterwards standby, merge each gained supernatant and be homogenate extraction liquid；
(2) filter residue of step (1) collection is taken, by feed liquid mass ratio 1：10~30 mix with water, add the filter residue matter
The neutral proteinase (enzyme activity of the neutral proteinase is 2~100,000 u/g) of amount 0.3%~0.6%, 1 is digested in 30~50 DEG C
~2h, afterwards boiling water bath go out 8~15min of enzyme, be cooled to room temperature, be centrifuged, collect supernatant and be enzyme-extracting solution；
(3) step (1) gained homogenate extraction liquid is merged with step (2) gained enzyme-extracting solution, (milipore filter is retained for ultrafiltration
2~10KD of molecular weight), take filtered solution vacuum concentration (20~45 DEG C of thickening temperature, be generally concentrated into original volume 1/5), vacuum
Freeze-drying (usual 48~72h), obtains described Ramulus et Folium Mussaendae Pubescentis extract (yellow green).
In the present invention, described raw material white tea (Camellia sinensis cv.Fuding-dabaicha) is that Fujian is white
Spring of tea produces the leaf of a bud 2~3 through the tealeaves after existing preparation method conventional machining, preferably can then give birth to commercially available on the market
The organic longevity eyebrow kind in Fuding of product.
In step (1) of the present invention, the volume fraction is 60%~80% ethanol water, by edible ethanol and pure
Water allotment is formed.Described normal temperature homogenate extraction, temperature is low, and the time is short, can effectively prevent the oxidation of white tea aldehydes matter.
In step (2), the natural pH during neutral protease enzymolysis is 5.5 or so.
In step (3), the ultrafiltration can effectively remove insoluble phenols component, and the protein component not digested and water are not
Soluble polysaccharide macromolecular substances, make sample water-soluble more preferably.The evaporating temperature of the filtered solution vacuum concentration is controlled at≤45 DEG C,
In the case of vacuum and low temperature oxygen barrier, be conducive to keeping active material.Described vacuum freeze drying is conducive to white tea phenols, polypeptide etc.
Bioactivator is guaranteed the quality.
Water used is pure water in the present invention, without ion component, it is to avoid institute's metal ion is reacted with aldehydes matter
Discoloration.
Heretofore described normal temperature is 20~30 DEG C.
Obtained Ramulus et Folium Mussaendae Pubescentis extract of the invention can be applied to the preparation of natural white tea functional cosmetic water, the side of the preparation
Method is：
Described Ramulus et Folium Mussaendae Pubescentis extract is added into dissolved in purified water, the solution for being configured to 0.02wt%~0.1wt% is (limpid transparent
Yellow-green liquid), it is filling after through pasteurization (70~90 DEG C, 15~30min), obtain final product natural white tea functional cosmetic aquatic products
Product.
The beneficial effects of the present invention are：According to the anti-oxidant of white tea, anti-aging, anti-mutation, radioresistance, antibacterial, anti-inflammatory
The innate efficacies such as removing toxic substances, to active ingredients such as polyphenol therein (tea bag includes polyphenol, catechin, flavones ingredient), albumen, polypeptides
Separation and Extraction is carried out, in the case of without various chemical addition agents, a kind of suitable various adult's skin quality is prepared into and is used
Natural white tea function toner, and with moisturizing, whitening, moisten, anti-ageing various functions of waiting for a long time.
(4) illustrate
Fig. 1：The preparation technology flow chart of natural white tea functional cosmetic water of the invention；
Fig. 2：The moistening effect figure of 0.03% white tea functional cosmetic water in embodiment 1；
Fig. 3：The moistening effect figure of 0.05% white tea functional cosmetic water in embodiment 2；
Fig. 4：The DPPH clearance rates of the white tea feature extract prepared in embodiment 3；
Fig. 5：The hydroxyl radical free radical clearance rate of the white tea feature extract prepared in embodiment 3；
Fig. 6：The tyrosinase inhibition rate of the white tea feature extract prepared in embodiment 3；
Fig. 7：The moistening effect figure of white tea functional cosmetic water in embodiment 3.
(5) specific embodiment
Below by specific embodiment, the present invention is further illustrated, but protection scope of the present invention is not limited in
This.
Embodiment 1：
(1) raw material white tea (manufacturer is taken：Fujian Ding Hao Tea Co., Ltd；The name of an article：Shou Mei；Credit rating：It is superfine；Only
Content 600g/ bags, In Aluminium Foil Packing, carton external packing) 1kg, pulverize and sieve to 50 mesh, by feed liquid mass ratio 1：20 with 20L volumes
Fraction is 60% ethanol water mixing, and normal temperature is flash, and (pilot scale type dodges and carries instrument, JHBE-100A, and Henan gold tripod development in science and technology has
Limit company) 2min is extracted, rotating speed 5000r/min centrifugations are removed slag, and (table-type low-speed centrifuge, TD5A-WS turn with 851400 levels
Son, Xiang Yi centrifuges Instrument Ltd.), 4000r/min (3040xg), collect centrifuged supernatant, filter residue is flash carry 1 time again after
Collect standby, merge each gained supernatant and be homogenate extraction liquid；
(2) filter residue of step (1) collection is taken, 20L pure water is added, is mixed in the stainless steel cool-bag of 50L, then added
Enter 3g neutral proteinases (enzyme activity is 100,000 u/g, Ningxia jade of the He family Bioisystech Co., Ltd), under natural pH5.5, in 40 DEG C of enzymes
Solution 1.5h, afterwards boiling water bath go out enzyme 10min, be cooled to room temperature, rotating speed 5000r/min centrifugations are removed slag, and are collected supernatant and are enzyme
Solution extract solution；
(3) step (1) gained homogenate extraction liquid is merged with step (2) gained enzyme-extracting solution, ultrafiltration (Cogent M1
Semi-automatic ultrafiltration system, the milipore filter film bag of Pellicon 2, Biomax 5, molecular cut off 5KD, Merck Millipore Corp.), take
Filtered solution is concentrated in vacuo to original volume 1/5 (temperature concentrated in vacuo≤45 DEG C, Rotary Evaporators, RE-5210A, Shanghai Asia Rong Shenghua
Instrument plant), (pilot scale type vacuum freeze drier Pilot7-12T, the rich doctor's health laboratory apparatus in Beijing is limited for vacuum freeze drying 72h
Company), white tea feature extract (yellow green) 226.4g is obtained, yield is 22.6%, and detection, external work(are analyzed afterwards
Can evaluate.
(4) pure water 30L is added to dissolve gained white tea feature extract 9g, the solution for being configured to 0.03wt% is (limpid
Transparent yellow-green liquid), after aluminium pot spray filling (350ml), through pasteurization (80 DEG C, 20min), obtain natural white tea work(
Energy property cosmetic water spraying agent, carries out sense organ and functional evaluation, shelf-life experiment afterwards.
Coherent detection and evaluation result are as follows：
The Major Components result of white tea feature extract under the present embodiment preparation condition of table 1
Under the present embodiment preparation condition, white tea feature extract with the increase of concentration, DPPH and hydroxyl radical free radical
Removing linearly increases trend, and in 60 μ g/ml, the average clearance rate of DPPH free radicals is up to 91.4%, the hydroxyl in 100 μ g/ml
The clearance rate of base free radical is up to 86.7%.Illustrate the white tea functional cosmetic water after white tea feature extract and its dissolving
(0.03%, i.e. 0.3mg/ml) has preferably anti-oxidant and potential effect of anti-aging.
With the increase of concentration, tyrosinase inhibition rate is in dramatically increase trend to white tea feature extract, in 0.3mg/
During ml, tyrosinase inhibition rate is up to 65.3%.Illustrate the white tea functional cosmetic water after white tea feature extract and its dissolving
(0.03%) with the potential effect of preferable whitening.
The moisture-keeping function contrast of 0.03% white tea functional cosmetic water and blank (pure water) as shown in Fig. 2 by
Fig. 2 understands that 0.03% white tea functional cosmetic water has preferable moisture-keeping function.
The Micro biological Tests result of white tea functional cosmetic water under the present embodiment preparation condition of table 2
* exceptionally, other are in terms of dry for above-mentioned content water removal.
As shown in Table 2, the Micro biological Tests result of the white tea functional cosmetic water under the present embodiment preparation condition meets phase
Close national product sanitary standard.Accelerated condition preserves (37 DEG C, 40 DEG C, 45 DEG C) and products measure, it is determined that natural white tea function
Property toner spray products shelf-life be 1 year.
Embodiment 2：
(1) raw material white tea (manufacturer is taken：Fujian Ding Hao Tea Co., Ltd；The name of an article：Shou Mei；Credit rating：It is superfine；Only
Content 600g/ bags, In Aluminium Foil Packing, carton external packing) 1kg, pulverize and sieve to 60 mesh, by feed liquid mass ratio 1：30 with 30L volumes
Fraction is 70% ethanol water mixing, and normal temperature is flash, and (pilot scale type dodges and carries instrument, JHBE-100A, and Henan gold tripod development in science and technology has
Limit company) 3min is extracted, rotating speed 5000r/min centrifugations are removed slag, and (table-type low-speed centrifuge, TD5A-WS turn with 851400 levels
Son, Xiang Yi centrifuges Instrument Ltd.), 4000r/min (3040xg), collect centrifuged supernatant, filter residue is flash carry 1 time again after
Collect standby, merge each gained supernatant and be homogenate extraction liquid；
(2) filter residue of step (1) collection is taken, 20L pure water is added, is mixed in the stainless steel cool-bag of 50L, then added
Enter 4g neutral proteinases (enzyme activity is 100,000 u/g, Ningxia jade of the He family Bioisystech Co., Ltd), under natural pH5.5, in 40 DEG C of enzymes
Solution 2h, afterwards boiling water bath go out enzyme 15min, be cooled to room temperature, rotating speed 5000r/min centrifugations are removed slag, and are collected supernatant and are enzymolysis
Extract solution；
(3) step (1) gained homogenate extraction liquid is merged with step (2) gained enzyme-extracting solution, ultrafiltration (Cogent M1
Semi-automatic ultrafiltration system, the milipore filter film bag of Pellicon 2, Biomax 5, molecular cut off 5KD, Merck Millipore Corp.), take
Filtered solution is concentrated in vacuo to original volume 1/5 (temperature concentrated in vacuo≤45 DEG C, Rotary Evaporators, RE-5210A, Shanghai Asia Rong Shenghua
Instrument plant), (pilot scale type vacuum freeze drier Pilot7-12T, the rich doctor's health laboratory apparatus in Beijing is limited for vacuum freeze drying 72h
Company), white tea feature extract (yellow green) 273.2g is obtained, yield is 27.3%, and detection, external work(are analyzed afterwards
Can evaluate.
(4) pure water 10L is added to dissolve gained white tea feature extract 5g, the solution for being configured to 0.05wt% is (limpid
Transparent yellow-green liquid), after aluminium pot spray filling (350ml), through pasteurization (80 DEG C, 30min), obtain natural white tea work(
Energy property cosmetic water spraying agent, carries out sense organ and functional evaluation, shelf-life experiment afterwards.
Coherent detection and evaluation result are as follows：
The Major Components result of white tea feature extract under the present embodiment preparation condition of table 3
* exceptionally, other are in terms of dry for above-mentioned content water removal.
Under the present embodiment preparation condition, white tea function extract with the increase of concentration, DPPH and hydroxyl radical free radical it is clear
Except linearly increasing trend, in 60 μ g/ml, the average clearance rate of DPPH free radicals is up to 97.1%, the hydroxyl in 100 μ g/ml
The clearance rate of free radical is up to 95.8%.Illustrate the white tea functional cosmetic water after white tea feature extract and its dissolving
(0.05%, i.e. 0.5mg/ml) has preferably anti-oxidant and potential effect of anti-aging.
With the increase of concentration, tyrosinase inhibition rate is in dramatically increase trend to white tea function extract, in 0.5mg/ml
When, tyrosinase inhibition rate is up to 79.5%.Illustrate the white tea functional cosmetic water after white tea function extract and its dissolving
(0.05%) with the potential effect of preferable whitening.
The moisture-keeping function contrast of 0.05% white tea functional cosmetic water and blank (pure water) as shown in figure 3, by
Fig. 3 understands that 0.05% white tea functional cosmetic water has preferable moisture-keeping function.
The Micro biological Tests result of white tea functional cosmetic water under the present embodiment preparation condition of table 4
As shown in Table 4, the Micro biological Tests result of the white tea functional cosmetic water under the present embodiment preparation condition meets phase
National product sanitary standard is closed, accelerated condition preserves (37 DEG C, 40 DEG C, 45 DEG C) and products measure, it is determined that natural white tea function
Property toner spray products shelf-life be 1 year.
Embodiment 3：
(1) raw material white tea (manufacturer is taken：Fujian Ding Hao Tea Co., Ltd；The name of an article：Shou Mei；Credit rating：It is superfine；Only
Content 600g/ bags, In Aluminium Foil Packing, carton external packing) 1kg, pulverize and sieve to 60 mesh, by feed liquid mass ratio 1：25 with 25L volumes
Fraction is 70% ethanol water mixing, and normal temperature is flash, and (pilot scale type dodges and carries instrument, JHBE-100A, and Henan gold tripod development in science and technology has
Limit company) 3min is extracted, rotating speed 5000r/min centrifugations are removed slag, and (table-type low-speed centrifuge, TD5A-WS turn with 851400 levels
Son, Xiang Yi centrifuges Instrument Ltd.), 4000r/min (3040xg), collect centrifuged supernatant, filter residue is flash carry 1 time again after
Collect standby, merge each gained supernatant and be homogenate extraction liquid；
(2) filter residue of step (1) collection is taken, 20L pure water is added, is mixed in the stainless steel cool-bag of 50L, then added
Enter 5g neutral proteinases (enzyme activity is 100,000 u/g, Ningxia jade of the He family Bioisystech Co., Ltd), under natural pH5.5, in 40 DEG C of enzymes
Solution 1.5h, afterwards boiling water bath go out enzyme 15min, be cooled to room temperature, rotating speed 5000r/min centrifugations are removed slag, and are collected supernatant and are enzyme
Solution extract solution；
(3) step (1) gained homogenate extraction liquid is merged with step (2) gained enzyme-extracting solution, ultrafiltration (Cogent M1
Semi-automatic ultrafiltration system, the milipore filter film bag of Pellicon 2, Biomax 5, molecular cut off 5KD, Merck Millipore Corp.), take
Filtered solution is concentrated in vacuo to original volume 1/5 (temperature concentrated in vacuo≤45 DEG C, Rotary Evaporators, RE-5210A, Shanghai Asia Rong Shenghua
Instrument plant), (pilot scale type vacuum freeze drier Pilot7-12T, the rich doctor's health laboratory apparatus in Beijing is limited for vacuum freeze drying 72h
Company), white tea feature extract (yellow green) 262.8g is obtained, yield is 26.3%, and detection, external work(are analyzed afterwards
Can evaluate.
(4) pure water 10L is added to dissolve gained white tea feature extract 5g, the solution for being configured to 0.05wt% is (limpid
Transparent yellow-green liquid), after aluminium pot spray filling (350ml), through pasteurization (80 DEG C, 30min), obtain natural white tea work(
Energy property cosmetic water spraying agent, carries out sense organ and functional evaluation, shelf-life experiment afterwards.
And carried out main component, external functional evaluation and the cosmetic of natural white tea function of gained white tea feature extract
The Micro biological Tests of water, moisture-retention function of skin are determined, and concrete outcome is as follows：
(1) white tea feature extract main component measurement result is as shown in table 5：
The Major Components result of white tea feature extract under the present embodiment preparation condition of table 5
* exceptionally, other are in terms of dry for above-mentioned content water removal.
(2) oxidation resistance
From Fig. 4, Fig. 5, white tea feature extract with the increase of concentration, the clearance rate of DPPH and hydroxyl radical free radical
Trend is linearly increased, the average clearance rate of DPPH free radicals is up to 95.6%, the hydroxyl in 100 μ g/ml in 60 μ g/ml
The clearance rate of free radical is up to 92.3%.
Illustrate white tea functional cosmetic water (0.02%~0.1%, i.e., 0.2 after white tea feature extract and its dissolving
~1mg/ml) there is preferably anti-oxidant and potential effect of anti-aging.
(3) casein enzyme activity inhibition
It will be appreciated from fig. 6 that increase of the white tea feature extract with concentration, tyrosinase inhibition rate is in dramatically increase
Gesture, in 0.8mg/ml, tyrosinase inhibition rate is up to 78.7%.Illustrate the white tea function after white tea function extract and its dissolving
Property toner (0.02%~0.1%, i.e. 0.2~1mg/ml) have the potential effect of preferable whitening.
(4) functional cosmetic water moistening effect
As shown in Figure 7,0.02%~0.1% white tea functional cosmetic water is made with respect to blank with good moisturizing
With, and with the increase of cosmetic water concentration, moistening effect increases.
(5) functional cosmetic water microbiology inspection
The Micro biological Tests result of white tea functional cosmetic water under the present embodiment preparation condition of table 6
It is above-mentioned test result indicate that, the Micro biological Tests result of white tea functional cosmetic water meets concerned countries product health
Standard.
(6) the toner shelf-life
Accelerated condition preserves (37 DEG C, 40 DEG C, 45 DEG C) and products measure, determines the guarantor of white tea function water aerosol can product
The matter phase is 1 year.
Analysis in the present invention is detected and the method for external functional evaluation is：
(1) analysis detection：Feature extract after lyophilized, has the original fragrant of obvious white tea on sense organ；First
Calculated extracting yield, moisture then to extract, ash content, total phenol, general flavone, gross protein, total free amine group
Acid, polypeptide, total reducing sugar main component are detected；
1. extract yield is calculated：(extract weight/white tea raw material weight after lyophilized) × 100%
2. determination of moisture：Using the measure of moisture in GB 50093-2010 national food safety standard food, convection drying
Method is determined.
3. ash determination：Using the measure of ash content in GB 5009.4-2010 national food safety standard food, in food
The measure of ash content.
4. determining total phenol：Using Forint phenol method.Take appropriate extract to be dissolved with 80% methyl alcohol, extract solution adds 200 μ l
The forint phenol reagent of 0.1N and 2000 μ l ultra-pure waters, place 3min after vibration, add 1000 μ l sodium carbonate (20%, w/v) molten
Liquid, light absorption value is determined after vibration under room temperature avoid light place 1h, 765nm, if distilled water is blank.Results expression is every gram white
The a great deal of (mg GAE/g dry weights) of gallic acid contained by tea raw material dry weight.
5. Determination of Total Flavonoids：Using aluminum nitrate-colorimetric method for determining.The ultrapure water dissolves of appropriate extract are taken, is added
0.3ml5%NaNO2Solution, concussion places 6min after shaking up, and adds 0.6ml 10%AlCl3·6H2O, concussion is put after shaking up
5min is put, then adds the 2ml 1N NaOH aqueous solution, plus ultra-pure water to determine light absorption value under 510nm to cumulative volume 10ml, if distilled water
It is blank.Results expression is a great deal of (mg RE/g dry weights) of rutin contained by every gram of white tea raw material dry weight.
6. total protein content is determined：Kjeldahl's method.
7. total free amino acid is determined：Using the assay method of GBT 8314-2013 tea free amino acid total amounts.
8. determining content of peptides：2.5ml extract sample solutions are taken, the trichloroacetic acid of 2.5ml 10% (W/V) is added
(TCA) aqueous solution, in being well mixed on whirlpool mixed instrument, stands 10min, and 15min then is centrifuged under 4000r/min, will be upper
Clear liquid is transferred completely into 50ml volumetric flasks, and TCA with 5% is settled to scale, shakes up.Then the above-mentioned solution of 6.0ml is taken to put
In another test tube, biuret reagent 4.0ml (sample liquids are added：Biuret reagent=3:2, V/V) it is, equal in mixing on whirlpool mixed instrument
It is even, 10min, 2000r/min centrifugation 10min are stood, supernatant is taken in measure OD values, reference standard curve (Gly- under 540nm
The poly saccharide peptide standard products of Gly-Tyr-Arg tetra-) try to achieve peptide concentration C (mg/ml) in sample solution, and then polypeptide contains in can trying to achieve sample
Amount.
9. total sugar determination：Determined using phend-sulphuric acid, with glucose as reference substance.
(2) external functional evaluation：Anti-oxidant evaluation (DPPH, hydroxyl radical free radical removes function), (tyrosinase presses down for whitening
Function processed)
I, anti-oxidant evaluation：
1. DPPH free radicals are removed：The DPPH reagents of 0.2mM are prepared, extract solution and 3.9ml that 0.1ml diluted is taken
DPPH reagents mix, and 1h is reacted in the dark, and light absorption value is determined under 515nm.80% methanol solution using 0.1ml as reference,
Water is used as blank.DPPH free radical scavenging activities are calculated as follows.
Clearance rate (%)=[1- (Aj-Ai)/Ac] × 100%
Wherein measure group is Ai, reference group is Aj, blank is Ac,
2. scavenging hydroxyl：By salicylic acid Fenton reaction assays.
Certain density prepare liquid is added in 10ml test tubes, 0.3ml 6mmol/LFeSO are subsequently adding4, 1.5ml
6mmol/L salicylic acid pure water polishings 4.8ml shakes up, and is eventually adding 0.3ml 6.0mmol/L H2O2Start reaction, stand
10min mensuration absorbances at 510nm.In view of extract solution light absorption value in itself, sample blank is done, measure deduction sample empty
White absorbance, while determining the absorbance of blank liquid.
Clearance rate (%)=(A0–A1+A2)/A0× 100%
Wherein：A0The absorbance of solution during not add extract solution；A1For plus extract solution after solution absorbance；A2To extract
The absorbance of liquid.
II, whitening (tyrosinase inhibitory activity)
Tyrosinase is main rate-limiting enzyme in melanin forming process, and the active size of the enzyme decides that melanin is formed
Quantity.Melanocyte mainly generates melanin with tyrosine as raw material under the oxidation of rate-limiting enzyme tyrosinase.Skin
Tyrosine in skin in collagen forms melanin after being acted on by tyrosinase, is skin darkening, the main cause of color spot generation.Junket
Propylhomoserin enzyme inhibitor can have potential treatment abnormal pigment disorderly, and skin-whitening additive is can be applied in cosmetic industry.
Take said extracted thing plus appropriate amount of purified water and prepare the μ L of sample liquid 100, plus pH 6.8 the μ L of phosphate buffer 56 with
The tyrosine solution 94 μ L of 2.0mmol/L, it is to be mixed it is uniform after the μ L (50U) of tyrosinase solution 50 are rapidly added in ice bath,
Mix, 37 DEG C of incubation 10min, the mensuration absorbance at 490nm.
Tyrosinase inhibition rate computing formula is：I%=[(A-B)-(C-D)]/[A-B] × 100%
A：It is not loaded the absorbance that the enzyme-added mixed liquor of product is surveyed；
B：It is not loaded the absorbance that the also not enzyme-added mixed liquor of product is surveyed；
C：The absorbance that the mixed liquor of sample-adding product and enzyme is surveyed；
D：Sample-adding product and absorbance that not enzyme-added mixed liquor is surveyed.
(3) functional cosmetic water moistening effect is evaluated
According to light industry professional standard QB/T 4256-2011《Cosmetics moisture-keeping efficacy evaluation guide》Method carry out reality
Test room self-inspection.Determine using the skin moisture content value (Moisture Measurement Value, MMV) after certain hour,
MMV is 0~150 numerical value.If pure water is blank.
(5) organoleptic test
According to light industry professional standard QB/T 2660-20《Toner》Sensory evaluation method.
(4) natural white tea function cosmetic water microorganism inspection
According to food medicine supervision general bureau of country《Cosmetics safety technical specification (version in 2015)》Chapter 5, Micro biological Tests
Natural white tea function cosmetic aquatic products have been carried out total plate count, heat-resisting coliform, pseudomonas aeruginosa, golden yellow by method
Staphylococcus, the laboratory self-inspection of yeast and mold.
Claims (4)
1. a kind of Ramulus et Folium Mussaendae Pubescentis extract, it is characterised in that described Ramulus et Folium Mussaendae Pubescentis extract is prepared as follows obtaining：
(1) raw material white tea pulverizes and sieves, by feed liquid mass ratio 1：15~30 is water-soluble with the ethanol that volume fraction is 60%~80%
Liquid mixes, normal temperature 2~4min of homogenate extraction, centrifugation, collects supernatant, and filter cake is flash to carry that to collect filter residue for 1~3 time afterwards standby again,
Merge each gained supernatant and be homogenate extraction liquid；
(2) filter residue of step (1) collection is taken, by feed liquid mass ratio 1：10~30 mix with water, add the filter residue quality
0.3%~0.6% neutral proteinase, the enzyme activity of the neutral proteinase is 2~100,000 u/g, digest 1 in 30~50 DEG C~
2h, afterwards boiling water bath go out 8~15min of enzyme, be cooled to room temperature, be centrifuged, collect supernatant and be enzyme-extracting solution；
(3) step (1) gained homogenate extraction liquid and step (2) gained enzyme-extracting solution is merged, with molecular cut off for 2~
The milipore filter of 10KD carries out ultrafiltration, takes filtered solution in being concentrated in vacuo at 20~45 DEG C, vacuum freeze drying afterwards, obtains described
Ramulus et Folium Mussaendae Pubescentis extract.
2. Ramulus et Folium Mussaendae Pubescentis extract as claimed in claim 1, it is characterised in that in step (1), the volume fraction is 60%~
80% ethanol water is formed by edible ethanol and pure water allotment.
3. application of the Ramulus et Folium Mussaendae Pubescentis extract as claimed in claim 1 in natural white tea functional cosmetic water is prepared.
4. application as claimed in claim 4, it is characterised in that the preparation method of the natural white tea functional cosmetic water is：
Described Ramulus et Folium Mussaendae Pubescentis extract is added into dissolved in purified water, the solution of 0.02wt%~0.1wt% is configured to, it is filling after through Pasteur
Sterilizing, obtains final product natural white tea functional cosmetic aquatic products.
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| CN108703240A (en) * | 2018-05-02 | 2018-10-26 | 贵州芳瑞堂生物科技有限公司 | A kind of Ramulus et Folium Mussaendae Pubescentis extract of high polyphenol content |
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| CN115844977B (en) * | 2022-12-30 | 2024-03-29 | 福建农林大学 | Antibacterial herbal composition capable of being used for dry eye SPA atomization and preparation method thereof |
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